Alternatives to Animal Use in Research, Testing, and Education February 1986 NTIS order #PB86-183134 Recommended Citation: U.S. Congress, Office of Technology Assessment, Alternatives to Animal Use in Research, Testing, and Education (Washington, DC: U.S. Government Printing Office, OTA-BA-273, February 1986). Library of Congress Catalog Card Number 85-600621 For sale by the Superintendent of Documents U.S. Government Printing Office, Washington, DC 20402 Foreword With an estimated 17 million to 22 million animals used in laboratories annually in the United States, public interest in animal welfare has sparked an often emotional de- bate over such uses of animals. Concerns focus on balancing societal needs for continued progress in biomedical and behavioral research, for toxicity testing to safeguard the pub- lic, and for education in the life sciences with desires to replace, reduce, and refine the use of laboratory animals. In 1985, Congress enacted three laws that dealt with labora- tory animals, including amendments to the Animal Welfare Act. In this assessment, OTA analyzes the scientific, regulatory, economic, legal, and ethical considerations involved in alternative technologies in biomedical and behavioral research, toxicity testing, and education. Included is a detailed examination of Federal, State, and institutional regulation of animal use, and a review of recent developments in 10 other countries. The report was requested by Sen. Orrin Hatch, Chairman of the Senate Com- mittee on Labor and Human Resources. The report illustrates a range of options for congressional action in seven principal areas of public policy regarding animals: using existing alternatives, developing new alter- natives, disseminating research and testing information, restricting animal use, count- ing the numbers and kinds of animals used, establishing a uniform policy for animal use within Federal agencies, and amending the Animal Welfare Act. OTA was assisted in preparing this study by an advisory panel of individuals and reviewers selected for their expertise and diverse points of view on the issues covered in the assessment. Advisory panelists and reviewers were drawn from animal welfare groups, industrial testing laboratories, medical and veterinary schools, Federal regula- tory agencies, scientific societies, academia, and the citizenry at large—in short, from representatives of all parties interested in laboratory-animal use and its alternatives. Written comments were received from 144 reviewers on the penultimate draft of the assess- ment. In addition, at the study’s inception, OTA solicited information and opinions from more than 600 interested groups and individuals. OTA gratefully acknowledges the contribution of each of these individuals. As with all OTA reports, responsibility for the content of the assessment is OTA’s alone. The assessment does not necessarily constitute the consensus or endorsement of the advi- sory panel or the Technology Assessment Board. Director . . .Ill Advisory Panel Alternatives to Animal Use in Research, Testing, and Education Arthur L. Caplan, Panel Chair The Hastings Center Hastings-on-Hudson, NY Perrie M. Adams The University of Dallas TX Melvin W. Balk Texas Health Science Center Charles River Breeding Laboratories, Inc. Wilmington, MA Earle W. Brauer Revlon Research Center, Inc. Edison, NJ David J. Brusick Litton Bionetics Kensington, MD G. Gilbert Cloyd Norwich Eaton Pharmaceuticals, Inc. Norwich, NY W. Jean Dodds New York State Department of Health Albany, NY Kurt Enslein Health Designs, Inc. Rochester, NY Alan M. Goldberg The Johns Hopkins School of Public Health Baltimore, MD Richard M. Hoar Findley Research, Inc. Fall River, MA Peter Barton Hutt Covington and Burling Washington, DC Connie Kagan Animal Political Action Washington, DC Ronald Lamont-Havers Massachusetts General Boston, MA John E. McArdle Humane Society of the Washington, DC Robert A. Neal Committee Hospital United States Chemical Industry Institute of Toxicology Research Triangle Park, NC J. Wesley Robb University of Southern California Los Angeles, CA Andrew N. Rowan Tufts University Boston, MA Jeri A. Sechzer The New York Hospital Center White Plains, NY Henry Spira Coalition to Abolish the LD50 and Draize Tests New York, NY OTA appreciates and is grateful for the valuable assistance and thoughtful critiques provided by the Advisory Panel members. The views expressed in this OTA report, however, are the sole responsibility of the Office of Technologv Assessment. iv OTA Project Staff Alternatives to Animal Use in Research, Testing, and Education Roger C. Herdman, Assistant Director, OTA Health and Life Sciences Division Gretchen S. Kolsrud, Biological Applications Program Manager Gary B. Ellis, Project Director and Analyst Judy K. Kosovich, Principal Analyst Lisa J. Raines, Legal Analyst Timothy J. Hart, Project Director l Gregory A. Jaffe, Research Assistantz Marcia D. Brody, Resarch Assistantz James A. Thomas, Research Assistant3 Thomas M. Bugbee, Research Assistant4 Jeffrey S. Stryker, Research Analyst5 Support Staff Sharon K. Smith, Administrative Assistant Elma Rubright, Administrative Assistant3 Linda S. Ray ford, Secretary/Work Processing Specialist Barbara V. Ketchum, Clerical Assistant Contractors Linda Starke (Editor), Washington, DC Battelle—Columbus Laboratories, Columbus, OH Leonard M. Chanin, Washington, DC Eileen M. Cline, Springfield, VA Paul N. Craig, Shady Side, MD Arthur H. Flemming, University of Chicago Gordon G. Gallup, Jr., State University of New York at Albany Gilbert S. Greenwald, University of Kansas Medical Center Anne M. Guthrie, Arlington, VA Health Designs, Inc., Rochester, NY Henry R. Hertzfeld and Thomas D. Myers, Washington, DC Meyer, Failer, and Weisman, P.C., Washington, DC Roland M. Nardone and Lucille Ouellette, Catholic University, Washington, DC Bennie I. Osburn, University of California, Davis Stephen P. Push, Washington, DC ‘Through December 1984. ‘Through July 1985, 3Through August 1984. 4Through June 1984. Through January 198.5 , CONTENTS Chapter Page 1. Summary, Policy Issues, and Options for Congressional Action . . . . . . . , . . . 3 2. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 3. Patterns of Animal Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 4. Ethical Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 5. The Use of Animals in Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 6. Ahernatives to Animal Use in Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 7.The Use of Animals in Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 8. Ahernatives to Animal Use in Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175 9. Animal Use n Education and the Alternatives . . . . . . . . . . . . . . . . . . . . . . . . . 199 10.Information Resources and Computer Systems . . . . . . . . . . . . . . . . . . . . . . . . 219 11. Economic Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . , . . . . . . . . . . . . . . . . 243 12. Public and Private Funding Toward the Development of Alternatives . . . . . 259 13. Federal Regulation of Animal Use ..., . . . . . . ., . . . . . . . . . . . . . . . . . . . . . . . 275 14. State Regulation of Animal Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305 15. Institutional and Self-Regulation of Animal Use . . . . . . . . . . . . . . . . . . . . . . . . 335 16. Regulation of Animal Use in Selected Foreign Countries. . . . . . . . . . . . . . . . . 359 Appendix Page A. Testing Guidelines. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 383 B. Regulation of Animal Use Within Federal Departments and Agencies. . . . . . 386 C. Public Health Service Policy . . . . . . . . . . ..., . . . . . . . . . . . . . . . . . . . . . . . . . . 395 D. Laboratory-Animal Facilities Fully Accredited by the American Association for Accreditation of Laboratory Animal Care . . . . . . . . . . . . . . . . . . . . . . . . . . 401 E. International Agreements Governing Animal Use . . . . . . . . . . . . . . . . . . . . . . . 412 F. List of Working Papers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 418 G. Acknowledgments. ..., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 419 H. Glossary of Acronyms and Terms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 423 Index . . . . . . . . . . . . . . . . ..., . . . . . . . . . . . . . . . . ., . . . . . . . . . . . . . . . . . . . . . . . . 433 vii Chapter I Summary, Policy Issues, and options for Congressional Action CONTENTS Page Definition of Terms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 How Many Animals Are Used? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Ethical Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 Alternatives in Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 Ahernatives infesting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 Alternatives in Education . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 Computer Simulation and Information Resources . . . . . . . . . . . . . . . . . . . . . . . . . . 11 Economic Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 Funding for the Development of Alternatives. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Regulation of Animal Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Federal Regulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 State Regulation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 Institutional and Self-Regulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 Regulation Within Federal Agencies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 International Regulation , . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 Policy Issues and Options for Congressional Action. . . . . . . . . . . . . . . . . . . . . . . . . 17 Issue: Should steps be taken to encourage the use of available alternatives in research, testing, or education? ., . . . . . . . . . . . . . . ,. . . . . . . . . . . . , . , . . 19 Issue: Should the more rapid development of new alternatives in research, testing, or education be stimulated? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 Issue: Should improvements be made in information resources to reduce any unintentionally duplicative use of animais in research and testing?. . . . . 23 Issue: Should animal use in research, testing, or education be restricted? . . . . 26 Issue: Should more accurate data be obtained on the kinds and numbers of animals used in research, testing, and education? . . . , . , . . . . . . . . . . . . . 29 Issue: Should Federal departments and agencies be subject to minimum standards for animal use? . . . . . . , . . . . . . . . . . . . . . . . . . . . . . . . , . . . . . . . 31 Issue: Should the Animal Welfare Act of 1966 be further amended, or its enforcement enhanced? . . . . . . . . . . . . . . . , . . . . . . . . . . . . . . . . . . . . . . . . . 32 List of Tables Table No. Page l-l. Animal Use Reported to the U.S. Department of Agriculture, 1983 . . . . . . . . 5 l-2. National Laws for the Protection of Animals in Selected European Countries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 l-3. Policy Issues Related to Alternatives to Animal Use and Options for Congressional Action . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 Figure Figure No. Page l-1. Chronological Sequence of Chick Embryo Chorioallantoic Membrane Assay... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 Chapter 1 Summary, Policy Issues, and Options for Congressional Action G G G G G G A former high school teacher in New York organizes demonstrations and advertising campaigns opposing the use of rabbits and rodents in two product-safety tests. Industry responds by giving several million dollars in grants to university scientists searching for alternatives to animal testing. Researchers induce seizures in rats, draw their cerebrospinal fluid, and use it to quell seizures in other rats; the anticonvulsant substance produced during seizures could bear on the under- standing and treatment of epilepsy. Industrial toxicologists in New Jersey adopt refined methods of testing potentially poisonous chemicals, reducing by 48 percent the number of animals used in acute toxicity studies and cutting the cost of compliance with government regulations. A Virginia woman donates $1,250,000 to the University of Pennsylvania to establish the Nation first endowed professorship in humane ethics and animal welfare. One of the goals of the chair is to investigate alternatives to animal experiments for medical research. Members of the Animal Liberation Front break into a biomedical research laboratory in Califor- nia and remove dogs being used in a cardiac pacemaker experiment. Veterinary students in Washington study principles of physiology without recourse to the tra- ditional dog dissection. Instead, they use a computer simulation of canine physiology. These recent events illustrate the complex po- litical, ethical, and economic issues raised by the use of animals in research, testing, and education. Concern about the continued use of animals has led to public calls for development of alternatives. The popular debate over animal use has been taken up by proponents holding a wide spectrum of views, ranging from belief in abolition of animal use on moral and ethical grounds to belief in free rein on the use of animals in research, testing, and education. An increasing number of groups are taking a middle ground. In the mid-1980s, it is misleading—and often impossible—to character- ize many vocal groups either as simply “pro-animal” or “pro-research. ” In light of requests for “a scientific evaluation of alternative methods to animal research, experi- mentation, and testing” from the Chairman of the Senate Committee on Labor and Human Resources, Senator Orrin G. Hatch (R-UT), and from Senator Alan Cranston (D-CA), this assessment examines the reasons for seeking such alternatives and the prospects for developing them. It describes ani- mal and nonanimal methods used by industry, academia, and government agencies; explains the roles and requirements of government regulation and self-regulation of animal use; and identifies policy issues and options that the debate over alter- natives places before Congress. The report covers three kinds of animal use: research in the biomedical and behavioral sci- ences; testing of products for toxicity; and edu- cation of students at all levels, including the advanced life sciences, and medical and veteri- nary training. The use of animals in these three situations—research, testing, and education-dif- fers considerably, and each has different prospects for development of alternatives. The assessment excludes examination of the use of animals in food and fiber production; their use in obtaining organs, antibodies, and other biologi- cal products; and their use for sport, entertain- ment, and companionship. Such purposes include numbers of animals generally estimated to be many multiples greater than the numbers used for pur- poses described in this report (see ch. 3). Issues of animal care, such as feeding and maintenance, are also beyond the scope of this assessment. 3 — 4 G Alternatives to Animal Use in Research, Testing, and Education DEFINITION In this report, animal is defined as any non- human member of the five classes of verte- brates: mammals, birds, reptiles, amphibians, and fish (see ch. 2). Within this group, two kinds of animals can be distinguished—warm-blooded animals (mammals and birds) and cold-blooded ani- mals (reptiles, amphibians, and fish). Other crea- tures customarily included in the animal kingdom, such as invertebrates (e.g., worms, insects, and crustaceans), are excluded by this definition. The use of human subjects is not examined in this assessment. The concept of alternatives to animal use has come to mean more than merely a one-to-one substitution of nonanimal methods for animal tech- niques. For alternatives, OTA has chosen a def= inition characterized by the three Rs: replace- ment, reduction, and refinement. Scientists may replace methods that use animals with those that do not. For example, veterinary students may use a canine cardiopulmonary -resus - citation simulator, Resusci-Dog, instead of living dogs. Cell cultures may replace mice and rats that are fed new products to discover substances poi- sonous to humans. In addition, using the preced- ing definition of animal, an invertebrate (e.g., a horseshoe crab) could replace a vertebrate (e.g., a rabbit) in a testing protocol. Reduction refers to the use of fewer animals. For instance, changing practices allow toxicolo- gists to estimate the lethal dose of a chemical with as few as one-tenth the number of animals used in traditional tests. In biomedical research, long- lived animals, such as primates, may be shared, assuming sequential protocols are not deemed in- humane or scientifically conflicting. Designing ex- perimental protocols with appropriate attention to statistical inference can lead to decreases (or to increases) in the numbers of animals used. Or several tissues may be simultaneously taken from a single animal as a result of coordination among investigators. Reduction can also refer to the mini- mization of any unintentionally duplicative exper- OF TERMS Resusci-Dog, Canine Cardiopulmonary Resuscitation Simulator Photo credit: Charles R. Short, New York State College of Veterinary Medicine, Cornell University Resusci-Dog, a plastic mannequin linked to a computer, can simulate an arterial pulse, and pressure can be applied to its rib cage for cardiac massage or cardio- pulmonary resuscitation. Resusci-Dog has replaced about 100 dogs per year in the training of veterinary students at the New York State College of Veterinary Medicine. iments, perhaps through improvements in infor- mation resources. Existing procedures may be refined so that ani- mals are subjected to less pain and distress. Refine- ments include administration of anesthetics to ani- mals undergoing otherwise painful procedures; administration of tranquilizers for distress; hu- mane destruction prior to recovery from surgical anesthesia; and careful scrutiny of behavioral in- dices of pain or distress, followed by cessation of the procedure or the use of appropriate analgesics. Refinements also include the enhanced use of non- invasive imaging technologies that allow earlier detection of tumors, organ deterioration, or meta- bolic changes and the subsequent early euthana- sia of test animals. Pain is defined as discomfort resulting from in- jury or disease, while distress results from pain, anxiety, or fear. Pain may also be psychosomatic, resulting from emotional distress. Although these are subjective phenomena, pain and distress can —. Ch. l—Summary, Policy Issues, and Options for Congressional Action G 5 sometimes be identified and quantified by observ- objectives of procedures. Professional ethics re- ing an animal’s behavior. Pain is relieved with quire scientists to provide relief to animals in pain analgesics or anesthetics; distress is eased with or distress, unless administering relief would inter- tranquilizers. Widely accepted ethical standards fere with the objective of the procedure (e.g., when require that scientists subject animals to as little the objective is a better understanding of the mech- pain or distress as is necessary to accomplish the anisms of pain). HOW MANY ANIMALS ARE USED? Estimates of the animals used in the United States each year range from 10 million to upwards of 100 million. OTA scrutinized a variety of surveys (see ch. 3), including those of the National Research Council’s Institute for Laboratory Animal Resources and the Animal and Plant Health Inspection Serv- ice (APHIS) of the U.S. Department of Agriculture (USDA). Indirect estimates of animal use were also based on data such as Federal funds spent on ani- mal research and sales revenues of the Nation’s largest commercial breeder of laboratory animals. All these data are unreliable, No data source ex- ists, for example, to enumerate how many institu- tions do not report animal use. In addition, non- reporting institutions may not be similar enough to reporting institutions to justify extrapolation. Thus every estimate of animal use stands as a rough approximation. With this caveat in mind, the best data source available--the USDA/APHIS census —suggests that at least 17 million to 22 million animals were used in research and testing in the United States in 1983. The majority of ani- mals used—between 12 million and 15 million— were rats and mice. Current data permit no state- ment about any trends in animal use through re- cent years. Animal use in medical and veterinary education amounted to at least 53)000 animals in the school year 1983-84. The Animal Welfare Act of 1966 (Public Law 89- 544), as amended and presently enforced, requires research and testing facilities to report to USDA their annual use of dogs, cats, hamsters, rabbits, guinea pigs, and nonhuman primates (see ch. 13). (About two-thirds of the reporting institutions also volunteer the number of rats and mice used ). For fiscal year 1983, the USDA reporting forms indicate the facilities used nearly 1.8 million of these six kinds of animals (see table 1-1). Table I-l.—Animal Use Reported to the U.S. Department of Agriculture, 1983a Number used Animal in 1983 Dogs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182,425 Cats. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55,346 Hamsters . . . . . . . . . . . . . . . . . . . . , . . . . . 454,479 Rabbits . . . . . . . . . . . . . . . . . . . . . . . . . . . . 509,052 Guinea pigs , . . . . . . . . . . . . . . . . . . . . . . . 521,237 Nonhuman primates . . . . . . . . . . . . . . . . . 59,336 Total . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1,781,875 aTotalS do not include rats or mice, two species that together r@ Present the majority of animals used. SOURCE” Office of Technology Assessment. USDA reports are of limited utility because: G G G G the Department counts only six kinds of ani- mals that together account for an estimated 10 percent of the total animals used (report- ing of rats, mice, birds, and fish is not re- quired); the annual summary report does not tabulate reports received after December 31st of each year, resulting in a 10-to 20-percent underes- timation of laboratory use of regulated species; ambiguities in the reporting form ask respond- ents to add figures in a way that can cause animals to be counted twice; and terms on the reporting form are undefined (e.g., the form has room for voluntary infor- mation about “wild animals, ” but does not specify what animals might be included). In the absence of a comprehensive animal census, the USDA reports will continue to provide the best data. Imprecise as they are, these reports can iden- tify major changes in the numbers of dogs, cats, hamsters, rabbits, guinea pigs, and nonhuman pri- mates. (It is important to note that any change in the total number of animals used may reflect not only the adoption of alternative methods, but changes in research and testing budgets as well.) 6 G Alternatives to Animal Use in Research, Testing, and Education ETHICAL CONSIDERATIONS At one end of a broad spectrum of ethical con- cerns about animal use is the belief that humans may use animals in any way they wish, without regard for the animals suffering. At the other ex- treme is the notion--epitomized by the slogan ‘(ani- mals are people, too’’ —that each animal has the right not to be used for any purpose that does not benefit it. Each view is anchored in a school of phi- losophical thought, and people considering this is- sue can choose from a variety of arguable posi- tions (see ch. 4). Prominent within the Western philosophic and religious tradition is the view that humans have the right to use animals for the benefit of human- kind. This view is predicated on the assumption that human beings have special intrinsic value and thus may use natural animate and inanimate ob- jects, including animals, for purposes that will en- hance the quality of human life. Yet this tradition suggests that because animals are intelligent and sentient beings, they should be treated in a hu- mane manner. Current policies and trends within the scientific community have reinforced this con- viction by advocating that pain and suffering be minimized when animals are used in research, test- ing, or education. Advocates of what generally is called animal wel- fare frequently question the objectives of animal use, as well as the means. They point out that ani- mals can experience pain, distress, and pleasure. Drawing on the utilitarian doctrine of providing the greatest good for the greatest number, some animal welfare advocates weigh animal interests against human interests. In this view, it might be permissible to use animals in research to find a cure for a fatal human disease, but it would be unjust to subject animals to pain to develop a prod- uct with purely cosmetic value. ALTERNATIVES In research, scientists often explore un- charted territory in search of unpredictable events, a process that inherently involves un- certainty, missteps, and serendipity. Some bio logical research requires-and in the foresee Some animal rights advocates carry this concern a step further and do not balance human and ani- mal rights. They generally invoke the principle of inalienable individual rights. They believe that ani- mal use is unjustified unless it has the potential to benefit the particular animal being used. Ani- mal rights advocates refer to the denial of animal rights as a form of “speciesism,” a moral breach analogous to racism or sexism. Animals, by this reasoning, have a right not to be exploited by people. People throughout the spectrum find common ground in the principle of humane treatment, but they fail to agree on how this principle should be applied. Society does not apply the principle of humane treatment equally to all animals. A cat may evoke more sympathy than a frog, for exam- ple, because the cat is a companion species and possesses apparently greater neurological sophis- tication than a frog, endowing it with both favored status and a familiarity that suggests to humans that they can interpret its behavior. Even within a species, all individuals are not treated consist- ently. Pet rabbits in the home and pest rabbits in the garden, like human friends and strangers, are treated differently. The improvements in public health and safety made possible through the use of ani- mals in research and testing are well known. But these questions remain” Do these advances justify animal use? How much of the improve= ments were actually dependent on the use of animals? Debate on these and other questions is bound to continue, but most parties agree that con- sideration of replacing, reducing, and refining the use of animals is desirable. IN RESEARCH able future will continue to require-the use of live animals if the study of the complex in- teractions of the cells, tissues, and organs that make up an organism is to continue. Knowledge thus gained is applied to improving the health and — Ch. l—Summary, Policy Issues, and Options for Congressional Action G 7 well-being of humans and of animals themselves, and it may lead to the development of methods that would obviate the use of some animals. Some nonanimal methods are becoming available in biomedical and behavioral research (see ch. 6). As more develop, animal use in research will likely become less common. It is important to note, how- ever) that even if animals cannot be replaced in certain experiments, researchers can at- tempt to reduce the number used and also to minimize pain and distress. Most alternatives to current animal use in re- search fall into one of four categories: G G G G Continued, But Modified, Use of Animals. This includes alleviation of pain and distress, substitution of cold-blooded for warm-blooded vertebrates, coordination among investiga- tors, and use of experimental designs that pro- vide reliable information with fewer animals than were used previously. Living Systems. These include micro-organ- isms, invertebrates, and the in vitro culture of organs, tissues, and cells. Nonliving Systems. These include epidemio- logic databases and chemical and physical sys- tems that mimic biological functions. Computer Programs. These simulate biologi- cal functions and interactions. The many fields of research—ranging from anatomy to zoology—use animals differently, and each thus has different prospects for de- veloping and implementing alternatives. To de- termine the prevalence of animal and nonanimal methods in varied disciplines of research, OTA sur- veyed 6)000 articles published between 1980 and 1983 in 12 biomedical research journals and 3 be- havioral research journals (see ch. 5). Research dis- ciplines were distinguished by their characteris- tic patterns of animal use, as measured by the percentages of published reports showing animal use, no animal use, and use of humans. Animal methods predominated in most of the journals sur- veyed, including the three behavioral research journals. The exceptions in the overall survey were cell biology, which used primarily nonanimal meth- ods, and cardiology, which used primarily human subjects. Using alternative methods in biomedical re- search holds several advantages from scientific, economic, and humane perspectives, including: G G G G G G G G G reduction in the number of animals used; reduction in animal pain, distress, and exper- imental insult; reduction in investigator-induced, artifactual physiological phenomena; savings in time, with the benefit of obtaining results more quickly; the ability to perform replicative protocols on a routine basis; reduction in the cost of research; greater flexibility to alter conditions and vari - ables of the experimental protocol; reduction of error stemming from interindi- vidual variability; and the intrinsic potential of in vitro techniques to study cellular and molecular mechanisms. Many of these alternative methods are accom- panied by inherent disadvantages, including: G G G G G G G G G reduced ability to study organismal growth processes; reduced ability to study cells, tissues, and or- gan systems acting in concert; reduced ability to study integrated biochem- ical and metabolic pathways; reduced ability to study behavior; reduced ability to study the recovery of damaged tissue; reduced ability to stud-y interaction between the organism and its environment; reduced ability to study idiosyncratic or species-specific responses; reduced ability to distinguish between male- and female-specific phenomena; and a handicap to probing the unknown and phe- nomena not yet identified. Behavior encompasses all the movements and sensations by which living things interact with both the living and nonliving components of their envi- ronment. Since one of the chief goals of behavioral research is an understanding of human behavior, there are obvious advantages to the use of human research subjects. There are also advantages to using animals, including the following: G Laboratory research on animals offers a greater opportunity to control variables such 8 . Alternatives to Animal Use in Research, Testing, and Education G G G as genetic background, prior experience, and environmental conditions, all of which affect behavior and can obscure the influence of the factor under study. The short lifespans of certain animals allow scientists to study behavior as it develops with age and across generations. Some animal behavior is less complex than hu- man behavior, facilitating an understanding of basic elements and principles of behavior. The behavior of certain animals holds particu- lar interest for humans. These animals include companion species, farm animals, and agri- cultural pests. Although behavior is a biological phenomenon, behavioral research differs substantially from bio- medical research in that researchers have fewer opportunities to study mechanisms isolated from living organisms. There is little prospect, for ex- ample, of using in vitro cultures to look at aggres- sion, habitat and food selection, exploration pat- terns, or body maintenance activities—all topics studied by behavioral scientists. Yet in each of these disciplines, reduction or refinements of animal use may be possible. It is the continued, but modi- fied, use of animals that holds the most prom- ise as an alternative in the field of behavioral research. ALTERNATIVES IN TESTING Several million animals are used each year in testing substances for toxicity and establish- ing conditions for safe use. The resulting data— together with information about use and ex- posure, human epidemiologic data, and other information—are used in assessing and man- aging health risks. As a reduction in the number of animals is a prin- cipal alternative, proper statistical design and anal- ysis in testing protocols play an important role (see ch. 7). The total number of animals needed for sta- tistically significant conclusions depends on the incidence of toxic effects without administration of the test substance, the degree of variation from animal to animal for the biological effect that is of interest, and the need to determine a quantita- tive relationship between the size of the dose and the magnitude of the response. Statistical analy- sis plays a similarly important role in research. One of the oldest and, perhaps for that reason, least sophisticated tests is the LD50 (“lethal dose” for “50” percent of the test animals). In this short- term, or acute, test, a group of animals, usually rats or mice, are exposed to a single substance, and the measured end point is death (although other observations may be made). The LD50 is the dose at which half the test animals can be expected to die. A range of doses is administered to some 30 to 100 animals and the LD50 is calculated from the results. Tests providing the same informa- tion have recently been developed using as few as 10 animals, i.e., a 3- to 10-fold reduction, The LD50 is used to screen substances for their relative toxicity and mode of toxic action. Scien- tists and animal welfare advocates have criticized it in recent years, in part because it cannot be ex- trapolated reliably to humans, and in part because the imposition of a highly toxic or lethal dose seems particularly inhumane. Another often-criticized acute toxicity assay is the Draize eye irritancy test. This involves plac- ing a test substance into one eye of four to six rab- bits and evaluating its irritating effects. Results are used to develop precautionary information for sit- uations in which exposure of the human eye to the substance is possible. Substances with certain properties-e.g., a caustic pH-could be assumed to be eye irritants and not tested. Draize proce- dures may also be modified to reduce pain, and in vitro methods to test for irritancy are under development. A promising new bioassay for tis- sue irritancy makes use of the chorioallantoic mem- brane of the chick embryo (see fig. 1-1). Other common tests include those for long-term chronic effects, carcinogenicity, reproductive and developmental toxicity, skin irritancy, and neuro - toxicity. In addition to such descriptive toxicology (i.e., tests that focus on the response of the organ- Ch. l—Summary, Policy Issues, and Options for Congressional Action “ 9 Figure l-l.— Chronological Sequence Embryo Chorioallantoic Membrane of Chick Assay Day O / Day 3 Y I Ii Day O: Fertile eggs are incubated at 3 7oC. Day 3: The shell is penetrated in two places: A window is cut at the top, and 1.5 to 2 milliliters of albumin is removed with a needle and discarded. The chorioallantoic membrane forms on the floor of the air space, on top of the embryo. The window is taped. Day 14: A test sample is placed on the embryonic membrane and contained within a plastic ring. Day 17: The chorioallantoic membrane is evaluated for its response to the test substance, and the embryo is discarded. SOURCE J. Leighton, J, Nassauer, and R, Tchao, “The Chick Embryo in Toxicol- ogy: An Alternative to the Rabbit Eye,” Food Chem. Toxicol. 23:293-298. Copyright 1985, Pergamon Press, Ltd. ism as a whole), testing may also be done to deter- mine the mechanisms by which a substance is metabolized or excreted, and the chemical re- actions by which toxic effects are produced. Such studies of mechanistic toxicology aid in the selec- tion and design of descriptive tests. The Federal Government plays a major role in this area, both through laws that directly or Chick Embryo Chorioallantoic Membrane Assay Photo credit: Joseph Leighton, Medical College of Pennsylvania Typical reaction seen when certain concentrations of household products are placed on the 14-day-old chorio- allantoic membrane and examined 3 days later on 17-day - old membranes. The thin white plastic ring has an internal diameter of 10 millimeters (0.4 inch). The area of injury occupies the entire plastic ring. Damaged blood vessels appear within the ring as an elaborate branching structure of pale, white, dead vessels of various sizes. The severity of the reaction is gauged by measuring the diameter of the injury, in this instance spanning the entire ring indirectly require testing and through guide- lines that influence testing procedures. The greatest amount of testing is done under laws administered by the Food and Drug Administra- tion (FDA) requiring that products be safe and ef- fective and that labeling claims be substantiated. The Environmental Protection Agency (EPA) re- quires testing to support pesticide registrations and in certain other cases. For substances other than pesticides, EPA relies largely on published literature and EPA-sponsored testing. Other agen- cies that use animal testing data include USDA, the Consumer Product Safety Commission, the Oc- cupational Safety and Health Administration, the Department of Transportation, the Federal Trade Commission, and the Centers for Disease Control. Although most laws do not explicitly require animal testing, requirements of safety im- plicitly require that the best available means for determining safety be used. Thus, alterna- tives are not likely to be used widely until they can be shown to be at least as valid and relia- ble as the tests being replaced. Meeting these criteria is probably not overly difficult with some 10 . Alternatives to Animal Use in Research, Testing, and Education alternatives that involve reduction or refinement, but it maybe harder to replace whole-animal test- ing totally with in vitro methods. Reductions in the number of animals used can be brought about by using no more animals than necessary to accomplish the purpose of the test, by combining tests in such a way that fewer ani- mals are needed, and by retrieving information that allows any unintentional duplication of earlier work to be avoided (see chs. 8 and 10). Refinements include increased use of anesthetics and analgesics to ameliorate pain and tranquilizers to relieve dis- tress. Replacements may involve human cell cul- tures obtained from cadavers or in surgery, animal cell cultures, invertebrates, or micro-organisms. For example, the use of an invertebrate in place of a vertebrate, as in the case of substituting horse- shoe crabs for rabbits in testing drugs for their production of fever as a side effect, is increasingly accepted as a replacement. The most promising in vitro methods are based on an understanding of whole-organ or organism responses that can be related to events at the cel- lular or subcellular level. Cells manifest a variety of reactions to toxins, including death, changes in permeability or metabolic activity, and damage to genetic material. ALTERNATIVES IN EDUCATION Although far fewer animals are used in edu- cation than in either research or testing, ani- mal use in the classroom plays an important role in shaping societal attitudes toward this subject. As educational goals vary from level to level, so does the use of animals and therefore the potential for alternatives (see ch. 9). In elementary schools, live animals are gener- ally present solely for observation and to acquaint students with the care and handling of different species. Although the guidelines set by many school boards and science teachers’ associations limit the use of living vertebrates to procedures that nei- ther cause pain or distress nor interfere with the animals’ health, these guidelines are not observed in all secondary schools. Science fairs are an addi- tional avenue for students to pursue original re- search. The Westinghouse Science Fair prohibits the invasive use of live vertebrates, whereas the International Science and Engineering Fair has no such prohibition. In the college classroom and teaching laboratory, alternatives are being developed and implemented because they sometimes offer learning advantages, are cheaper than animal methods, and satisfy ani- mal welfare concerns. As a student advances, ani- mal use at the postsecondary level becomes in- creasingly tied to research and skill acquisition. As graduate education merges with laboratory re- search and training, animal use becomes largely Finalist, 1985 Westinghouse Science Talent Search Photo credit: Gary B. Ellis Louis C. Paul, age 18, Baldwin Senior High School, Baldwin, NY, with his research project, “Effect of Temperature on Facet Number in the Bar-Eyed Mutant of Drosophila melanogaster. ” a function of the questions under investigation. In disciplines such as surgical training in the health professions, some measure of animal use can be helpful but is not universally viewed as essential. Many alternative methods in education are already accepted practice (see ch. 9). Replace- ments include computer simulations of physiolog- ical phenomena and pharmacologic reactions, cell culture studies, human and animal cadavers, and audiovisual materials. Clinical observation and in- struction can also replace the use of animals in some laboratory exercises in medical and veteri- Ch. l—Summary, Policy Issues, and Options for Congressional Action G 1 1 nary schools. Reduction techniques include the use of classroom demonstrations in place of individ- ual students’ animal surgery and multiple use of each animal, although subjecting an animal to mul- tiple recovery procedures may be viewed as in- humane and counter to refined use. Refinements include the use of analgesics, euthanasia prior to recovery from surgery, observation of intact ani- mals in the classroom or in their natural habitats, and the substitution of cold-blooded for warm- blooded vertebrates in laboratory exercises. Humane education aspires to instill positive attitudes toward life and respect for living ani- mals. Instruction in proper care and handling of various species may be complemented by exposure to the principles of animal use in research and test- ing and to alternative methods. This type of edu- cation promotes attitudes conducive to the devel- opment and adoption of alternatives. COMPUTER SIMULATION AND INFORMATION RESOURCES Recent advances in computer technology hold some potential for replacing and reduc- ing the use of animals in research, testing, and education (see chs. 6, 8) 9, and 10). Inmost cases, however, research with animals will still be needed to provide basic data for writing com- puter software, as well as to prove the validity and reliability of computer alternatives. In research, scientists are developing computer simulations of cells, tissues, fluids, organs, and or- gan systems, Use of such methods enables less use of some animals. Limitations on the utility of com- puter simulations are due to a lack of knowledge of all the parameters involved in the feedback mechanisms that constitute a living system, which means the information on which the computer must depend is incomplete. In testing, computers allow toxicologists to de- velop mathematical models and algorithms that can predict the biological effects of new substances based on their chemical structure. If a new chem- ical has a structure similar to a known poison in certain key aspects, then the new substance also may be a poison. Such screening can thus preempt some animal use. ment and the effects of extraneous variables, help- ing students concentrate on a lesson’s main point. Aside from their direct use in research, testing, and education, computers also could reduce ani- mal use by facilitating the flow of information about the results of research and testing. Scien- tists routinely attempt to replicate results of ex- periments to ensure their accuracy and validity and the generality of the phenomenon. Uninten- tional duplication, however, can waste money and animal lives. To avoid such situations, the scien- tific community has established various modes of communication. Research and testing results are published in journals, summarized by abstracting services, discussed at conferences, and obtained through computer databases. One way any existing unintentional duplication might be ended, and thus animal use reduced, is to establish or refine existing computer-based regis- tries of research or testing data. The National Can- cer Institute and the National Library of Medicine (NLM) developed a limited registry in the late 1970s, but it failed: The Laboratory Animal Data Bank (LADB) had few users, as it did not serve user needs. Any new registry should contain descriptions In education, computer programs simulate class- of the methods of data collection and the labora - room experiments traditionally performed with tory results for both experimental and control animals. The most advanced systems are video- groups of animals. Inclusion of negative results disks that combine visual, auditory, and interac - (which are seldom reported in journals) could 40 tive properties, much as a real classroom experi- reduce animal use, Entries should undergo peer ment would. Computer simulations can eliminate review before inclusion in the registry; that is, both the detailed work of conducting an experi- studies should be scrutinized to judge the validity 12 . Alternatives to Animal Use in Research, Testing, and Education and reliability of the data. A registry along these lines would probably be 3 to 15 times as complex and costly as the unsuccessful LADB. As alternative methods are developed and im- plemented, a computerized registry of informa- tion about these novel techniques might serve to speed their adoption. In 1985, the NLM incorpo- rated “animal testing alternatives” as a subject head- ing in its catalogs and databases, which help users throughout the world find biomedical books, arti- cles, and audiovisual materials. In amending the Animal Welfare Act in 1985, Congress directed the National Agricultural Library to establish a serv- ice providing information on improved methods of animal experimentation, including methods that could reduce or replace animal use and minimize pain and distress to animals. ECONOMIC CONSIDERATIONS The total dollar cost of the acquisition and main- tenance of laboratory animals is directly related to the length of time animals stay in the labora- tory. With no accurate source of data on various species’ length of stay, it is impossible to calculate the actual total dollar cost of animal use. Reduc- ing the number of animals used can lower acqui- sition and maintenance costs. Yet, the overall sav- ings will not be proportionate to the smaller number of animals used, as the overhead costs of breeding and laboratory animal facilities must still be met. Animal use carries with it both great expense and major economic and health benefits (see chs. 5, 7, and 11). Nonetheless, it is difficult to ex- press many of the costs and benefits monetarily. What price does society put on the pain and dis- tress of an animal used in research, for example, or on the life of a person saved by a new medical treatment that was made possible by the use of animals? In research, there is no way of knowing when a particular result would have been obtained if an experiment had not been done. Thus, it is im- possible to predict many of the costs related to the use of alternatives in research. Attempts to do so are likely to result in economic predictions with little basis in fact. The primary reason a company conducts ani- mal tests is to meet its responsibilities to make safe products under safe conditions. For pharmaceu- ticals, the need extends to the assurance of product effectiveness. In testing, animal methods gener- ally are more labor-intensive and time consuming than nonanimal methods, due to the need, for ex - ample, to observe animals for toxic effects over lifetimes or generations. Testing can cause delays in marketing new products, including drugs and pesticides, and thus defer a company’s revenue. Rapid, inexpensive toxicity tests could yield ma- jor benefits to public health. There are more than 50,000 chemicals on the market, and 500 to 1)000 new ones are added each year. Not all must be tested, but toxicologists must expand their knowl- edge of toxic properties of commercial chemicals if human health is to be protected to the extent the public desires. Rapid and economical testing would facilitate the expansion of that knowledge. Government regulatory practices can be read as promoting animal testing although the laws and practices appear flexible enough to accept alternatives when such tests become scientifi- cally acceptable. To date, regulatory practices have not, in fact, provided a basis for companies to expect that acceptance of alternative methods will be an expedient process. In addition to re- sponding to regulatory requirements, companies conduct animal tests to protect themselves from product liability suits. Here, the necessary tests can exceed government requirements. Because of the great expense and long time re- quired for animal research and testing, priority in research results has considerable value to in- vestigators and testing results bear considerable proprietary value for industry. Some data are made public by statute, and various arrangements can be made for sharing testing costs. Yet many data are held in confidence, for example, by the com- pany that generated them. Ch. l—Summary, Policy Issues, and Options for Congressional Action G 1 3 FUNDING FOR THE DEVELOPMENT OF ALTERNATIVES The Federal Government does not explicitly fund the development of alternatives to animal use per se. Because research on and development of alternatives is founded on a broad base of disci- plines, it is difficult to ascertain the dimensions of the effective level of support. No category of research funds, for example, distributed by the National Institutes of Health (NIH) or the National Science Foundation is earmarked for the develop- ment of alternatives. Yet despite this lack of iden- tifiable, targeted funding, Federal dollars do sup- port areas of testing and research that generate alternatives. In biomedical and behavioral research, it is not clear whether targeted funding efforts would pro- duce alternatives faster than they are already being devised. The research areas most likely to result in useful alternatives include computer simu- lation of living systems; cell, tissue, and organ culture technology; animal care and health; and mechanisms of pain and pain perception. Funding to improve animal facilities can result in healthier, less stressed animals and can free research from confounding variables bred by a less well defined or inferior environment. Some Federal agencies, notably the National Toxi- cology Program and FDA, conduct in-house re- search on alternatives to animal testing, as do some corporations. Industry has also committed funds to university researchers seeking alternatives. Rev- lon has given $1.25 million to the Rockefeller University to support research on alternatives to the Draize eye irritancy test. The Cosmetic, Toi- letry, and Fragrance Association and Bristol Myers Company have given $2.1 million to the Center for Alternatives to Animal Testing at The Johns Hop- kins University, which funds research into test- ing alternatives, especially in vitro methods. Alternatives to animal use in education gener- ally build on techniques developed in research and funded by research monies. Some Federal support for research in science education addresses the development of alternatives, particularly in the area of computer simulation. In 1985, the enact- ment of Public Law 99-129 authorized the Secre- tary of the Department of Health and Human Serv- ices to make grants to veterinary schools for the development of curriculum for training in the care of animals used in research, the treatment of ani- mals while being used in research, and the devel- opment of alternatives to the use of animals in re- search. Colleges and universities may offer courses re- lated to humane principles or principles of experi- mentation. In addition, animal welfare groups are active sponsors in the areas of humane education and attitudes about animals. A number of humane societies and animal welfare groups fund research on alternatives in research, testing, or education. Several private foundations, notably the Geraldine R. Dodge Foundation, des- ignate support for research in animal welfare as among their funding missions. REGULATION OF ANIMAL USE Several Federal and State laws, regulations, Care and Use of Laboratory Animals by Awardee guidelines, and institutional and professional so- Institutions (revised in 1985; see app. C). cieties’ policies affect the use of animals in research and testing (see chs. 13, 14, and 15; app. B). Chief among these are the Animal Welfare Act, the Health Federal Regulation Research Extension Act of 1985 (Public Law 99- 158), rules on good laboratory practices established Prompted by publicity about pet dogs used in by FDA and EPA, the NIH Guide for the Care and research, Congress passed the Animal Welfare Act Use of Laboratory Animals (revised in 1985), and to halt the use of stolen pets in experimentation. the Public Health Service (PHS) Policy on Humane Enacted in 1966 and amended in 1970, 1976, and 74 G Alternatives to Animal use in Research, Testing, and Education 1985, the statute also contains provisions for the care and treatment of certain animals used in ex- periments. The act defines “animal” as: . . . any live or dead dog, cat, monkey (nonhuman primate animal), guinea pig, hamster, rabbit, or such other warm-blooded animal, as the Secre- tary [of the Department of Agriculture] may de- termine is being used, or is intended for use, for research, testing, experimentation or exhibition purposes . . . USDA, empowered to identify other mammals and birds to be regulated, has done so only for marine mammals. In fact, in 1977, USDA promul- gated a regulation excluding birds, rats, mice, and horses and other farm animals from coverage by the Animal Welfare Act. The use of rats and mice, the most common laboratory animals, is therefore not regulated. The act does not cover facilities that use none of the regulated species. Facilities that use regu- lated species but that receive no Federal funds and maintain their own breeding colonies also fall out - side the act’s coverage. The Animal Welfare Act regulates housing, feeding, and other aspects of animal care but bars USDA from regulating the design or performance of actual research or testing. A facility need only report annually that the provisions of the act are being followed and that professionally acceptable standards are being followed during actual experi- mentation. Facilities must also describe procedures likely to produce animal pain or distress and pro- vide assurances that alternatives to those proce- dures were considered. The Food Security Act of 1985 (Public Law 99- 198) amended the Animal Welfare Act (amend- ments effective December 1986) to strengthen standards for laboratory animal care, increase en- forcement of the Animal Welfare Act, provide for the dissemination of information to reduce unin- tended duplication of animal experiments, and mandate training for personnel who handle ani- mals. For the first time, the Department of Health and Human Services is brought into the enforce- ment of the Animal Welfare Act, as the Secretary of Agriculture is directed to “consult with the Sec- retary of Health and Human Services prior to the issuance of regulations” under the act. Each research facility covered by the Animal Welfare Act—including Federal facilities—is re- quired to appoint an institutional animal commit- tee that includes at least one doctor of veterinary medicine and one member not affiliated with the facility. The committee shall assess animal care, treatment, and practices in experimental research and shall inspect all animal study areas at least twice a year. Many groups concerned about animal welfare want the act and its enforcement strengthened, They criticize USDA’s exclusion of rats and mice, the level of funding for enforcement, and the choice of USDA’s Animal and Plant Health Inspection Serv- ice as the enforcement agency. Inspectors, whose primary concern is preventing interstate transport of disease-carrying livestock and plants, spend about 6 percent of their time enforcing the re- search provisions of the Animal Welfare Act. Ad- ditional criticism is leveled at the act’s failure to offer guidance in research practices during experi- mentation. A 1982 report by the Humane Society of the United States indicates that USDA regula- tions and guidelines failed to provide “information sufficient to demonstrate that researchers have used pain-relieving drugs ‘appropriately’ and in accord- ance with ‘professionally acceptable standards ’.” The Health Research Extension Act of 1985 mandates the establishment of animal care com- mittees at all entities that conduct biomedical and behavioral research with PHS funds. [t requires all applicants for NIH funding to submit assurances that they are in compliance with the law’s provi- sions for the operation of animal care committees and that all personnel involved with animals have available to them training in the humane practice of animal maintenance and experimentation. The NIH Director is empowered to suspend or revoke funding if violations of the act are found and not corrected. In essence, the act puts the force of Federal law behind certain elements of the PHS Policy. The act also directs the NIH Director to estab- lish a plan for research into methods of biomedi- cal research and experimentation that do not re- quire the use of animals, that reduce the number of animals used, or that produce less pain and dis- tress in experimental animals than methods cur- rently in use. Ch. l—Summary, Policy Issues, and Options for Congressional Action G 1 5 FDA and EPA both established rules on good lab- oratory practices to ensure the quality of toxicity data submitted by industry in compliance with the agencies’ regulations. Because proper animal care is essential to good animal tests, these rules in- directly benefit animals. The NIH Guide for the Care and Use of Labora- tory Animals prescribes detailed standards for ani- mal care, maintenance, and housing. It applies to all research supported by NIH and is in fact used by most animal facilities throughout the public and private sector. The Department of Defense (DOD) has been crit- icized for its use of animals in weapons research and in training for treatment of wounds, In 1973, Congress prohibited DOD from using dogs for re- search and development of chemical or biological weapons. In 1983, publicity caused an uproar about the use of dogs, pigs, and goats to train mili- tary surgeons in the treatment of gunshot wounds. The furor led to congressional action that pro- hibited DOD from using dogs and cats in such train- ing during fiscal years 1984 and 1985. State Regulation Most State anticruelty statutes forbid both ac- tive cruelty and neglect (see ch. 14). Many of these laws incorporate vague terms, and alleged offend- ers offer a variety of defenses. Enforcement may be delegated to humane societies, whose members are not well trained to build criminal cases skill- fully and are underfunded for the task. Twenty States and the District of Columbia reg- ulate the use of animals in research to some ex- tent. As in the case of the Federal Animal Welfare Act, most State laws address such matters as procurement rather than the actual conduct of experiments. All 50 States and the District of Columbia allow some form of pound animal use for research and training. In some States, laws permitting or requir- ing research and teaching facilities to purchase stray dogs and cats from pounds and shelters have been the targets of repeal efforts. To date, 9 States prohibit in-State procurement (although not im- portation from out-of-State) of pound animals for research and training. Of these, Massachusetts will in October 1986 prohibit the use of any animal obtained from a pound. Institutional and Self-Regulation Opponents of increased government regulation of research assert that investigators and their in- stitutions are best suited to determine what con- stitutes appropriate care and use of animals. To regulate animal use at this level, the scientific com- munity relies on a variety of policies and adminis- trative structures (see ch. 15). Taken together, the requirements for institu- tional animal committees contained in the Ani- mal Welfare Act (as amended), the Health Re- search Extension Act of 1985, and the PHS policy bring the overwhelming majority of experimental-animal users in the United States under the oversight of a structured, local re- view committee. Institutions that receive funds from PHS for re- search on warm-blooded laboratory animals must have committees that oversee the housing and rou- tine care of animals. NIH reports that about a quar- ter of these animal care and use committees cur- rently review research proposals to determine whether experimental procedures satisfy concerns about animal welfare. Committees with such re- sponsibility are not unique to research with ani- mals: For 15 years, similar groups have been weigh- ing ethical issues raised by the use of human research subjects, and these committees have served as models in the development of animal care and use committees. Committees usually have included the institu- tion’s attending veterinarian, a representative of the institution’s administration, and several users of research animals. Some committees also have nonscientist members, or lay members not affil- iated with the institution. Nonscientist and lay seats have been filled by clergy, ethicists, lawyers, hu- mane society officials, and animal rights advocates. Animal care and use committees at PHS-sup- ported facilities are today required to consist of not less than five members, and must include at least: G one Doctor of Veterinary Medicine with training or experience in laboratory ani- 76 G Alternatives to Animal Use in Research, Testing, and Education mal science or medicine, who has respon- sibility for activities involving animals at the institution; G one practicing scientist experienced in re- search using animals; G one member whose primary concerns are in a nonscientific area; and G one individual who is not affiliated with the institution in any way. The minimum committee structure required by the PHS policy is thus more rigorous than that man- dated by Federal law. The Animal Welfare Act and the Health Research Extension Act do not require, for example, that the committee veterinarian be trained in laboratory-animal medicine. The acts require a minimum committee of three individ- uals, whereas the PHS policy requires five. Institutional regulation generally entails compli- ance with some type of minimum standards for an animal facility, usually those of the NIH Guide for the Care and Use of Laboratory Animals. Com- pliance can be checked in-house or through ac- creditation by the American Association for Ac- creditation of Laboratory Animal Care (AAALAC), a voluntary private organization. As of April 1985, a total of 483 institutions had received AAALAC accreditation, which requires site visits that include interviews, inspection of facilities, and review of policies and records. Accredited institutions in- clude hospitals, universities, facilities of the Vet- erans’ Administration (VA), and pharmaceutical manufacturers (see app. D). A number of scientific and professional socie- ties, universities, and corporations have promul- gated statements of policy concerning their mem- bers’ or employees’ standards of conduct in animal use. These policies generally require: G G G G humane care and use of animals, minimization of the number of animals used, alleviation of pain and suffering, and supervision of animal use by qualified personnel. Twelve of fifteen such policies reviewed by OTA encourage or require consideration of the use of alternatives. But only 3 of the 15 include enforce- ment provisions or mention sanctions against vio- lators. Regulation Within Federal Agencies Six Federal departments and four independent agencies use laboratory animals intramurally and account for approximately one-tenth of the animal use in the United States. Beginning in December 1986, Federal facilities in those departments and agencies using animals will be required by the 1985 amendments to the Animal Welfare Act to install institutional animal committees. Each committee shall report to the head of the Federal agency con- ducting the experimentation. Most Federal agencies that use animals in re- search or testing have formal policies and admin- istrative structures to ensure that the animals re- ceive humane treatment. At the request of the Executive Office of Science and Technology Pol- icy, the Interagency Research Animal Committee developed a 450-word policy statement, Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Education, to be followed by all Federal agencies supporting ani- mal use (see ch. 13). No one Federal agency policy on animal care and use has all the characteristics needed toad- dress all issues adequately. Combining certain aspects from each would produce an effective uniform Federal policy. Almost all policies today require adherence to the NIH Guide and the Ani- mal Welfare Act. Most agencies also require an attending veterinarian and an animal care and use committee at each facility. The committees gen- erally review research protocols to ensure that animals are not used in excessive numbers, that adequate provisions are made for animal care and pain relief, and that alternatives are used when- ever possible. Most committees and attending veterinarians have little enforcement power, and those who have such power rarely use it. Some agencies’ policies have features that would be considered advantageous by animal welfare ad- vocates. NIH and the National Aeronautics and Space Administration have laypeople on their ani- mal care and use committees. The VA requires all its animal facilities to acquire AAALAC accredita- tion. The Department of Defense has a separate policy and committee for nonhuman primates. The Air Force has solicited evaluation of its policies by a panel of independent experts and plans to im- plement the group’s recommendations. Ch. l—Summary, Policy Issues, and Options for Congressional Action 17 International Regulation OTA surveyed laws controlling use of experi- mental animals in 10 foreign nations, including countries of Western Europe (see table 1-2) and Australia and Canada. Comparative analysis of reg- ulation of animal use abroad can yield lessons from foreign regulatory experiences, models for regu- lation, and models for funding of alternatives. A review of foreign laws, especially those revised or instituted in the last decade, indicates three trends of note in government control of animal research (see ch. 16): G Attention is shifting away from intentionally or negligently “cruel” treatment and toward the avoidance of pain and suffering. This change in perspective raises the difficulty of defining prohibited conduct, and disagree- ment arises over the definition of animal pain and suffering. Newer statutes rely on author- ized reviewers who check experimental plans in advance and apply their own sensibilities to satisfy themselves—and thereby the pub- lic interest–that pain and suffering are not being inflicted without justification. G There is increasing emphasis on finding alter- natives. The old method of justifying animal research by reference to its potential for pro- viding new knowledge is being enhanced by the greater burden of demonstrating that no less painful method is available to achieve the same result. Increasingly, animals are being viewed as having an interest in not being hurt. Countries with comprehensive reporting sys- terns (e.g., the United Kingdom) have found that fewer animals are now being used in ex- periments. The data are insufficient to deter- mine the reasons for these reductions or what the effect may be on the production of new information. These trends indicate a growing interest in Western Europe in replacing, reducing, or re- fining the use of animals through legislation. It is not clear whether the tighter control found in some West European countries can be applied in the United States. Most West Euro- pean nations are more homogeneous than is this country of federated States. In geographical dis- persal and size, the research enterprises in those countries are small—there are fewer than 300 in- vestigators using animals in Denmark, for exam- ple. The British system functions well, despite its complexity, because it has been refined over the course of a century. New scientists are weaned on it, and the inspector is a familiar sight in the laboratory. The British system’s enforcement is based more on advice and negotiation than on con- frontation. P O L I C Y I S S U E S A N D O P T I O N S F O R C O N G R E S S I O N A L A C T I O N Seven policy issues related to alternatives to ani- mal use in research, testing, and education were identified during the course of this assessment. The first concerns the implementation of alterna- tives and examines options that might encourage the research, testing, and education communities to adopt currently available methods of replacing, reducing, and refining their use of animals. The second issue explores options for promoting re- search and development leading to more and bet- ter alternatives. Both recognize that scientifically valid alternative methods can make positive con- tributions to research, testing, and education and might therefore be promoted. The five additional policy issues examined are: disseminating information about animal experi- mentation, restricting animal use, counting ani- mal use, establishing a Federal animal use policy, and changing the implementation of or amending the Animal Welfare Act. Although these policy is- sues do not explicitly address either the implemen- tation or development of alternative methods, they are inextricably linked to the replacement, reduc- tion, and refinement of animal use. Associated with each policy issue are several op- tions for congressional action, ranging in each case from taking no specific steps to making major Table 1=2.—National Laws for the Protection of Animals in Selected European Countries Federal Republic Provisions Denmark of Germany Netherlands Norway Sweden Switzerland United Kingdom Species protected . . . Vertebrates All animals Vertebrates, native Vertebrates Vertebrates Vertebrates Distinctions among species . . . Should use lowest Better to use rank; dogs, cats, invertebrates or cold- monkeys purpose-bred blooded vertebrates Alternatives must be used if available . . . . . . . . .Yes Anesthetics, analgesics, or approval required for painful experiments . . . . . . Except for minor or transient pain Educational uses. . . . Higher education, technique Ban on animal use for more than one painful experiment .All dogs, cats, monkeys; most experiments License/permit for dealers, facilities, and investigators ., .All facilities, head investigators Review of experiments . . . . . . . Most experiments need approval by national Board Administration. . . . . . Centralized, government/ nongovernment board iicensee is responsible Animal welfare representation ...3 nominees to national Board Reporting . . . . . . . .Annual report Yes if pain, suffering, or injury likely High school and above No multiple surgeries on vertebrates Dealers, facilities, investigators Not needed; proposed that facility’s animal welfare officer review States enforce and administer (proposed that facilities have animal welfare officer) Being considered in-house recordkeeping species Vertebrates better protected Vertebrates If injury or pain likely University and vocational Rarely reused because of pain requirements Dealers (dogs and cats), facilities Head of institute reviews Central enforcement and reporting; administration by institute Not required, but facility reports are public Annual report Vertebrates, crustaceans Monkeys, dogs, cats better protected Yes if pain is possible (unless Board approves) Professional training Only one experiment allowed per animal Investigators or facilities licensed investigator or facility (licensee) review Central coordination, some functions delegated to licensees Not required Annual report Should use lowest Should use lowest rank rank; all purpose-bred Alternatives promoted Surgery on mammals unless committee approves Allowed, but restricted Rarely reused because of pain requirements Breeders, facilities Notification/application; tiered system Central coordination with oversight by facility head and committee On all committees; being reconsidered Government recordkeeping Yes Slight pain or anxiety; if too painful, must forgo Not allowed Only reused if pain was slight Breeders, facilities 2 State committees review Central coordination, administered by States Members of national commission in-house recordkeeping . - SOURCE: Office of Technology Assessment Primates, dogs, cats, equidae preferred; no stray dogs Alternatives encouraged Statute does not specify, but certificate may require Some demonstration; not for practicing if anesthetized or because of pain requirements Facilities registered, investigators licensed Home Office and Advisory Committee Centralized, shared by Head Office, Advisory Committee, Royal Society Advisory Committee Annual reports Ch. l—Summary, Policy Issues, and Options for Congressional Action 19 changes. The order in which the options are pre- sented should not imply their priority. Further- more, the options are not, for the most part, mutu- ally exclusive: Adopting one does not necessarily disqualify others in the same category or within any other category. A careful combination of op- tions might produce the most desirable effects. In some cases, an option may suggest alterations in more than one aspect of alternatives to using animals, It is important to keep in mind that changes in one area have repercussions in others. Some of the options involve direct legislative ac- tion. Others are oriented to the actions of the ex- ecutive branch but involve congressional oversight or encouragement. Congress can promote alterna- tives in at least three ways, It can provide incen- tives through tax policies, grants, or educational assistance. It can mandate the adoption or devel- opment of alternatives by means of appropriations or legislation. And it can provide encouragement via oversight or resolutions. Table 1-3 summarizes the seven policy issues and associated options de- rived from this assessment. ISSUE: Should steps be taken to encourage the use of available alternatives in research, testing, or education? Alternatives to animals become accepted prac- tice in the research, testing, and educational com- munities as methods are developed through re- search, validated by independent measurements, gradually accepted by the scientific community, and implemented as they come to be relied on or required. Several alternatives to the use of animals are in the validation or implementation phase to- day; for the most part, these methods are based on reductions and refinements. Approaches that replace the use of animals have generally not been completely validated and accepted. Instead, these represent possibilities for the longer term. (An ex- ception may be educational simulations of living systems where an adequate range of physiologi- cal variables is known. ) The processes of valida- tion and gradual implementation are certain to con- tinue, and they could be accelerated. Analysis of alternatives in research (see ch. 6), testing (see ch. 8), and education (see ch. 9) dem- onstrates differing availability both among and within these three areas. In research, for exam- ple, animal methods can be complemented by com- puter models, and experiments may be designed to provide the desired information with fewer ani- mals. Dissemination of information within the re- search community may reduce any instances of unintentional duplication, thereby lowering the number of animals used. In testing, the LD50 pro- tocol has in many cases been modified to use fewer animals. And eye irritancy can be assumed—with- out testing—for substances exhibiting strong skin irritation or having a strongly acid or alkaline pH. In educational settings, exercises not involving ani- mals may be substituted to teach the scientific method or to introduce biological concepts. In other instances, animals are destroyed humanely following a single surgery in a teaching session, rather than experiencing multiple recovery pro- cedures. Four options address the implementation of alternatives such as these. Option I: Take no action. As alternatives are developed and validated, they are likely to continue being implemented at an un- even pace, influenced by factors largely external to Congress. Science and technologies will continue to evolve, and as nonanimal methods emerge from research and validation, they may or may not be accepted and implemented by the scientific com- munity. This course does not necessarily pass judgment on the value of adopting alternatives per se. Nor does it mean that alternatives will not be imple- mented. It would merely indicate that Congress has decided against encouraging or forcing the im- plementation of alternatives beyond its direction in 1985 to NIH to establish a plan to develop and assess alternatives in biomedical research (Public Law 99-158). This option might illustrate the be- lief that external political, ethical, economic, and scientific factors are sufficient to govern the im- plementation of alternatives. Further congressional action toward implemen- tation might be judged unnecessary because vari- ous other sources are already acting to implement alternatives. For example, EPA has defined circum- stances where the LD50 test can be replaced by a limit test (see ch. 8), and FDA has stated that it does not require data derived from the LD50 test; industry is watching to gauge the practical effects . . Table 1-3.-Policy Issues Related to Alternatives to Animal Use and Options for Congressional Action Policy issue Using existing Developing new Disseminating Restricting Counting animals Establishing a Federal Changing Animal alternatives alternatives information animal use used animal-use policy Welfare Act Options for congressional action Take no action Take no action Charge a Federal entity charge a Federal with coordinating the entity with implementation of coordinating the alternatives development of Encourage alternative alternatives methods in Federal Fund development of testing requirements alternatives Ban procedures for which alternatives are available aAnimal and plant Health Inspection ServiCe. SOURCE: Office of Technology Assessment. Take no action Mandate easy access to federally funded testing and research data Promote greater use of testing data submitted to Federal agencies Require literature searches Create new data- bases Translate foreign literature into English Take no action Restrict use of certain kinds of animals Restrict use of certain protocols Restrict acquisition of animals from certain sources License animal users for certain protocols and/or kinds of animals Prohibit animal use Take no action Eliminate APHISa census Correct inadequacies in present APHISa reporting system Expand APHISa census to include rats and mice Establish independent census Take no action Establish intramural Federal policy of minimum standards Take no action Eliminate funding for enforcement Increase funding for enforcement Amend to expand coverage to include experimentation Amend to realign enforcement authority Amend to preempt State and local laws — Ch. l—Summary, Policy Issues, and Options for Congressional Action G 2 1 of these statements. Also, members of the soap and detergent industry have implemented modifi- cations of the LD50 test. Noteworthy, too, is the important role of institutional animal care and use committees in all phases of animal experimenta- tion. In education, medical schools are conduct- ing some laboratory exercises with computer simu- lations or video demonstrations in lieu of live animals. Medical students in some instances by- pass experiments and training involving animals, proceeding from cadavers to people. Activities such as these are likely to continue without new con- gressional action. Additional congressional steps may be deemed inappropriate because implementation of alterna- tives may be judged unimportant. Some people do not object to animal use, for example, in toxico- logical testing. They believe the status quo brings the comforts and health benefits of new products and technology and protects them from hazards. Option 2: Require a new or existing Federal en- tity to coordinate the validation and im- plementation of alternatives. This action is based on the assumption that vali- dation and implementation of alternatives would occur more rapidly with enhanced Federal coordi- nation. Along this line, an information service at the National Agricultural Library on improved methods of animal experimentation was mandated by Congress in 1985 (Public Law 99-198). A clear- inghouse for resources required to implement alternatives would further hasten their adoption. This entity might, for example, be a central source for computer software or cell culture material. Existing Federal entities that might be assigned such responsibilities include some component of the National Institutes of Health (e.g., the Division of Research Resources), the National Toxicology Program, or the National Center for Toxicological Research. Coordinating activities could include symposia, workshops, newsletters, scholarships, grants, and the issuance of model protocols or guidelines. The coordinating body could monitor both public and private initiatives. In 1985, Con- gress took a step toward coordination of the use of alternatives in biomedical research conducted by or through NIH. It directed NIH to disseminate information about alternatives found to be valid and reliable to those involved in animal experimen- tation (Public Law 99-158). Educational programs play a central role in this type of effort. Training scientists in replacement methods and raising awareness about reductions and refinements is likely to increase the implemen- tation of alternatives. This type of education is closely allied with the teaching of principles of hu- mane care and use (see ch. 9). Animal care and use committees at individual institutions might function as a relay between Fed- eral coordination efforts and individual investiga- tors (see ch. 15). The institutional animal care and use committee might be required to suggest alter- native methods as part of its review of animal care and use. Linked in this way to a Federal implemen- tation effort, these committees would both feed into and draw on the resources of the Federal entity. A different type of coordination, particularly in research, would be the attachment of provisions to Federal grants regarding the implementation of alternatives. Research grant applications using alternative methods could be awarded higher pri- ority scores in the grant evaluation process or be otherwise favored. This strategy would require sufficient flexibility to ensure that valuable, state- of-the-art scientific proposals that may not involve alternatives are not handicapped. Funding mech- anisms could also be used to encourage coordina- tion between laboratories. The responsibility for overseeing the implementation of alternatives via funding mechanisms could be borne by each source of Federal funding (see ch. 12). Option 3: Encourage regulatory agencies to re- view existing testing guidelines and re- quirements and to substitute alterna- tives whenever scientifically feasible. Through oversight or legislation, Congress could encourage or require Federal agencies to evalu- ate existing alternatives in testing, to participate in their validation, to adopt them where appro- priate, and to report to Congress on their prog- ress in implementing alternatives, as the NIH has been asked to do (Public Law 99-158). Such agency review would have to be a periodic or continuing effort, given rapid advances in the state of the art. Some review of testing guidelines now occurs in keeping requirements up to date, although the pur- pose of that review is to improve the science rather than to protect animals per se. Formal agency re- view of international testing guidelines, such as 22 G Altematives to Animal Use in Research, Testing, and Education those of the Organization for Economic Coopera- tion and Development, could also be encouraged (see ch. 7 and app. E). The costs of agency review should be moderate, entailing input from agency experts, comment from outside experts, and pub- lication. If Federal laboratories were involved in the validation of alternative testing methods, ad- ditional costs would be incurred. Such a policy could encourage industry to develop alternatives because the barriers to acceptance would be reduced. Option 4: Ban procedures for which alternatives are available, or give a Federal agency authority to ban procedures as valid alternatives become available. This option recognizes that prohibitions can be used to force technological change. Prohibiting procedures for which scientifically acceptable alternatives are already available would acceler- ate the implementation of such alternatives. Ex- isting reductions and refinements in animal use include the greater use of analgesics in research, the use of fewer animals in the LD50 and Draize eye irritancy tests, and reliance on videotaped dem- onstrations and computer simulations in edu- cation. A ban could not only force implementation of existing alternatives, but, over time, help focus the development of new techniques (as discussed in the next section) and allow considerable flexibil- ity in achieving the desired end. A disadvantage of banning a specified procedure is that the replace- ment, or the process of developing one, may be even more politically unacceptable (e.g., the in vitro culture of human fetal nerve cells). A prohibition also takes no account of the question of judging the scientific acceptability of an alternative. In pursuing this option or the preceding one, it is important to appreciate that the swiftest adop- tion of alternatives may come about if regulatory agencies avoid mandating specific testing require- ments. Requiring specified tests might actually serve as a strong inhibitor to the implementation (and development) of alternative methods. Greater flexibility is achieved when testing requirements are defined in a manner that allows judgment and encourages use of alternate methods, Viewed from this perspective, the adoption of alternatives might be best stimulated by regulatory requirement for evaluation of a potential toxic response, such as mutagenicity, rather than requirement of a speci- fied test for mutagenicity. ISSUE: Should the more rapid development of new alternatives in research, testing or education be stimulated? Alternatives are currently being developed in many phases of animal use. It is worth noting that development of many of these techniques, espe- cially their validation, cannot occur without ani- mals being used (unless humans are used instead). In addition, many replacement systems will never be fully divorced from animal research and test- ing, and therefore they will serve to reduce but not eliminate animal use. Certain research and testing methods now be- ing developed, such as in vitro culture of animal components, bear great promise as alternatives. Similarly, the growing capabilities of computer modeling, for example biological simulation (see ch. 6) and pharmacology (see ch. 8), may reduce the number of animals needed. Development of an enhanced ability to detect and relieve pain can help refine animal use. Research that spawns alternatives usually takes place across traditional disciplinary lines—princi - pally within the life sciences–but also in applied mathematics, statistics, engineering, physics, and chemistry. The principal support for such research comes from Federal funds, predominantly NIH and the National Science Foundation. In general, there is little incentive for private investment in meth- odologies at a stage so remote from commerciali- zation and, in the case of testing, so governed by regulation. Some private concerns, however, spe- cifically fund research into alternative testing methods (see ch. 12). Clearly, research and development require money. Determining the optimum level of fund- ing, however, and the best way to distribute funds remains elusive, Nonetheless, the promotion of such research is likely to increase the number of alternatives available for implementation; in turn, increased implementation is likely to spur research in this area. Option 1: Take no action. If Congress takes no specific steps beyond its recent charge to NIH to establish a plan for the Ch. l—Summary, Policy Issues, and Options for Congressional Action G 2 3 development of alternatives in biomedical re- search, the development of alternatives will con- tinue to be a function of ethical, political, economic, and scientific factors. That alternatives are being developed in the ab- sence of direct legislation is best illustrated by re- search centers at Rockefeller University and The Johns Hopkins University funded by corporate and private donations (see ch. 12). In addition, corpo- rations are undertaking work in-house or sponsor- ing it in universities, often in response to scien- tific, economic, animal welfare, and public relations considerations. An uncertain pace of development marks the chief disadvantage of this option. Although alter- natives may emerge, changing research priorities in both the public and private sectors will affect the rate of development. From another perspec- tive, this is an advantage: It permits researchers to respond to changing needs and priorities with minimal Federal interference. Option 2: Require a new or existing Federal en- tity to coordinate the development of alternatives. Implementation of this option would have great symbolic value within the scientific and animal wel- fare communities and could lead to more rapid development of alternatives. A central clearing- house for the development of alternatives could compile and maintain records of all federally funded research and development (R&D) on alter- natives. Information on R&Din the private sector would be a valuable component of the coordina- tion effort, though it may prove difficult to obtain. Coordination could involve identifying research areas likely to lead to new alternatives and review- ing Federal support for those areas across agency lines. The latter responsibility might preclude hous- ing this entity within an existing Federal agency involved in funding R&D on alternatives to avoid either a real or apparent conflict of interest. As in the implementation of alternatives (see pre- ceding issue), education plays a central role in the development of such approaches. Coordination of efforts aimed at informing investigators and stu- dents about animal research (see ch. 9) could be among the responsibilities of this Federal entity. Option 3: Provide intramural and extramural Fed- eral funding for the development of alternatives. An effective mechanism for encouraging R&D on alternatives is funding. Small pilot programs might assess whether or not targeted development is effective. Development of alternatives in testing within the Federal Government is a natural offshoot of and closely allied with toxicological research. The agen- cies most likely to produce alternatives in response to new Federal funding are the National Cancer Institute and NIH. Because testing is so closely tied to regulation, funding could also be directed to FDA, EPA, the Consumer Product Safety Commis- sion, and the National Institute for Occupational Safety and Health. Regulatory agencies could be required to develop alternatives to specified tests or to spend funds generally toward their devel- opment. To stimulate extramural R&D, granting agencies reviewing applications could be required to assign priority to those that contain research with prom- ise for the development of new alternatives. Post- doctoral training programs could be established, along the lines of NIH’s National Research Service Awards, to ensure a steady supply of young re- searchers schooled in traditional disciplines, rang- ing from molecular biology to animal behavior, with applications in the development of alter- natives. Financial incentives to private groups develop- ing alternatives could take the form of tax incen- tives—perhaps tax credits in addition to those al- ready in place for R&D. Such groups could also be eligible for a new program (analogous to the Small Business Innovation Research program) that would target the development of alternatives (see ch. 12). ISSUE: Should improvements be made in infor- mation resources to reduce any unin- tentionally duplicative use of animals in research and testing? Science is able to advance rapidly because infor- mation about what has been done is disseminated (see ch. 10). If attempts to find prior work are in- adequate or prior work is not sufficiently accessi - 24 G Alternatives to Anima/ use in Research, Testing, and Education ble, unintentional duplication may occur. Such unnecessary repetition of experiments must be distinguished from replication of experiments to demonstrate the reproducibility of a method or to confirm the validity of results. The amount of unintentional, largely duplica- tive research and testing that occurs today is un- known. Investigations into the amount and cir- cumstances of unintentional duplication would be valuable in determining whether it results in sub- stantial waste of animals or funds. Moreover, consultations with potential users of any new in- formation resources would be essential in imple- menting certain options addressing this issue. Although the storage and retrieval of data are costly, there are clear benefits to making infor- mation that reduces unintentional duplication readily available. Among these benefits are sav- ings in the expense and time associated with ani- mal research and testing. Other benefits are sav- ings in animal lives and the additional work that might be done if resources are not wasted (see ch. 11). Option 1: Take no action. By making the National Agricultural Library the focus of a service to provide information on im- proved methods of animal experimentation (Pub- lic Law 99-198), Congress in 1985 indicated its intention to facilitate the dissemination of infor- mation about alternatives and to prevent unin- tended duplication of animal experimentation. Even if no further improvements in information resources are made specifically for the sake of avoiding unintentionally duplicative animal use, general improvements in information resources will proceed as a matter of course. Many resources already exist. The National Library of Medicine, the National Toxicology Program, and other Fed- eral entities maintain large databases that contain information or citations to published sources. Ma- jor commercial databases exist as well. National libraries and information centers provide the full text of articles and reports. The National Techni- cal Information Service (NTIS) catalogs, stores, and distributes on request many unpublished Federal reports. Improvements in these resources can be expected, either to fill needs for which the bene- fits justify the costs or to achieve other informa- tion policy goals, such as openness in government or advancement of science. Option 2: Require that results of all federally funded research and testing be conven- iently accessible. By means of oversight authority or legislation, all Federal entities could be required to provide convenient access to the results of all federally funded animal research and testing. Implementa- tion could be largely through mechanisms already available—publishing in the scientific literature; circulating published reports or depositing them with NTIS, NLM, the National Agricultural Library, or other entity; or entering the results in a pub- licly available database. New databases might also be established. Requirements that results be made conveniently accessible could apply to Federal em- ployees, contractors (through contract terms), and grantees (as a condition of awards). Contractors and grantees, however, may not be enthusiastic about assuming the burden of publicizing their results and responding to requests for information. This option recognizes that much research and some testing using animals is federally funded, that dissemination of research and testing results could be more comprehensive, and that better dissemi- nation might reduce any unintentional duplication. Because publication and information dissemina- tion are normally much less costly than obtaining original data, the benefits of enhanced communi- cation extend beyond saving animal lives. It is important to note that most federally funded work, indeed the vast majority of significant work, is already accessible, although access comes with different levels of convenience. And the results of federally funded work (except some grants) are available under the Freedom of Information Act (FOIA). Requiring that all results be conveniently accessible may burden databases and libraries with inconclusive results or other information that will not be used. Option 3: Promote greater use of animal testing data submitted by industry to Federal agencies, except where confidentiality protections apply. Industry must submit data to regulatory agen- cies before it can market certain products or some- times in response to reporting requirements. Stat- utory and regulatory provisions already exist that make some of this information publicly available, thus theoretically avoiding unintentional duplica- tion. In addition, information that is voluntarily Ch. l—Summary, Policy Issues, and Options for Congressional Action G 2 5 submitted and not claimed as confidential is avail- able under FOIA. Using oversight authority or legislation, greater use of nonconfidential information could be pro- moted, for example, by requiring that it be put into databases, compiled in reports, or summarized in newsletters. Industry could bear the cost of in- formation dissemination, and any data submission to the Federal Government would have to be ac- companied by evidence of intent to publish non- confidential testing data. Industry may be unen- thusiastic about such a procedure, because in some cases nonconfidential data provide direct clues to confidential data. Nevertheless, greater availabil- ity of nonconfidential data could aid in avoiding unintentionally duplicative testing. The extent to which researchers who need such data already know how to obtain them is not known. The needs of those engaged in animal test- ing must be carefully gauged prior to considera- tion of this option. A further consideration is the willingness of those who generate the data to en- courage others to benefit from their investment. Option 4: Require comprehensive literature searches to ensure that federally funded research or testing involving animals is not duplicative. A literature review is normally conducted by an investigator in the course of preparing a grant ap- plication, contract proposal, or data submission. In addition, the reviewers of such proposals are expected to be familiar with work that has already been done. Implementation of this option would require proof of a literature search through, for example, a companion document in any proposal to conduct federally funded research or testing. The funding entity would presumably have to judge the appropriateness of the literature search. Both the investigator’s act of searching the litera- ture and the funding agency’s certification of the search may reduce any unintentional duplication. To make a mandatory literature search palatable to investigators, free access to some or all of the necessary information resources may have to be provided. An alternative strategy is to require a literature search by the funding agency, or other entity, prior to the release of any funds. The disadvantages of requiring a comprehensive literature search be- fore work could be funded include the delay that an additional step would cause, the cost of the search itself to the Federal Government, and pos- sibly part of the cost of developing new informa- tion resources. Option S: Create new databases designed to re- duce unintentional duplication of ani- mal use in research and testing. New computerized databases might play an important role in reducing any unintentionally duplicative animal use. There are at least three types that could contribute to this end: G G G Unpublished Results, Including Negative Results. Such a database would disseminate results that are otherwise distributed narrow- ly or not at all. The major problem with un- published information is that its quality is dif- ficult to evaluate because it is rarely subjected to peer review. Another problem is that the most useful unpublished data are owned by industry and would not be disclosed because of their proprietary value (although provision could be made for voluntary submissions). A category of special interest, particularly from the standpoint of duplicative testing, is nega- tive results (e.g., showing the absence of toxic effects). Few journals are willing to publish negative testing results. Dissemination of neg- ative results could spare any unintentional duplication, direct investigators away from fruitless paths, or suggest improvements in methodologies. Data From Untreated, or Control, Animals. Data pertaining to the health or behavior of animals not given a test substance could be used in choosing the best species for experi- mentation (e.g., a species most likely to yield unambiguous results). This information might obviate the need to use more than one species or might allow smaller control groups in some experiments (see ch. 7). Compiling the data- base could be both difficult and costly because the necessary data are often not published (see ch. 10). Experimental Protocols and Results. This database could be as narrow as abbreviated listings of methods and results, perhaps ar- ranged by species, or as comprehensive as the on-line full text of all published scientific liter- ature. (The full text of a scientific report in- cludes not only protocol and results, but also discussion and interpretation of the results, tables, figures, and bibliography. At present, 38-750 0 - 86 - 2 26 G Alternatives to Animal Use in Research, Testing, and Education the full text (minus figures and images) of a few dozen scientific journals is available on- line.) The greatest obstacle to the successful creation of a database of this size is catering to the diverse needs of animal users. In its fullest incarnation, this would cost hundreds of millions of dollars to start and maintain. Most important, the extent to which any of these databases would be used is unknown. Within the Federal Government, the NLM has the greatest ex- pertise in establishing and operating large data- bases, and implementation of any form of this op- tion is likely to build on the experience and existing resources of that library. Option 6: Facilitate the use of foreign data by pro- viding translations of foreign journals. An often-overlooked source of published data is foreign-language literature, although most im- portant scientific work is routinely published in or translated into English. The advantages of pro- viding translations of additional work are thought by many experts to be quite limited and economi- cally unjustifiable. English translation costs for the four principal languages of science (French, Ger- man, Russian, and Japanese) range from $40 to $88 per thousand words. An estimated $4 billion to $5 billion would be required, for example, to translate the current foreign-language holdings of the NLM into English, with an ongoing yearly trans- lation cost of $150 million (see ch. 10). Copyright protections might involve costly inconvenience as well. The impact of this option is uncertain, as Eng- lish abstracts are today available for most foreign journals, and translations can be obtained on an ad hoc basis by those interested in a particular report. ISSUE: Should animal use in research, testing, or education be restricted? The use of animals for research, testing, and educational purposes is not closely restricted in the United States. Only four types of constraints can be identified. The Animal Welfare Act requires humane handling, care, and treatment of nonhu- man primates, dogs, cats, rabbits, guinea pigs, and hamsters. However, any regulation of these ani- mals within an actual experimental protocol is spe- cifically excepted by the Animal Welfare Act (see ch. 13). Second, at the State and local levels, cru- elty to animals is generally proscribed, although such statutes are generally not applied to animal use during experimentation (see ch. 14). Third, self- regulation takes place at individual institutions and facilities through the implementation of Federal policies. These call for assessment of animal care, treatment, and practices in experimentation by institutional animal care and use committees. Fourth, the Department of Defense was prohibited in fiscal years 1984 and 1985 from expending any funds for training surgical personnel by treating in dogs and cats wounds that had been produced by weapons (see app. B). The few existing restrictions on animal use illus- trate two phenomena. First, they show that pri- mates and pets have a privileged position in pub- lic policy. The Animal Welfare Act names only six kinds of animals, omitting the rats and mice that together constitute approximately 75 percent of the animals used in research, testing, and educa- tion. It requires exercise for dogs and a physical environment adequate to promote the psychologi- cal well-being of primates. In the case of the DOD appropriation, dogs and cats were named, while goats and pigs (also used in surgical wound train- ing) were not. Second, the restrictions demarcate the long- standing frontier of legislative province over ani- mal use—the laboratory door. The actual conduct of experiments stands largely outside of any spe- cific mandatory provisions of law. (In contrast, Brit- ish investigators are licensed to carry out speci- fied procedures using specified animals and face inspection visits to the laboratory bench by gov- ernment officials; see ch. 16.) Solely in the case of the prohibition of DOD expenditures is one use of two particular species addressed. Considering the issue of restriction of animal use may require the resolution of four difficult questions: G Are there some kinds of animals on which experimentation is inherently inappropriate? G Are some methods or procedures beyond the realm of societal acceptability? G Should some sources of animals be deemed off limits for animal use in research, testing, or education? G Should licensed investigators alone be per- mitted to engage in animal experimentation? The resolution of these questions turns on sci- ence, law, politics, and, to a large degree, ethics. Ch. I—SummaryJ Policy Issues, and Options for Congressional Action G 2 7 Six options for congressional action have been iden- tified. Option 1: Take no action. In the absence of new restrictions, animal use in research, testing, and education will continue to be governed loosely at the Federal level. Like the American system of education, control of ani- mal use can be largely a local issue, and institu- tional animal care and use committees stand as the arbiters of community standards. One draw- back of a minimal Federal role is the possible de- velopment of conflicting or confusing State and local policies. Maintenance of the status quo would reaffirm that Congress concurs that no methods or proce- dures are beyond the realm of societal acceptabil- ity (except the training of military personnel in sur- gical techniques on wounded dogs and cats in fiscal years 1984 and 1985). Maintenance of the status quo would leave unaffected the acquisition of ani- mals for research, testing, and education: Sources of animals today include breeders, dealers, pounds, and in-house breeding. Some States will continue to bar the acquisition of pound animals for research (see ch. 14). Finally, in the absence of a licensing scheme, investigators and their areas of inquiry will remain wholly a function of available resources and individual interests. Option 2: Restrict the use of certain kinds of animals. Some people feel it is wrong to use particular animals in research, testing, or education. This be- lief may stem from respect for apparent intelli- gence, and animals most closely related to humans, such as nonhuman primates, may be considered off limits for investigation or manipulation. Simi- larly, attachment to companion animals such as dogs and cats or to pet species such as hamsters may lead to a desire for their legislated immunity from experimentation. A restriction of this nature is likely to have sev- eral consequences. The restricted species would be protected while investigators faced, at a mini- mum, an inconvenience until new methods are developed. Development of new model systems would likely necessitate the generation of new fun- damental data about the characteristics of the model system, while the existing base of data— which could be large—about the restricted animal is set aside because it is no longer useful. In some cases, new methods would lead to a substitution of a less favored species for the restricted one. Per- haps the most important consequence would be that where the restricted species (e.g., monkey or dog) is the most scientifically appropriate model for research or testing, a prohibition on the use of that species may affect the ability to extrapo- late results to humans. Given that few, if any, kinds of animals are ex- clusively used in testing, research, and education, a restriction of this nature would be difficult to impose. How, for example, might a restriction dis- tinguish between primates under behavioral ob- servation in a field colony and those observed by tourists at a safari-style game preserve? Restric- tion of the use of particular kinds of animals may be inconsistent with the popular treatment and use of those same animals (e.g., circus, zoological park, sport, hunt, or farm) throughout the United States. Combining this option with the next one—to restrict the use of a species in a certain protocol— would yield a more limited, more practicable form of restriction than a blanket prohibition on use of a species. Option 3: Restrict the use of particular protocols. Some people feel that it is inhumane to manipu- late animals in certain ways, irrespective of the motivation for the procedure. Such concerns usu- ally focus on procedures that cause the animal pain or are painful for humans to watch. The Draize eye irritancy test is such a procedure, as are in- flictions of blunt head trauma in neurology re- search and of bullet wounds in surgical training. In research, blanket prohibitions either of a par- ticular animal’s use (the preceding option) or of a specified procedure entail a risk of being overly inclusive. They could have unintended or un- foreseen consequences, especially in the face of incomplete knowledge about how animals are used and in what protocols and what the results might portend. One risk of such a restriction would be the elimination of the use of animal models that may be the best available or the sole method of studying conditions present in humans but that 28 G Alternatives to Animal Use in Research, Testing, and Education do not lend themselves to systematic study in hu- mans (see ch. 5). In testing, procedures like the Draize test and the LD50 are used in part because investigators be- lieve that Federal regulatory agencies, such as FDA and EPA, require the results of these tests in data submissions (see ch. 7). Exercise of oversight au- thority could induce Federal regulatory agencies to make explicit their disinterest in data derived from objectionable tests and to demonstrate their ready acceptance of data obtained through alter- nate means. Such oversight action, coupled with active research into alternative methods, would probably end most use of the targeted procedures. It is likely that review of protocols by commit- tee, particularly a committee with expertise in bio- ethics, laboratory animal science, and anesthesia, would effectively restrict procedures to those that are generally accepted as humane. In both research and testing, banning animal use for a specific pur- pose would reflect the judgment that knowledge gained via that procedure could never justify the cost in animal suffering or lives. Option 4: Restrict the acquisition of animals from particular sources. For several decades, States and municipalities have wrestled with the issue of the release of dogs and cats from pounds to research and educational institutions (see ch. 14). Some people feel that the release of pound animals for experimentation is wrong, because the animals are former pets or are too unhealthy to be proper subjects for study. In some jurisdictions, research and educational institutions are barred from acquiring pound ani- mals, while other jurisdictions require that pound animals be released to researchers after a certain number of days in captivity. As pound animals are usually sold at low cost (see ch. 11), banning their sale would lead to higher procurement costs as the pound animals were re- placed with animals that are purposely bred for experimentation. (Some animals are already pur- pose-bred because certain pound animals are not suitable candidates for experimentation.) The pur- poseful breeding of such animals for experimen- tation in parallel with routine euthanasia of pound animals would probably work out to a net increase in dogs and cats being killed. Option 5: License animal users (e.g., for specified uses or for particular kinds of animals). Animal users could be granted licenses specify- ing the procedures they are authorized to perform or the animals with which they may work. Such a system is in place in the United Kingdom under the auspices of the Home Office (see ch. 16). Given that at least five to six times as many animals are used in the United States annually (17 million to 22 million) as in the United Kingdom (3 million to 4 million), achieving and maintaining licensure here would be a considerably larger and more costly enterprise than now exists in any country. Implementation of this option would require a Federal licensing body with inspection and enforce- ment capability. If the British system is the model, licenses would be legally enforceable personal doc- uments. A license to perform a particular experi- ment or a series of experiments or to work with a particular species would be nontransferable. Confidentiality would be guaranteed in order to protect, for example, an investigator’s claim to pri- ority in research results. Comprehensive annual reporting by licensees and auditing by an over- sight body—both integral parts of the British system—would be necessary. It is noteworthy that in the United Kingdom this system allows every animal experiment to be logged (see ch. 16). The British system works. It relies heavily on a tradition of cooperation between experimenter and Home Office inspector. The feasibility of such a system in the United States is difficult to predict because the dimensions of animal use are so poorly characterized. Hence, the number of licensees and the resources required for monitoring are un- known. perhaps most important, the extent to which the parties involved would cooperate is un- certain. Option 6: Prohibit the use of animals in research, testing, and education. No other country and no jurisdiction in the United States has completely banned animal use in research, testing, or education. In Switzerland, a binding referendum of this nature was presented to the public for a vote in December 1985, but it was defeated (see ch. 16). Action to ban animal use fully is the most ex- treme of the six options related to the issue of re- Ch. l—Summary, Policy Issues, and Options for Congressional Action G 2 9 striction. It would undeniably provide great impe- tus towards implementing alternatives. Indeed, the alternatives of reduction and refinement of ani- mal use would be immediately and completely achieved. However, the development of many re- placements to animal use depends itself on ani- mals. A ban would, for example, eliminate the use of organ cultures, nonhuman tissue cultures, and cell cultures, except for those self-perpetuating ones already in existence. Replacements would have to be drawn from among human and veteri- nary patients, micro-organisms, plants, chemical and physical systems, and simulations of living sys- tems. The development of new computer simula- tions would faker, with new data from animal sys- tems being unavailable. The ability to verify new simulations or proposed replacements would also come to a halt. Implementation of this option would effectively arrest most basic biomedical and behavioral re- search and toxicological testing in the United States. Education would be affected, too, although per- haps not as severely as research and testing. In the advanced life sciences and in medical and veterinary training, students might be handi- capped, although not to as great a degree as once thought. Some medical schools today, for exam- ple, use no animals in physiology curricula (see ch. 9). The economic and public health consequences of a ban on animal use are so unpredictable and speculative that this course of action must be con- sidered dangerous. Caution would demand, for example, that any new products or processes have substantial advantages over available ones to merit the risk of using them without animal testing. ISSUE: Should more accurate data be obtained on the kinds and numbers of animals used in research, testing, and education? Accurate data on the kinds and numbers of ani- mals used in research, testing, and education in the United States do not exist (see chs. 3 and 9). The best numbers now available on the use of cer- tain species (nonhuman primates, dogs, cats, rab- bits, guinea pigs, and hamsters) are produced by the Animal and Plant Health Inspection Service of the USDA. The APHIS Animal Welfare Enforce- ment Report submitted to Congress each year is best viewed as a rough estimate of animal use. It records approximately 10 percent of all animals used annually; omitted are rats, mice, birds, fish, reptiles, and amphibians. Estimates of animals used yearly in the United States range to 100 million and more. Although the development and implementation of alterna- tives do not require an accurate count, public pol- icy formation would be helped by better data. Reg- ulating animal use, for example, or funding the development or validation of alternatives to a par- ticular procedure, may depend on how many ani- mals are used and what fraction of the total this represents. Trends in animal use have similar ap- plications. In the United Kingdom, the exact ani- mal use records kept since 1876 have influenced policymakers (see ch. 16). Some animal welfare advocates suggest that the moral and ethical issues surrounding animal use are independent of the precise number of animals used. Others question whether the value of the data obtained is worth the cost of obtaining ac- curate numbers. A rough estimate based on mini- mal data may be all that is necessary to put the relevant issues into context. Selecting among the following options will depend, therefore, on judg- ment of how important it is to know the number and kinds of animals used, who uses them, and what trends exist. Option 1: Take no action. The primary advantage of this option is that no additional funding would be required, since noth- ing within the system would change. Continued funding of current APHIS activities would keep yielding rough estimates of the use of six kinds of animals that account for about 10 percent of total animal use. The major disadvantage of maintaining the sta- tus quo is that an inaccurate and ambiguous report- ing system would be perpetuated, yielding mar- ginally useful analysis of animal use in the United States, The APHIS counting system is ineffective because of problems with ambiguous reporting forms and a failure to audit the forms that are returned. Funding for the APHIS survey has been derived from the approximately $5 million allocated an- nually in recent years to APHIS to enforce the Ani- mal Welfare Act. Depending on the uses to which 30 . Alternatives to Animal Use in Research, Testing, and Education data on animal use are put, maintaining the status quo may be adequate, an unnecessary expense, or not nearly enough. Option 2: Eliminate the APHIS reporting system. If the value of the information obtained by the APHIS system is not justified by the money allo- cated for its collection, the APHIS reporting sys- tem could be terminated. In adopting this option, Congress would signal a willingness to rely on esti- mates produced by nongovernment organizations and individuals without the benefit of reports or inspections. Option 3: Correct inadequacies in the present APHIS system of reporting use of ani- mals mandated by the Animal Welfare Act. To gain a more accurate picture of the use of nonhuman primates, dogs, cats, rabbits, guinea pigs, and hamsters in the United States, oversight authority could be used to require that APHIS alter its present practices in one or more of the follow- ing ways: G G G G G correct its reporting form to eliminate am- biguities; change the reporting deadline or publication schedule for the annual Animal Welfare En- forcement Report, so that fewer institutional reports are excluded; audit or spot-check the “Annual Report of Re- search Facility” forms and facilities; strictly enforce the regulation requiring that all institutions within the United States using mandated species register with APHIS and complete the “Annual Report of Research Fa- cility” forms as required by law; or allocate more of APHIS’ resources for enforce- ment of the Animal Welfare Act to reporting. These changes would require little additional government funding or expenditure by regulated entities, although it could affect how they allocate their resources. Adoption of this option would bring APHIS closer to delivering the information it is obliged to deliver under the Animal Welfare Act. Option 4: Alter the APHIS system to count addi- tional kinds of animals (e.g., rats and mice). Rats and mice account for approximately 75 per- cent of the animals used in research, testing, and education in the United States. They go uncounted because a USDA regulation under the Animal Wel- fare Act excludes them from its definition of ani- mals. There is, however, some voluntary report- ing of the use of these species on the APHIS “Annual Report of Research Facility” forms. Data on rats and mice (or other currently un- regulated animals) could be obtained in either of two ways. Congressional oversight of the Secre- tary of Agriculture could lead to a requirement that the use of rats and mice be reported. This would require additional funding for APHIS, be- cause the number of facilities under the act’s reg- ulations would increase. On the other hand, the counting mechanism is already in place, and only minor changes would be needed. Expanding the APHIS animal counting require- ment to include rats and mice would raise costs for some members of the research and testing corn- munities. Accurate counting of these species, in- cluding categorization of experiments for pain and pain relief, is a labor-intensive activity and hence costly. Such costs will be of exceptional concern to institutions using large numbers of rats and mice, and these users can be expected to question whether accounting needs for policy evaluation require the extra expense. A broadening of the APHIS census to include rats and mice would still leave some uncounted. The Animal Welfare Act’s definition of research facil- ity covers any institution that uses primates, dogs, cats, rabbits, guinea pigs, hamsters, or other warm- blooded animals, as the Secretary of Agriculture may determine are used in experimentation, and that either purchases or transports animals in com- merce or receives Federal funds for experiments. Thus, a facility that breeds all its animals in- house—most likely rats or mice—falls outside the scope of the Animal Welfare Act and accompany- ing USDA regulations. The number of facilities breeding and using rats and/or mice exclusively is unknown. Some toxicological testing laboratories are likely to fall into this group. Option 5: Establish an independent census of ani- mal use, either on a one-time or peri- odic basis. Ch. l—Summary, Policy Issues, and Options for Congressional Action . 31 Fundamental changes could be made in the ways animals are counted. An animal census could be periodic-e.g., occurring every 2, 5, or 10 years. An organization other than APHIS, such as the pri- vate Institute for Laboratory Animal Resources (ILAR) of the National Research Council, could do the counting. In 1986, ILAR will undertake another in its series of surveys of laboratory-animal facil- ities and resources in the United States. (The last survey was conducted in 1978.) ILAR will survey the use of two classes of vertebrates—mammals and birds-at approximately 3,000 facilities. Another approach to gathering information on the kinds and numbers of animals used would be to conduct a comprehensive, one-time study of re- search, testing, and education. Such a study could survey all species acquired or bred for research, testing, and education; count the number of ani- mals actually used in experimentation; record the length of stay in animals in the facility; and catego- rize the purposes of the experimental-animal use. Such a comprehensive survey would not merit repetition every year—the purposes of animal use in research, for instance, do not change that quickly. A different way to count animals used would be to obtain figures from breeders on the num- ber of animals bred for experimentation. This would not take into account the percentage of ani- mals bred that are never used in experimentation, or animals bred within a laboratory, but it would yield a valuable index of animal use. Yet another source of information would be to count the num- ber of facilities or individuals using animals for specified activities. It is noteworthy that the revised PHS Policy on Humane Care and Use of Laboratory Animals by Awardee Institutions (effective Dec. 31, 1985) re- quires listing the average daily inventory, by spe- cies (with none excepted), of each animal facility, as part of each institution’s annual report to the NIH Office for Protection from Research Risks. Thus, PHS-supported facilities are now required to report more complete census data to NIH than facilities covered by the Animal Welfare Act re- port to APHIS. Consequently, a portion of animal use in research (e.g., NIH-supported animal re- search) and testing (e.g., FDA-supported animal testing) is about to become more closely censused. The choice among census types under this op- tion will depend on the ways in which the infor- mation is to be used, the resources available for obtaining it, and the utility of the new census re- quired by PHS. ISSUE: Should Federal departments and agen- cies be subject to minimum standards for animal use? The Federal Government has six cabinet depart- ments and four independent agencies involved in intramural animal research or testing (see ch. 13 and app. B). These departments and agencies ac- count for at least 1.6 million animals for intramural research (see ch. 3). Federal agencies have gener- ally followed the existing PHS policy and as of De- cember 1986 will be required to operate institu- tional animal committees (Public Law 99-198). Many departments and agencies also follow the NIH Guide for the Care and Use of Laboratory Ani- mals. Yet there is no stated, detailed policy of min- imum standards for animal use within the Fed- eral Government. Therefore, this issue has just two options: either maintaining the present system or establishing a minimum policy for intramural ani- mal use. Financial considerations are not a major factor because funds will be needed either to con- tinue the present system of variable policies or to implement and enforce a minimum, government- wide policy. Option 1: Take no action. The advantages of the present system are its flex- ibility and minimal bureaucratic structure, The policies mentioned previously, along with the In- teragency Research Animal Committee’s Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training, allow each agency or department to have policies and mech- anisms unique to its situation. The disadvantages are the potential for conflicting policies and the lack of a neutral enforcement authority. Option 2: Establish minimum standards for all in- tramural animal use in Federal depart- ments and agencies. This option would require that a policy be de- veloped and perhaps that an organizational entity be established to oversee its implementation and enforcement. This could be accomplished by an interagency committee or by a designated agency. 32 Alternatives to Animal Use in Research, Testing, and Education Setting minimum standards would still give each agency and department the flexibility to tailor spe- cific policies to unique situations, yet it would estab- lish a Federal model for standards of animal care in experimentation and ensure humane proce- dures in Federal facilities. A Federal intramural policy might incorporate policies and procedures that address facility ac- creditation and institutional review of research proposals. A composite, minimum Federal policy could reflect the most progressive parts of vari- ous current agency standards. It is noteworthy that this type of action has been taken to protect human research subjects. A Model Federal Policy for the Protection of Human Re- search Subjects involved in research conducted, supported, or regulated by Federal departments or agencies is now in draft form. The policy will be implemented through routine policy and pro- cedural channels of the departments and agencies. The advantage of minimum standards is that all concerned parties know the policy and can im- mediately and permanently put in place the appro- priate organizational structure and facilities to guarantee adherence. ISSUE: Should the Animal Welfare Act of 1966 be further amended, or its enforcement enhanced? One criticism of the Animal Welfare Act is the lack of coverage of practices other than anesthe- sia and analgesia during actual experimentation. Although the most recent amendments to the act, in 1985, direct institutional animal committees to assess practices in experimentation and require that professionally acceptable standards are fol- lowed during experimentation, the act at the same time forbids any regulation related to the design or performance of experiments. Additional com- plaints concern the adequacy of resources for its enforcement, the enforcement structure, the choice of APHIS as the primary enforcement agency, and the cumbersome recordkeeping. In considering whether the act should be strength- ened, some related issues must be kept in mind. First, a change in authority may require funding for implementation and enforcement. Second, any change must take into account the present re- sources of those affected and their ability to achieve compliance without compromising other objec- tives. Thus, an important consideration is whether or not regulated institutions have sufficient institu- tional and independent veterinary resources to ef- fect meaningful compliance with a strengthened law and still meet their testing or research objec- tives. Finally, strengthening the Animal Welfare Act in the face of differences within the scientific and animal welfare communities will carry con- siderable symbolic value. Option 1: Take no action. By maintaining the status quo, Congress would give a strong signal to all concerned parties that it is satisfied with the present regulatory structure for animal use in the United States and that no change is deemed necessary. More specifically, selection of this option would imply that current enforcement efforts are sufficient and that it is not necessary to regulate rats and mice used in experimentation. Option 2: Eliminate funding for enforcement of the Animal Welfare Act. Elimination of funding for enforcement of the Animal Welfare Act by APHIS would save the Fed- eral Government approximately $5 million annu- ally. Without these funds, there would be no in- spections of facilities (including exhibitors, dealers, and research institutions) using nonhuman pri- mates, dogs, cats, rabbits, guinea pigs, or hamsters and no annual census of these six kinds of animals. Action taken by APHIS against violators would cease. Therefore, the objective of the Animal Wel- fare Act—to safeguard the humane care and treat- ment of certain animals—would no longer be met. Option 3: Increase funding for enforcement of the Animal Welfare Act. Increased funding for the enforcement of the Animal Welfare Act would bolster enforcement of the present law. Additional funds could be used to: G G G increase the training of inspectors; increase the number of enforcement agents in the field, so as to raise the number of in- spections; oversee consistent interpretation of existing regulation by inspection and enforcement agents in the field; and/or Ch. l—Summary, Policy Issues, and Options for Congressional Action . 33 G replace voluntary assurances and simple cer- tifications of compliance with more rigorous procedures. Additional funding could help stimulate the present passive regulatory situation to become a more ac- tive, aggressive regulatory environment. Such a transition would rest on APHIS’ level of enthusiasm for enforcing the Animal Welfare Act. Option 4: Expand the jurisdiction of enforcing agencies to include standards of care, treatment, and use during the actual conduct of experimentation. The Animal Welfare Act exempts the treatment of animals while they are actually involved in ex- perimentation, except for a requirement for ap- propriate anesthesia or analgesia and the use of professionally acceptable standards in the care, treatment, and use of animals. The original law exempted actual experimentation because Con- gress did not want to interfere with the conduct of the scientific process (see ch. 13). Animal care and treatment are essentially regulated only be- fore and after a scientific procedure. Implemen- tation of this option would broach the design and execution of experimental protocols and would require statutory change. Such action would in- crease the responsibility of APHIS and its enforce- ment would require additional funding. A deter- rent to implementation of this option is APHIS’ lack of expertise in reviewing experimental protocols. Option 5: Realign existing and any new responsi- bilities for enforcement among Federal departments and agencies. APHIS spends little of its resources, either mone- tary or personnel, enforcing the Animal Welfare Act (see ch. 13). It was selected by Congress in 1966 to enforce the act because it had some expertise in animal issues but did not have the conflict of interest that an entity such as NIH or DHHS might have. Enforcement power could be changed by trans- ferring enforcement authority for violations of the Animal Welfare Act from USDA (APHIS) to DHHS. This would set up a potential conflict of interest: A single department would both sponsor animal experimentation and have oversight authority. In addition, many of the regulations in the Animal Welfare Act affect areas in which DHHS has no expertise (e.g., animal use by exhibitors). In amending the Animal Welfare Act in 1985, Congress mandated that the Secretary of Agricul- ture consult with the Secretary of Health and Hu- man Services prior to issuing regulations under authority of the act. The implementation of this provision may lead to DHHS having increased in- fluence on the enforcement of the act, Option 6: Amend the Animal Welfare Act to pre- empt State and local laws concerning animal use in areas not already covered by the Animal Welfare Act. Although the Edward Taub case in Maryland (see ch. 14) did not decide the preemption question, it did bring up the issue of whether the Animal Welfare Act could preempt a State statute. Con- gress may wish to examine its authority to preempt State anticruelty statutes and may then wish to specify for the judiciary whether it intended its law to supersede any State or local laws on this issue. In doing so, Congress could remove uncer- tainty in the law by making clear whether it in- tends the Animal Welfare Act to be a comprehen- sive, exclusive system of control over the use of animals in experimental facilities and activities in interstate and foreign commerce. Without such clarification, the possibility exists for local crimi- nal prosecution, seizure of animals, injunctions to close facilities, and cessation of animal investi- gations. Current State and local efforts to assure humane treatment have been criticized for several reasons. Compliance schemes are overly complex, training and resources are inadequate, and existing laws are not specific enough in their standards for care, treatment, and use. If Federal preemption is not exercised, then State and local laws will be con- sidered concurrent and complementary to exist- ing Federal laws. It is important to note that Federal preemption means that the administrative system for moni- toring, including on-site inspection, should be made adequate to ensure continued compliance with na- tional standards for humane treatment. Otherwise, State-level organizations with a sincere and rea- sonable concern about the care of animals will be 34 . Alternatives to Animal Use in Research, Testing, and Education justified in demanding local enforcement and sur- question of whether the Federal Government has veillance of research, testing, and education in- the authority to assert itself into areas tradition- volving animals. ally regulated by the States (e.g., pound animal use) Finally, it should be recognized that if Federal may well land in the courts. preemption is deemed necessary, the constitutional Chapter 2 Introduction Donahue: What doesn’t feel pain? When do you stop feeling pain? Does a frog feel pain? McArdle: Yes. Donahue: Frogs feel pain? . . . now what about laboratory high school? You remember, you had to dissect the frog? . . . Should we eliminate that? How about fishing? . . . how about baiting a hook with a worm? IS that fair? In other words, where do we stop? McArdle: You bring up fishing and I think that's a good point. I used to wonder whether or not the nonvertebrate animals would feel pain. A few years ago they found en- dorphins, which are substances that handle chronic pain, in earth worms. So, earth wvrms may in fact be subject to chronic pain when you’re putting them on that hook. Phil Donahue with John E. McArdle, Humane Society of the United States Donahue (transcript #02065) February 1985 Although the highest standard of protection must be applied to all animals, we acknowl- edge that it is right to pay special attention to the companions of man [non-human primates, cats, dogs, and equidae] for whom there is the greatest public concern. Scientific Procedures on Living Animals, Command 9521 British Home Office May 1985 CONTENTS What is an Animal? . . . . . . . . . . . . . . . . What is an Alternative? . . . . . . . . . . . . . Biological Models . . . . . . . . . . . . . . . . . . Chapter 2 References . . . . . . . . . . . . . . . . Page . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..,,., 39 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 Table Table No. Page 2-I. Some Types of Living Organisms Used in Research, Testing, and Education . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. ...38 Chapter 2 Introduction This report assesses the state of the art and the potential for alternatives to using animals in three contexts: biomedical and behavioral research, test- ing of products for toxicity, and education. Dis- tinguishing among these three areas is important because both the patterns of animal use and the potential for alternatives vary among them. Re- search develops new knowledge and new technol- ogies; although prediction of results is one goal, unpredictable results may prove even more sig- nificant. Testing relies on standardized procedures that have been demonstrated to predict certain health effects in humans or animals. It entails the measurement of biological phenomena, such as the presence or absence of cancer or of skin irri- tation, or the concentration of certain substances in tissue or in bodily fluids. Education involves teaching students in the life sciences, health profes- sionals and preprofessionals, and research scien- tists, as well as the cultivation of humane attitudes toward animals at all levels. Alternatives in each of these three areas consist of procedures that re- place animals with nonanimal methods, that re- duce the number of animals used, or that refine existing protocols to make them more humane. In addition to evaluating alternatives in three areas, the assessment also examines ethical con- cerns regarding the use of animals, economic con- siderations of their use and the alternatives, funding for the development of alternatives, and current regulation of animal use, Most important, this re- port delineates seven major public policy issues (and associated options for congressional action) in relation to alternatives (see ch. 1). With a focus on the prospects for alternatives to animal use in research, testing, and education, this assessment necessarily excludes certain re- lated topics and treats others only in brief. The role of animals in food and fiber production falls outside the scope of this study, as does the role of animals in the commercial production of anti- bodies and other biological materials. In addition, OTA has not evaluated the use of animals for compan- ionship, sport, or entertainment. Although laboratory animals are an integral part of this assessment, OTA did not examine contemporary standards of their care (e.g., cage size, sanitation, ventilation, feeding, and watering). Lastly, the use of human subjects is not considered in this assessment. WHAT IS AN ANIMAL? In any biological definition of the word “animal,” Political and scientific discussions often incor- all vertebrate and invertebrate organisms are in- porate other subdivisions for the term “animal.” eluded and plants and unicellular organisms are Although not strictly part of the definition in this excluded. For the purposes of this report, how- report, the terms “lower” and ‘(higher” are used ever, an “animal” is defined as any member in many discussions of alternatives that refine ex - of the five classes of vertebrates (nonhuman isting animal procedures or that replace certain mammals, birds, reptiles, amphibians, and animal species with other ones. In these contexts, fish). These five classes of vertebrates can be fur- the substitution of “lower” animals for ‘(higher” ther divided into two major groups, cold-blooded animals usually refers to using cold-blooded ver- vertebrates (reptiles, amphibians, and fish) and tebrates instead of warm-blooded vertebrates. In warm-blooded vertebrates (mammals and birds). addition, within the class of mammals, “lower” is Invertebrates, therefore, are not discussed as generally used to designate, for example, rodents, animals. while “higher” refers to primates, companion ani- 37 38 G Alternatives to Animal Use in Research, Testing, and Education mal species (e.g., dogs, cats, or rabbits), and do- and education. It indicates the laboratory species mestic farm animals (e.g., horses, cattle, or pigs). falling within this assessment’s definition of an ani- — Table 2-1 is a classification of the principal liv- mal and the species that can be classified as “alter- natives.” ing organisms that are used in research, testing, Table 2.1.—Some Types of Living Organisms Used in Research, Testing, and Education Alternatives: 1. Prokaryotes (any living organism without a nuclear membrane) A. Bacteria 1. Escherichia coli 2. Salmonella 3. Streptococcus 4. Bacillus B. Fungi—e.g., yeast Il. Eukaryotes (any living organism with a nuclear membrane) A. Plants B. Invertebrates 1. Protozoa a. Paramecium b. Amoeba 2. Porifera—e.g., sponges 3. Coelenterates—e. g., Hydra and Jellyfish 4. Flatworms—e.g., Planaria 5. Segmented worms a. Earthworms b. Leeches c. Annelids 6. Nematodes—e.g., Caenorhabdiitis elegans 7. Molluscs a. Gastropods—e.g., snails and Aplysia b. Pelecypods—e.g., mussels c. Cephalopods—e.g., squids and octopuses 8. Arthropods a. Lirnulus (horseshoe crabs) b. Arachnids (1) Spiders (2) Ticks (3) Mites (4) Scorpions c. Crustaceans (1) Daphnia (2) Brine shrimp (3) Crayfish d. Insects (1) Crickets (2) Cockroaches (3) Drosophila (fruit flies) (4) Lice (5) Beetles (6) Moths (7) Butterflies 9. Echinoderms a. Sea urchins b. Sand dollars c. Sea cucumbers Animals: C. Vertebrates 1. Cold-blooded vertebrates a. Fish (1) Jawless fish–e.g., lampreys SOURCE: Off Ice of Technology Assessment. (2) Cartilaginous fish–e.g., sharks (3) Bony fish b. Amphibians (1) Frogs—e.g., Rana (2) Toads–e.g., Xenopus (3) Salamanders c. Reptiles (1) Turtles (2) Crocodiles (3) Alligators (4) Snakes (5) Lizards 2. Warm-blooded vertebrates a. Birds (1) Quail (2) Chickens (3) Pigeons (4) Doves (5) Ducks b. Mammals (1) Bats (2) Rodents (a) Mice (b) Rats (c) Gerbils (d) Guinea pigs (e) Hamsters (f) Squirrels (3) Marine mammals (a) Dolphins (b) Whales (c) Seals (d) Sea lions (4) Rabbits (5) Armadillos (6) Carnivores (a) Dogs (b) Cats (c) Ferrets (7) Ungulates (a) Cattle (b) Sheep (c) Horses (d) Pigs (e) Miniature pigs (f) Goats (g) Donkeys (h) Burros (8) Primates (a) Baboons (b) Capuchins (c) Chimpanzees (d) Macaques, Cynomolgous (e) Macaques, Pig-tailed (f) Macaques, Rhesus (g) Marmosets (h) Squirrel monkeys Ch. 2—introduction 39 WHAT IS AN ALTERNATIVE? Defining the word “alternative” is in a sense al- ways doomed to failure: Regardless of how accom- modating or strict the definition, many will fault it. The term evolved in the political arena, coined by animal welfare activists and for the most part nonscientists, and yet it has direct implications for scientists using laboratory animals. Its meaning varies greatly, depending on who uses it and the context in which it is used. The definition of “alternatives” employed by OTA obviously affects this entire assessment: It defines the scope of the study. Too narrow a definition would dispose of the need for this report, while too broad a definition would render it unmanage- able. Defining alternatives as the nonuse of ani- mals, as some would have it, would restrict the bounds of the study to the consideration of inver- tebrate organisms, chemicals, plants, and comput- ers. On the other hand, stretching the definition to include humans, for example, would create a whole new series of issues that would be virtually impossible to address within one assessment. With these concerns in mind, OTA chose to define “alternatives” as encompassing any subjects, protocols, or technologies that ‘(replace the use of laboratory animals altogether, reduce the number of animals required, or refine existing procedures or techniques so as to minimize the level of stress endured by the animal” (4; adapted from 5). Some examples of alternatives under this defi- nition include computer simulations to demon- strate principles of physiology to medical students, the use of the approximate lethal dose methodol- ogy in acute toxicity studies, and the increased use of anesthetics with pain research subjects. The “re- duction” part of the definition indicates that the increased use of cultured cells, tissues, and organs instead of whole animals is also an alternative. A very broad interpretation of alternatives might also include the substitution of cold-blooded for warm- blooded vertebrates. BIOLOGICAL MODELS When animals-or alternatives—are used in re- search, testing, and education, it is because they possess a simpler or more accessible structure or mechanism in comparison with the object of pri- mary interest (which is often the human) or are themselves the object of primary interest, or be- cause certain procedures cannot be carried out on humans. Viewed from this perspective, both animals and alternatives stand as models. In the broadest sense, a biological model is a surrogate, or substitute, for any processor organism of ulti- mate interest to the investigator. It is a represen- tation of or analog to some living structure, orga- nism, or process. In addition to analogy, biology has another ana- lytical tool at its disposal—homology, which is cor- respondence in structure and function derived from a common evolutionary origin (i.e., a com- mon gene sequence). The most closely related spe- cies are generally presumed to offer the best homo- logs. Relationships between species are not always known in detail, however, and unresolved ques- tions about evolutionary events and pathways are numerous. Care must therefore be used in evalu- ating the degree of homology and the extent to which it relates to analogy (3). Some biological mechanisms, such as the cod- ing of genetic information and the pathways of metabolism, arose early in evolution. These mech- anisms have been highly conserved and are widely shared by organisms, including humans, at the cel- lular and molecular levels. Thus, good models for these fundamental molecular mechanisms in hu- mans can be found in a wide array of organisms, some of which, such as bacteria, have structures and functions far less complex than those of mam- mals (3). Several characteristics are important in choos- ing a model for research, testing, or educational purposes. The most important is the model’s dis- crimination-the extent to which it reproduces the particular property in which the investigator is interested. With greater discrimination, the pre- 40 G Alternatives to Animal Use in Research, Testing, and Education dictability between the model and the property under study increases. After the discrimination or predictability of a model, certain other criteria stand out as being necessary for a good biological model (1)2). A model should: G G G G G G G G G G G G accurately reproduce the disease or lesion un- der study; be available to multiple investigators; be exportable from one laboratory to another; be large enough to yield multiple samples; fit into available facilities of most laboratories; be capable of being handled by most investi- gators; survive long enough to be usable; exhibit the phenomenon under study with relative frequency; be of defined genetic homogeneity or heter- ogeneity; possess unique anatomical, physiological, or behavioral attributes; be accompanied by readily available back- ground data; and be amenable to investigation with available, sophisticated techniques. CHAPTER 2 1. Leader R,A,, and Padgett, G.A., “The Genesis and Vali- dation of Animal Models) ’’Azn. J. Pathol. 101:s11-s16, 1980. 2. National Research Council, Mammalian Models for Research on Agl”ng (Washington, DC: National Acad- emy Press, 1981). 3. National Research Council, Models for Biomedical Research: A New Perspective (Washington, DC: Na- tional Academy Press, 1985). Depending on the type and needs of the investiga- tion, certain of these criteria might be more im- portant than others. Overall, a model with more of these characteristics will have higher discrimi- nation and stronger predictive ability, In research, testing, and education, a small num- ber of species have achieved prominence as experi- mental tools because they have been extensively studied from a number of perspectives and thus provide well-understood paradigms that have been described in detail in terms of genetics, biochem- istry, physiology, and other aspects. These organ- isms include the laboratory rat, laboratory mouse, fruit fly, and bacterium Escherichia coli. Yet taxo- nomic breadth is also required in research and testing, since it is often impossible to predict what species will lend themselves particularly well to the study of specific problems. In biological mod- eling, concentration on selected species and taxo - nomic diversity are not mutually exclusive; both play a role in the establishment of a maximally use- ful matrix of biological knowledge (3). REFERENCES 4. Rowan, A. N., Of Mice, Models, & Men: A Critical Evaluation of Animal Research (Albany, NY: State University of New York Press, 1984). 5. Russell, W.M.S., and Burch, R. L., Principles of Humane Experimental Technique (Springfield, IL: Charles C. Thomas, 1959). chapter 3 Patterns of Animal Use Twenty million rats, rabbits, cats, dogs, mice, and monkeys are killed each year in the name of science. And the number has quadrupled in recent years . . . 150 living creatures are sacrificed every minute. Paul Harvey Radio broadcast of April 30, 1985 Each minute around the clock, 150 creatures are sacrificed ., . a total of 70 million a year. Included are 25,000 primates . . . and nearly 500,000 dogs and cats. parade, January 13, 1985 Each year in the United States, almost 100 million animals are used in scientific research. Nearly a million are dogs and cats. Ed Bradley CBS News, 60 MINUTES October 14, 1984 OTA ignores the fact that more than one-half of all research goes unreported because unfunded. Secondly, funded researchers consistently understate the number of animals used for several reasons I won ‘t enumerate. My personal guess is that 120-150 million animals is the right ballpark figure. Sidney Gendin Eastern Michigan University The Research News 36(3-4):17, 198.5 CONTENTS Page The Federal Government’s Use of Animals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 Federal Departments and Agencies Using Animals in Research . . . . . . . . . . . . . 44 Patterns of Federal Animal Use...... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Animal Use in the United States . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 Limitations of Animal-Use Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . SO Critical Evaluation of Animal-Use Estimates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 Calculating Rat and Mouse Usage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57 Summary and Analysis of Estimates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 Future Animal Censuses. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65 Summary and Conclusions . . . . . . . . . . . . . . . ..........+.. . . . . . . . . . . . . . . . . . 65 Chapter p references. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 List of Tables Table No. Page 3-1. Research-Animal Use in the Federal Government, by Major Department and Division for Fiscal Year 1983 . . . . . . . . . . . . . . . . . 50 3-2.Total Numbers of Animals Used in Federal Government Facilities as Reported to Congress in APHIS Animal Welfare Enforcement Reports, 1978-83 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 3-3.Reliabilityof Various Data Sources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 3-4. Estimates of Rat and Mouse Usage in laboratories, 1978, 1982, 1983 . . . . . . 59 3-5. Various Estimates of the Number of Animals Used in the United States... . 60 3-6. USDA/APHIS Data, Changes 1982-83 . . . . . . . . . . . . . . . . . . . . . . . . ~ . . . . . . . . . 64 3-7. Animal Use Reported to the U.S. Department of Agriculture, 1982 and 1983 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66 List of Figures Figure No. Page 3-1. USDA/APHIS “Annual Report o fResearch Facility’’ . . . . . . . . . . . . . . . . . . . . . . 48 3-2. Example A of APHIS”Annual Report of Research Facility” . . . . . . . . . . . . . . . 61 3-3. Example B of APHIS ’’Annual Report of Research Facility” . . . . . . . . . . . . . . . 62 3-4.Example C of APHIS ’’Annual Report of Research Facility” . . . . . . . . . . . . . . . 63 Chapter 3 Patterns of Animal Use Humans “use” animals in several different ways. In addition to animal use in research, testing, and education, animals are involved in food and fiber production, the production of biological products, sports, and entertainment. Animals can also be kept as pets for the purpose of companionship. It has been roughly estimated that 2 billion to 4 billion animals are used in food and fiber production every year and that Americans have approximately 75 million dogs and cats as household pets. The uses not considered in this assessment therefore account for many times more animals than the esti- mated 17 million to 22 million animals used annu- ally in research, testing, and education. There are no easily obtainable data in the United States allowing an accurate estimate of animal use for research, testing, and education that satisfies all interested parties; estimates range over a full order of magnitude, from approximately 10 mil- lion to 100 million animals. These estimates have all been prepared by different people or institu- tions with different data sources under different standards (e.g., different time periods or defini- tions). Comparison of the various estimates is dif- ficult and, in many cases, impossible. The issue of numbers is important to any dis- cussion of animal use in research, testing, and edu- cation. Most basically, a number is needed from which to consider arguments to decrease or elim- inate animal use. In addition, comparing absolute numbers in different years would provide some idea of whether laboratory-animal use is increas - ing or decreasing in the United States; these num- bers are powerful and important to many people. A high overall total, or high numbers of certain species (such as nonhuman primates or companion species), supports the claims of interest groups hop- ing to restrict or ban such experimentation. On the other hand, a low number indicates the issue is not as important as some claim. In addition, a decreasing trend in animal use supports the posi- tion that the present system will lower animal use on its own. For this assessment, some idea was needed of the scope of animal use in terms of both the num- bers of particular species used and the different major users. In addition, an analysis of different data sources helps put the various estimates of ani- mal use into some comparative perspective. It pro- vides the context in which to discuss alternatives and how much effect they might have. Although it is true that the development of alternatives and alternative methods does not require a perfectly accurate estimate of usage, the planning of public policy certainly should be based on firm data. By looking critically at the different data sources and coming up with possible estimates of labora- tory-animal use in the United States, this assess- ment attempts to base discussions on a realistic, factually backed range of figures. Without such an analysis, any discussion or decisions on policy issues and possible solutions lack an important per- spective. THE FEDERAL GOVERNMENT’S USE OF ANIMALS To document the scope and extent of animal use about animal use in each department or agency. for research by Federal departments and agen- Together, the information illustrates: cies, information was obtained from the Animal and Plant Health Inspection Service (APHIS) an- G nual reports for Federal research facilities for 1983, the Animal Welfare Enforcement Reports for fis- G cal years 1978 through 1983 (both obtained from the U.S. Department of Agriculture (USDA)), and G personal communications or written material the extent of animal use in different de- partments, the amount and type of animals being used in the Federal Government, the experimental conditions under which most animal experiments are carried out, 43 44 G Alternatives to Animal Use in Research, Testing, and Education the general purpose for which animal re- search and testing is carried out in different departments and agencies, and how much research and testing for the Fed- eral Government is conducted intramurally (i.e., within Federal facilities). Federal Departments and Agencies Using Animals in Research Six departments and four independent Federal agencies conduct intramural research or testing involving animals. Uses of animals range from combat-casualty-care investigations in the Depart- ment of the Army, to acute toxicity studies by the Consumer Product Safety Commission of poten- tially hazardous substances, to National Aero- nautics and Space Administration research on pro- tecting the health of American astronauts. (For additional information on the use of animals within the Federal Government, see chs. 7 and 13 and app. B.) Department of Agriculture USDA performs biomedical research using ani- mals under the authority of the Animal Welfare Act in order to improve animal breeds, food, and fibers. Most of the research is conducted in- tramurally by the Agricultural Research Service, although some extramural research (i.e., research supported by USDA, but conducted in non-USDA facilities) is contracted out by the Cooperative State Research Service. Some of this USDA animal re- search involves farm animals, however, which are largely excluded from Government regulatory pol- icies and are exempt from the Animal Welfare Act and APHIS regulations (44). Department of Commerce The Department of Commerce conducts a small amount of intramural research with animals and lets some extramural contracts that involve ani- mal studies. There are no specific Commerce guide- lines or policies governing the humane treatment and appropriate veterinary care for laboratory ani- mals (33). Department of Defense The divisions within the Department of Defense (DOD) that conduct experimental research on ani- mals are the Air Force, the Army, the Navy, the Uniformed Services University of the Health Sciences, the Defense Nuclear Agency, and the Armed Forces Institute of Pathology; the first three of these account for most of the research. To- gether, all the divisions have approximately 40 re- search facilities that conduct animal experimen- tation. The Aerospace Medical Division (AMD) of the Air Force accounts for about 95 percent of that service’s use of animals. Of this, 84 percent is due to intramural research (9). AMD research and de- velopment projects fall within the following areas: humans in space, chemical defense and threat countermeasures, safety and environment, logistics and technical training, air combat training, human components of weapons systems, and personnel and force management. The safety and environment program uses the most animals, while those on human components of weapons systems and chemical defense also have some animal use (50). The Army does medical research to protect the soldier by the authority in the mission of the US. Army Medical Research and Development Com- mand. Medical research and development (R&D) are carried out in five areas: infectious diseases (tropical disease and biological warfare defense), combat casualty care, combat systems, dental re- search (facial injuries), and chemical defense. About one-third of the research is done in-house and two-thirds is contracted out (38). The Navy in fiscal year 1985 allocated $58 mil- lion for the life sciences or biomedical research. Of this, $37 million (64 percent) is for extramural research while the remainder is for intramural use. The two main branches of the service doing research involving animals are the Naval Medical Research and Development Command and the Of- Ch. 3—Patterns of Animal Use . 45 fice of Naval Research (ONR). The Naval Medical Research and Development Command does re- search in: G submarine and diving medicine, G electromagnetic radiation, G aviation medicine/human performance, G fleet health care systems, . infectious diseases, and G oral and dental health. ONR conducts research using animals in four ma- jor areas: molecular biology, neurophysiology/ physiology, cellular biosystems, and psychologi- cal sciences (45). Department of Energy The Department of Energy has no intramural research facilities and so contracts out all its re- search (47). The primary research objective within its Office of Health and Environmental Research is to study the health and environmental effects of energy technologies and programs. To do this, in the past, the Department contractor used dogs. Recently, though, there has been a gradual shift from whole animals to cellular and molecular re- search and a much greater emphasis on rodents as opposed to companion species or primates (12). Department of Health and Human Services Intramural animal research or testing is carried out by four components of the Department of Health and Human Services’ Public Health Serv- ice: the National Institutes of Health (NIH), the Food and Drug Administration (FDA), the National In- stitute on Drug Abuse (NIDA) (a part of the Alco- hol, Drug Abuse, and Mental Health Administra- tion), and the National Institute for Occupational Safety and Health (NIOSH) (a part of the Centers for Disease Control). NIH is the largest research institution in the Fed- eral Government and uses more animals than any other department or agency. The mission of NIH is to uncover new knowledge that will lead to bet- ter health (51). It does this by both intramural and extramural research. Approximately 88 percent of the NIH budget is spent on extramural programs while 10 percent goes to intramural research and 2 percent is used for NIH administration. Some 44 percent of the research awards go to research involving animals (28). Research in the FDA is mission-oriented, with the principal objective being to provide data to sup- port regulatory decisions. Research is conducted to determine the safety of human and animal foods; detect contaminants in human and animal foods; determine the safety and efficacy of human and animal drugs, biological products, and medical de- vices; reduce unnecessary exposure to artificial radiation; and increase fundamental understand- ing of the toxicological effects of chemicals. Ninety percent of the dollar budget for FDA research is allocated to intramural research studies while the other 10 percent goes to extramural research (5). Department of the Interior The Department of the Interior does more than 95 percent of its research in-house (31). Most ani- mal research is performed by the U.S. Fish and Wildlife Service to support its mission “to provide the Federal leadership to conserve, protect, and enhance fish and wildlife and their habitats for the continuing benefit of people. ” This involves maintenance of relevant research and education programs in cooperation with other State and pri- vate organizations to enhance fish and wildlife re- source management (53). Department of Transportation The Department of Transportation conducts ani- mal research under the authority of the Hazard- ous Transportation Act of 1974 to determine the level at which substances become Class B poisons (see ch. 7). Most of the research involving animals is conducted extramurally (42). The Department also performs animal research under the author- ity of the National Traffic and Motor Vehicle Safety Act of 1966 (10). 46 G Alternatives to Animal Use in Research, Testing, and Education Consumer Product Safety Commission The Consumer Product Safety Commission (CPSC) both relies on data provided by manufacturers and conducts its own testing to determine the toxic potential of consumer products. Animals are used by CPSC’s Directorate for Health Sciences in de- terminations of substances’ acute oral toxicity, their potential for skin and eye irritation, and their combustion toxicity (16). Environmental Protection Agency The Environmental Protection Agency (EPA) per- forms research involving animals under the stat- utory and regulatory authority of the Toxic Sub- stances Control Act and the Federal Insecticide, Fungicide, and Rodenticide Act. The general pur- pose of this research fits into one of three catego- ries: methods development to assess potential haz- ards to the environment, dose-response data for risk assessment, or low dose to high-dose data for risk assessment. EPA has two major research fa- cilities, one in Cincinnati, OH, and the other in Re- search Triangle Park, NC. In addition to the intra- mural research done in these facilities, EPA does contract extramural research. The amount done outside the agency varies from year to year and depends on the program, but it usually does not exceed 40 percent of total research (48). National Aeronautics and Space Administration The National Aeronautics and Space Adminis- tration (NASA) has three facilities that maintain or conduct research with animals, although ap- proximately 65 percent of NASA’s Life Sciences research is conducted extramurally. About 12 per- cent of the life sciences budget was used to fund animal research in fiscal year 1984 (37). The general purpose of NASA’s research is to acquire knowledge that can be used to protect and ensure the health of American astronauts, both during their missions in space and after their re- turn to Earth. National Science Foundation The National Science Foundation awards grants for scientific research involving animals but per- forms no intramural research. Veterans’ Administration The Veterans’ Administration (VA) has 174 fa- cilities, 91 of which have the ability and authori- zation to do animal research. The VA’s mandate to do research that may involve animals comes from part of the agency’s defined mission to un- derstand health maladies better, with a special em- phasis on those that affect veterans. The VA uses animals in its research and development divisions and its education programs, which are located in many of its local facilities. All research funded by the VA is done intramurally, and some of the re- search done by the VA is funded by other agen- cies, such as NIH (29). Research and development within the VA has three elements: the Medical Research Program, Rehabilitative R&D, and Health Services R&D. The Medical Research Program includes research basic to disease and deformities, while Rehabilitative R&D includes studies on artificial appliances or substances for use in restoring structure or func- tion of parts of the human body. Finally, Health Services R&D includes research toward improve- ment, replacement, or discontinuance of health care delivery systems (32). Thus, the VA’s man- date for research and development is extremely broad and holds the potential to use animals in many programs. Patterns of Federal Animal Use APHIS is the agency within the U.S. Department of Agriculture responsible for administering and enforcing the Animal Welfare Act of 1966 (Public Law 89-544) and its amendments (see ch. 13). The act defines research facility as any individual, in- stitution, organization, or postsecondary school that uses or intends to use live animals in research, tests, or experiments and that purchases or trans- ports live animals in commerce or that receives Federal funds for research, tests, or experiments. It defines “animal”to include “any live or dead dog, cat, monkey (nonhuman primate mammal), guinea pig, hamster, rabbit, or such other warm-blooded animal, as the Secretary [of the Department of Agri- culture] may determine is being used, or is intended for use, for research, testing, experimentation, or exhibition purposes, or as a pet .“ The act excludes horses not used for research purposes and other Ch. 3—Patterns of Animal Use G 4 7 Primate Involved in Behavioral Research i Photo credit: David Hathcox C), 1935 farm animals intended for use as food or fiber. Under this definition, dead frogs used in biology classes or animals killed prior to usage are not in- cluded. Rats, mice, and birds were specifically ex- cluded from the act coverage by regulations pro- mulgated in 1977 by the Secretary of Agriculture (9 C.F.R. 1.l(n); 42 FR 31022); reporting the use of these animals is voluntary. The regulations that APHIS enforces require that each research facility fill out an Annual Report of Research Facility (see fig. 3-1) by December 1 on the preceding Federal fiscal year (October 1- Sep- tember 30). Elementary and secondary schools are exempt, as are facilities using only exempt species (rats, mice, or birds). In addition, any facility that does its own in-house breeding and does not re- ceive Federal funds does not have to file a report. Although Federal research facilities are not re- quired to register with APHIS, many of them do fill out the annual reporting forms. Each year, APHIS reports to Congress on the data collected from these forms in its Animal Welfare Enforce- ment Report. Since 1982, two lines on the Annual Report of Research Facility have listed rats and mice under column A, ‘(Animals Covered by the Act” (which is therefore no longer an accurate heading). Al- though not legally required, many respondents who used mandated species filled in the number of rats and mice anyway, either not realizing that reporting on these species is voluntary or elect- ing to report their use, Thus, for many institutions a usage figure for rats and mice is given. In other cases, though, facilities reporting on mandated spe- cies omitted data on rats and mice. Table 3-1 details the total reported animal use by research facilities within the Federal Govern- ment broken down by departments, major divi- sions, and agencies for fiscal year 1983. The An- nual Report of Research Facility requires not only that total animals used be reported, but that the animals used be categorized as being used in re- search, experiments, or tests: 1) involving no pain or distress; 2) where appropriate anesthetic, anal- gesic, or tranquilizer drugs were administered to avoid pain or distress; or 3) involving pain or dis- tress without administration of appropriate anes- thetic, analgesic, or tranquilizer drugs (see fig. 3-1). Several qualifications are necessary on the num- bers reported in table 3-1, which are based on the annual reports obtained from APHIS: G G G G The 131 research reports include only intra- mural Federal research done at Federal fa- cilities. The 131 facilities are not all the Federal facil- ities that might have used animals in 1983; at least 25 facilities did not file a report for that year. The numbers obtained were tabulated from each report. The reports were checked and corrected for improper coding of information and inaccurate addition. In many cases, these changes reflected substantial differences in the number of animals used for specific insti- tutions. The numbers for mice and rats are included from any institution that reported them volun- tarily. Several facilities, however, specifically mentioned that they were not required to sub- mit these data and did not do so. In addition to these general limitations on over- all numbers, some specific qualifications for indi- vidual departments and agencies are also war- ranted: G For FDA, table 3-1 does not include its primary research facility, the National Center for Tox- icological Research (NCTR), since no report 48 . Alternatives to Animal Use in Research, Testing, and Education I I F O R M A P P R O V E O OMB NO. 05 79-0036 L -- ——-—- - — — . — I 5. Dogs I I I I 1 — — — 1 — . 6 Cats 7 Guinea Pigs 8 Hamsters — 9. Rabbits 10. Primates * 11. Rats 12, MICe Wild Animals (specify ) I 3. . — I 4. — — i 5. CE RTI FIC ATl ON BY ATTENO 1 NG v ETIZR IN AR IAN FOR R E P O R T I N G FA CI L I TY O R IN ST IT UTi ONA L COMMI T T E E I [w.) hereby certify that the type G nd G mount Of G fU190$lc, anmthetlc, G nd tranqUlll Zln9 drugs used on Jnlmal$ dUrln9 actual research, t~stln9 Of G xDeflm6n- tatlon Includlnq Oost.oDeratlva and post-procedural care was deemed G pgroprlate to relieve pain and distress for the subject animal. 16. SIGNATURE OF ATTENDING VETIERINARIAN 1 7 . T I T L E . — I S . D A T E S I G N E D 1 1 t 9 . S I G N A T U R E O F C O M M I T T e e M E M b e R -. 2 0 . T I T L E 2 1 D A T E S I G N E D 2 0 . S I G N A T U R E O F C O M M I T T E E M E M B E R 2 3 . T I T L E 2 4 . O A T E S I G N E D [ 1 CERTIF ICATION BY HEADQUARTERS RESEARCH FACIL ITY OFFICIAL I cortlfy that tho G OOvO Is trUe, Correct, G nd COmOIOtO G nd that orofosslonally G cc.ptablo standardt govefnlng tfse csro, trsatmont, and um of animals Includlng G PPfOPrlJt* u~ Of G n*sth*tlc. G naKf@c. G nd tranwlll~lm drum durlnif G ctual research, tostlns, or .xosrlmentatlon Includlng po8t-op4ratlve G nd post-procedural are G ro b.lng follow.d by tho G bovo rcs.arch faCll RlOS or sites (7 U.S.C. Ssctlon 2143). X 8 . S I G N A T U R E Q P R E S P O N S I B L E O F F I C I A L 2 6 . T I T L E 2 7 . O A T E S I G N E D 4 VS FORM 18-23 (AUG 81) SOURCE: Animal and Plant Health Inspection Service, U.S. Department of Agriculture. Ch. 3—Patterns of Animal Use G 4 9 was filed for 1983. This probably excludes a substantial number of animals since the fis- cal year 1984 annual report for NCTR reported the use of 8 dogs, 334 rabbits, 29 primates, 14)621 rats, and 11,744 mice. G The VA has 81 facilities accredited by the American Association for Accreditation of Laboratory Animal Care (AAALAC) yet only 63 reports were filed for 1983. Therefore, there is a strong possibility that the numbers for the VA are underreported. Bearing in mind all the limitations and qualifica- tions of the data used to generate table 3-1, OTA estimates that a minimum of 1.6 million ani- mals are used annually by the Federal Gov- ernment in intramural research. The Depart- ment of Defense, the Department of Health and Human Services, and the Veterans) Ad- ministration together account for 96 percent of reported Federal animal use. DHHS alone reported 49 percent of the total. Among the six kind of animals whose inclusion in annual reporting forms is mandated by the Ani- mal Welfare Act, guinea pigs are used most often— twice as frequently as hamsters or rabbits (the sec- ond and third most used species). Overall, about the same number of dogs and primates are used, while far fewer cats are involved in Government experiments. Finally, table 3-1 suggests that cer- tain agencies do research on specific species. For example, the VA uses a disproportionately large number of dogs and the Department of the Interior is the major user of wild animals. Reports of Federal facilities indicate that most animal use falls into the experimental situation categorized as involving no pain or distress. Sixty- three percent of the animals used were in this cat- egory while 32 percent were given drugs to avoid pain or distress and only 5 percent experienced pain or distress without receiving anesthetics, anal- gesics, or tranquilizers. The largest user of drugs in experiments was the VA (62 percent of the ani- mals in this category), whereas the largest user of animals experiencing pain or distress was the Department of Defense (84 percent of the animals in this category). The latter figure maybe inflated, however, by the fact that DOD has reported mice and rats voluntarily under these categories in many cases and has listed in this column all animals dy- ing in infectious and neoplastic disease studies, which many Federal agencies may not do (43). Table 3-2 shows the trends in animal use for Fed- eral agencies as a group from 1978 to 1983, accord- ing to the Aninal Welfare Enforcement Reports submitted by APHIS to Congress (49). As with the numbers from the 1983 Annual Reports of Re- search Facilities, these data do not tell the whole story. Most important, these data do not include rats and mice, which together make up a majority of the animals used. Second, only reports that have been received by December 31 each year (the re- ports are due December 1) are included (26). It has been estimated that between 10 percent and 20 percent of the total reporting institutions fail to report by December 31 and are therefore not included in the Animal Welfare Enforcement Re- ports (17). (Thus, the 1983 data are lower in table 3-2 than in 3-1, which included all available an- nual reports.) The data are difficult to interpret due to the dif- ferent numbers of research facilities included each year. Therefore, no conclusions can be drawn about whether the trend in animal use is increas- ing or decreasing. This is also the case for trends in the use of individual species. The 1983 data do indicate, however, that no more than 8 percent of animals used in Federal programs reported here have experienced pain or distress in an experiment since 1978. The percentage of animals experienc- ing no pain or distress has remained between 50 and 60 percent, while drugs have been used to alleviate pain or distress for 30 to 40 percent of the animals. ANIMAL USE IN THE UNITED STATES OTA surveyed the available data concerning the corrected for methodological deficiencies, and numbers of laboratory animals used for research, evaluated for their statistical reliability. As a final testing, and education. These were summarized, step, estimates were made of current levels of an- 50 “ Alternatives to Animal Use in Research, Testing, and Education Table 3-1.—Research=Animai Use in the Federai Government, by Major Department and Division for Fiscai Year 1983 Department of Department of Defense Health and Animals used USDA Commerce Misc. Air Force Army Navy Total Human ServicesFDA NIDA Facilities reporting . . . . . . . . 11 1 3 6 20 10 39 1 1 Dogs . . . . . . . . . . . . . . . . . . . . 25 0 994 635 827 344 2,800 113 51 ‘/0 row. . . . . . . . . . . . . . . . . . . <1 0 11 7 9 4 31 1 <1 Cats . . . . . . . . . . . . . . . . . . . . 39 0 491 61 214 36 802 0 84 ‘/0 row. . . . . . . . . . . . . . . . . . . 1 0 18 2 8 1 29 0 3 Guinea pigs . . . . . . . . . . . . . . 6,105 0 1,601 586 26,695 609 29,491 0 98 0/0 row. . . . . . . . . . . . . . . . . . . 9 0 2 1 41 1 46 0 <1 Hamsters. , . . . . . . . . . . . . . . 7,487 0 627 1,352 4,822 417 7,218 0 ‘/0 row. . . . . . . . . . . . . . . . . . . 21 0 2 4 14 1 21 : o Rabbits . . . . . . . . . . . . . . . . . 1,047 0 1,863 703 3,731 264 8,581 0 0 ‘/0 row. . . . . . . . . . . . . . . . . . . 4 0 6 2 13 1 23 0 0 Primates ., . . . . . . . . . . . . . . 0 418 527 676 219 1,840 0 0 ‘/0 row. . . . . . . . . . . . . . . . . . . : o 6 7 9 3 25 0 0 Rats . . . . . . . . . . . . . . . . . . . . 7,862 0 25,259 10,570 55,057 4,243 95,128 0 312 ‘/0 row. , . . . . . . . . . . . . . . . . . 2 0 6 2 13 1 22 0 <1 Mice . . . . . . . . . . . . . . . . . . . . 30,625 0 72,085 6,140 143,503 42,094 263,822 0 600 ‘/0 row. . . . . . . . . . . . . . . . . . . 3 0 7 1 14 4 26 0 <1 Wild animals . . . . . . . . . . . . . 24 43 1,377 34 2,762 479 4,652 0 ‘/0 row. . . . . . . . . . . . . . . . . . . < 1 < 1 10 <1 19 3 32 : o Total . . . . . . . . . . . . . . . . . . . . 53,214 43 104,715 20,806 238,287 48,705 412,315 113 1145 ‘/0 row. . . . . . . . . . . . . . . . . . . 3 <1 6 1 15 3 25 <1 <1 KEY: USDA-United States Department of Agriculture; FDA-Food and Drug Admlnistration; NIDA-National Institute on Drug Abuse; NIH-National Institutes of Health; CDC-Centers for Disease Control; NIOSH-National Institute for Occupational Safety and Health; DOT-Department of Transportation; EPA-Environmental Protec- tion Agency; NASA-National Aeronautics and Space Administration; VA-Veterans’ Administration; CPSC-f.kmsumer Product Safety Commission. Percentages may not add up to 100 due to rounding. SOURCE: Office of Technology Assessment, from 1963 APHIS Annual Reports of Research Facilities (Form 18-23); CPSC data from K.C. Gupta, Deputy Director, Divi- sion of Health Sciences Laboratory, Directorate for Health Sciences, U.S. Consumer Product Safety Commission, Washington, DC, personal communica- tion, Sept. 24, 19S5. nual animal use in the United States. The purpose of this exercise was to examine numbers on ani- mal use and compare the reliability of estimates from different data sources. The figures published in this assessment on the number of animals used are not abso- lute. They are only as accurate as the data from which they were obtained. All publicly available information on past and current animal use was collected from a variety of sources, often through personal contacts. Data from the two most reliable sources, the Institute of Laboratory Ani- mal Resources (ILAR) of the National Research Council and the USDA’s Animal and Plant Health Inspection Service, were corrected to take into ac- count the actual years of reporting and the omis- sion of certain data that were not received before a deadline. Laboratory-animal use was then estimated and projected using statistical techniques where appro- priate. For this purpose, the corrected ILAR and APHIS data were used, as well as more indirect means based on National Institutes of Health fund- ing, National Cancer Institute (NCI) usage, and NIH total usage as a function of NIH intramural use. Although the number of animals bred should lead to good estimates of animals used in the labora- tory, the larger laboratory-animal breeders would not confirm or deny sales figures that had appeared in the news media and literature. Therefore, esti- mates based on such reports are of uncertain relia- bility. Limitations of Animal-Use Study Two types of limits on this study exist: intrinsic and extrinsic. The major intrinsic limitations were funding constraints and a limited time span dur- ing which the study could be performed. This pro- hibited the collection of raw data and required that OTA rely on existing data sources. The extrinsic Ch. 3—Patterns of Animal Use G 5 1 Table 3-1 .—Research.Animal Use in the Federal Government, by Major Department and Division for Fiscal Year 1983 (Continued) Department of Health and Human Services Animals used NIH CDC/NIOSH Total Interior DOT EPA NASA VA CPSC Total Facilities reporting . . . 3 Dogs . . . . . . . . . . . . . . . 756 0/0 row. . . . . . . . . . . . . . 8 Cats . . . . . . . . . . . . . . . 503 0/0 row. . . . . . . . . . . . . . 18 Guinea pigs ... , . . . . . 23,973 0/0 row. . . . . . . . . . . . . . 37 Hamsters. . . . . . . . . . . 14,003 0/0 row. . . . . . . . . . . . . . 40 Rabbits . . . . . . . . . . . . 8,783 0/0 row. . . . . . . . . . . . . . 30 Primates . . . . . . . . . . . 4,452 0/0 row. . . . . . . . . . . . . . 61 Rats . . . . . . . . . . . . . . . 196,458 0/0 row. . . . . . . . . . . . . . 45 Mice . . . . . . . . . . . . . . . 533,094 0/0 row. . . . . . . . . . . . . . 52 Wild animals . . . . . . . . 2,787 0/0 row. . . . . . . . . . . . . . 19 Total . . . . . . . . . . . . . . . 784,809 % row. . . . . . . . . . . . . . 48 2 0 0 0 0 0 0 10 0 30 <1 287 4 3,750 <1 1,120 <1 0 0 5,197 <1 7 920 10 587 21 24,071 37 14,013 40 8,813 30 4,739 65 200,520 46 534,814 52 2,787 19 791 ,264 49 2 0 0 0 0 0 0 0 0 0 0 0 0 900 <1 923 <1 4,228 29 6,051 <1 1 30 <1 0 0 0 0 0 0 0 0 0 0 150 0 4,552 <1 0 0 4,732 <1 4 2 <1 0 0 978 2 1,723 5 842 3 33 <1 0 0 0 0 0 0 3,578 <1 2 14 <1 40 1 58 <1 0 0 74 <1 184 3 3,936 1 622 <1 232 2 5,160 <1 63 5,187 58 1,304 47 3,747 6 4,732 14 11,508 39 461 6 122,872 ‘ 28 188,560 18 2,393 17 340,764 21 1 0 0 0 0 0 0 0 0 600 2 0 0 2,080 <1 0 0 0 0 2,680 <1 131 8,978 100 2,772 100 84,450 100 35,173 100 29,445 100 7,257 100 433,449 100 1,023,918 100 14,358 100 1,619,801 100 KEY: USDA-United States Department of Agriculture; FDA-Food and Drug Administration; NIDA-National Institute on Drug Abuse; NIH-National Institutes of Health; CDC-Centers for Disease Control; NIOSH-National Institute for Occupational Safety and Health; DOT-Department of Transportation; EPA-Environmental Protec- tion Agency; NASA-National Aeronautics and Space Administration; VA-Veterans’ Administration; CPSC-Consumer Product Safety Commission. Percentages may not add up to 100 due to rounding. SOURCE: Office of Technology Assessment, from 1983 APHIS Annual Reports of Research Facilities (Form 18-23); CPSC data from K.C. Gupta, Deputy Director, Divi- sion of Health Sciences Laboratory, Directorate for Health Sciences, U.S. Consumer Product Safety Commission, Washington, DC, personal communica- tion, Sept. 24, 1985. Table 3=2.—Total Numbers of Animals Used in Federal Government Facilities as Reported to Congress in APHIS Animal Welfare Enforcement Reports, 1978-83 Fiscal year 1978 1979 1980 1981 1982 1983 Federal facilities included in reports. . . . 188 150 118 131 131 Dogs . . . . . . . . . . . . . . . . . 20,128 15,605 13,153 13,930 6,369 Cats. . . . . . . . . . . . . . . . . . 5,354 4,709 3,388 3,183 1,940 Primates . . . . . . . . . . . . . . 7,286 5,031 3,459 3,081 6,907 Guinea pigs . . . . . . . . . . . 65,009 40,425 25,402 33,495 45,972 Hamsters . . . . . . . . . . . . . 45,291 25,213 17,830 32,367 35,220 Rabbits , . . . . . . . . . . . . . . 43,867 32,205 21,631 21,962 16,209 Wild animals . . . . . . . . . . 5,537 4,137 3,209 2,007 7,618 Total animalsa . . . . . . . 192,472 127,325 88,052 110,025 120,235 aT~tala fjo not lnCIU& rats or mice, two species that together account for the majority of animals used. 6 , 6 ; 1,825 1,837 36,033 18,992 16,355 8,037 89,747 SOURCE: Office Technology Assessment, from APHIS Animal Welfare Enforcement Reports, 1978-1983. 52 . Alternatives to Animal Use in Research, Testing, and Education limitations include various information deficien- cies, such as: G G G G G inadequacies of information on most of the survey and data collection methodologies, difficulties with definitions, problems with categorizing areas of use, reporting requirements of sources, and animals under different data an inability to verify completeness of data sources. For example, there is often a discrepancy in the definition of the term “use.” In some cases, the term reflects the number of animals acquired; in other cases, it corresponds only to those used in labora- tory experiments. This distinction is frequently obscured in the data sources, and only after care- ful reading of the documents (and, sometimes, per- sonal inquiry) was the definition used in each case clarified. This leads to large differences in num- bers, since not all animals acquired are used in experiments. It also makes any comparative anal- ysis between surveys very unsound. In addition to this problem of the difference be- tween production and use, the extrinsic problem of the number of animals not used in a procedure because they do not fit the proper criteria comes into any extrapolation of animal use from labora- tory-animal market share data, A substantial pro- portion of the animals bred for research die or must be discarded because they do not meet pro- tocol specifications (age, sex, weight, general health). The number has been estimated as be- tween a few percent of those acquired to almost 50 percent. In general, the unused proportion of a species is inversely related to the cost of the ani- mals. In other words, the more expensive the ani- mal, the less likely it will be unused, once bred or purchased. Thus, nonhuman primates are much less likely to go unused than are mice or rats; in some cases 50 percent of a rodent species may go unused. Using only one sex of a rodent species in a given experiment, for example, would account for 50 percent of the animals going unused. This information must be borne in mind when compar- ing “production” with “use” and when estimating animal use. Overall, these limitations reflect on the accuracy of the data and any projections based on them. The limitations are such that the only reasonably credible source for current use and projections is APHIS, particularly its institutional data sheets (the Annual Report of Research Facility discussed earlier). Only the detailed APHIS institutional data sheets for fiscal years 1982 and 1983 were used in this assessment, though those for earlier years were also available (although they would not have had any data on mice or rats, which were not even listed on the form until 1982). Consequently, the APHIS data are less reliable for the years before 1982 inasmuch as these are based on reports to Congress that did not contain late-reporting insti- tutions. (The Animal Welfare Enforcement Reports to Congress underestimate use of the mandated species by 10 to 20 percent due to the cutoff date and do not treat data from Federal institutions con- sistently (17).) For some species, such as fish and birds, only rough estimates of use could be ob- tained, due to the diffuse nature of use and the fact that they are not included in the APHIS data. Critical Evaluation of Animal-Use Estimates In evaluating the reliability of various data sources, the following parameters were con- sidered: G G G G ability to trace the methodology used in pro- ducing the numbers, including the survey technique; ability to extrapolate to nonreporting institu- tions, which implies that there is a clear state- ment as to which institutions did or did not report data; method of data collection, whether some for- mal manner or through a few interviews, re- sulting in broad estimates; and ability to determine the fraction of animals . reported as being actually used in lab experi- mentation, as contrasted to, for example, ani- mal husbandry. These parameters were chosen because meet- ing these criteria permits extrapolation of the limited data to the entire population of institutions. Ch. 3—Patterns of Animal Use G 5 3 In general, if the numbers cannot be justified through some rational process (such as the above), too much significance should not be attached to them. These four criteria were used to assign a confi- dence rating to each data source. The confidence categories are: ‘(excellent)” “good,” “fair,” ‘(poor, ” and “indeterminate .“ (These ratings refer only to the published numbers, not to their usefulness as a predictive tool.) Such a confidence rating is nec- essarily subjective; the categories are comparative and should not be viewed as absolute. Upon reviewing all the data sources avail- able for predicting the laboratory-animal use in the United States, it is clear that no source accurately portrays the number of animals being used. Each has methodological prob- lems that prevented it from accurately count- ing all users of animals. What follows is an anal- ysis of the available data sources and how they rank in comparison with the other surveys in terms of confidence and reliability. USDA Animal and Plant Health Inspection Service (APHIS) The 1982 and 1983 data were analyzed on a case- by-case basis. Copies of the original report forms were obtained from USDA; they were sorted by institution type, checked, coded, and entered into a computer database. Comparing the 1982 and 1983 APHIS data (see table 3-5, in the ‘(Summary and Analysis of Estimates” section) with the USDA Animal Welfare Enforcement Report for 1980 (the APHIS 1980 data in table 3-5) reveals a large dis- crepancy. The USDA reports invariably contain lower numbers for all species, as the data sheets received after the December 31 cutoff date are not included in reports in either the current or the next fiscal year. It is estimated that between 10 and 20 percent of the reports are not used to compile the report to Congress in a given year (17). This limitation does not apply to the results con- tained in the present compilation for 1982 and 1983, since all data for a given year were used no matter when received. The assumption is made that copies of virtually all of the data sheets re- ceived by USDA in the 1982 and 1983 are used in this study. No verification was made of which institutions did not report. The number of institutions reporting to APHIS has hovered around 1,000 since 1972. The num- bers for 1982 and 1983 (shown in table 3-6, in the “Summary and Analysis of Estimates” section) were tabulated from the actual summary data sheets provided to APHIS by the institutions and include all possible reports. Even these figures—1, 127 for 1982 and 1,146 for 1983—are probably low, as not all institutions submit reports. (The total number of institutions registered by APHIS was 1,113 in 1982 and 1,166 in 1983; this excludes Federal agen- cies, which are not required to register.) Some of the institutions may not report because they have not used any animals that year, or because they have only used exempt species. For the six required species listed on the form (dogs, cats, guinea pigs, hamsters, rabbits, and pri- mates), the numbers reported provide a very close approximation of the animals actually used. Thus these data were assigned a confidence rating of ‘(excellent .“ (For a summary of all the data sources discussed in this section and their confidence rat- ings, see table 3-3.) For exempt species (primarily rats and mice), it is possible to estimate the num- ber of unreported rats and mice by extrapolating from the numbers reported (see the section on ‘(Es- timate Using Corrected APHIS Data”). Some com- mentators (1)3)2 7) claim, however, that a certain number of exempt animals go unreported—and would be missed in an extrapolation-because they are purchased directly by the user and not re- ported to the central facility. This contention could not be confirmed. Therefore, the voluntarily re- ported data on rats and mice on the 1982 and 1983 APHIS annual reports received a confidence rat- ing of “good. ” ILAR Surveys The Institute for Laboratory Animal Resources, a component of the National Research Council, periodically surveys users of laboratory animals (18,19,20,21,22,23), although it is generally more concerned with facilities and personnel than with quantity of animals used. The ILAR data repre- sent the number of animals “acquired by own 54 G Alternatives to Animal Use in Research, Testing, and Education Table 3-3.-ReIiabiiity of Various Data Sources Years Confidence Source covered rating Strength(s) Limitation(s) USDA/APHIS: Mandated species 1982-83 Excellent Required by law. Data available by institution, thus extrapolation to nonreporters is possible Mandated species 1972-81 Fair Required by law. Data by 10 to 20 percent of institutions institution available, but not not included in reports to Con- used gress. Totals not consistent (some years include Federal agencies, others do not) Data by institution available. Not required by law Rats and mice were on the form so anyone who reported probably provided an accurate number. Many did not realize that these were voluntary since they were listed on form. Extra- polation possible Exempt species 1982-83 Good ILAR Surveys of 1965-71 Poor Of some use in establishing Laboratory Animal Use trends for that period 1968 Survey 1967 Fair Statistically sound survey. Possibility of extrapolating to other institutions 1980 Survey 1978 Fair Thorough and statistically solid. Extrapolation to non- reporting institutions possible Old data. Cannot extrapolate to missing data Limited to 683 Federal-grant- eligible institutions Primary attention given to nonprofit Federal-grant-eligible institutions. Not required by law to be filled out W.B. Saunders & Co. 1965 Indeterminate Company defunct, survey methodology unclear; no evaluation possible Foster D. Snell 1975 Indeterminate Data appear to be based on in- terviews with two breeders Methodology unclear. Person- nel no longer available SOURCE: Office of Technology Assessment. breeding and from commercial sources,” not nec- essarily the number actually used in experimen- tation. The ILAR and APHIS surveys are so different in their organization and methodology that it is not meaningful to compare the two sources, even in years for which data from both are available. It is also difficult to point out significant changes within this data source because the ILAR method- ology varied over time and could not be verified adequately, so changes in numbers are difficult to substantiate. ILAR Surveys of Laboratory Animal Use (20) con- sist of tables summarizing the results of question- naires on the number of animals used for research. As ILAR personnel cannot discern who was sur- veyed and who responded, extrapolation for miss- ing data is impossible. The surveys could, how- ever, be of some use in assessing trends between 1965 and 1971. A “poor” confidence rating was given. The 1968 Survey of Laboratory Animal Facilities and Resources (21) appears to have been a very thorough and statistically sound survey including all known users of laboratory animals. The results shown, however, are only for the 683 organiza- tions eligible for Federal grants that responded be- cause of the interest of the survey sponsor (NIH). It is possible, however, to normalize for missing data based on the reported biomedical research expenditures for these 683 organizations of $920 Ch. 3—Patterns of Animal Use G 5 5 Table 3-3.–Reliabiiity of Various Data Sources (Continued) Years Confidence Source covered rating Strength(s) Limitation(s) Alex Brown & Sons 1981 Poor At the time, it was thought to Data based on a few inter- represent best estimate for lab views, and mostly broad animals in U.S. market estimates Andrew N. Rowan 1985 Poor Data distinguishes between Broad analysis with many production, acquisition, and assumptions. Based mainly on actual use one breeding facility Amphibians: Emmons 1989 indeterminate Giobal estimates Culley 1981 Indeterminate Many assumptions Nace 1974-81 Fair Fair detail for basis of Difficult to know actual estimates numbers due to large number of users Various, on fish 1983 Fair Data consistent Global estimates only usage Various, on bird 1983 Poor Good detail by institutions Uncertainty about nonreporting usage institutions, and fraction of fowl used by lab experimentation Data on animal trends: Wadsworth Center, NY 1980-83 Poor Good detail of different Difficult to predict any trends species used Johns Hopkins, MD 1975-85 Poor Limited data that are impossible to analyze SOURCE: Office of Technology Assessment. million in fiscal year 1967. (The results for all re- spondents, while not mentioned in the report, were compiled and reported for comparison purposes in the ILAR 1980 survey,) The confidence rating was “fair. ” The ILAR National Survey of Laboratory Ani- mal Facilities and Resources (22) also appears to be a thorough and statistically solid report, al- though the data (for fiscal year 1978) are now 8 years old. Since it also was funded by NIH, pri- mary attention was given to nonprofit biomedical research institutions eligible for Federal grants. In addition, data were received from Federal orga- nizations, commercial research labs, and the phar- maceutical industry. Seventy-two percent of the 2,637 questionnaires were returned; 47 percent of those were acceptable, thus providing 1,252 re- spondents (including 992 nonprofit Federal-grant- eligibles, 137 commercial laboratories, 25 compo- nents of the DOD, 21 units of NIH, and 77 compo- nents of other Federal agencies). Although the individual identities of the respondents are un- known, the biomedical research expenditures of the nonprofit organizations are known. Since their data are reported separately from all respondents, an extrapolation to the unknown cases can be at- tempted based on the known national (meaning “all use in the United States”) biomedical research expenditures. This source was assigned a confi- dence rating of ‘(fair.” W.B. Saunders & Company W.B. Saunders&Company (41) surveyed the lab- oratory animal market in 1965 and projected fig- ures for 1970. The survey and its estimates are widely quoted as one of the first estimates of ani- mal use. The survey methodology is unclear and the company no longer exists, so these data fall under the “indeterminate” category. Foster D. Snell, Inc., for Manufacturing Chemists Association A study performed by Foster D. Snell, Inc., for the Manufacturing Chemists Association (25) esti- mated that 35 million mice and 40 miIlion rats were 56 G Alternatives to Animal Use in Research, Testing, and Education produced domestically in the United States in 1975, and that 20)000 monkeys were imported from India. The report’s authors could not be located and the methodology is unclear, thus making it impossible to validate. It appears that the data are based on interviews with personnel at two ani- mal breeding facilities (Charles River Breeding Labs, Inc., and White Eagle Farms) and perhaps a few other people in industry, academia, and gov- ernment. As it is difficult to give any credibility to such data, the source was assigned a confidence rating of “indeterminate. ” Alex Brown & Sons An Alex Brown&Sons (2) report on Charles River Breeding Labs, Inc., stated that the company pro- duces 22 million animals annually worldwide, spe- cializing in mice, rats, guinea pigs, hamsters, and primates. It did not give any breakdown by spe- cies, nor do any other analyses of Charles River. The number was primarily a guess based on a few interviews and so its value must be questioned. The confidence rating of this source was “poor.” Andrew N. Rowan In a 1984 book, Of Mice, Models, & Men:A Criti- cal Evaluation of Animal Research, Andrew N. Rowan estimated that approximately 71 million laboratory animals are used each year, including 45 million mice and 15 million rats (39), These figures were obtained by looking at all the availa- ble data sources for animal use in the United States, especially information on Charles River breeding production. In 1985, Rowan revised these estimates to distinguish between production, acquisition, and actual use. The new estimates on animals used sug- gest that between 25 and 35 million animals are used per year (40). As these are based on a very broad analysis with many assumptions, they have been given a confidence rating of “poor.” Surveys and Estimates on Amphibians, Fish, and Birds There is little good survey information on labora- tory use of amphibians, fish, or birds. Use of these animals is not required to be reported on the USDA/ APHIS annual reports. Therefore, the only sources of estimates are personal communications with experts in these fields. The most recent assessments of amphibian use were the ILAR surveys of 1965-71, which indicated the use of 3.37 million amphibians in 1971. As men- tioned earlier, however, it is not known how to normalize for institutions that did not report, so the usefulness of these data are questionable and the confidence rating is “indeterminate.” Several individuals who use or produce amphib- ians were surveyed, yielding a wide range of esti- mates. A former general manager of a major sup- plier of amphibians estimated that approximately 9 million frogs were shipped by suppliers in 1969 for educational and teaching purposes (13). This is a global estimate and so its confidence rating was considered “indeterminate. ” An amphibian researcher at Louisiana State University did a sur- vey of the use of bullfrogs that estimated that 150,000 bullfrogs and 200,000 tadpoles” could have been used in 1981 (a decrease since 1971, he found). He then assumed that bullfrogs represent roughly 10 percent of amphibian use and estimated that about 1 million frogs and 2 million tadpoles were used in the United States for teaching and research in 1981 (8). The assumptions in this method are very general and so the value of this estimate is questionable; an “indeterminate” rating was as- signed. Finally, George Nace (34,35) estimated that about 9 million frogs were shipped by suppliers in 1971, but that this dropped to roughly 4.5 mil- lion in 1981 and stabilized at that level in 1984, with 90 percent of the usage educational and 10 percent research. There is fairly good detail for the basis of the estimates, but it is difficult to con- firm the totals due to the large number of users. This source was given a confidence rating of “fair.” Reliable data on fish used in laboratories were particularly difficult to obtain. Estimates were received from commercial and institutional (includ- ing Government) users in the field. For fish over half an inch long, the yearly use appears to range between 500,000 and 1 million. For smaller fish, the best estimate is that 2 million to 3 million are used yearly. Most are used for toxicological studies. Although the numbers are fairly consistent from source to source, they are only global estimates and so were given a confidence rating of “fair. ” These numbers apply only to laboratory use. They do not include fish that are used in the wild in propagation, contamination, feeding, and other ecological studies. Ch. 3—Patterns of Animal Use G 5 7 For birds, many of those completing the APHIS data sheets voluntarily reported bird use under the “wild animal” category. According to these data, at least 33,910 birds were used in fiscal year 1982 and 29,781 in fiscal year 1983. Of these, the Univer- sity of Maryland used 17)915 birds in 1982, and 12,305 in 1983 (46). Since this one institution used such a large fraction of the reported total, inquiries about other large possible users indicated that many of the poultry research institutions (mostly land-grant universities in the East and South) did not report birds on their APHIS forms, The largest of these, in terms of poultry research, is North Carolina State University, from whom it was learned that approximately 41,000 birds were used for poultry science and 1,100 in veterinary schools (7). Checking the APHIS data sheets for other land- grant institutions showed that most had reported bird usage. In addition, discussions with research- ers at several institutions established that only 80 to 85 percent of the poultry science usage is in laboratories with the remainder mostly in feed- ing, management, and breeding studies. Therefore, although there is good detail for many institutions on bird use, there is uncertainty in the APHIS data about nonreporting institutions and about the pro- portion of fowl used in actual experimentation. Several individuals have estimated bird use in the United States. James Will of the Animal Re- source Center at the University of Wisconsin in Madison, WI, estimated that 25,000 to 100,000 avian individuals are used for laboratory experi- mentation (54). Andrew N. Rowan of Tufts Univer- sity School of Veterinary Medicine in Boston, MA, estimated that at least 500,000 birds are used in biomedical research (0o). Both of these figures are based on very weak data and so are assigned a confidence rating of “poor. ” Thus, using these esti- mates and the APHIS bird data, an annual use of between 100,000and 500,000 birds is as accurate an estimate as can be made. Data on Trends in Animal Use Several limited data sources exist that suggest trends in animal use in the past several years. At Wadsworth Center for Laboratories and Research, New York State Department of Health (Albany, NY), the use of mandated species decreased 40 percent from 2,925 in 1980 to 1,754 in 1983. The use of rats and mice also decreased substantially (22 per- cent), from 72,796 in 1980 to 56,681 in 1983, at a time when total research dollars available con- tinued to increase (11. At The John Hopkins School of Hygiene and Public Health in Baltimore, MD, the daily census of animals decreased from over 8,000 in 1975 to approximately 2,000 in 1985 while animal care personnel dropped from 10 to 4 and research expenditures more than doubled (14). These data sources are limited in scope, use differ- ent counting mechanisms, and can be considered anecdotal in nature. They were assigned a confi- dence rating of “poor. ” Calculating Rat and Mouse Usage Using these same data sources, estimates for an- nual laboratory use of rats and mice in the United States were calculated. The criteria and scales de- scribed earlier were also applied to assign confi- dence ratings to the estimates. To gauge annual laboratory-animal use, minimum average costs of $4 per rat and $2 per mouse (6,15,24,30,36,55) were assumed to represent conservative prices for a typical research subject. This permitted extrap- olations based on price to represent an expected maximum of animals that could be purchased. Three different methods were used to estimate the use of rats and mice in the United States. The first involved using indirect means for the calcu- lations, while a second method used 1978 ILAR data. The third, and most reliable, method relied on corrected USDA/APHIS data and involved cal- culations using regression equations. Indirect Estimates Possible methods for estimating rat and mouse usage under this category involve extrapolations from data based on NIH funding, NCI usage, NIH total use as a function of intramural use, and ani- mal breeder information. For example, an estimate based on NIH funding involves the following steps and assumptions: G NIH funds 37 percent of all national biomedi- cal research expenditures (52). G In 1983, NIH awarded $582,571)000 in direct costs to 5,011 extramural projects utilizing rats and other species (4). If it is assumed that all 38-75o 0 - 86 - 3 58 . Alternatives to Animal Use in Research, Testing, and Education G G G G G expenditures went to projects that used only rats, an upper limit can be extrapolated for rats purchasable using NIH funds. Twelve percent of direct costs of NIH- sponsored research funds go toward the pur- chase of supplies, glassware, chemicals, re- search animals, and items listed as expenda- ble (55). If it is assumed that half of the supply funds went toward the purchase of animals, then $34,954,260 would be available for the pur- chase of rats. At $4 a rat, 8.7 million rats could be purchased. In 1983, NIH awarded $531,519)000 in direct costs to 4,080 projects using mice. At an aver- age cost of $2 per mouse, 16 million mice could be purchased with NIH funds. Assuming that NIH supports 37 percent of ani- mal use in the country, then the potential num- ber of these two species purchasable in the United States is estimated at 23.6 million rats and 43.1 million mice. This indirect method (whether it uses NIH data or NCI data or ani. mal breeder information) involves many as- sumptions, limited data sources, and cannot be considered very reliable. It was assigned a “poor” confidence rating. Estimate Using Corrected ILAR Data, 1978 The results of the 1978 National Survey (22) per- mit approximation of animal use for all users with techniques that fill in the missing data of non- respondents based on a method such as the fol- lowing: G G G The NIH-grant-eligible nonprofit biomedical research organizations responding to the sur- vey reported biomedical research expendi- tures of $2,2 billion for 1978. Total national biomedical research expendi- tures are estimated at $6.27 billion for 1978 (52). If it is assumed that animal use (in numbers) is proportional to the dollar amount spent on research utilizing them and that the usage rate of animals by all institutions is proportional to that of nonprofit institutions, national usage equals (nonprofit ILAR 1980) X 6.27/2.2. This yields an estimate of 16 million mice and 5.6 million rats used in 1978. Such methods do in- volve some assumptions not easily justifiable and so the confidence rating is somewhat lower than for the ILAR data on which they are based. In addition, they are based on in- formation already 8 years old. Estimate Using Corrected APHIS Data About two-thirds of the institutions completing APHIS annual reports for 1982 and 1983 volun- teered information on the number of rats and mice used. Regression equations based on those insti- tutions reporting the specific species on the An- nual Report of Research Facility forms were used to estimate the numbers of rats and mice for those institutions not reporting these species (17). The estimates obtained using these regression equa- tions and then simply applying the mean value for reporting institutions to the nonreporters are shown in table 3-4 (which summarizes all the esti- mates discussed). These regression equations yield estimates of 8.5 million mice and between 3.4 mil- lion and 3.7 million rats used annually in 1982 and 1983; applying the mean value for reporting insti- tutions to those that did not report yields higher estimates. Given the fairly detailed database to which the regression equations were applied, these estimates received a confidence rating of “good. ” The estimates generated from these corrected APHIS data are likely the most accurate that can be obtained with data currently available. Summary and Analysis of Estimates Table 3-5 summarizes the various estimates on animal use discussed in this chapter. Several fac- tors reduce the usefulness of these data, however: APHIS’s definition of animal (which excludes rats, mice, and birds) and the exemption from regula- tion of research facilities that do in-house breed- ing and receive no Federal funds. These limitations may cause the numbers generated from the APHIS data to be underestimations of the total animal use in the United States for research, testing, and edu- cation. For example, the Directory of Toxicology Testing Laboratories published by the Chemical Specialties Manufacturers Association, Inc., lists 110 facilities in the United States. In checking these against the list of APHIS registered research facil- Ch. 3—Patterns of Animal Use . 59 Table 3-4—Estimates of Rat and Mouse Usage in Laboratories, 1978, 1982, 1983 Mice Rats (millions) (millions) Confidence Basis of estimation 1978 1982 1983 1978 1982 1983 rating Indirect means—NIH funding . . . . — — 43.1 — — 23.6 Poor Corrected ILAR data: Nonprofit funding share. . . . . . 16.0 — — 5.6 — — Fair Corrected APHIS data: Regression . . . . . . . . . . . . . . . . . — 8.5 8.5 — 3.4 3.7 Good Average . . . . . . . . . . . . . . . . . . . . — 10.2 11.2 — 4.1 4.6 Good SOURCE: Off Ice of Technology Assessment. ities, 40 percent were not registered and so would not file a report. Any animals used in those facil- ities would not be reported in the APHIS data. The 1978 ILAR National Survey of Laboratory Animal Facilities and Resources stated that 35 percent of mice and 19 percent of rats acquired for research were bred in-house by the researchers (22), so these too might not appear on the the APHIS data sheets. Thus, all these limitations mean the APHIS data may be underestimations of total animal use, but it is impossible to estimate if the difference is significant. Ideally, the results based on APHIS data could be compared with estimates based on animal breeder numbers. However, since informa- tion on distribution of costs per animal is proprie- tary, such an analysis is impossible. Therefore, al- though the data contained in the APHIS reports are the most reliable, they do not include all possi- ble users of laboratory animals. Inspection of some 150 institutional Annual Re- port of Research Facility forms raises several other doubts as to the accuracy of the data collected by APHIS. In general, the form seems to lack any in- struction to the individual institutions on how it should be filled out. As a result, there is no con- sistency in the ways in which forms are completed. The reliability of the data on the forms today is in question. Figures 3-2, 3-3, and 3-4, which exem- plify the reporting problems, are actual forms re- turned to APHIS for 1983, although the institution names have been deleted. For example: G Some forms have an error that can lead to miscalculations of the number of animals used: Column F asks for the addition of columns B +C +D +E. The actual number desired is C +D +E. Thus, some reports have dou- G G G bled the number of animals used (since B =C +D +E) (see fig. 3-2). These types of mis- calculations, along with normal mathemati- cal errors, were corrected in the OTA esti- mate of animal use in the Federal Government. Thus, the numbers for Federal agencies in these two sections are different for the same APHIS institutional reports. (For Federal agen- cies, this difference is fairly small.) In many cases, respondents did not seem to understand how to classify the animals used in the different experimental categories. If the APHIS form is read literally, any animal given drugs to avoid pain or distress is also an animal that experiences no pain or dis- tress and could be counted in both catego- ries (See fig. 3-3). The answers to the category “wild animals” differed greatly. Some forms listed legitimate wild animals, such as seals, while others in- cluded as wild such animals as gerbils, cattle, sheep, and pigs (see fig. 3-4). In fact, the “wild animals” line was often filled in with farm ani- mals, which are exempt from being reported. The forms are now improperly labeled in that rats and mice are included under col- umn A, “Animals Covered by the Act, ” yet they are specifically exempted by USDA regulation from coverage by the Animal Welfare Act. Many institutions that filled out APHIS forms may have been unaware that re- porting rats and mice was voluntary. These examples serve to characterize the present system as lacking clarity and uniformity in def- inition and accurate reporting, Redesign and en- hanced explanation of the APHIS form would lead to collection of more accurate data on animal use. Table 3-5.—Various Estimates of the Number of Animals Used in the United States W. B. Saunders W. B. Saunders ILAR ILAR ILAR APHIS APHIS APHIS Health Designs (estimate) (projection) (estimate) Group Species 1965 1970 1967 1970 1978 1 9 8 0a 1982 b 1983 b 1983 1,371 1,523 1,252Number of reporting institutions . . . . . . . Rodents Total Mice Rats Hamsters Guinea pigs Other rodents Rabbits Total Carnivores Total Cats Dogs Other carnivores Ungulates Total Nonhuman primates Total Birds Total Amphibians Total Frogs and toads Other Total ALL ANIMALS TOTAL 975 1,127 1,146— — 13,175,716 8,500,000 3,700,000 454,479 521,237— 509,052 237,771 55,346 182,425 — — 59,336 Ioo,oooc 500,000C— 4,000,000 c’f 18,561,875 58,440,000 36,840,000 15,660,000 3,300,000 2,520,000 120,000 94,480,000 59,560,000 25,320,000 5,340,000 4,070,000 190,000 30,363,000 22,772,300 6,131,000 785,900 613,300 60,500 504,500 370,400 99,300 262,000 9,100 106,200 57,700 2,070,500 — — — 33,472,300 37,247,377 18,646,171 25,687,067 13,413,813 9,870,628 4,358,766 870,056 368,934 737,899 426,665 81,727 79,993 494,591 439,986 247,310 242,961 56,646 54,908 182,728 183,063 7,936 4,990 95,636 144,595 54,437 30,323 667,263 450,352 2,039,490 — 2,022,755 — 601,663d — 41.667.767 19,956,386 828,216 10,530,685 — 6,889,744 — 2,725,814 405,826 417,267 422,390 497,860 — — 12,156,377 7,913,167 3,269,494 454,479 521,237 — 1,560,000 2,520,000 471,297 547,312 509,052 257,265 254,628 68,482 59,961 188,783 194,867 — — 237,771 55,346 182,425 — — — — — — — — — — 58,024 54,565 59,336— — — — — — — — — — — — — — 4 9 , 1 0 2e —— 60,000,000 97,000,000 1,661,904 11,387,390 12,964,536. . aDatq obtained from A“/ma/ we/fare Enforcement Ffepoti to Congress for 19S0. They do not include any numbers for rats and mice bData ~ompild by H=lth @signs, lnc, (RWhester, Ny) with all available Annual Reports of Re~arch Facilities The data for rats and mice are from volunta~ reporting of the use of these SpeCieS. cEStimateS Stated are highest value Of a rough ran9e. ‘Marine mammals, fish, and reptiles. ‘Wild animals. ‘Fish. SOURCE: Office of Technology Assessment. Ch. 3—Patterns of Animal Use . 61 Figure 3-2.—Example A of APHIS “Annual Report of Research Facility” U N ITEo ST A T ES OEPA RT M E N T o r A G RICU L T u R E I 1. OAT E O F R E P O R TA N I M A S - A N O P L A N T H E A L T H I N S P E C T I O N S E R V I C E 1 . “ ” w ! - . . ” ” ”v E T E R I N A R Y S E R V I C E S ?larch 7, 1984 -. - - -—. Th IS report Is rectutrect by law ( 7 USC 2 143). Fa!l r I report accoralng to the regulations cdn result In an oraer to cease dna ales, st and to be sub 3 penalt Ies as provlaea for tn Section 2150. \ [ ‘ a ’ ” ” ‘ n - - - E D i UMtl NU. 0579-0036 I 2. HE AOQU A RTERS R ESEA RC M FACILITY f ,A’r2m@ .& A ddres.c, as WgSS. I tared wt(h L’SLl . \ , i n c l u d e Z IP C o d ? ) ANNUAL REPORT OF RESEARCH FACILITY (’Required For Ldc/I Rcportl)tf Fucllif}, 11’/terc .4 ntmu/s ,ire ffeid .4 d .-111 A trelldl~ig [ ‘ercn~]urla!; Has Respmlstbility) INSTRUCTICsNS Repssrtmg FacIIItv - complete Itcms 1 ckough 24 and submit to vour Headquarters FacIII:>’ Attach additional sheets If ncccssary. S . R E G I S T R A T I O N N O. 4 R E P O R T I N G F Acl L I T V f \“ome a n d A d d r e s s , i n c l u d e Z IP C o d e / I Headquarters Facdity - complete ]tcms 25 though 27 and submit on or before December 1 of’ each year for the precedin~ Fedcr~! fiscal }“ear (October 1, to Septcmbcr 30’ to rhc Veterinarian In Ch.irgu fur the Sutc where t hc research facd~t~ hcadqu~rtcrs IS rcgtstcrcd. R E P O R T o F A N tM A LS uSEO IN ACTUAL RESE A RCfi , TESTING, O R E X P E R I M E N T A T I O N - S S C t l O n 2.28 O f A n i m a l w e l f a r e R 0 9 U l a t t 0 n S r S @ U l r e S a P P r @ Dr Ia te use of anesthetics, analgesics, ana tranau,1 Izlng arugs aur{ng re;eafch, test lng, or exDerlmentat Oon. Exsrertments !nvolvtng pain or atstress w!thout use of tnese aruqs must De feoortea and a br !ef statement explatnnnq the researcn. A. la. / c. ) D. }E. I F, An&mals COvereC! B Y A c t Number of animals used Number of animals usea in research, exper!mants, New Num Ber of an!rnals usea !a :ase.c:ci-l, :;:perfm, erots, o. tests where agtwotsriate or tests !nvolvtng pain or An!mals !n research. exDerlmer, ts, distress without aamtn(s. T O T A L N O . Aadeo th, $ or tests /rrvolv!rsg no pa~n i anesthctlc, G nalgestc, or tra!lon of apDr09rlate Of An#mals tranau!ltzer drugs were Year or rrlstress. anesthetic, analgeslc, or ( C o k . B + C + D + k’) aamlnfsterea to a v o f d pain or alstress. tranq ”ulozer drugs, fA ttach brwf exp/anatlon} I 5. Dogs ~ 610 I I Io I 610 I o 1220 - — 7. Guinea Pigs 1 0 0 r! ~. iidmstcrs o 0 1 (-) I 9. Kabblts ?4 24 0 0 48 I 10. Pr imates o 0 0 I f-l I n I 11. Rats 4500 0 4500 0 9000 12. Mice 2000 1200 800 0 4000 Wii’&RKi?tT3i3 (specz~y) ~ 3 . G ~ ~ b i l ~ 50 0 0 .- 0 50 i<. sheep 8 1 7 16 ~ 5 ’ C a t t l e 10 0 10 0 20 CERTIF ICATION BY ATTENDING VETERINARIAN FOR REPORTING FACIL ITY OR INSTITUTIONAL COMMITTEE I (we) hareby certify that the ;YIM wsa amount of G sa19esIc. anestfsetlc, G nd tranaulllzln9 drum used on an~mals durm9 G ctual research, te3tln9 or •xoer~mem tatlOn tnCJ;aln9 POSt -ODOratlva G nd O,OSt-OrOCddUral CW8 was deefnea G fsss?ooriate to relieve Daln Jnd dlstreas for tfso subject G nimal. 1 6 . S I G W A T U R E O F A fis?NDING U ETJ?Rl N4Rt AN s 7. ~STLE I a . O A T C S t G U E O . 3 / s / P ~ / A T E S l G f 4 = D 22. w GN ATU R h q C O M MI T T E E M E M B E R 23. f C 4 T L E “ C E R T I F I C A T I O N B Y H E A D Q UA R T E R S R E S E A R C H P A c I L I T Y OF F I CI A L I CertlfY that the G bove 15 true, COrreCt, and COm Olete G nd that Professlorrally G cceptable stana?rds governtng the care, tr~tment, G nd uee of G mmals Includlng G rsgroertatc use of anesthetic, dfIa19eSW ana tranQu Nlzln9 arugs, durlnq actual raeezrch, testing. or axperlmwstat!on including post-operative and post-procedural ~re are bsdnq followea by the above research fac!llt tes Or Sites ( 7 U.S. c. Section 2 143). 2 S S I G N A T U R E O F R E S P O N S I B L E O F F I C l A L 2 6 . T I T L E 2 7 , D A T E S I G N S I D b VS FORM 18-23 PreL,Ious edtl!on obsolete {AUG 81) SOURCE: Animal and Plant Health Inspection Service, U.S. Department of Agriculture. 62 G Alternatives to Animal Use in Research, Testing, and Education Figure 3=3.-Example B of APHIS “Annual Report of Research Facility” Thk reoort Is retaulrotl DY law ( 7 USC 2143). Failure to rWJrt acC0rd~n9 to tne ro91Jl~t10ns Can -esult in G n order to ceJse G nd desist and to b. subject to penaltles as cwovidetl f In SectIon 2150. UN 17=0 ST ATSSS OSS?ARTMCNT O F AG RICU LTU RC 1. D A T E O F R E P O R T A N I M A L A N O G L A N T H E A L T H I N S P E C T I O N S S S R V I C C 10/17/8s I FOR M APPROVED V C T E R I N A R V S E R V I C E S OMB NO. 0579-0036 I ANNUAL REPORT OF RESEARCH FACILITY (Required For Each Reporting Facifity Where Animals Are Held Artd An Attertdirtg Veterinarian MU Responsibility) INSTRUCTIONS Reporting Facility . complete items 1 through 24 and submit to your Headquarters Facility. Attach additional sheets if necessary. Headquarters Facility . complete items 25 through 27 and submit on or before December 1 of each year for the preceding Federal fucal year (October 1, to September 30) to the Veterinarian in Charge for the State where the research facility headquarters is rcuistered. R E P O R T O F A N I M A L S U S C O I N A C T U A L R S S S E A R C H , T C S T I NC. OR E X P prlate use of anesthetics, ana19eslcs, G nd tranqutlizlng drugs during research, tOSl these drWS must be rePOrted G nd a brief statement explaining thsj rsaearch. 4 Z . H E A O Q U A RTE R S R E S E A R C H FACI L I T V ( N a m e & A d d r e 6 & w r e i ? u - tered with USDA, melude Zip Code) . -- , 3. REGISTRATION N O 4. R E P O R T I N G F A C I LITY (Name and A d d r w a , i n c l u d e Z a p C o d e ) II MC NT AT ION - Sect Ion 2.28 of Animal Welfare Regulations retruires aocwm ), or experimentation. Experiments involvlng pain or dlstreas without uee of A . 8 . c . t D . E. F. Number of animals U i e d Numbw of G nimals used In research, G xperlmonts, In research, experlmcnts, New Number of G nimals used Animals Covered or tests where G ppropriate or tests involving pain orAnimals in research, G xperiments, distress without arYminls- TOTAL NO. B Y A c t Added this or tests involvlng no pain anestlwtic, analgesic, or tratlon of G ppropriate Of An!mals Year transrullizer drugs were or distress. anesthetic, analgesic, or (cola. + C + D + E) G dmlnktered to G void Pain or distress. tran~ullizer drugs. (Attach brwf explanation) 5. Dogs I 14 I 14 6. Cats 6 6 12 7. Guinea Pigs 18 18 8. Hamsters I 36 I 9. Rabbits 130 130 260 10. Primates il.. Rats 20 20 40 12. Mice 250 250 500 Wild Anisnda (specify)13. -14. i5.- CERTIF ICATION BY ATTENDING VETERINARIAN FOR REPORTING FACIL ITY OR INSTITUTIONAL COMMITTEE I (W.) hwcby csrtlfy that the typo G nd G mount of •nalgssl~ G toathot.i~ G fid trMIqulllZlra9 drugs used on G tlmms during G ctual rsaserch, t4Stln9 or G Xparhrrem tition Including peat-operathre and post-procedural c4re was desmed G ssprowlate to r.lleV4 PSln G nd dlatmaa fOr the subject G fSl_l. 16. SS* A TU W C O r ~ N O I N G V C T K R I N A R I A N s 7. TITLE 8 S . DA7C S IGNCD 19. s i ~ U R E ~ /o/4/1~3 M C M B C R 2 0 . T I T L E 2t . DATE S#eNCO . I I 22. SIG NATURC Of COMMITTCX M B . E R 23. TITLE 24. OATC SIGNCD I I CERTIF ICATIC$N BY HEADQUARTERS RESEARCH FACIL ITY OFFICIAL I certify that the G bove Is true, COrrSCt, G nd complete and that profosslonally G cceptable standards 9overning the ~re, trsatmont, G nd use of wslmals Irrcludlng aPPrOPrlatO use of G nOSttS6tlC, G na194Sk, and tranquilizing strugs, durlrw G ctual reasarch, testing, or sxparlmontatlon Inclsrdlng peat.opwatlw G nd post-procedural u r e ~ rch facllltles or sltea (7 U.S. C. Sactlon 2143). 28. SI+NATUR* OF 27. DATE SIGNED 1- 1 1 VS FORM 18-23 Rsviorw G ditfon obookte (AUG 81) SOURCE: Animal and Plant Health Inspactlon Service, U.S. Department of Agriculture. Ch. 3—Patterns of Animal Use “ 63 Figure 3=4.—Example C of APHIS C’Annual Report of Research Facility” &I&C& &w,r6 DI .w(7 USC21431. F . , lUreto , . ,o r ta .co~d , .g tot .e reg . , . t iomcan result in G n order to cease ana desl$t ana to be sublect tO Penaltles as Drovlded fOr In section 2150. T ATCS DEPARTMENT O F AG R ICV LTU RE DEG&wT&fh 1. OAT E O F RcPO RT O G LA NT H E A LTH INSPECTION SE RV ICf? I FORM APPROVED V E T E R I N A R Y s E R V I C E S 30 NOV 83 OMB NO. 0579-0036 2. HE AOQU ART E R S R ESE A RCU FACI LIT Y 1,\aMC A AOareSS, a W17U- w . m l & . B tered with t)SD,l, Include X IP Code) ANNUAL REPORT OF RESEARCH FACILITY (Required For 12ZCII Reporflrrg Facility It’here Animak Are Held Artd An .~ttendtng l’ctermurwt Ha-s Responsibility) [ INSTRUCTIONS I 3. R E G I S T R A T I O N N O . Roportmg Faclltty - complete Items 1 through 24 and subm]t to your 1 4 R E P O R T I N G F AC! L I T + ( A a m c a n d A d d ? e s $ , ! n c ) u d e Z w C O d e ) Headquarters Facihty. Atuch addmonal sheets If necessary. Headquarters Facilny . complete Items 25 through 27 and submit on or before December 1 of each year for the preceding Federal fual yczr (Octobe: 1, to September 30) to the Vcceruuran m Charge for the State where the research facilltv headquarters K reg~stered. REPORT o F A NtM A LS USEO I N ACTU A L RESEARCH, TESTING, O R E X PE RI M IS NTAT ION - Sect ton 2.28 Of Animal We! fare Reguiat ,On S rCOU lreS aPOrG prlate use of Jnesthetlcs, analgesics, and tranou[l Izlng drugs CIurlng research, teStln9, or 8x Perlmentatmn. Exper{ment$ tnvolvtng patn or dtstress wlrnout use of these Orugs KIU51 be fCPOrterJ and a or !el statement exolaonlng the rS!sear Ch. A . ifs. I c . I o. Ic. 1 F. Number of G n!mals used Numtser of G nimals used In reuarcn, G xpeflments, on fesearcn, exoer$ments, N e w Number of animals used An8mals Covered or tests where aopromiate or tests lnvolvtng Pain or An!mals 1 In research, exver oments, d,strem without aamlnts. T O T A L N O . B Y A c t I AcS~g?rthlS anestnetlc. analgeslc, or Of An8mals or rests Involvong no patn tfanaulllzer drugs were tratlon of G oprorwmte or Olstress. anestnetlc. analgesic, or (Cols. X G C G D + L“) G dm!nlsterest to G vo!d pain or cststress. t rancaual tzer dru9s. (A ttoch bwrf G xaslanation) 5. Dogs I 44 I 12 189 0 I 201 I 16. C&:s o { o 1 0 I o 0 7. Guinea Pigs I 383 I 137 186 0 323 8. Hamsters ~ 207 0 207 [ o I 207 9. Rabbits I 638 ~ 40 598 0 638 1 10. Primates I o I 12 i 119 0 I 131 11. Rats 4357 1940 1444 I o 3384 12. Mice 1373 363 739 0 1102 Wild Animals (spec:fy) 13. TURTLES 293 0 293 0 293 14. DOMESTIC PIGS 609 0 609 0 609 15 . GERBILS 100 0 100 0 100 CERTIF ICATION By ATTENDING VETERINARIAN FOR REPORTING FACIL ITY OR INSTITUTIONAL COMMITTEE I (We) hereby certlfY that th. tYPe G nd G mount of G nalgeslc, anesthetic, ~nd trantsulllzlng drugs used on G nimals dtsrln9 actual research, tssattng or G xpertmen. tation #nclurSlng POSt-OP*rative G nd. oost-orocodural-r~ wasKteemeO G ptxom!atc to relleve oaln G nd distress for the subject G nimal. ts. sIC3NATu RE OF ATT fsNOING V =TEfelN4~~ — f 7. TITLE $8. OATE SIGNEO 30 NOV 83 t e. SIGNATURE OF C O SAM IT T C SS MS@fBER 2 0 . T I T L E 2 1 . O A T E S I G N E D 30 NOV 83 2 2 . S I G N A T U R E O F C O M M I T T E E & - . E M B E R 2 3 . T I T L E 2 4 O A T E S I G N E D 1 I CERTIF ICATION BY HEADQUARTERS RESEARCH FACIL ITY OFFICIAL correct, and complete and tfsat rxofesslonally G cceptable stanaards governing the care, treatment, and usc of animals Inclualng G pwopriate use of anesthet analgeslc, and trancrutlazlng Orugs. Ourting G ctual rosearcn, testtng, or ex$aertmentataon Including post-osserat!vo arm post- Proc@aural above researcn facll It les or sites ( 7 U.S. C. section 214 3). 25. S IGN ATU RE O & 2 6 . T I T L E 2 7 . O A T E S I G N E D I 30 NOV 83b , VS FORM 18-23 Previous edat:on obsolete [AUG 81) SOURCE: Animal and Plant Health Inspection Service, U.S. Department of Agriculture. 64 . Alternatives to Animal Use in Research, Testing, and Education Even with these limitations and qualifications, the numbers generated by the APHIS data pro- vide a range that can be used in discussions of ani- mal use. The totals include: 1.8 million mandated species, 100,000 to 500,000 birds, 100)000 to 500,000 amphibians, 2.5 million to 4.0 million fish, and 12.2 million to 15.25 million mice and rats. Therefore, it appears that between 17 million and 22 million animals are used in united States laboratories annually. The largest group is represented by mice and rats. For reporting institutions, mice represent 60.8 percent of all animals used, and rats 25.1 percent. In addition, for the mandated species, certain in- stitutions use specific species disproportionately to their percentage of overall total use (see table 3-6). Fifty percent or more of all cats and dogs are used by universities and medical schools. Guinea pigs are used mostly by the pharmaceutical indus- try, whereas hamsters are used more often in bio- medical research, and to a lesser extent in univer- sities, medical schools, and the pharmaceutical firms. Sixty-two percent of rabbits are used in universities, medical schools, and the pharmaceu- tical industry, as are 75.6 percent of the primates. For rats and mice, the trends indicated in table 3-6 are clouded by the fact that there was more reporting of rat and mouse usage in 1983 than in Table 3-6.–USDA/APHlS Data, Changes 1982-83 (reporting Institutions only) Institution type Univers i t ies Hospi ta ls Bio- Toxicology Pharmaceu- State & Food, feed & medical nonuni- medical testing Chemical tical, device local & miscel. Federal schools versity research labs companies & d iagnosis government Ianeous agencies Total ..— – rear Rats: 1982 1983 0/0 change Mice: 1982 1983 0/0 change Dogs: 1982 1983 %O change Cats: 1982 1983 0/0 change Guinea pigs: 1982 1983 0/0 change Hamsters: 1982 1983 0/0 change Rabbits: 1982 1983 ‘/0 change Primates: 1982 1983 0/0 change lnstitutions reporting: 1982 1983 0/0 change 1,079,208 1,234,864 14 1,678,300 1,951,466 16 98,983 90,001 - 9 34,555 32,535 –5 82,198 64,554 –21 151,365 115,483 –23 173,716 158,058 - 9 23,353 22,201 - 4 410 402 - 2 86,472 108,430 23 203,768 222,080 8 13,622 12,605 - 7 2,716 2,265 –16 6,104 7,195 17 5,501 5,472 – 0.5 15,171 15,042 -0.8 557 1,059 90 129 140 8 343,915 408,938 18 1,579,664 1,512,424 –4 22,291 21,483 –3 7,697 6,788 -12 25,225 30,696 21 65,146 169,272 159 63,863 64,626 1 13,543 13,272 –2 167 159 –4 97,237 144,162 48 431,464 495,087 14 3,457 5,003 44 137 172 25 35,145 28,753 -18 12,954 11,922 –7 60,785 55,785 - 8 2,577 5,809 125 : : 8 176,874 114,215 -35 161,659 158,752 –1 2,194 1,591 –27 115 44 –61 18,182 14,722 – 19 3,180 612 –80 20,970 22,034 5 144 25 –82 27 31 14 558,630 778,425 39 1,669,629 2,021,157 21 37,604 38,311 1 9,073 8,624 –4 272,405 297,849 9 131,227 112,618 – 14 177,289 159,276 - 1 0 7,709 9,376 21 145 155 6 11,299 30,378 168 200,150 477,250 138 322 436 35 87 72 -17 9,044 10,090 11 8,401 3,193 –61 2,102 1,948 –7 329 243 –26 19 18 - 5 12,700 14,355 13 6,247 3,632 –41 3,698 3,400 - 8 2,040 2,092 2 1,504 930 –38 23 22 –4 1,862 2,504 34 66 82 24 26 32 23 359,479 439,729 22 958,863 1,071,339 11 12,696 9,595 –24 3,541 2,774 –21 48,053 66,448 38 39,490 35,885 - 9 31,554 29,779 –5 6,287 7,269 15 125 123 –2 2,725,814 3,269,494 19 6,889,744 7,913,167 14 194,867 182,425 –6 59.961 55,346 - 7 497,860 521,237 4 417,267 454,479 9 547,312 509,052 –6 54,565 59,338 8 1,127 1,146 2 SOURCE: Office of Technology Assessment. Ch. 3—Patterns of Animal Use G 6 5 1982. So, although it appears from table 3-6 that the usage increased, this was in fact not so (as can be seen from table 3-4). Data for all institutions from the regression equations show no change in mice and a small increase in the use of rats. How- ever, since the same pattern of increase by institu- tional group reporting can be seen from table 3-6, there has likely been no increase or decrease in use of these two species between 1982 and 1983. In table 3-6, the number of reporting institutions includes those that reported any number for any species, whether these included rats or mice or not. Few significant changes occurred as a func- tion of institution type for the 2 years surveyed. No trend in animal use can be identified be- tween 1982 and 1983, and the available data provide no justification for predicting either increases or decreases in future years. It would have been possible to examine the 1981 APHIS data sheets and determine whether, on the basis of 3 years’ data, a trend for the mandated species existed, but the 1981 data sheets would not indicate trends for rats and mice. The other methods of estimating laboratory-animal use do not match the reliability of the APHIS data, and thus do not lend much credence to the numbers reported in the past. Future Animal Censuses The major limitation with this estimate of an- nual laboratory-animal usage was the need to de- pend on available data sources, with all the limita- tions just described. Although the APHIS data sheets were of considerable value, they still do not substitute for an appropriately designed stratified random sampling of all possible users. Only then would all possible institutions be represented. The APHIS scheme depends on institutions to request certification. Some may be operating and not re- porting to APHIS. Still, with considerable further effort, a post-hoc stratification could be done based on the APHIS data. Estimates could be improved by two major ap- proaches. The first, and least expensive, would in- volve the use of all annual APHIS reporting forms— following an imperative redesign of the form—as well as thoroughly determining which registered institutions in each year did and did not report. Then appropriate statistical estimation techniques could be used on an institution-type and year- specific basis to correct for missing data. The sec- ond, and more ambitious, approach would be to conduct a stratified random sample study of all possible users. The stratification would be by type of institution, size of institution, and species of ani- mals. From such a sample, appropriate statistical techniques could be used to project to the entire population of user institutions. In 1985, the National Research Council’s Insti- tute of Laboratory Animal Resources announced plans for another in its series of surveys of experi- mental animal usage. The 1986 census will include mammals and birds, but omit fish, amphibians, and reptiles. SUMMARY AND CONCLUSIONS A rough analysis of the number of laboratory the individual annual reports furnished by each animals used is important to provide some non- registered facility for 1982 and 1983 were evalu- text in which to discuss alternatives to using ani- ated. Generally, it was found that great dispari- mals, evaluate progress toward the goal of using ties existed among the different sources. No sin- fewer animals, and judge the effect that alterna- gle data source presents an accurate count of the tives might have. OTA therefore evaluated exist- number of laboratory animals used in the United ing data on the number of laboratory animals used States since not one includes all potential users. each year in the United States. In addition, it is impossible to compare data among The data sources considered included various sources due to the inadequacy of information on reports and surveys published by the National Re- survey and data collection methodologies, defini-tions, areas of use, reporting requirements, andsearch Council’s Institute of Laboratory Animal Resources, various market surveys, and the an- the inability to justify completeness of the data. nual reports submitted to USDA’s Animal and Plant In a comparative analysis of data sources, it was Health Inspection Service. For the latter source, found that the most useful data were the APHIS . 66 G Alternatives to Animal Use in Research, Testing, and Education data sheets completed by every institution that uses laboratory species regulated under the Animal Welfare Act, APHIS requires that registered insti- tutions report all use of dogs, cats, guinea pigs, hamsters, rabbits, and nonhuman primates. Even with this requirement, though, it seems that APHIS does not receive animal-use information from all possible users. The data from these forms were found to be more accurate than the Animal Wel- fare Enforcement Report, a summary submitted annually by APHIS to Congress. This report usu- ally neglects 10 to 20 percent of the annual reports (those submitted late, usually after December 31) and so underestimates the actual number of dogs, cats, guinea pigs, hamsters, rabbits, and nonhu- man primates used. For fiscal years 1982 and 1983, the numbers of these kinds of animals used, according to the APHIS data sheets, are shown in table 3-7. For other lab- oratory species—mice, rats, birds, amphibians, and fish–the ability to obtain accurate estimates of the number used is impaired by a lack of reliable data sources. The best estimates are that 100,000 to 500,000 birds, 100,000 to 500,000 amphibians, 2.5 million to 4.0 million fish, and 12,2 million to 15.25 million rats and mice were used. (Animal use in medical and veterinary education is estimated to beat least 53,000 animals per year and is discussed in ch. 9.) Total animal use in the United States, therefore, is estimated as between 17 million and 22 million a year. The great discrepancies in data sources meant no trends could be observed overtime and among different types of institution. Even within the APHIS data for six kinds of animals, no clear trends Table 3-7.–Animai Use Reported to the U.S. Department of Agriculture, 1982 and 1983 Number used Number used Animal in 1982 in 1983 Dogs . . . . . . . . . . . . . . . . . . 194,667 182,425 Cats . . . . . . . . . . . . . . . . . . 59,961 55,346 Hamsters . . . . . . . . . . . . . . 417,267 454,479 Rabbits. . . . . . . . . . . . . . . . 547,312 509,052 Guinea pigs . . . . . . . . . . . . 497,860 521,237 Nonhuman primates. . . . . 54,565 59,336 Total . . . . . . . . . . . . . . . . 1,771,832 1,781,875 aTOtalS d. not lncl~e ratg or mice, two species that together represent the majority of animals used. SOURCE: Office of Technology Assessment. were found. Indeed, the most important finding was that no accurate source exists on the num- bers of animals used annually in the United States. A stratified random sample of all possible user in- stitutions done with a correct statistical analysis would probably be the best way to estimate labora- tory-animal use in the United States. In the Federal Government, six departments and four agencies use animals for intramural research and testing. These investigative efforts range from uncovering new knowledge that will lead to bet- ter health (within the National Institutes of Health), to evaluating hazardous substances in consumer products (within the Consumer Product Safety Commission’s Directorate for Health Sciences), to protecting the health of American astronauts (within the National Aeronautic and Space Admin- istration’s Life Sciences Division). OTA used the APHIS Annual Report of Research Facility forms to track animal use within the Fed- eral Government itself by department (and by di- vision within departments) and by species. In this way, it was possible to identify what portion of the estimated 17 million to 22 million animals used yearly were used within Federal facilities. In 1983, the Federal Government used at least 1.6 million animals, largely rats and mice. Ninety-six percent of the 1.6 million animals were used by DOD, DHHS, and the VA. Of the total, about 9 percent were dogs, cats, hamsters, rabbits, guinea pigs, and nonhuman primates. The APHIS forms require that all experiments be categorized as: 1) involving no pain or distress; 2) involving appropriate anesthetic, analgesic, or tranquilizer drugs to avoid pain or distress; or 3) involving pain or distress without administration of appropriate anesthetic, analgesic, or tranquilizer drugs. Sixty-three percent of the animals used within Federal departments and agencies were in the experimental situation categorized as involv- ing no pain or distress while 32 percent were given drugs and only 5 percent experienced pain or distress. The APHIS reporting system lacks clear defini- tions and uniform reporting, If accurate data are to be obtained, the forms must be revised and bet- ter explanations of how to complete them must be provided. Ch. 3—Patterns of Animal Use G 6 7 CHAPTER 3 REFERENCES 1. Abood, L. G., Professor, Center for Brain Research, University of Rochester, Rochester, NY, personal communication, 1984. 2. Alex Brown & Sons, Inc., Charles River Breeding Laboratories, inc., Market Smnrey (Baltimore, MD: 1981). 3. Bier, R., Bio-Research Laboratories, Ltd., Senner- vil]e, Quebec, Canada, personal communication, 1984. 4. Bohrer, D., Division of Research Grants, National Institutes of Health, Bethesda, MD, personal com- munication, 1984. 5. Borsetti, A., Staff Scientist, Office of Science Coordi- nation, Food and Drug Administration, U.S. Depart- ment of Health and Human Services, Rockville, MD, persona] communication, 1984. 6. Charles River Breeding Laboratories, Inc., price list, Wilmington, hlA, 1983. 7. Cook, R., North Carolina State Llniversity, Raleigh, NC, personal communication, 1984. 8. Culley, D., Louisiana State University, Baton Rouge, LA, persona] communication, 1984. 9. David, T., Staff Veterinarian, Aerospace Medical Division, Air Force, U.S. Department of Defense, Brooks Air Force Base, TX, personal communica- tion, Jan. 18, 1985. 10. Digges, K., Deputy Associate Administrator for Re- search Development, National Highway Traffic Safety Administration, U.S. Department of Trans- portation, Washington, DC, personal communica- tion, March 1985. 11. Dodds, W.J., Division of Laboratories and Research, New York State Department of Health, Albany, NY, personal communication, April 1985. 12. Edington, C., Associate Director, Office of Health and Environmental Research, Office of Energy Re- search, U.S. Department of Energy, Washington, DC, personal communication, Nov. 16, 1984. 13. Emmons, M. B., ‘(Secondary and Elementary School Use of Live and Preserved Animals, ’’Am”mals in Edu- cation, H. McGriffin and N. Browrdey (eds.) (Wash- ington, DC: Institute for Study of Animal Problems, 1980). 14. Goldberg, A. M., Director, The Johns Hopkins Cen- ter for Alternatives to Animal Testing, Baltimore, MD, personal communication, April 1985. 15. Guilloud, N., Ortho Pharmaceutical Corporation, Raritan, NJ, personal communication, 1984. 16. Gupta, K. C., Deputy Director, Division of Health Sciences Laboratory, Directorate for Health Sci- ences, U.S. Consumer Product Safety Commission, Washington, DC, personal communication, Sept. 24, 1985. 17. Health Designs Inc., ‘(Survey and Estimates of Lab- oratory Animal Use in the United States, ” contract report prepared for the Office of Technology As- sessment, U.S. Congress, July 1984. 18. Institute of Laboratory Animal Resources, Animal Facilities in Medical Research (Final Report), A Re- port of the Committee on the Animal Facilities Sur- vey (2 parts) (Washington, DC: National Academy of Sciences, 1964). 19. Institute of Laboratory Animal Resources, Animal Facilities in Medical Research: A Preliminary Studv (Washington, DC: National Academy of Sciences, 1962). 20. Institute of Laboratory Animal Resources, Annual Surveys of Animals Used for Research Purposes (Washington, DC: Nationa] AcademY of Sciences, 1965-71). 21. Institute of Laboratory Animal Resources, “Labora- tory Animal Facilities and Resources Supporting Bio- medical Research, 1967 -68,” Lab Anim. Care 20: 795-869, 1970. 22. Institute of Laboratory Animal Resources, National Survey of Laboratory Animal Facilities and Re- sources (Washington, DC: National Academy of Sciences, 1980). 23. Institute of Laboratory Animal Resources, Non- Human Primates: Usage and A~~ailabilituv for Bio- medical Programs (Washington, DC: National Acad- emy of Sciences, 1975). 24. Kellogg, C., Department of Psychology, University of Rochester, Rochester, NY, personal communica- tion, 1984. 25. Manufacturing Chemists Association, Studuy of Po- tentialEconomic Impacts of the Proposed Toxic Sub- stances Control Act as Illustrated by Senate BillS 776 (Florham Park, NJ: Foster F. Snell, Inc., 1975). 26. Matchett, A., Animal Care Staff, U.S. Department of Agriculture, Washington, DC, personal commu- nication, 1984. 27. McArdle, J. E., Director, Laboratory Animal Wel- fare, Humane Society of the United States, Wash- ington, DC, personal communication, 1984. 28. McCarthy, C., Director, Office for Protection from Research Risks, National Institutes of Health, Pub- lic Health Service, U.S. Department of Health and Human Services, Bethesda, MD, personal commu- nication, October 1984. 29. Middleton, C., Chief Veterinary Medical Officer, Veterans’ Administration, Washington, DC, per- sonal communication, Oct. 3, 1984. 30. Miller, T., McNeil Laboratories, Ft. Washington, PA, personal communication, 1984. 31. Moorehouse, K., Division of Wildlife Service, Fish . 68 . Alternatives to Animal Use in Research, Testing, and Education and Wildlife Service, U.S. Department of the In- terior, Washington, DC, personal communication, October 1984. 32. Moreland, A., “Animal Research Protocol Review Within the Veterans’ Administration” (draft), Gaines. vil]e, FL, 1984. 33. Mosed, R., Executive Director for Oceanic and At- mospheric Administration, National Oceanic and Atmospheric Administration, U.S. Department of Commerce, Rockville, MD, personal communica- tion, September 1984. 34. Nace, G. W., Professor of Zoology, University of Michigan, Ann Arbor, MI, personal communication, 1984. 35. Nace, G. W., Response to a questionnaire on the use of frogs in the United States, questionnaire prepared by Rolf Martin, Department of Chemistry, Brooklyn College of the City University of New York, Brook- lyn, NY, 1984. 36, New A., Director, Laboratory Animal Science, Na- tional Cancer Institute, Public Health Service, U.S. Department of Health and Human Services, Be- thesda, MD, personal communication, 1984. 37. Nicogossian, A., Director, Life Sciences Division, Na- tional Aeronautics and Space Administration, Wash- ington, DC, personal communication, 1984. 38. Roberts, C., Director, Division of Veterinary Medi- cine, U.S. Department of Defense, Washington, DC, personal communication, September 1984. 39. Rowan, A. N., Of Mice, Models, & Men: A Critical Evaluation of Animal Research (Albany, NY: State University of New York Press, 1984). 40. Rowan, A. N., Assistant Dean for New Programs, Tufts University School of Veterinary Medicine, Bos- ton, MA, personal communication, 1985. 41. W.B. Saunders and Company, “Market Survey)” Inform. Lab. Anim. Res, 9(3):10, 1965. 42. Schultz, C., Chief, Sciences Branch, Technology Di- vision, Office of Hazardous Materials Regulation, Materials Transportation Bureau, Research and Special Programs Administration, U.S. Department of Transportation, Washington, DC, personal com- munication, September 1984. 43. Simmonds, R., Director of Instructional and Re- search Support, Uniformed Service University of the Health Sciences, Bethesda, MD, personal com- munication, April 1985. 44. Stewart, W., Senior Veterinarian, Animal and Plant Health Inspection Service, U.S. Department of Agri- culture, Hyattsville, MD, personal communication, November 1984. 45. Taylor, J., Staff Veterinary Officer, Naval Command, U.S. Department of Defense, Washington, DC, per- sonal communication, November 1984. 46. Thomas, O., Professor of Poultry Science, Univer- sity of Maryland, College Park, MD, personal com- munication, 1984. 47. Thomas, R., U.S. Department of Energy, Washing- ton, DC, personal communication, October 1984. 48. Ulvedal, F., Acting Director, Water and Toxic Sub- stances Health Effects Research Division, U.S. Envi- ronmental Protection Agency, Washington, DC, personal communication, September 1984. 49. U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Am”mal Welfare Enforce- ment: Report of the Secretary of Agriculture to the President of the Senate and the Speaker of the House of Representatives (annual) (Washington, DC: 1972 through 1983). 50. U.S. Department of Defense, Air Force Division, Aerospace Medical Division Animal Use Review Panel Meetings (Washington, DC: May 1984). 51. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, 1983 NIH Almanac (Bethesda, MD: 1983). 52. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, NIH Data Book (Bethesda, MD: 1983). 53. U.S. Department of the Interior, U.S. Fish and Wild- life Service, Service Managment Plan (draft) (Wash- ington, DC: October 1984). 54. Will, J. A., Director, Research Animal Resources Center, University of Wisconsin, Madison, WI, per- sonal communication, 1984. 55. Willett, J. D., Project Officer, Biological Models and Materials Resource Section, Animal Resource Pro- gram, Division of Research Resources, National In- stitutes of Health, Public Health Service, U.S. De- partment of Health and Human Services, Bethesda, MD, personal communication, 1983. chapter 4 To use interests Ethical Considerations either human or non-human animals for purposes that are not in their own is both ethically unjustifiable and, in the long run, counter-productive. Alex Pacheco People for the Ethical Treatment of Animals appropriately March 15, 198.5 concerned for theFortunately there are many who, while deeply and compassionate treatment of animals, recognize that human welfare is and should be our primary concern. One cannot intelligently assess vivisection areas of human life: for food, furs, leather, Frederick A. King Yerkes Regional Primate Research Center Psychology Today, September 1984 in isolation f-rem animal exploitation in other in so-called sports, in movies, in the wild. Vivisection, properly seen, is simply one variation on the cultural theme of animal sacrifice. Michael A. Giannelli The Fund for Animals, Inc. March 10, 1985 The use of any particular animal-say, a sheep—in medical research is more important than its use as lamb chops. The The Research Carl Cohen University of Michigan News 35(1()- 1 2):9, 1984 CONTENTS The Religious and Philosophical Traditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Ethical Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Moral Standing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Moral Constraints. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Summary Chapter4 and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Page 72 75 76 79 82 84 Chapter 4 Ethical Considerations The range of opinion on the rights and wrongs of using animals to satisfy human needs is as broad as the political spectrum itself. At one extreme, animals are thought to be entitled to at least a por- tion of the respect, individual freedom, and dig- nity that are considered to be basic human rights. Some say that animals should be recognized as be- longing to a community that includes humans. At the other extreme, humans are thought to have broad and absolute authority over the lives and interests of animals. From this perspective, expe- diency alone, not morality, dictates what we may do with animals. To illustrate the distance between these extremes, a recent legal brief for animal rights can be con- trasted with a televised interview with three scien- tists who perform animal experimentation. Ac- cording to the brief (43): If being alive is the basis for being a moral ob- ject, and if all other interests and needs are predi- cated upon life, then the most basic, morally rele- vant aspect of a creature is its life. We may correlatively suggest that any animal, therefore, has a right to life. The scientists, in a televised exchange with Harvard philosopher Robert Nozick, were asked whether the fact that an experiment will kill hundreds of animals is ever regarded by scientists as a reason for not performing it. One answered: “Not that I know of .“ When Nozick asked whether the ani- mals count at all, one scientist replied, “Why should they?” while another added that he did not think that experimenting on animals raised a moral is- sue at all (45). People at both extremes would probably agree that, given a choice between experiments equiva- lent in cost and scientific value, one that does not require the destruction of animals would be prefer- able to one that does. This consensus, however, would probably evaporate if animal experimen- tation produced greater scientific validity or the technique that used animals had significant cost advantages. In morals, as in politics, most people tend to shun extremes. However, a middle view is at once the most defensible and the most difficult to defend, Pitted against extreme or esoteric positions, the numbers on its side create a presumption in its favor. Yet a presumption given only by the weight of opinion will not amount to a moral justification. A belief is not shown to be true simply by count- ing the votes of those who accept it. Some basis for an opinion, independent of it being accepted, must be found. Adoption of a middle view is hazardous in two respects. First, it runs the risk of inconsistency. Propositions located at polar extremes will usu- ally contradict one another, and a position that seeks to incorporate both may find itself embrac- ing a contradiction. In the case of toxicity testing, for example, it may not seem possible to respect the interests of experimental animals and yet use them as tools for enhancing human health and safety. The second risk is that consistency will be se- cured at a price too high, by way of a theoretically unattractive ad hoc device. In principle, two con- tradictory propositions can be reconciled simply by making one an exception to the other. It could, for example, be stipulated that the general rule against harming animals does not hold when they are used to test for toxicity. But it is one thing to say this and another to give a reason for it. Com- plex rules, introduced for no reason other than to remove a particular inconsistency, muddy a point of view without shedding any light on the hard moral cases it must address. More important, they are arbitrary. 71 72 G Alternatives to Animal Use in Research, Testing, and Education T H E R E L I G I O U S A N D P H I L O S O P H I C A L T R A D I T I O N S Interest in the moral status of animals is by no means modern. The ancient religions had much to say about the place that animals were to occupy in the cosmic scheme of things. Oriental creeds were, as a rule, reluctant to draw a sharp distinc- tion between humans and other species. All ani- mal lives were judged worthy of protection and some were thought to be sacred. The doctrine of transmigration left still more room for caution— any animal body might house a soul entitled to spe- cial care. The various forms of the doctrine of trans - migration share the thesis that a single, continu- ous, immaterial individual may pass from one body to another, which maybe of the same or a differ- ent species. If the latter, its conduct in the earlier incarnation may determine the kind of body it in- habits next. For such reasons, the prescribed die- tary regimen in the Orient was frequently vegetar- ian. Modern influences have relaxed, but not wholly removed, the grip of these beliefs. In the West, a different tradition took root, one that seems to have assigned value to animals only as they serve human purposes. Judeo-Christian doctrine appears to have condoned an indiffer- ent, if not openly exploitative, attitude toward non- human animals (38,45). (For an opposing view, see refs. 6 and 49.) The Genesis account suggests that humans are the last and most perfected of God’s creatures. Humans alone, of all living things, bear the likeness of God, and receive the divine com- mission to exercise “dominion over the fish of the sea, and over the birds of the air, and over the cattle, and over all the earth, and over every creep- ing thing that creeps upon the earth” (Genesis 1:26- 28; all Biblical references and quotations are to the Revised Standard Version). After the flood, God rewarded Noah and his sons with this blessing: “Every moving thing that lives shall be food for you; and as I gave you the green plants, I give you everything” (Genesis 9:3). A brief Talmudic story indicates that Judaic practice was to the same ef- fect: “A calf was being taken to the slaughterer, when it broke away, hid his head under the Rabbi’s skirts, and lowed in terror. ‘Go,’ said he, ‘for this wast thou created’ “ (12). These passages do not warrant the inference that humans are permitted to treat animals in any way they please. Even when suffering is inflicted as a means to some human end, humans are subject to the condition of using the animal. Wanton cru- elty would not be allowed. Nor is it clear just what human dominion includes, until the terms of a model ruler-subject relation are spelled out. Hu- mans must presumably rule well, and the good ruler does not take authorized but unjustified ac- tions. Much depends, too, on whether human sov- ereignty over nature is to be thought absolute or limited by a divine will that may have set some value on animals in addition to their utility for humans. In a number of passages, the scriptures seem to place a rein on the use of animals. Genesis con- firms that God had already judged the world as good–that is, possessed of some value–before hu- mans were created (Genesis 1:3,10,13,18,21). And on several occasions in the later books of the Old Testament, humans are expressly directed to show kindness to the animals under their control. Thus, “you shall not muzzle an ox when it treads out the grain” (Deuteronomy 25:4). And, “a righteous man has regard for the life of his beast, but the mercy of the wicked is cruel” (Proverbs 12:10). The most persuasive evidence for restraint may lie in the role-model of the good shepherd, often cited in both testaments. At one point, by report of the prophet Ezekiel, God becomes annoyed (Ezekiel 34:2,4): Ho, shepherds of Israel who have been feeding yourselves! Should not shepherds feed the sheep? . . . The weak you have not strengthened, the sick you have not healed, the crippled you have not bound up, the strayed you have not brought back, the lost you have not sought, and with force and harshness you have ruled them. God’s own rule is often compared with the con- cern that shepherds should have for their flocks (Ezekiel 34:11-13; John 10:11; Luke 15:4-7). De. signed to show that God stands to humans as they stand to animals—a kind provider even if there are no duties to provide for them—that simile would fail if the shepherds could wholly disregard the welfare of their animals. Ch. 4—Ethical Considerations . 73 Aside from this figurative guide, the New Testa- ment is spare in its references to handling animals. Saint Paul’s discussion of the proscription against muzzling the ox suggests a human benefit: “Is it for oxen that God is concerned? Does he not speak entirely for our sake? It was written for our sake, because the plowman should plow in hope and the thresher thresh in hope of a share in the crop” (I Corinthians 9:9-10). Thus, the thresher was to let the ox feed from the corn being worked, not so much for the good of the ox, but because a well- fed animal would yield a larger return. This passage suggests a shift in sentiment from the old to the New Testament. For Christians, the paramount practical concern is the condition and future of the immortal soul possessed by human beings. Animals are not believed to have immor- tal souls, nor be repositories for human souls. In the Christian world view, then, animals are left without the one thing that has special value in itself—a soul. An animal’s welfare is a good thing only as it is good for the human being. The letter containing Paul’s reading of the Old Testament rule was written only a generation af- ter Christ’s death, when Christianity was still anew faith. The distinction between humans and other animals hardened as the creed acquired the trap- pings of theory, but in such a way as to raise new questions about its real source. The legacy of Greek philosophy exercised such a pervasive influence over Christian theology in its formative years that the distinction could be traced to Athens as easily as to Jerusalem. It might be said that in theology all roads lead back to Augustine or Aquinas. On the subject of animals, the Augustinian position finds expression in his critique of a competing doctrine, which, on the premise that animals also had souls, would not allow killing them. Augustine cited the conduct of Christ as a lesson to the contrary (7): Christ himself shows that to refrain from the killing of animals and the destroying of plants is the height of superstition, for judging that there are no common rights between us and the beasts and trees, he sent the devils into a herd of swine and with a curse withered the tree on which he found no fruit. If Christ could use animals for his own purposes, then so apparently could we. Augustine’s view, however, was tempered in two respects. First, he denied that animals were mere instruments of hu- mans. As creatures made by God, they also pos- sessed a good of their own (7)8). Second, animals’ utility was the use to which human intelligence might put them, not the convenience or incon- venience that they might present. Augustine did not hold that humans were to treat animals accord- ing to their own pleasure or displeasure (8). Aquinas ’view of animals was more sophisticated and less sympathetic. Every natural being that underwent development had an end or perfected state that God had created it to achieve. God made humans, however, as free and rational agents, with control over their actions. People’s lives took their objectives from their designs. Being neither free nor rational, an animal was merely a means to an end existing outside it (in the form of some pur- pose that a rational individual might have for it). Thus, the nonhuman animal was ordered, by na- ture and providence, to the use of humans (l). From Aquinas’ perspective, the Old Testament concern for animals had been appropriately char- acterized by Saint Paul. People should avoid mis- treating animals not because this would be best for the animals, but because cruelty could be harm- ful to humans. Strictly understood, disinterested charity towards animals was impossible, since there was no common fellowship between humans and them (2). In its essentials, this view prevails within the Catholic Church today. Its implications for research in the life sciences have not gone unnoticed. Writ- ing at the turn of this century, Father Joseph Rick- aby, the English Catholic moral theorist, denied that the suffering of animals was an obstacle to biological inquiry (42): Brutes are as things in our regard: so far as they are useful to us, they exist for us, not for them- selves; and we do right in using them unsparingly for our need and convenience, though not for our wantonness. If then any special case of pain to a brute creature be a fact of considerable value for observation in biological science or the medi- cal art, no reasoned considerations of morality — 74 Ž Alternatives to Animal Use in Research, Testing, and Education can stand in the way of man making the experi- ment, yet so that even in the quest of science he be mindful of mercy. Protestantism retains the thesis that humans en- joy a rightful hegemony over other animals, but suggests a shift towards a “stewardship” interpre- tation of that role. John Calvin, the 16th-century Reformation theologian, maintained that when God placed animals “in subjection unto us, He did it with the condition that we should treat them gently” (13). They were brute beasts, to be sure, but for Calvin as for Augustine they were also crea- tures of God. Calvin went a step further, however, in making this fact about animals a limitation on humans’ use of them. Here humans would seem to be less the sovereigns of nature than deputies appointed to manage God’s earthly estate. Every creature would still be subject to God’s ownership and control. A person was still worth more than any number of sparrows, yet ‘(no one of them will fall to the ground without your Father’s will” (Mat- thew 10:29). Thus, Karl Barth, a leading modern Protestant theologian, urged that people possess the right to use and sometimes to kill animals, but only because God has so authorized it in order that humans might live (9). There have been a few distinguished Judeo-Chris- tian defenders of a position much closer to the oriental view. Saint Francis and Albert Schweitzer both pressed for a principle of reverence toward every living thing. But their ideal has been received as just that: a norm perhaps for saints, and some- thing all should desire, but not binding on imper- fect individuals in less-than-ideal circumstances. In the absence of mainstream philosophical sup- port, the intellectual authority of the reverence- for-all-life rule is thought to be outweighed by the personal prestige of its practitioners (21). Until 1600, the philosophical mainstream was Aristotelian. Using a much broader conception of the soul than the current one, Aristotle distin- guished living from nonliving beings by the pres- ence or absence of some form of a soul, or life- giving power. Its function might be nutrition, sensation, desire, locomotion, or thought. The first of these, but not the rest, was found in plants. All animals had sensation and desire as well, and most also had locomotion. Humans alone had the power of thought (4). This advantage made humans nat- urally suited to rule over other living beings and made animals natural slaves. Aristotle reached this conclusion by generalizing from phenomena al- ready at work within humans: Those with greater rationality exhibited an internal mastery of rea- son over desire and an external mastery over those who, because they lacked the mental equipment to tend to more than their bodily needs, required direction from others (5). This resulted in leader- ship by those most competent to rule. Natural fitness implied that nature worked toward certain ends that together formed a master plan. The significance of the 17th-century scientific rev- olution lay not so much in its overthrow of church authority in the empirical realm as in its discov- ery of a method and a subject matter (i.e., mechan- ics treated as a branch of physics) that dispensed with the hypothesis that nature had purposes. Na- ture became simply the sum of matter in motion, mathematically describable without reference to goals that phenomena might serve. The philosophical foundations for the new world view were supplied by Rene Descartes, who rec- ognized only two kinds of existence, material and mental. Bodies were extended in space and time and divisible into parts, with properties of size, shape, and weight. Minds contained beliefs, emo- tions, and intentions, but no physical properties. The human was a composite being-the only one— with both a body and a mind (18). Animals did not fit comfortably into the Carte- sian scheme. They obviously had bodies, but did they not also have sensations and desires? Des- cartes answered that in a sense they did, but that their behavior could be duplicated by a machine, while human behavior could not. In their use of language and thought, humans revealed a capac- ity to respond to stimuli in a variety of ways, whereas animals would respond in only one, “ac- cording to the arrangements of their organs” (19). For all their differences, the Aristotelian and Cartesian theories joined hands in making the activ- ities that required reasoning the distinctive mark of humanity. Both defined the human being as a rational animal. That thesis was not questioned until the following century, when British empiri- cists criticized it as inflated claims for the power of reason. The Scottish skeptic David Hume con- Ch. 4—Ethical Considerations G 7 5 curred with Descartes that the human mind was capable of creatively entertaining a variety of pos- sible conclusions from a given body of experience. But this, Hume argued (29), was nothing more than a habit of inference formed from repeated obser - vations, something that dogs could do as well (30): 'Tis necessary in the first place, that there be some impression immediately present to their memory or senses, in order to be the foundation of their judgment. From the tone of voice the dog infers his master’s anger, and foresees his own punishment. From a certain sensation affecting his smell, he judges his game not to be far distant from him. Secondly, the inference he draws from the pres- ent impression is built on experience, and on his observation of the conjunction of objects in past instances. As you vary the experience, he varies his reasoning. Make a beating follow upon one sign or motion for some time, and afterwards upon another; and he will successively draw different conclusions according to this most re- cent experience. The issue dividing Descartes and Hume survives, still unsettled, in current controversies over artifi- cial intelligence and animal cognition. Recent decades have witnessed an explosion of empirical inves- tigations into the behavior of nonhuman animals (26,35,50). Among these, various efforts to teach higher primates how to use a nonverbal language have captured the public’s imagination. Inferences drawn from such studies, however, encounter two obstacles. First, to argue that chimps consciously use gestures in the same way that human deaf- mutes do is to assume a certain theory about the relation between bodily behavior and mental oper- ations. No consensus on mind-body relations exists today. The same difficulty, it is worth noting, af- fects various efforts to use similarities in brain structure and function as evidence for similari- ties in thought. Even if such matters could be resolved, a greater conceptual hurdle would remain: what is the con- nection between language and thought? Language requires combining terms into well-formed sen- tences using rules of grammar and meaning. Lin- guistic mastery includes the capacity to create novel sentences in situations not precisely like those already encountered and the resources to express thoughts indifferent modalities (as descrip- tions, questions, commands, and so on) (48). It also seems to require recognition that something said is true, false, or uncertain (17,24). Although no one knows whether other primates will ever approach human beings in linguistic per- formance, it would be a mistake to focus on that issue. Evidence is mounting that animals can rec- ognize visual patterns, remember where their food is located, learn how to perform nonmechanical tasks, and foresee where a moving prey will even- tually be positioned, even if they cannot master a language (26,50). In this sense, animals exhibit intelligence as defined by ability to adapt to envi- ronmental conditions. From a Darwinian (evolu- tionary) perspective, humans do not hold a privi- leged status over animals. Humans are not more highly evolved than other animals; all have evolved to fill their respective niches. Neither linguistic nor nonlinguistic findings hold all the answers. The moral issue is not simply whether animals have some and lack other abili- ties that human beings possess, but whether the differences between them make for differences in how humans and animals should be treated. Sometimes the differences matter, common sense might say, and sometimes they do not. T H E E T H I C A L Q U E S T I O N S HOW, if at all, should animals be used in research, testing, and education? Before this can be answered, a preliminary question must be asked (14,15,44,47): What moral standing does an animal have? Is it the kind of being to which humans could possibly have moral duties and obligations? Taking one side or another on the question need not include any particular moral judgment. Whatever its resolu- tion, the separable moral issue remains: what con- straints, if any, regulate humans’ use of animals? These constraints might be weaker if animals lack moral standing, but not necessarily absent al- together. 76 . Alternatives to Animal Use in Research, Testing, and Education Moral Standing Modern moral theory operates under a “law con- ception” of ethics (3). It judges particular human actions as right (lawful) or wrong (unlawful) as they comply with or violate some universal principle of conduct. In this, it departs from the classical theory of the virtues, which makes individual char- acter the unit of evaluation and does not attempt to reduce ethics to a system of rules. Under the law conception, moral standing also goes to per- sons, but it is not conferred by an individual, insti- tution, or community. From this point of view, an individual counts as a person because of some in- herent characteristic. This is the chief reason why it is within the moral domain to speak of the natu- ral duties and the natural rights of a person. A le- gal system can, of course, recognize natural duties and rights. For obvious reasons, no one has ever argued that animals can have moral duties (40). That would require that they freely choose to act among alter- natives they judge to be right or wrong—a skill as demanding as full-blown linguistic competence would be. Nevertheless, it is possible to take the view that animals have moral standing but do not have rights. There are two broad theoretical approaches to the subject of rights. The first, sometimes called the will theory, would discourage efforts to attrib- ute rights to animals. In its classical form, as given by Emmanuel Kant, it would define a right as a capacity to obligate others to a duty. Possession of a right carries with it an authorization to use coercion to enforce the correlative duty (3 I). This, in turn, implies that the right-holder’s capacity is a power of discretion, either to enforce or waive the right. A right is therefore something that a right-holder may choose to exercise or not. The choice itself will be an act of will. H.L.A. Hart, a leading contemporary defender of the will theory, treats a right as a choice that gives the right-holder authority to control the ac- tions of someone else. The possessor of a moral right has a moral justification for limiting the free- dom of another, not because the action the right- holder is entitled to require has some moral qual- ity, but simply because in the circumstances a certain distribution of human freedom will be maintained if the right-holder has the choice to determine how that other shall act (28). The will theory helps to avoid confusion between claims of right, and other, separable requirements to promote or secure some valued state of affairs (e.g., to assist someone in need). Since animals could not be said to have waived or exercised the rights they had, all references to animal rights could sim- ply be translated into talk of human duties. Those who would assign rights to animals have embraced the alternative interest theory of what it means to have a right. A right, in their view, is a claim to the performance of a duty by someone else, but the right -holder need not be in a position or possess the competence to make this claim by an act of will. It is enough that the right-holder has interests that can be represented (by others) in a normative forum (20). These interests will in- clude things that are intrinsically good and things in which the right-holder “takes an interest, ” self- ish or not (40). To have a right, then, will be sim- ply to have interests that can be affected by some- one else. The interest theory surfaces in Peter Singer’s Animal Liberation, among the first contemporary theoretical statements of the case for animals. In that work, Singer uses the term “right” to describe any claim that individuals may make to have their interests equally considered with those of others. It implies, therefore, nothing more than a capac- ity for suffering, which both humans and animals possess (45). The modest measure of animal awareness that such a test demands has been one source of its appeal. It has not, however, been free of contro- versy. Some have objected that animals cannot have interests because interests require beliefs and animals cannot have beliefs in the strict sense (24, 36). This criticism suggests that pain-avoidance is not an “interest” because it is not a “belief ,“ a dis- tinction that seems more semantical than useful. Nevertheless, a more serious charge remains. As stated, the interest theory shows only that having interests is a necessary condition for having rights, not that it is sufficient. Singer himself has since abandoned the attempt to show sufficiency and, Ch. 4—Ethical Considerations 77 accordingly, recanted his earlier references to the language of rights (46): I could easily have dispensed with it altogether. I think that the only right I ever attribute to ani- mals is the “right” to equal consideration of inter- ests, and anything that is expressed by talking of such a right could equally be expressed by the assertion that animals’ interests ought to be given equal consideration with the interests of humans. Singer effectively acknowledges Hart’s charge that the notion of a right has lost its distinctive func- tion in this context because it no longer refers to the discretionary control that one individual has over the conduct of another. There is one very general consideration that ap- pears to weigh against the will theory, if not en- tirely in favor of the interest theory. It underlies a form of argument so ubiquitous in the animal- rights literature that it deserves a name. The con- sistency argument is exemplified in the following passage from an essay on vegetarianism by Tom Regan. Rejecting rationality, freedom of choice, and self-consciousness as conditions for having a right to life, Regan adds (41): It is reasonably clear that not all human beings satisfy them. The severely mentally feeble, for example, fail to satisfy them. Accordingly, if we want to insist that they have a right to life, then we cannot also maintain that they have it because they satisfy one or another of these conditions. Thus, if we want to insist that they have an equal right to life, despite their failure to satisfy these conditions, we cannot consistently maintain that animals, because they fail to satisfy these condi- tions, therefore lack this right. Another possible ground is that of sentience, by which I understand the capacity to experience pleasure and pain. But this view, too, must en- counter a familiar difficulty—namely, that it could not justify restricting the right only to human beings. In short, given that some human beings (infants, mental defective, and senile adults) lack such ca- pacity as well, Regan points to the inconsistency of holding both that this capacity is a condition of having a right and that all humans and only hu- mans have moral rights. Any less burdensome test, however, will presumably admit animals as possi- ble right-holders (33)45). (For an opposing perspec- tive, see refs. 22 and 24.) This reasoning appears to overlook a significant difference between an incompetent human being and an animal. In most cases, human beings have the capacity for rationality, freedom of choice, and self-consciousness, whereas in all cases animals do not. If most humans have these characteristics, it might be appropriate (or at least convenient) to treat humans as a homogeneous group, even though some members lack these characteristics. If all animals lack certain characteristics, it maybe sim- ilarly appropriate to treat them as a group, re- gardless of whether some humans also lack these characteristics. Furthermore, if rights do not imply present pos- session of the qualifying condition (as suggested by the way that people treat those who are men- tally incapacitated only for a time), then babies who have yet to mature and people who have become incapacitated after a period of competence will still have rights. The animal, as far as can be ascer- tained, has never met and will never meet this qual- ification. The rare human being whose deficiency is complete over a lifespan is nevertheless differ- ently situated from the animal. The condition is a disability—the loss of some skill the person would normally be expected to have. The animal’s con- dition is not disabling, even though it lacks the same skill. The very fact that the human has been de- prived of an ability implies that the person has been harmed; a human’s failure to acquire an ability means that person is in need of help. The condi- tion of the animal does not call for either infer- ence. This difference, to be sure, makes no men- tion of rights. Yet it creates a special duty to meet the human need that would not extend to animals. Because the animal without a will has not lost what it was biologically programmed to possess, it “needs” a will only as a human might ‘(need” to fly. In nei- ther case does the condition give rise to a moral demand for assistance. Ironically, the consistency argument contains a basic inconsistency. On the one hand, the argu- ment asserts that humans are not superior to ani- mals; animals should therefore be treated like hu- mans. On the other hand, the very nature of the moral argument is promotion of morally superior behavior: Humans should refuse to exploit other species, even though the other species exploit each other. 78 . Alternatives to Animal Use in Research, Testing, and Education The consistency argument nevertheless succeeds to the extent that it shows that the genera] reason for moral concern in the cases discussed cannot be limited to humans. Other things being equal, the fact that a condition is harmful or threatens harm to an individual—human or animal-creates amoral reason to intervene. That reason need not take the form of a duty owed to the victim, with a correlative right that this would entail. It need not always be a duty of any sort. The highest ap- proval is often reserved for the good deed that, like the good samaritan’s, goes beyond what duty strictly requires. There is a spectrum of possible positions, be- ginning atone end with a strict prohibition against the cruel infliction of suffering, moving to a still powerful requirement to lend help when the indi- vidual alone is in a position to provide it for someone in great need, and then to the milder requirements of charity and generosity when the individual can provide them without great personal sacrifice (even if others can do the same), and finally, at the other extreme, to the highly praised but not binding act of genuine self-sacrifice that distin- guishes the moral saint. The moral vernacular cov- ers this spectrum with a single term. The act in question is called the ‘(humane” thing to do, and sometimes failure to perform it is labeled in- humane. The term itself refers to the actor, not the recip- ient. Humane treatment, following the Oxford Eng- lish Dictionary, is “characterized by such behavior or disposition toward others as befits a man.” This meaning, which dates back to the 18th century, applies to conduct marked by empathy with and consideration for the needs and distresses of others, which can include both human beings and animals. This does not mean that animals will generally command the same degree of affection and atten- tion as humans. The attitude of empathy, which is the psychological spring for humane treatment, consists in ‘(feeling like” the object of sympathy, and the basis for this response must be a certain understanding of what it is like to be in the other’s position. Other human beings are much more ac- cessible in this respect, not only because they are structurally and functionally like each other, but because they can communicate their feelings in ways that animals can scarcely approach. In such areas as the capacity for experiencing pain, how- ever, the differences across species are by no means so great as to make empathetic identifica- tion impossible. Here the mark of the humane in- dividual will be the extent to which sympathy jumps the barrier between species (11). There are differences among animals, too, in the capacities they have, the things they do, and the relations they have with humans, all of which af- fect the moral weight that humane considerations will have. A gorilla will gather more sympathy than a trout, not so much because it is more intelligent as because it exhibits a range of needs and emo- tional responses to those needs that is missing al- together in the trout, in which evidence of pain can barely be detected. Predatory animals and wild rodents rarely elicit affection because their char- acteristic activities do not mark them as helpless and in need. Even within one species, the regard an animal may receive will rise with the social ties and responsibilities that human beings have de- veloped with it. As a possible recipient of humane treatment, the garden-pest rabbit will stand to the pet rabbit much as the stranger does to an ac- quaintance. Each of the morally significant differences among animal recipients of humane treatment builds on an analogy to the human case. Thus, whatever the merits of the consistency argument on the score of rights, it applies here because the humane treatment principle crosses the species border. Mary Midgeley has put the point eloquently in another context (33): [Animals] can be in terrible need, and they can be brought into that need by human action. When they are, it is not obvious why the absence of close kinship, acquaintance or the admiration which is due to human rationality should entirely can- cel the claim. Nor do we behave as if they obvi- ously did so. Someone who sees an injured dog lying writhing in the road after being hit by a car may well think, not just that he will do something about it, but that he ought to. If he has hit it him- self, the grounds for this will seem stronger. It is not obvious that his reasons for thinking like this are of a different kind from those that would arise if (like the Samaritan) he saw an injured hu - Ch. 4—Ethical Considerations 79 man being. And he too may feel about equally jus- tified in both cases in being late for his uncle’s party. Humane treatment is the most commonly cited standard in Federal legislation concerning animals. Its wide range of application due to its lack of pre- cision, however, leads to a temptation to dismiss it as a pious but essentially vacuous sentiment. A theory of moral constraints is needed to determine whether this or some other standard is sufficiently precise to serve as a guide for legislation regulat- ing the use of animals. Moral Constraints A rule that allows an individual to do whatever that person wished would not be a moral rule. Morality by its very nature operates as a check on the tendency to go wherever desire leads. The constraints it imposes can be applied prospectively, contemporaneously, or retrospectively. Prospec- tive analysis looks ahead to the possible conse- quences, while retrospective analysis may restrict the results it is permissible to promote (37). Be- fore the action is taken, it can be said that the ac- tion that morally ought to be performed is the one with the best consequences. An individual succeeds in this objective to the extent that an action pro- duces as much benefit and as little harm as possi- ble. During the course of the action, conditions concerning the intention of the individual and the consent of the recipient may have to be met be- fore a moral license to pursue the best conse- quences is granted. The fact that a lie will produce more benefit than the truth will not necessarily make it the right thing to do. Moral theories divide according to the weight they give to one or the other kind of constraint. In its purest form, the prospective approach holds that an action or policy is right if it has better con- sequences, for everyone affected by it, than any available alternative. The language here is care- fully drawn. “Better” does not mean “morally bet- ter.” A good consequence is simply an outcome that someone finds desirable. If an action gives pleasure to someone, the enjoyment is a good thing; if it causes pain, the person’s suffering would be a bad thing. It is not necessary to ask whether the pleasure or pain is morally fitting. Intuition will ideally play no part in determin- ing an outcome. One consequence will count as better than another if, after assigning positive nu- merical values to its good elements and negative values to its bad ones, the sum of positive values exceeds that of negative values (10). Better for whom? The utilitarian principle, still the most influential formulation of the forward- looking approach, holds that actions and policies are to be evaluated by their effects, for good or ill, on everyone, not just the individual alone or some select group of individuals. Between an in- dividual’s own good and the good of others, “utili- tarianism requires him to be as strictly impartial as a disinterested and benevolent spectator” (10,34). The interests of each affected individual are to count equally. Any two experiences that are alike except that they occur indifferent individuals are to be given the same value. Among utilitarians, en- joyment is a good and suffering an evil, and so every animal with the capacity for such experiences will also count as one individual. Sentience suffices for possessing this value, even if it does not confer rights. “The question, ” as Bentham once put it, “is not, Can they reason? nor Can they talk?, but Can they suffer?’’. Because it extends the scope of moral concern to animals without committing itself to a vulner- able theory of animal rights, utilitarianism has be- come the theory of choice among those who would press for more constraints on humans’ treatment of animals. Singer derives the credo that all ani- mals are equal from the utilitarian conception of equality (45). If the principle of utility requires that suffering be minimized, and if some kinds of suffer- ing are found in animals as well as humans, then to count human suffering while ignoring animal suffering would violate the canon of equality. It would make a simple difference of location-in one species rather than another—the basis for a dis- tinction in value. Like racism, such "speciesism ” enshrines an arbitrary preference for interests simply because of their location in some set of in- dividuals (45). (For arguments that speciesism is not immoral, see refs. 16,23,51,52.) As a general moral principle, utilitarianism is sub- ject to several objections, the most serious being that its standard of equality is much too weak to 80 G Alternatives to Animal Use in Research, Testing, and Education satisfy the demands of justice (25,37,39). Since it only requires that individuals with interests be given the same consideration, but in its summa- tion of interests allows the claims of any one indi- vidual to be overridden by the sheer weight of numbers on the other side, it seems to sanction a tyranny of the majority that permits violations of individual rights. This may not, however, under- mine the utilitarian case for animals if animals have doubtful standing as right-holders. Some commentators have suggested that there may be an acceptable double standard in morals, consisting of a nonutilitarian principle for agents with standing as persons and a utilitarian rule for handling individuals with interests but not rights (21,37). The use of different rules for different kinds of individuals is already well established. Rules that would be objectionably paternalistic if applied to adults are admissible if restricted to chil- dren. The dangers are that inconsistent standards might hold for the same individual or that differ- ences between the two classes of individuals might be arbitrary. The suggestion that the adult-child and human- animal distinctions are comparably rational and justifiable (21) is superficial for two reasons. First, it does not seem to be arbitrary to distinguish be- tween the adult and the child, because human soci- ety understands that children may be intellectually and experientially unable to make wise choices. Thus, society can choose for children that which society believes is in their best interests. The prob- lem with the human-animal distinction is that an animal may in fact be able to make and communi- cate a decision that expresses the animal’s self- interest: It wants no part of any scientific proce- dure that results in pain or distress. Even if the animal could not make or communicate a decision, it may be arbitrary to distinguish between such animals and humans who are similar in their in- ability to make such decisions (the profoundly men- tally handicapped), allowing society to use the former but not the latter as research subjects. The second difference between the adult-child and human-animal distinctions relates to the pur- pose for distinguishing between two groups. The first distinction is permissible because it allows so- ciety to protect the interests of the child, while the purpose of the human-animal distinction is to allow society to ignore, or at least diminish, the interests of the animal. The device of a double standard is often used to explain the sharp differences in the constraints governing the treatment of animals and humans as experimental subjects. For animals the stand- ard is humane treatment, which forbids unneces- sary suffering but otherwise allows experiments that harm and even kill the animal. That same rule, proposed for human subjects, is generally consid- ered unethical. There are many experiments in which perfectly reliable results can only be ob- tained by doing to a human what is now done to an animal. Nevertheless, without the subject’s in- formed consent—indeed, sometimes even with it— such experiments are absolutely impermissible, no matter how beneficial the consequence might be. They would violate the rights of the human subject. The proscription against unnecessary suffering is best understood as a corollary of the principle of utility. Since suffering is a bad consequence, there is an initial utilitarian onus against behavior that would produce it. Such treatment calls for justification. To meet this burden, a bare appeal to some offsetting good consequence will not be sufficient. The principle of utility, as formulated, is comparative. It requires that an action or policy have better consequences than any available alter- native. Among the alternatives will be uses that do not involve animal suffering. If one of them has consequences at least as good as or better than the one proposed, the suffering will be unneces- sary. Other things being equal, then, it should prove harder to establish necessity than the contrary, since the former must rule out all the alternatives while the latter need find only one. Necessity is a relation between a means (an ac- tion or policy) and an end (its objective). Restricted necessity takes the end as given—that is, not sub- ject to evaluation—and asks only whether the course of action suggested is an indispensable means to that end, For example, in an LD50 test for toxicity that uses 40 rats as subjects (see chs. 7 and 8), if no alternative procedure using fewer or no rats could get the same results with the same reliability, that test would be necessary in the re - Ch. 4—Ethical Considerations G 8 1 stricted sense. In unrestricted necessity, the end is open to assessment on utilitarian grounds: G G How likely is the objective to be met, in compari- son with other possible goals? If the LD50 test yields unreliable results, its necessity in the unrestricted sense would be open to challenge. Assuming that the objective will be met, how beneficial will it be? Suppose, for instance, that an LD50 test were to be run on a new cosmetic not significantly different from those already on the market. The test may be considered unnecessary because the objective is unnec- essary. Unrestricted necessity is more difficult to prove, because it always includes restricted necessity and more. Thus, a stringent standard of necessity, one that lets fewer procedures through, would require that a procedure be necessary in the unrestricted sense. In addition, since necessity is more difficult to establish than the possibility of substitution, the burden of proving both the existence of necessity and the absence of alternatives could be placed on those who would use the procedure. A more lenient test could invert these priorities by pre- suming that the procedure is necessary and that alternatives are lacking unless shown otherwise. This approach would not expect the user to show beforehand that no other alternative was avail- able; it is generally followed when a research pro- posal is reviewed by a scientist’s peers or an insti- tutional animal care and use committee (27). Nonutilitarian positions on the use of animals have one feature in common: Although virtually none ignores consequences, they unite in deny- ing that a course of action can be justified wholly by appeal to the value of its consequences (39). This leaves room for substantial variation, with the differences traceable to the considerations they would add in order to complete amoral assessment. Ironically, both extremes in the animal treatment debate are nonutilitarian. The hard line support- ing unlimited exploitation of animals builds from the premise that animals lack moral standing. With- out rights, they cannot be recipients of a duty owed to them. On some theories of value, moreover, en- joyment does not count as a good thing in itself, nor is suffering per se an evil. Kant, for example, thought that the only unconditional good was a will whose choices are undetermined by desire for enjoyment or fear of punishment (31). Not hav- ing a will, animals could not have this value. Morally, they were indistinguishable from inani- mate tools—mere means to be used for the pur- poses of beings who do have a will. Like Aquinas, however, Kant did acknowledge an indirect duty of kindness, given that “tender feelings toward dumb animals develop humane feelings toward mankind” (32). The indirect duty theory stumbles in the attempt to explain why there should be any empirical con- nection at all between people’s feelings for animals and their feelings for other humans. Some simi- larity must be seen in the objects of the two senti- ments if one is to influence the other; yet the the- ory says that there is no such likeness in reality. Thus either a person’s motive is proof by itself that humans have a direct moral interest in animals, in which case the theory is mistaken; or the the- ory is correct and the individual has misunder- stood it, in which case the person will be free, once educated in the theory, to abuse animals without fear that this will tempt abuse of human beings. Kant cannot have it both ways: He cannot require individuals to act on a belief that his own theory alleges to be false (33). The Kantian position could be turned on its head if animals had moral standing after all. In The Case for Anhnal Rights, Regan gives the most cogent defense to date for that view. He concedes that animals are not moral agents: Since they are un- able to choose freely among impartially determined moral alternatives, they cannot have any moral duties. At least some animals, however, have be- liefs, desires, memory, a sense of the future, prefer- ences, an identity overtime, and an individual wel- fare of their own (41). In these respects, they are indistinguishable from human infants and men- tal defective, who also fail to qualify as moral agents. Nevertheless, these animals possess an in- herent value, independent of the value that their experiences may have, that gives them standing as “moral patients” — that is, as individuals on the receiving end of the right and wrong actions of moral agents. They have this value equally, and equally with moral agents (40). Inherent value in turn gives them a claim, or right, to certain treatment. 82 . Alternatives to Animal Use in Research, Testing, and Education Regan’s major thesis is that, as moral patients, animals enjoy a presumptive right not to be harmed. He considers this principle a radical alternative to utilitarianism. But once the reference to rights is filtered out, the utilitarian might find Regan’s theory quite congenial. Both Regan and utilitarians would hold that harm to animals is a bad conse- quence and so it would be wrong, in the absence of an overriding consideration, to harm them. The conflict between the two theories, therefore, lies in the kind of justification that each theory would permit to overturn this presumption. Regan offers two guiding principles (40). By the first, when the choice is between harm to a few and harm to many and when each affected indi- vidual would be harmed in a comparable way, then the rights of the few ought to be overridden. As Regan acknowledges, the utilitarian commitment to minimize suffering would have the same result. By the second principle, when the choice is be- tween harm to a few and harm to many, if a mem- ber of the affected few would be worse off than any member of the affected many, the rights of the many ought to be overridden. This “worse- off” rule parts company with utilitarianism in set- ting aggregate consequences aside and protecting minority interests. In view of this possibility, it is surprising to find that Regan calls for a blanket prohibition against the use of animals in research and toxicity test- ing. That conclusion would follow only if his two rules for defeating the right not to be harmed could never be successfully invoked in these areas. Re- gan is apparently drawn to this result by a con- straint he attaches to the rules: They hold only for harms suffered by innocent victims. Animals are always innocent, in the sense that Regan gives to that term (41). But human patients will be, too, and at least sometimes human agents will also be. Regan would have to show that these occasions can never arise in research, testing, or education, or that, if they do, the human agent/patient never faces the greater harm. His analysis does not show this. This difficulty aside, Regan’s theory can be read as holding, first, that the necessity standard can- not be applied until the innocence of all parties has been established and, second, that when it does apply, the worse-off rule should replace the util- ity principle in cases where they diverge. It is unclear whether the worse-off rule is prefer- able to the utilitarian principle for the purposes of animal use. But the notion of innocence, with its judicial implications, appears to have no place in the issue of experimental-subject rights for three reasons. First, the notion that animals are always innocent because they cannot be otherwise is prob- lematic. Innocence makes sense only when guilt does, because innocence means that one has done no wrong though doing wrong was an option. If animals are not rational decisionmakers, if they cannot choose between right and wrong, then the concept of innocence has little meaning. Second, most human subjects are probably innocent in the sense that Regan uses the term, so that the con- cept does little to advance the theory that ex- perimenting on humans is preferable to experi- menting on animals. Finally, even a guilty person may have certain rights. While a person guilty of a crime against society maybe imprisoned or other- wise punished, society holds that the guilty have a right to avoid cruel and inhumane punishments. Bioethics similarly rejects the involuntary use of guilty prisoners in medical experiments. SUMMARY AND CONCLUSIONS The present debate over animal use in research, testing, and education is marked by a cacophony of voices. A critical survey of the religious and philo- sophical backgrounds to the debate yields some hope that, if the competing voices were muted by reflection, they would begin to coalesce as varia- tions around a single theme. That theme would be the standard of humane treatment, extended to animals as well as to humans. Much has been made of the historical contrast between Western and Oriental religious views on animals. The biblical and theological texts in the Judeo-Christian tradition do not give us a princi- ple of unconditional respect for animals. Humans alone are accorded inherent value as being cre- ated in the image of God, and this gives them a license to use animals for their own purposes. Not, however, to abuse them. Cruelty and callous in- Ch. 4—Ethical Considerations . 83 difference to the needs of animals find no scrip- tural support, and virtually all religious thinkers condemn them. If God is a good shepherd, treat- ing humans kindly without being bound to, hu- mans can be as much to the animals in their care. The Christian position thus amounts to a synthe- sis of two elements in tension. On the one hand, animals are inferior in worth to humans, as the body of a person is inferior to the soul. On the other hand, they are not so inferior that their own welfare cannot stand in the way of unbridled use of them. Modern religious and philosophical patterns of thought are branches of the same ancestral trunk. It should not be surprising, then, that the philo- sophical tradition exhibits the same tension on the subject of animals. Humans have standing as per- sons—that is, as individuals who can assume duties and enjoy rights. To join them, animals must at least be capable of possessing rights. But they can- not assume duties and do not have the power of discretion that gives rights a distinctive role in morals. Consistency suggests rights should be ascribed to animals once rights are given to infants and mentally handicapped humans who also lack discretion. Yet it would be inconsistent to assert that humans are not superior to animals while sug- gesting that humans should refuse to exploit other species, even though other species exploit each other. Even if animals are not moral persons, however, it does not follow that they are mere things, morally indistinguishable from machines. They are suffi- ciently like humans in one morally relevant re- spect—their capacity for suffering in basic forms— to generate a moral claim on humans. It would be inconsistent to hold that, other things being equal, human suffering ought to be relieved, but animal suffering ought not. Because it extends the scope of moral concern to animals without committing itself to a vulner- able theory of animal rights, utilitarianism has be- come the theory of choice among those who would press for more constraints on humans’ treatment of animals. If the principle of utility requires that suffering be minimized, and if some kinds of suffer- ing are found in animals as well as humans, then to count human suffering while ignoring animal suffering would violate the canon of equality. It would make a simple difference of location-in one species rather than another—the basis for a dis- tinction in value. Like racism, such “speciesism” enshrines an arbitrary preference for interests simply because of their location in some set of in- dividuals. The rule that suffering ought to be relieved, in humans or animals, is the principle of humane treatment. It covers a large and heterogeneous range of situations; the most germane, for the de- bate over animal use, are those in which someone inflicts suffering on someone else. The humane treatment principle establishes a presumption against doing this, but that presumption can be overcome—always in the case of animals, and sometimes even in the case of a human—by show- ing that the harm done is necessary. Necessity here is not bare utility, but necessity overall. The harm must not only be a means to a good end, it must be the only means. A broader definition of neces- sity might also require that the harm be a means to an end whose value is considered in light of the degree of harm necessary to achieve that end. In addition, necessity always implies a comparison with available alternatives. Animal use in research, testing, and education creates a conflict of interests between the liberty that humans have to use animals for human ends (knowledge, health, safety) and the need that ani- mals have to be free of suffering. There is no rea- son why either one of these broad interests should always prevail over the other. The fulcrum on which they are balanced is the necessity standard itself. That is, when the suffering inflicted on ani- mals is not necessary to satisfy a desirable human objective, the animal interest will prevail. And when the suffering is unavoidable, the human in- terest will be controlling. Animals are morally en- titled to be treated humanely; whether they are entitled to more than that is unclear. 84 G Alternatives to Animal Use in Research, Testing, and Education Reprinted with permission from CHEMTECH 15(1):63, Copyrlgnt 1965, American Chemical Society. CHAPTER 4 REFERENCES 1. Aquinas, T., Summa Contra Gentiles, tr. English Do- minican Fathers (New York: Benzinger Brothers, 1928). 2. Aquinas, T., Summa Theological, tr. English Domini- can Fathers (New York: Benzinger Brothers, 1918). 3. Ancombe, G. E. M., “Modern Moral Philosophy )’’Phi- losophy 33:1-19, 1958. 4. Aristotle, De Anjma/, tr. K. Foster and S. Humphries (New Haven, CT: Yale University Press, 1954). 5. Aristotle, PoJitics, tr. E. Barker (Oxford, England: The Clarendon Press, 1952). 6. Attfield, R., The Ethics of Environmental Concern (New York: Columbia University Press, 1983). 7. Augustine, The Catholic and Manichean Ways of Life, tr. D.A. Gallagher and I.J. 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Thomas, K., Man and the Natural World (New York: Pantheon Books, 1983). 50. Walker, S., Animal Thought (London: Routledge & Kegan Paul, 1983). 51. Watson, R. A., “Self-Consciousness and the Rights of Nonhuman Animals and Nature, ” Environ. Ethics 1:99-129, 1979. 52. Williams, M., “Rights, Interest, and Moral Equal- ity, ” Environ. Ethics 2:149-161, 1980. — chapter 5 The Use of Animals in Research I know that half of what I teach as fact, ” said a wise medical pedagogue, “will be proved false in 10 years. The hard part is that I don’t know which half. ” His statistics may not be exact, but the notion is right enough. No one knows which half, and it is impossible to know except in retrospect. That is what research is—the reason for the prefix. The half that is wrong is at least as important as the half that is right, because the new questions come in ferreting out the errors-and new answers too. Kenneth L. Brigham Vanderbilt University School of Medicine N. Engl. J. Med, 312:794, 1985 CONTENTS Page The Role of Animals in Biomedical Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 Nonhuman Primates in Biomedical Research. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 Experimental Animals’ Contribution to Coronary Artery Bypass Graft Surgery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Use of Multiple Species in Biomedical Research . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Choice of Species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98 The Role of Animals in Behavioral Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99 What is Behavioral Research? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99 Why Are Animals Used in Behavioral Research? . . . . . . . . . . . . . . . . . . . . . ....101 Methods of Behavioral Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ........102 Use of Multiple Species in Behavioral Research . . . . . . . . . . . . . . . . . . . . .. ....103 Pain and Distress in Research Animals . . . . . . . . . . . . . . . . . . . . . ..............103 Animal and Nonanimal Protocols in Biomedical and Behavioral Research Reports . . . . . . . . . . . . . . . . . . . . . ...................105 Survey Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ......106 Survey Limitations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .........106 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. ...108 Chapter 5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..109 List of Tables Table No. Page 5-1.Some Uses of Nonhuman Primates in Research on Human Health and Disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 5-2. Some Anatomical, Physiological, and Metabolic Similarities and Differences Between Humans and Various Laboratory Animals . . . . . . . . . . . 97 5-3. Classification of Research Experiments and Procedures According to the Degree of Pain or Distress for the Animal . . . . . . . . . . . . . . . . . . . . . . .105 5-4. Classification of Published Research Protocols in OTA Survey of 15 Journals . . . . . . . . . . . . . . . . . . . . . . ...................106 5-5. Percentage of Papers (Average, 1980-83) Using Animal, Nonanimal, and Human Subjects in 15 Biomedical and Behavioral Research Journals Surveyed by OTA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..................107 Figure Figure No. Page 5-1. Steps in Biomedical Research That Preceded Successful Coronary Artery Bypass Graft Surgery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Chapter 5 The Use of Animals in Research Research, as the word denotes, is an ongoing search—a search for new information and for novel ways to apply existing information. Re- search assumes a multitude of directions in a wide variety of disciplines. It is not surprising, then, that the use of animals in research—and the po- tential for alternatives to using them—mirrors the multifaceted nature of research itself. Viewed broadly, almost any research investi- gation involving members of the animal kingdom, including humans, and sometimes even members of the plant kingdom, can be categorized as bio- medical research. In this sense, biomedical re- search covers a long list of disciplines: anatomy, anesthesiology, behavioral biology, biochemistry, biomedical engineering, biophysics, cardiology, cell biology, dentistry, developmental biology, en- docrinology, entomology, epidemiology, genetics, gerontology, histology, immunology, metabolism, microbiology, molecular biology, neurology, nu- trition, oncology, parasitology, pathology, phar- macology, physiology, psychology, radiology, re- productive biology, surgery, teratology, toxicol- ogy, veterinary science, virology, and zoology. When considering animal use—and alternatives to animal use—in research, it is useful to isolate behavioral research from the broader category of biomedical research. Behavioral research is a part of biomedical research, yet is distinguished from the larger topic by the nature of the exper- iments, the identity of the researchers, and the kinds of alternatives available (see ch. 6). This chapter defines and describes animal use in biomedical and behavioral research. Also in- cluded are the results of a brief survey done by OTA of the use of animal and nonanimal meth- ods in published research reports in selected dis- ciplines of biomedical and behavioral research. T H E R O L E O F A N I M A L S I N B I O M E D I C A L R E S E A R C H To discuss alternatives to using animals in bio- medical research, it is important to review the context in which animals are presently included. A comprehensive review of this subject (see ref. 41) is beyond the scope of the present assessment. However, animals’ broad role in contemporary biomedical research can be at least partly delin- eated by considering: G the manifold contributions to biomedical re- search of a single group of animals—non- human primates; G the role of experimental animals in the de- velopment of a single medical procedure, namely coronary artery bypass surgery; and G the reasons multiple species are used in bio- medical research. These perspectives illustrate two fundamental principles of animal use in biomedical research. First, a single species or group of animals often serves a multitude of purposes in widely varying research enterprises. Second, a single advance in applied research often represents results gener- ated from many species. Nonhuman Primates in Biomedical Research Primates—humans, monkeys, and apes—share a common genetic basis and anatomical, physio- logical, biochemical, and behavioral traits that provide unique research opportunities. As a con- sequence, humans and other primates are sus- ceptible to many of the same diseases and have many of the same disease-fighting capabilities. Re- viewing the use of nonhuman primates is also appropriate because they are relatively expensive” research animals (e.g., a rhesus monkey costs from $600 to $2,000) and the object of much pub- lic interest. Two recent reports describe the role of primates in biomedical research (29,45). Those 89 38-750 0 - 86 - 4 ¼iG Alternatives to Animal Use in Research, Testing, and Education reports are summarized in table 5-1; highlights mans had been made by the early 1900s, but the of the studies follow. cause of the disease was still unknown. A break- through occurred in 1908, when scientists exper- Polio imentally transmitted the poliovirus to monkeys for the first time. Studies in rhesus and cynomolgus The development of the polio vaccine exempli- monkeys and in chimpanzees followed isolation fies the key role of primates in the research lab- of the virus, but a vaccine remained elusive. Af - oratory. Many thorough studies of polio in hu- ter nearly a half-century, researchers were able Table 5-1.—Some Uses of Nonhuman Primates in Research on Human Health and Disease Human health concern Primate experimental model Acquired immune deficiency syndrome (AIDS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Chimpanzee, African green monkey Atherosclerosis . . . . . . . . . . . . . . . . . . . . . . . . . . Cynomolgus monkey Balding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Stumptail monkey Cancer from solid tumors . . . . . . . . . . . . . . . . . Chimpanzee Cholesterol gallstones . . . . . . . . . . . . . . . . . . . .Squirrel monkey Circadian rhythms . . . . . . . . . . . . . . . . . . . . . . . .Squirrel monkey Cornea transplant . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey, African green monkey, Stumptail monkey, Patas monkey, Cynomolgus monkey Dental implants . . . . . . . . . . . . . . . . . . . . . . . . . . Pig-tailed monkey Diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Celebes black macaque Dietary fats and heart disease . . . . . . . . . . . . .Cynomolgus monkey Embryo transfer . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey, Cynomolgus monkey Eye damage from ultraviolet radiation . . . . . . . Rhesus monkey Eye disorders in children . . . . . . . . . . . . . . . . . . Rhesus monkey Fetal alcohol syndrome . . . . . . . . . . . . . . . . . . . Pig-tailed monkey Fetal surgery . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey Genital herpes . . . . . . . . . . . . . . . . . . . . . . . . . . .African green monkey Gilbert’s syndrome . . . . . . . . . . . . . . . . . . . . . . . Bolivian squirrel monkey Glaucoma . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey Hearing impairment. . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey Hepatitis B . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey, Chimpanzee, African green monkey Herpes-virus-induced cancer . . . . . . . . . . . . . . . Owl monkey, Marmoset High blood pressure . . . . . . . . . . . . . . . . . . . . . . Cynomolgus monkey Hyaline membrane disease in newborns. . . . . Rhesus monkey, Pig-tailed monkey In vitro fertilization . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey, Chimpanzee, Baboon, Cynomolgus monkey Infertility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey Inflammatory bowel disease . . . . . . . . . . . . . . .Marmoset Laser surgery on damaged nerves . . . . . . . . . . Baboon Leprosy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Sooty mangabey Liver disorders . . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey Malaria. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chimpanzee, Owl monkey, Rhesus monkey Male and female behavior patterns . . . . . . . . .Rhesus monkey Male birth control . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey, Cynomolgus monkey Menopausal problems. ., . . . . . . . . . . . . . . . . . . Rhesus monkey, Stumptail monkey Mother-infant behavior . . . . . . . . . . . . . . . . . . . . Rhesus monkey Motion sickness . . . . . . . . . . . . . . . . . . . . . . . . . Squirrel monkey Nonhormonal fertility regulation . . . . . . . . . . . . Bonnet monkey, Chimpanzee Obesity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Baboon Parkinson’s disease , ... , . . . . . . . . . . . . . . . . . Rhesus monkey Polio. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey, Cynomolgus monkey, Chimpanzee Premature labor . . . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey, Baboon Rh factor disease . . . . . . . . . . . . . . . . . . . . . . . . Rhesus monkey Slow viruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Owl monkey, Squirrel monkey, Stumptail monkey Systemic lupus erythematosus . . . . . . . . . . . . . Cynomolgus monkey SOURCES: Adapted from “Toward Better Health: The Role of Primates in Medical Reaearch,” Primate News 21(1):1-24, 1984; and F. A. King and C. J. Yarbrough, “Medical and Behavioral Benefits From Primate Research,” Physlo/ogkt 28:75-87, 1985, Ch. 5—The Use of Animals in Research . 91 to grow the poliovirus in human tissue culture (15), and an effective vaccine became available to the public in 1955. When the vaccine was devel- oped, monkey kidney tissue was essential for pro- duction of pure virus in great quantities, and live monkeys were essential for safety and effective- ness testing. Today, noninfectious polio vaccine can be produced in continuously propagating cells without the need for monkeys, although monkeys are required to test for safety. The impact of the polio vaccine has been dramatic: In 1952, at the height of one epidemic, 58,000 cases of polio occurred in the United States; in 1984, just 4 cases were reported (39). Hepatitis B Hepatitis B is the most dangerous form of hepa- titis, a debilitating liver disease characterized by fever, weakness, loss of appetite, headache, and muscle pain. There are nearly 1 million hepatitis B virus carriers in the United States today, and the infection is estimated to cost $1 million per day in this country. Worldwide, there are some 200 million carriers, primarily in Asia and Africa. Up to 1 percent of those infected with hepatitis B die of the disease, and 5 to 10 percent become chronic carriers of the virus who can remain in- fectious indefinitely (21). Since there is no known treatment for hepatitis B infection, prevention is essential. Research with rhesus monkeys and chimpan- zees led to the development just a few years ago of a vaccine, derived from human plasma, against hepatitis B infection. In 1981, the Food and Drug Administration (FDA) licensed this vaccine for hu- man use. In 1984, recombinant DNA technology was used to prepare a hepatitis B vaccine from yeast cells (the first vaccine for human use so produced). Prior to its trial in 37 human volun- teers, this yeast recombinant hepatitis B vaccine was administered to African green monkeys in order to gauge its effectiveness (52). These new vaccines are expected to have a worldwide im- pact on the disease, and they may also reduce the incidence of hepatocellular carcinoma, a form of liver cancer associated with chronic hepatitis B infection (40). Herpes Estimates of the number of persons afflicted with recurrent genital herpes virus infections range from 5 million to 20 million worldwide (61). A new antiviral drug, acyclovir, was recently li- censed for use against human genital herpes in- fections and appears to yield antiviral and clini- cal benefits when taken orally (48). Acyclovir was extensively tested in African green monkeys. The opportunity to run such tests arose because of a natural outbreak of a virus closely related to that causing both chickenpox and shingles (i.e., herpes zoster) in humans. The infected monkey colony at the Delta Regional Primate Research Center in Louisiana enabled scientists to study the herpes disease process and test antiviral drugs. In 1984, researchers reported an in vitro model system for studying the herpes simplex virus, using human fetal nerve cells as the host. This in vitro model is expected to enable analysis of the state of the herpes virus as it establishes and remains latent in human nerve cells (62). High Blood Pressure High blood pressure, when untreated, increases the risk of stroke, heart disease, and kidney fail- ure. In most cases, the cause or causes of high blood pressure remain unknown, and the condi- tion is a public health problem of immense pro- portions. Data from the early 1980s indicate that fully one-third of Americans use medication to control blood pressure. From 1971 to 1981, visits to physicians for diagnosis and therapy of high blood pressure increased by 55 percent, while visits for all other causes decreased by approxi- mately 5 percent (32). Monkeys are used to examine mechanisms of high blood pressure because the natural hormone molecules controlling blood pressure (e.g., the kid- ney hormone renin) are identical in humans and other primates. In contrast, the renin molecules of humans and nonprimate species are dissimi- lar. In addition to using monkeys to study the ef - fects of diet and drugs on high blood pressure, researchers are examining the genetic transmis- sion of high blood pressure. One breeding colony of cynomolgus monkeys exhibiting high blood 92 “ Alfernatives to Animal Use in Research, Testing, and Education pressure has been monitored for 5 years; this per- mits the study of high blood pressure in parents, offspring, and future generations to analyze the tendency to inherit the condition. Parkinson's Disease Parkinson’s disease is a neurological disorder of older adults characterized by palsy and rigid muscles. Progress in understanding the cause and development of the disease and in refining meth- ods of long-term drug therapy has been ham- pered by lack of an adequate animal model. At- tempts to induce the disease in rats, guinea pigs, and cats either have failed to produce all the symptoms or have yielded symptoms that do not last long and so cannot be effectively researched. In 1983, the first animal model of Parkinson’s disease was developed. Scientists at the National Institute of Mental Health (NIMH) induced a form of parkinsonism in eight rhesus monkeys by giv- ing them a drug, l-methyl-4-phenyl-1,2,3)6-tetra- hydropyridine (MPTP), that selectively destroys specific cells in the substantial nigra, a region of the brain destroyed in humans by Parkinson’s dis- ease. The monkeys exhibited all the major clini- cal features of Parkinson’s disease in humans. They also responded dramatically to L-dopa, the standard medication for people with this disease (8). NIMH researchers have speculated that the availability of this new animal model may lead to understanding the reason Parkinson’s disease oc- curs in older adults, the course of the disease, and drug therapy and its side effects (30). In 1984, squirrel monkeys were used to shed further light on the mechanism of MPTP-induced parkinsonism. Pargyline, a drug currently pre- scribed for high blood pressure in humans (Eu- tonyl, Abbott Laboratories, North Chicago, IL), was used to prevent the neurotoxic effects of MPTP (31). These results in squirrel monkeys sug- gested that MPTP itself may not be the actual neu- rotoxic agent. Instead, attention is now focused on an MPTP metabolize and on the mechanism of MPTP metabolism in the brain (33). Baldness Like many men, stumptail monkeys become bald as they age. This trait has made the stump- tail monkey the animal of choice in baldness research. Although it is not life-threatening, bald- ness is a matter of concern for many people: Hair- combing patterns suggest that many men desire to have hair where there is none, and advertise- ments for hair restoration abound in the popular media. The public spends large sums on hair- restoration nostrums, and in 1985 the FDA pro- posed banning the sale of all nonprescription drug products sold to prevent or reverse baldness, hav- ing concluded there is no scientific evidence that such lotions and creams are effective (50 FR 2191). A drug originally developed to manage high blood pressure, minoxidil, has the unexpected side effect of causing thick hair growth from fol- licles that normally produce only fine, downy hair. To test the potential of minoxidil for hair growth, researchers applied it externally to the bald front scalp of stumptail monkeys. The results with monkeys showed promise, and clinical trials are now in progress with bald men across the United States. Monkey studies are continuing to assess the effects and safety of minoxidil as a means of counteracting hair loss. Menopausal Hot Flashes Of the 30 million postmenopausal women in the United States, as many as 75 percent have ex- perienced or will experience hot flashes brought on by increased blood flow to the skin. Hot flashes produce a feeling of warmth for several minutes, and they are often followed by sweating. These physical symptoms may be accompanied by ner- vousness, irritability, and depression. At present, physicians can treat the symptoms of menopause, but the causes of the symptoms remain unknown. Research into the mechanisms of menopause and the development of therapy for menopausal problems has been hampered by the difficulty of studying this condition in animals. This difficulty stems from three facts: only primates, and no other nonhuman species, have menstrual cycles; monkeys do not exhibit symptoms of menopause until at least age 25; and monkeys brought into the laboratory from the wild are rarely of meno- pausal age. Throughout the last decade, researchers have studied the menstrual cycle and its cessation in Ch. 5—The Use of Animals in Research G 9 3 ~ . Left: Bent, flexed posture and absence of movement exhibited by a rhesus monkey treated with the drug MPTP to induce Parkinson’s disease. Other symptoms in both monkeys and humans include tremor, eyelid closure, difficulty swallowing (drooling), and difficulty with vocalization and speech. Right: Reversal of abnormal posture and return of nor- mal movement following treatment with L-dopa. The right photograph was taken 2 hours after the left one. Photo credit: f?. Stanley Bums, National Institute of Mental Hea/th First animal model of Parkinson’s disease, developed in 1983 a limited number of rhesus monkeys reaching 25 and of hormones in the control of this problem. to 30 years of age (11). In 1984, hot flashes were Once the underlying mechanisms that produce described in another primate, the stumptail mon- hot flashes are better understood, more effective key (25). The aim of developing an animal model treatments may be developed for women who for hot flashes is to determine the role of the brain suffer from menopausal problems. 94 Ž Alternatives to Animal Use in Research, Testing, and Education Experimental Animals’ Contribution to Coronary Artery Bypass Graft Surgery A second way to describe the role of animals in biomedical research is to review the ways in which a single advance in applied biomedical re- search came about. As an illustration of this proc- ess, the development of the coronary artery by- pass graft operation, recently recounted (9,46), is summarized here. Coronary or arteriosclerotic heart disease, often caused by a narrowing or blocking of the arteries supplying blood to the heart, is the number one cause of death in the United States. In 1982, it was responsible for approximately 500,000 deaths (59). Coronary artery bypass graft surgery was introduced in the early 1970s. In this procedure, which has become the primary surgical approach to treatment of coronary artery disease, a grafted vessel is attached to the coronary artery to cir- cumvent the constricted portion. The graft im- proves the blood and oxygen supply to the heart muscle. The growth of the procedure has been quite rapid: Approximately 70,000 operations were performed in 1977; 160,000 in 1981 (7); and 191)000 in 1983 (38). Coronary artery bypass graft surgery is now the most commonly performed major operation in the United States (7). It is accepted as far more effective than medication in relieving the severe chest pain, or angina pectoris, associated with coronary heart disease (47). The long-term ben- efit of this procedure, in terms of mortality, varies among patient groups (60). The experimental steps leading to the success- ful coronary artery bypass graft operation are depicted in figure 5-1, in which the cardiac sur- geon stands at the summit of Mt. Coronary Ar- tery Bypass. In the early stages of research—that is, in the foothills of the mountain—there was a great deal of variability in the kinds of animals required. Studies in frogs, reptiles, horses, cats, dogs, sheep, and deer contributed to scientists’ understanding of the fundamental principles of circulation, blood pressure, and temperature reg- ulation. As problems became more specialized, the choices of animal species became more re- stricted. Dogs, chimpanzees, and, ultimately, hu- mans contributed to the later stages of research leading to the coronary artery bypass. Virtually every step up Mt. Coronary Artery Bypass re- quired initial stages of study on living animal models of various species. Today, in retrospect, the experimental steps leading to this surgical procedure appear as a sim- ple and logical progression. In this sense, figure 5-1 is deceptive. It is important to note that the first step was not predictive of the second step, the second not predictive of the third, and so on. The advance from each step involved uncertainty, missteps, and serendipity. All are inherent in the process of basic biomedical research. Moreover, only a poor understanding exists of the path lead- ing from basic to applied biomedical research. Al- though Mt. Coronary Artery Bypass stands as a bona fide illustration of the integration of data drawn from several species, it was formed with- out a blueprint. Use of Multiple Species in Biomedical Research The contributions of animals are an important part of the history of human health, disease, and medicine. It is noteworthy that animals have not only contributed to human welfare, but deterred from it as well. The benefits and detriments de- rived from animals involve numerous species. The number of animal diseases labeled zoo- noses-diseases transmissible from animals to humans–now stands at about 200. These exact a heavy toll of human morbidity and mortality on a worldwide scale. Research to combat zoo- noses logically focuses on the species that are the principal sources of the diseases. And the more species that are infected by a particular agent, the greater are the biological resources available for research to overcome it. Numerous animal vectors of an infectious agent provide increased opportunities for the study of variation among species in the incubation of, transmission of, and susceptibility to the infectious agent. Most of the threats to humans from animals—including ra- bies, tuberculosis, brucellosis, toxoplasmosis, an- thrax, and dengue fever—infect a sufficient va- Ch. 5—The Use of Animals in Research G 9 5 Figure 5-1.—Steps in Biomedical Research That Preceded Successful Coronary Artery Bypass Graft Surgery dog, human L : ’ Q J ~ : “ “ ~ Direct autograft - dog, human :., - M - $ 1 1 SOURCE: Redrawn from W, C Randall, ‘iCrises in Physiological Research,” Physiologist 26:351 -356, 1963, after J. H. Comroe, Jr., and R. D. Dnpps, “Ben Franklln and Open Heart Surgery,” Circ Res. 35:661-669, 1974 . Alternatives to Animal Use in Research, Testing, and Education riety of species that effective research on their control has been possible (19). Perhaps ironically, the same diverse mix of species that transmits dis- ease to humans forms the substrate for research to ameliorate human disease. Infection of multiple animal species has led to virtual control in industrial countries of the plagues just mentioned. Yet a paucity of animal vectors or models hampers control of certain other human infectious diseases. Leprosy, herpes, and gonorrhea (which are not zoonoses) have yet to be brought under control, owing partly to the lack of effective animal models. Recent discoveries of leprosy and herpes infections in primates, the culture of the leprosy bacillus in armadillos, and adaptation of the gonorrhea organisms to some species of laboratory animals offer promise that effective animal models will soon become avail- able for research (19). Yet research on other con- ditions of still-unknown etiology, such as Alz- heimer’s disease, remains impeded by the inability to identify an appropriate animal model. Additional impetus for employing a variety of species in the course of research comes from a consideration of the immune response, which rec- ognizes material that is foreign to the body. The immune system thus serves as an animal’s defense against infections due to viruses, fungi, or bacte- ria. When foreign proteins, or antigens, are in- troduced into an animal, the immune system re- sponds by manufacturing a protein of its own, an antibody, to counter the invader. This is the principle on which the development of vaccines is based: An antigen is injected, and it stimulates production of an antibody that combats the for- eign antigen. The strength of an immune response varies from species to species, and even within a spe- cies, according to the genetic constitution of the animal used. Researchers often cannot gain a full understanding of how to develop useful vaccines unless they test several species to examine sub- tle differences in immune responses. In this way, species differences in response to foreign antigens are found and can be exploited in the production of effective vaccines for humans and animals. It is this use of the immune system that has con- trolled most of the major infectious viral diseases, including smallpox, which was controlled through the use of the cowpox, or vaccinia, virus. No one animal species is the complete research model for the human. In general, nonhuman pri- mate species have the greatest anatomical, physio- logical, and metabolic similarities to humans. Yet, as table 5-2 indicates, much important biological information can be provided by using dissimilar organisms. (This table oversimplifies the use of various animals in studying human health and dis- ease because it does not rate the closeness of the similarity of the conditions between humans and animals (19).) It is important to establish any new biological principle or a new phase of understanding a dis- ease condition in as many species as possible in order to improve safely the extrapolation from one animal to another and to humans (19). Re- search results derived from multiple systems in varied species, such as those listed in table 5-2, complement each other to approximate human anatomy, physiology, and metabolism. Some biomedical research, collectively known as veterinary research, seeks to understand the life processes of animals and applies this knowl- edge to serve animals themselves, as well as hu- mans. Veterinary research addresses the normal structure and function of animals and the causes, diagnosis, prevention, and treatment of disease in experimental animals and clinical (i.e., patient) animals. Research on food- and fiber-producing domestic animals supports the utilization of plant and animal resources for human sustenance. Veterinary research plays a prominent role in controlling diseases of importance in food-produc- ing animals and, hence, of importance to humans. Veterinary research supports, and is closely al- lied with, veterinary medicine. Practitioners of veterinary medicine maintain and improve the health and well-being of animals. The profession concentrates on the health of animals important for food and fiber and on companion animals. other species receiving veterinary attention in- clude laboratory animals, fish and aquatic ani- mals, and zoo and wild animals, Thus, the ma- jority of veterinary medicine addresses 30 to 40 different species of economic, ecologic, and envi- ronmental importance. These include: G domestic animals (e.g., cats, cattle, chickens, dogs, donkeys, goats, horses, sheep, and turkeys); Ch. 5—The Use of Animals in Research G 9 7 Table 5-2.—Some Anatomical, Physiological, and Metabolic Similarities and Differences Between Humans and Various Laboratoy Animals Conditions, systems, or structures Animal Similarities to humans Differences from humans Cat . . . . . . . . . . . . . . . . . Splenic vasculature Sphenoid sinus in skull Liver Middle ear and ear drum Epidermis Cattle. . . . . . . . . . . . . . . Ascending colon Electrolyte excretion Chicken . . . . . . . . . . . . . Palate Chinchilla . . . . . . . . . . . Inner ear structures Dog . . . . . . . . . . . . . . . . Pituitary gland vasculature Renal arteries Splenic vasculature Sphenoid sinus in skull Superficial kidney vasculature Liver Epidermis Adrenal gland innervation Goat. . . . . . . . . . . . . . . . Embryonic blood circulation Guinea pig . . . . . . . . . .Spleen Immune system Horse. . . . . . . . . . . . . . . Pulmonary vasculature Bile duct Pancreatic duct Lung Mouse . . . . . . . . . . . . . . Senile hepatic changes Pig . . . . . . . . . . . . . . . . . Maturation of red blood cells Cardiovascular tree Teeth Adrenal gland Skin Penile urethra Retinal vessels Spleen Reaction to foreign protein Laryngeal structures Sweat glands Mediastinum (interior chest tissue) Development of embryonic gonads Sleep Heat regulation Digestion Plasma gamma globulins in newborn Sleep Heat regulation Vomiting Sweat glands Retinal vessels Lymphoid tissue in liver Pituitary gland Respiratory system Oviduct Reproductive system Acetate metabolism Intestinal circulation Anal sacs Sweat glands Pancreatic ducts Heat regulation Sleep Laryngeal nerves Mediastinum Stomach and digestion Heat regulation Sweat glands Vomiting Sleep Plasma gamma globulins in newborn Sweat glands Carotid body Spleen Cecum and colon Gall bladder Plasma gamma globulins in newborn Spleen Spleen Liver Plasma gamma globulins in newborn Sweat glands 98 G Alternatives to Animal Use in Research, Testing, and Education G G G G G G G G Table 5-2.—Some Anatomical, Physiological, and Metabolic Similarities and Differences Between Humans and Various Laboratory Animals (Continued) Conditions, systems, or structures Animal Similarities to humans Differences from humans Nonhuman primates . . Brain vasculature lnguinal canal Rabbit . . . . . . . . . . . . . Rat . . . . . . . . . . . . . . . . Sheep . . . . . . . . . . . . . Intestinal circulation Placenta Pancreatic duct Adrenal gland Innervation Nucleic acid metabolism Teeth and mandible Brain Larynx Kidney Reproductive performance Menstrual cycle Spermatozoa .Splenic vasculature Liver Spleen Sweat glands Immunity Lung elasticity Innervation Middle ear and ear drum . Spleen Cardiac circulation Senile splenic changes Abdominal circulation Senile pancreatic changes No gall bladder . Splenic vasculature Stomach and digestion Sweat glands Heat regulation Breeding Vomiting Sleep Plasma gamma globulins in newborn SOURCES: Adapted from B.M. Mitruka, H.M. Rawnsley, and D.V. Vadehra, Animals for Researclr, Mode/s for the Study of Human Disease (New York: John Wiley & Sons, 1976); and W.1. Gay and J.D. Wlllett, “The Spectrum of Biological Systems and the Selection of Models, ” in National Symposium on Imperatives in Research Animal Use: Scientific Needs and Anima/ Welfare, NIH Pub. No. 652746 (Bethesda, MD: National Institutes of Health, 1965). laboratory animals (e.g., mice, rats, guinea pigs, rabbits, hamsters, and ferrets); nonhuman primates (e.g., baboons, new- world monkeys, and old-world monkeys); exotic birds (e.g., parakeets, parrots, cock- atiels, and cockatoos); birds of prey (e.g., falcons, hawks, and eagles); freshwater and marine fish; marine mammals (e.g., porpoises and whales); large terrestrial mammals (e.g., deer, ante- lope, elk, lion, tigers, elephants, and llamas); and assorted reptiles and amphibians. Choice of Species The variety of animal species used in research spans the animal kingdom, and some species are used more often than others (see ch. 3). Various reasons exist for search: G G G G Some species using particular species in re- are more available than others. For example, certain primate species are in chronic short supply. Conversely, in the case of rats and mice, large numbers of commer- cial breeding businesses can supply particu- lar strains, ages, and sex on the purchaser’s demand. Existing databases and literature have been built on a particular species. Additional work, in order to contribute to the field in a direct way, needs to be based on the same species. For most research purposes, nonendangered, commercially available animals are preferred over endangered ones. Some species exhibit the physiology or be- havior of interest in a more vivid and robust form than do other species. For example, the desert-adapted kangaroo rat is the species of Ch. 5—The Use of Animals in Research Ž 99 G G G choice for studies of the kidney’s role in water conservation, Certain aspects of physiology or behavior are exhibited by only a limited number of spe- cies. For example, studies of echolocation are best done with bats, which emit sounds in radar-type fashion. The costs of acquisition vary widely among species. For example, a mouse costs approx- imately $2, a hamster approximately $5, and a guinea pig approximately $19. (The actual cost for a particular species varies with the sex, strain, weight, age, quantity ordered, method of shipping, and distance shipped.) Maintenance costs vary widely among spe- cies. Depending on the laboratory lifetime of the animal, maintenance expenses can quick- ly exceed acquisition costs. For example, T H E R O L E O F A N I M A L S I N Like all of biomedical research, behavioral re- search relies on animals to identify models for and aid in the understanding of human phenomena. Behavioral research has the further goal of un- derstanding the behavior of animal species of eco- nomic or intrinsic interest to people. Behavior encompasses all the movements and sensations by which organisms interact with both the living and nonliving components of their envi- ronment (2). The environment includes not only objects and events external to the organism, but internal events as well (e.g., visceral cues, moti- vations, and emotions). Behavior is not an object or a thing. It is a process that continues in most organisms until they die. Even sleep is a form of behavior. Unlike coloration or size, behavior is a dynamic property that functions primarily to enable an organism to adapt to changing environ- mental conditions. What is Behavioral Research? Classes of Behavioral Research There are several classes of behavioral re- search, each with a distinct focus: G Abnormal Behavior. In the broadest sense, abnormal behavior is any that deviates from G maintaining a mouse in a research laboratory costs approximately 5 cents per day, a ham- ster approximately 11 cents per day, and a guinea pig approximately 40 cents per day. (The actual per diem cost varies among differ- ent animal facilities, depending, for example, on accounting practices and local labor costs.) Results obtained from different species varv in their ability to be generalized, both among animals and between animals and humans. Generalizations are more readily made among species that are more closely related than among species that are less closely related. Attempts to identify alternatives to using animals in research are likely to be influenced by these considerations. BEHAVIORAL RESEARCH G G G G G normal patterns. Instances among animals in- clude seemingly suicidal, self-induced beach- ings by whales, phobic and neurotic prob- lems in pets, and various laboratory-induced animal models of human psychopathology (e.g., depression, drug addiction, or obesity). Aggression Aggression can be defined as an organism’s threatening to inflict, attempting to inflict, or actually doing physical harm to another organism. Animal Movements Animal movements rep- resent major changes in location over time and space, such as patterns of migration, herding, homing, navigation, orientation, and dispersal. Body Maintenance. Behaviors that function to provide body maintenance and homeo- stasis include hunger, thirst, respiration, ther- moregulation, excretion, grooming, preening, and parasite removal. Cognition Although this label has been used indiscriminately to encompass practically all aspects of learned behavior (36), the term is more strictly applied to instances of appar- entmentalistic activity in animals (e.g., con- sciousness, thinking, imagery, self-awareness, intention, or attribution). Communication. Communication consists of an exchange of information between two 92 G Alternatives to Animal Use in Research, Testing, and Education G G G G G or more organisms that results in a change in behavior. Instances of this range from those that are stereotyped and instinctive, such as the dance “language” of honey bees, to those that might appear to have a symbolic basis, as in the case of the recent attempts to teach chimpanzees various forms of sign language. Depending on species, communi- cation can involve visual, auditory, olfactory, or tactile cues. Exploration and Activity. In addition to in- stances of curiosity and exploratory behavior, patterns of activity included in this discipline are circadian rhythms, sleep, hibernation, roost-time restlessness, and different patterns of locomotion (e.g., swimming, swinging, or flying). Habitat and Food Selection. Habitat and food selection refer to the areas where ani- mals live under natural conditions (e.g., fresh- water streams, forests, or deserts) and the ways they exploit resources. Areas of inquiry by behavioral researchers include competi- tion between species and optimal foraging strategies. Learning Memory, and Problem Solving. These behaviors are represented by the ac- quisition and retention of new information that allows organisms to anticipate recurring environmental events, as well as changes in behavior that maximize or minimize certain outcomes. Included in this discipline is the cultural transmission of information from one generation to the next and imitation. Motivation and Emotion The study of moti- vation looks at mechanisms and manipulations that activate and sustain behavior. Emotion typically includes reactions that accompany different motivational states and is often asso- ciated, for example, with fear, anxiety, appre- hension, pleasure, and rage. Predator-prey Relations As a consequence of selective pressure associated with preda- tion, many prey species have developed an extensive and elaborate array of predator defenses couched in terms of sensory and/or behavioral adaptations, such as burrowing or voluntary immobility. Likewise, predators use a variety of behavioral strategies in prey identification and capture. G G G G Reproduction and Parental Care. Patterns of courtship, mate selection, copulatory be- havior, nest building, nurturing, and care of offspring all fall within this discipline. Sensation and Perception. Sensation and perception refers to the ways in which orga- nisms detect and interpret their environ- ment. Topics included in this discipline in- clude studies of sensory mechanisms, the development of search images, and highly specialized sensory mechanisms, such as echolocation. Social Behavior. Social behavior is defined by a situation in which the behavior of one organism serves as a stimulus for the be- havior of another, and vice versa. Instances of social behavior range from simple forms of aggregation to complex exchanges among individuals (e.g., dominance, cooperation, and reciprocal altruism). Spacing Mechanisms Spacing mechanisms are intimately tied to social behavior, and range from such topics as individual distance to the maintenance of territories. Behavioral v. Biomedical Research Distinctions between behavioral and biomedi- cal research, although they are commonly made (and are followed in this assessment), frequently break down. Behavior, in the final analysis, is a biological phenomenon, Behavior presupposes a living organism, and the way that organism be- haves is influenced in complex ways by its genetic makeup, hormonal status, physiology, and neuro- chemistry. Intervening between the input of envi- ronmental events and the output of behavioral events are complex neuroanatomical networks in- volving receptors, electrochemical reactions, nerve impulses, and effecter organs. Behavior does not occur in a vacuum. The biology of the organism provides the foundation that makes be- havioral events possible. It is increasingly apparent that many aspects of behavioral research must be viewed in conjunc- tion with biomedical research. Strong compo- nents of both behavioral and biomedical research are evident, for example, in the study of obesity, hypertension, drug addiction, headaches, aggres- sion, alcoholism, sexual dysfunction, brain dam- Ch. 5—The Use of Animals in Research G 101 age, epilepsy, schizophrenia, depression, learn- ing disorders, smoking, anorexia nervosa, stomach ulcers, mental retardation, and a variety of other psychological disorders. Why Are Animals Used in Behavioral Research? Control The use of animals under laboratory conditions enables the manipulation and control of a vari- ety of factors that in different settings would con- fuse, contaminate, and confound any attempt to interpret a behavioral outcome. Animal models also allow the control of genetic background, prior experience, temperature, humidity, diet, and previous social encounters. When these vari- ables are uncontrolled, observed behavioral re- sponses can be virtually impossible to interpret. objectivity Two prerequisites to any research are objec- tivity and impartiality. When humans study hu- mans, as can be the case in behavioral research, unique problems may arise. Not only can it be dif- ficult for the investigator to remain objective in interpreting behavioral phenomena, but a vari- ety of other complications can arise from the so- cial relationship among those conducting the re- search and those participating as subjects (50). The use of nonhuman species partially amelio- rates this problem. Developmental Effects Among many species behavior changes as a function of age. The problem this poses for hu- man research is one of time. Human development continues for many decades. To chart behavioral changes within the same persons would take many years, involving exhaustive followup studies and the ever-present danger of losing research subjects, for example, because of death or relo- cation. The alternative to such longitudinal work is to conduct cross-sectional studies, where simul- taneous samples are drawn from different age groups. A problem in this case is that sociologi- cal and cultural changes over time (e.g., 50 years ago, an eighth-grade education was the norm) confound apparent differences between people of different ages. Because a range of lifespans is available among laboratory species, the use of ani- mal models can minimize or circumvent al- together some problems associated with the study of behavior over time. Genetic Effects There is growing evidence of a variety of ge- netic effects on behavior (23). With animal models, selective breeding studies can establish, pinpoint, and quantify genetic effects on behavior. The op- portunity for human research in this area, apart from studies of identical twins, is limited. Methodology The fact that animals cannot talk seems at first to constitute a serious disadvantage to conduct- ing behavioral research with animals. Yet, the stark limits of trans-species communication help to keep human investigators unbiased in their work. The use of animal models forces the be- havioral scientist to develop objective, operational definitions and research techniques that may later be applied to humans. Lower Complexity Behavior, notably human behavior, can be ex- tremely complex. The use of animals that appear to be structurally and functionally less complex presents a way to identify some of the basic ele- ments and principles of behavior that might other- wise remain inextricably embedded in a mosaic of other factors. Species-Specific Behaviors Certain behavioral phenomena fall outside the realm of human sensory or motor abilities. For example, flight, echolocation, infrared detection, and homing require the use of nonhuman spe- cies as subjects for research purposes. Heuristic Value Research on the behavior of animals has been an important source of hypotheses about human behavior and an impetus to research on humans (35). Much of what is now known about the prin- 102 G Alternatives to Animal Use in Research, Testing, and Education ciples of learning, for example, was initially de- rived from research on animals. Likewise, a vari- ety of therapeutic techniques (e.g., desensitization) were derived from work with animals, Human studies were done to verify what was learned from animal research and to gauge the limits of extrapolation from animals to humans. Practical Application to Animal Species In addition to providing models of a variety of biomedical and psychological problems in hu- mans, research on animal behavior is in many in- stances focused on benefits to the animals them- selves. For example, an understanding of behavior has proved crucial for designing optimal captive environments for the protection and breeding of endangered species (55). Increased attention has also been paid to the behavior of farm animals. The study of mother-infant attachments, social behavior in groups, stress resulting from over- crowding and confinement, and habitat prefer- ences has led to important insights into farm- animal welfare and husbandry (13,27,54). It is also noteworthy in this context that knowl- edge gained about behavioral problems in humans through animal research is now being applied to animals. Effective treatments have been devel- oped for aggressive problems in cats (5) and fears and phobias in dogs (24)58). A knowledge of animal behavior has helped identify and solve ecologic problems. The discov- ery and subsequent synthesis of insect sex attrac- tants, or pheromones, has important implications for the control of agricultural pests. Rather than having to use toxic pesticides applied over vast areas, there is already some application and much future potential in baiting traps with specific pheromones, which precludes environmental contamination. One unique application of laboratory findings to the solution of ecologic problems involved stud- ies of taste-aversion conditioning in rats (18). Re- searchers paired unpleasant, chemical- or radia- tion-induced illness with different flavors. After just one or two trials, rats developed highly dura- ble aversions to the flavors paired with unpleasant stimuli. outside the laboratory, by pairing lithi- um-chloride-induced illness with the flesh of vari- ous prey species, it is now possible to control coy- ote attacks on sheep and turkeys (14). Indeed, one or two trials is sufficient to eliminate attacks on specific domestic farm animals but leave the coy- ote free to feed on alternative prey (22). This procedure has recently been extended to reducing crop damage by crows and even appears to have promise for dealing with cancer patients undergoing radiation therapy (l). (A frequent complication of radiation therapy has been un- pleasant gastrointestinal illness that the patient generalizes to all food; the patient may be una- ble to eat. Using the principles of conditioned taste aversion developed in rats, it is now possible to circumvent the problem by restricting patients to one particular kind of food during radiation treatment, so that the aversion that develops is specific to that food alone.) Individual Animals in the Service of Humans Behavioral research occasionally centers on a trait of a particular species that maybe especially well suited to assist humans. For example, using animals to help handicapped persons has required a knowledge of animal behavior. Seeing eye guide dogs, usually German shepherds or golden re- trievers, assist the blind (20), and trained capu- chin monkeys perform as aides for quadriplegics (63). Pet dogs and cats have been shown to have therapeutic value for psychiatric patients (10), the handicapped (12), and the elderly (49), and they may even hold promise for alleviating depression resulting from loss of a child (57). Methods of Behavioral Research The methods of behavioral research are as var- ied as the disciplines, but most fall into one of three general categories: field studies and natural- istic observation; developmental studies; and lab- oratory studies. Field studies represent an attempt to examine the behavior in question as it occurs under natu- ral circumstances, Such studies do not typically involve attempts to manipulate or control the con- ditions of observation. watching animals in nat- ural conditions has frequently been suggested as Ch. 5—The Use of Animals in Research 103 an alternative to using them in laboratory re- search (6,44). The following benefits and limita- tions of naturalistic observation have been rec- ognized: G G G G Naturalistic observation is frequently a start- ing point. Observation of animals in the field provides a base of descriptive information and serves as a source of hypotheses to be subsequently tested under laboratory con- ditions, Naturalistic observation can be used to com- pare behavior observed in the field with that occurring in the laboratory to assess the ex- tent to which an artificial environment may alter behavior, and whether the results can be generalized. Field studies can increase the efficiency with which animals are used by providing impor- tant information on natural species variables and biological constraints on behavior. The principal drawback to naturalistic obser- vation is the absence of control. Under nat- ural conditions, events frequently change in both important and spurious ways, often making it impossible to establish cause-and- effect relations (37). Behavioral research often requires study of one animal or a group through time, as development proceeds. Among many species the emergence of different patterns of behavior is a reflection of both maturational and experiential factors. De- velopmental variables have been identified as be- ing important in the expression of such diverse behaviors as aggression, communication, activity, learning, and social behavior. Laboratory studies undertake to manipulate and control the condition of observation so as to specify more precisely the variables and condi- tions that influence the behavior in question. Most laboratory studies of behavior can be subdivided into those that attempt to identify the environ- mental determinants of behavior and those con- cerned with the organic basis for behavior. Within the latter category are a number of ap- proaches involving attempts to identify the neu - roanatomical, neurochemical, endocrinological, and genetic underpinnings for behavior. Use of Multiple Species in Behavioral Research Many behavioral phenomena appear common to different species. Patterns of migration, for ex- ample, are common to such diverse groups as in- sects, fish, birds, and even some species of mam- mals. Much the same appears true for learning, motivation, and bodily maintenance. Yet, gen- eralizations about categories of behavior (e.g., parental care or hoarding) in unrelated species may be misleading, because the species evolved independently (34). Moreover, comparing the per- formance of different species on a simple task may have no bearing on larger issues such as in- telligence (26). What used to be seen as general principles and “laws” of learning, for example, now turn out to be specific to certain species un- der certain situations (3,4,53). There are some behaviors that are of limited scope across species but of profound importance in terms of their bearing on the question of hu- man behavior. For example, the capacity to rec- ognize one’s own reflection in a mirror has only been found in humans, chimpanzees, and orangu- tans, and much the same may apply to instances of intentional deception, gratitude, grudging, sym- pathy, empathy, attribution, reconciliation, and sorrow (17). P A I N A N D D I S T R E S S I N R E S E A R C H A N I M A L S There are two general kinds of animal ex- tion of pain and the monitoring of the responses perimentation in which pain may occur. First, to pain are usually integral parts of the experi- there are studies that investigate the nature of mental procedure. The goal is the prevention, pain itself and the anatomical, behavioral, chem- treatment, and amelioration of human and ani- ical, pharmacological, and physiological mecha- mal pain. The second, and much larger, class of nisms responsible for it. In such studies, the inflic- animal experimentation in which pain may occur 104 Ž Alternatives to Animal Use in Research, Testing, and Education consists of those studies in which pain is but a byproduct of the procedures used (28). When indices of pain are observed or antici- pated in living research animals as byproducts of an experimental protocol, the investigator is both informally and formally obliged to supply pain re- lief. (For a further discussion of the investigator’s responsibilities in this area, see chs. 4, 13, 14, 15, and 16.) Pain relief for a laboratory animal is usually ac- complished by one of three means. An analge- sic is an agent that relieves pain without caus- ing loss of consciousness. The most frequent use of analgesic drugs in laboratory animals is likely to be in the postoperative period. An anesthetic is an agent that causes loss of the sensation of pain, usually without loss of consciousness. An anesthetic may be classified as topical, local, or general, according to the breadth of its effect. Topical anesthetics find only limited use in ani- mal research, usually as components of ointments applied to minor injuries, whereas local anes- thetics are used for many minor surgical proce- dures. The use of local anesthetics requires post- surgical care, because anesthetized surfaces are particularly liable to accidental and self-inflicted damage (43). General anesthetic, either injected or inhaled, is widely used in research. A tran- quilizer is an agent that quiets, calms, and re- duces anxiety and tension with some alteration of the level of consciousness and without effect- ing analgesia. Tranquilizers are particularly use- ful in reducing distress and resistance to con- finement. The perception of pain is largely subjective. It is best described as an awareness of discomfort resulting from injury, disease, or emotional dis- tress and evidenced by biological or behavioral changes. A frequent companion to pain is dis- tress—the undesirable stress resulting from pain, anxiety, or fear (51). Distress can also occur in the absence of pain. An animal struggling in a re- straint device may be free from any pain, but it may be in distress. Despite the difficulty associated with objectively defining pain, it can usually be recognized. The most obvious sign is an animal’s behavior (16)42, 56), G G G G G G G Signs of pain include the following: Impaired activity. Animals may be rela- tively inactive or may remain completely im- mobile within their pen or cage. If they do move, it is often with an abnormal gait, such as limping or not using a leg. Change in personality. Pain may result in guarding behavior (attempting to protect or move away). Animals may also be uncharac- teristically aggressive. Restlessness. Animals may move about continually or may rise up and lie down repeatedly. Decreased intake. Food and water con- sumption are usually severely retarded, often to the extent that moderate or severe de- hydration can occur. Abnormal vocalization. Dogs may whine or whimper, rats and hamsters may squeak at a high pitch, and primates may scream or grunt. Abnormal posture. Dogs, cats, and rodents may tense the muscles of the back and ab- domen to effect a “tucked-up” appearance. Self-mutilation. Dogs and rodents may gnaw at the site of a lesion on their own flesh or, for example, remove their own tumor. In identifying pain, all these criteria must be considered in conjunction with the nature of the experimental procedure and the previous normal behavioral characteristics of the animal. Also, it should be noted that no one criterion is a wholly reliable indicator of pain. An experimental procedure probably involves pain if it includes, for example, induction of any pathological state, administration of toxic sub- stances, long-term physical restraint, aversive training, or major operative procedures such as surgery and induction of physical trauma. Vari- ous procedures employed in the research labora- tory can be compared, ranking each for the esti- mated degree of pain for the animal subject (see table 5-3). Educated estimates of pain perception in animals can be made by understanding animal behavior; by drawing analogies based on compar- Ch. 5—The Use of Animals in Research G 105 Table 5-3.—Classification of Research Experiments and Procedures According to the Degree of Pain or Distress for the Animal Level of pain/distress Examples of types of experiments Examples of procedures Absent or n e g l i g i b l e . G G Low G G G Moderate G G High Noninvasive behavioral testing Studies of migration or homing Dietary preference studies Determination of pain threshold Manipulation of blood chemistry Experiments carried out on anesthetized animals that do not wake up again Behavioral study of flight or avoidance reactions Operations carried out under anesthesia or analgesia, with the animal waking up or experiencing the cessation of the action of the painkiller (postoperative pain) Chronic stress studies Drug withdrawal studies Studies of certain infectious agents Experiments on mechanisms of pain in conscious animals Experiments on mechanisms of healing Studies of radiation toxicity G G G G G b G G G G G G b G G G G G G G G G G G G G Banding for identification or tracking Field observation Fecal examination Conditioned learning with food reward Flinch or jump response Injections Tube feeding Tattooing Administration of anesthetic Surgery under deep anesthesia and subsequent sacrifice —Removal of organs for histological or biochemical investigation —Culture of surviving organs Blood sampling Stimulation of unanesthetized animal Biopsies Implantation of chronic catheters Castration Mild electric shock Implantation of electrodes Central nervous system lesions Exposure of internal organs Food or water restriction for more than 24 hours Prolonged physical restraint Chronic sleep deprivation Intense electric shock Production of pain clearly beyond threshold tolerance Induction of burns or wounds Surgery on conscious animal SOURCE: Office of Technology Assessment. ative anatomy, physiology, and pathology; and by enced by laboratory animals—ranging from ab- basing inferences on subjective responses to pain sent or negligible to high-can provide a basis for experiences by humans. Careful attempts to esti- efforts to minimize the pain or distress caused mate and categorize the degree of pain experi- by research procedures. ANIMAL AND NONANIMAL PROTOCOLS IN BIOMEDICAL AND BEHAVIORAL RESEARCH REPORTS One way to measure the balance of animal and mal and nonanimal protocols in contemporary re- nonanimal methods in research is to survey the search. end-product of experimentation—the published literature. OTA examined approximately 6,000 re- Fifteen leading scientific journals were selected search reports published from 1980 through 1983 to represent disciplines within biomedical and be- in an effort to document the prevalence of ani- havioral research, These journals were chosen be- . 106 G Alternaives to Animal use in Research, Testing, and Education cause of their primary emphasis on research done in the United States by American scientists and because of the respect accorded them by scien- tists in each discipline. The editors of all 15 sub- ject manuscripts to independent peer review prior to publication. For each year from 1980 through 1983, OTA examined the first 100 research papers published in each journal; in a few cases, fewer than 100 papers were published in a given year. Short com- munications and review articles were not in- cluded. In each report, OTA checked whether ani- mals were used. Thus, the materials and methods employed in each article were categorized as ei- ther use of animals, no use of animals, or use of humans. ‘(Animal” is defined as described earlier (see ch. 2), any nonhuman vertebrate. “Use” of animals is defined conservatively as any use of an animal in an experiment. Table 5-4 lists some examples of the ways specific protocols were categorized by OTA. Survey Findings The results of this survey indicate that the re- search journals—and perhaps the disciplines they represent—fall into two categories: Table 5-4.—Classification of Pubiished Research Protocois in OTA Survey of 15 Journais Examples of protocols classified as use of animals: Whole animals used as experimental subjects Animals used to obtain cell, tissue, or organ of interest Animals used in the establishment of new cell, tissue, or organ cultures Extraction of protein or other biological molecule from animals Production of antibodies by whole animals or animal components Use of egg, sperm, or embryo from animal source Use of animal epidemiologic data Examples of protocols classified as no use of animals: Use of invertebrate organisms Use of computer systems Use of previously established cell lines Acquisition of biological molecules from a commercial manufacturer Use of physical or chemical systems Examples of protocols classified as use of humans: Use of living human subjects Use of cadavers Use of human placenta Use of human blood cells or components Use of human epidemiologic data SOURCE: Office of Technology Assessment. G journals representing disciplines that have al- ready largely incorporated nonanimal meth- ods into the research process; and . journals representing disciplines that either have not incorporated, or may not have avail- able, nonanimal methods. For most of the 15 journals (see table 5-5), pub- lished protocols fall predominantly into just one of the three categories of methods; in many cases, over 80 percent of the protocols are in one cate- gory. Only Cell, representing cell biology, had a majority of articles using nonanimal methods. The American Journal of Cardiology contained a ma- jority of articles using humans or human mate- rials as research subjects. All the remaining jour- nals except Developmental Biology and the Jour- nal of Biological Chemistry included a majority of articles using animals. Developmental Biology and the Journal of Biological Chemistry contained approximately equal percentages of articles em- ploying animal and nonanimal methods. The 12 biomedical research journals included in this survey cover a diverse array of disciplines under this one rubric. The differing patterns of animal, nonanimal, and human use make gener- alizations misleading at best and perhaps impos- sible. In the same way that biomedical research itself is not monolithic, the patterns of animal use among disciplines of biomedical research are not uniform. Perhaps not surprisingly, veterinary re- search, insofar as it is represented by two jour- nals, relies primarily on animals, a minimal per- centage of nonanimal methods, and no protocols with humans. The three behavioral research journals included in this survey registered a predominance of ani- mal methods—more than 90 percent of the pro- tocols in each case. The research reported in these journals involved minimal use of humans or nonanimal methods. Other behavioral research journals, for example those reporting on clinical psychology, largely publish reports of research with human subjects. Survey Limitations This attempt to gauge the implementation of nonanimal methods in selected areas of biomedi- cal and behavioral research had certain limita- Ch. 5—The Use of Animals in Research G 107 Table 5-5.–Percentage of Papers (Average, 1980-83) Using Animal, Nonanimal, and Human Subjects in 15 Biomedical and Behavioral Research Journals Surveyed by OTA Percentage of papers using: Journal Animals Nonanimals Humans to distinguish between different vertebrate spe- cies. Any alternative protocol, therefore, that tended to reduce or refine an existing animal pro- cedure was still categorized as “use of animals. ” An example of the problem of overestimation of animal use by a survey such as this exists in immunology. Today, antibodies, needed in most immunology research, can be obtained by inject- ing rabbits with foreign proteins, or antigens, and extracting the antibodies that the rabbit produced or by using mouse spleens and the monoclinal antibody technique to produce antibodies to an antigen. Each process requires animals. Once the monoclinal cells are in culture, however, there is a virtually unlimited supply of the needed anti- body, and there is essentially no further need for animals. Thus the monoclinal technique can de- crease animal use, as was the case in many of the most recent articles surveyed in the Journal of Immunology. These articles, though, were still coded under the “use of animals” category be- cause the primary methods and materials in- volved animals. But the total number of animals in a given experiment decreased, for they were used in just one aspect of the experiment instead of two. The monoclinal antibody technique is be- ing used as an alternative to the repeated use of rabbits, yet its impact is underestimated in a sur- vey such as this. The OTA scoring of protocols published in the Journal of Immunology did not reflect certain reductions that are currently be- ing implemented. Along with underestimating the implementation of nonanimal methods, the boundaries within which the OTA survey was carried out also tended to overestimate the use of animals as ex- perimental subjects. This was due principally to two factors included in the scoring procedure under animal use-epidemiologic studies and the study of biological molecules obtained from animals. Epidemiologic data are the primary sources in some articles in the American Journal of Cardi- ology, the Journal of the National Cancer Insti- tute, and in many veterinary studies. These pro- tocols were included under “use of animals,” yet they did not manipulate animals in any way as experimental subjects. Biomadical research: American Journal of Veterinary Research . . . . Journal of Animal Science . . . . . . . . . . . . . . . Endocrinology . . . . . . . . . . . American Journal of Physiology . . . . . . . . . . . . Anatomical Record . . . . . . . Proceedings of the Society of Experimental Biology and Medicine . . . . . . . . . . Journal of Immunology . . . Journal of the National Cancer Institute . . . . . . . . Developmental Biology . . . Journal of Biological Chemistry . . . . . . . . . . . . . Cell . . . . . . . . . . . . . . . . . . . . American Journal of Cardiology. . . . . . . . . . . . . Behavioral research: Behavioral and Neural Biology . . . . . . . . . . . . . . . Journal of Comparative and Physiological Psychology . . . . . . . . . . . . Phvsioloav and Behavior . . 96 4a o 4b 8 96 91 0 1 90 88 6 3 4 9 10 10 7 19 83 71 60 54 9 46 31 0 9 2 39 31 52 67 12 0 88 96 4 0 96 93 3 1 1 6 aprimarily virus research. bprimarily computer rn~d~lin~ or grain fermentation applicable to ruminant nutrition. SOURCE: Office of Technology Assessment. tions. The conservative scoring procedure tended to underestimate the use of alternatives to ani- mals as defined in this assessment (see ch. 2). For instance, if an experimental protocol used both animal and nonanimal methods, it was catego- rized under use of animals. If a study involved both nonanimal methods and humans, it was counted as use of humans. Further, if a study in- volved both animal methods and humans, it was counted as use of animals. The approach used to categorize protocols took into consideration only the replacement, not the reduction or refinement, of animal methods. Whether a protocol involved 1 or 100 animals, it still fell under the category of “use of animals, ” and all reports bore equal weight in determining percentage of protocols using humans or animal or nonanimal methods. In addition, there was no attempt to quantify the pain or stress of an animal in an experiment or 108 G Alternatives to Animal Use in Research, Testing, and Education Obtaining many biological molecules that are studied experimentally requires that they be ex- tracted from animals who produce them. Subse- quent experiments on these molecules themselves (reported, for example, in the Journal of Biologi- cal Chemistry) do not involve animals at all. In such cases, animals may be used in preparation for an experiment but are not actually involved in the experiment being performed. Therefore, protocols that involved animals as donors of bio- logical molecules (e.g., bodily fluids) for an exper- iment prior to its initiation were also included under use of animals, and this tended to overesti- mate the use of animals as research subjects. It is important to distinguish between the num- ber of published articles involving animal meth- ods and the actual number of animals used in re- search. The OTA survey provides no information on the latter. Some protocols may involve only a few animals, while others may employ tens or hundreds. Moreover, depending on the species and type of research, some subjects might be used in multiple experiments. In primate research, for example, it is not uncommon for animals to be used in a succession of either related or unrelated studies over a period of years; this would not be the case for rodents. SUMMARY AND CONCLUSIONS Biomedical and behavioral research center on the understanding of human health and disease and rely on animals to achieve this goal. They use animal subjects to understand not only human phenomena, but animal phenomena as well. The broad spectrum of enterprises involved in these fields of research includes disciplines ranging from anatomy to zoology. Although the varied dis- ciplines that make up biomedical and behavioral research have distinct foci, they often overlap. Animals are used throughout these disciplines to address an array of questions. Nonhuman pri- mates, for example, have contributed to an under- standing of polio, hepatitis B, high blood pressure, Parkinson’s disease, baldness, menopausal hot flashes, and other human conditions. Beyond the nonhuman primates, diverse species are used in biomedical research because of their anatomical, physiological, and metabolic similarities to or differences from humans. Principles and tech- niques developed in varied animal species (e.g., dog, horse, and sheep) may combine to support a single application to humans, as in the case of coronary artery bypass graft surgery. In be- havioral research, different animal species may also be used to learn about characteristics unique to the species under study, usually one of eco- nomic importance or intrinsic interest to humans. Animals may suffer pain or distress in the course of research on the mechanism of pain, or, more generally, as a byproduct of experimental procedures. In such cases, the investigator is obliged to supply pain relief to the animal or to justify withholding pain-relieving drugs as nec- essary to the experiment. Institutional animal care and use committees play an important role in overseeing this process (see ch. 15). Pain relief is usually effected by the administration of analge- sic, anesthetic, or tranquilizing agents. Indices of pain can usually be recognized in experimental animals, and experimental procedures can be ranked according to estimates of the degree of pain produced. Such a ranking provides a basis for efforts to minimize the pain caused by re- search procedures. An OTA survey of published research reports in 15 scientific journals documented the preva- lence of animal v. nonanimal protocols in contem- porary research. Each research journal, and per- haps the discipline it represents, can be identified by a characteristic balance of protocols using ani- mals, nonanimals, and humans. The data permit research journals to be classified as representing disciplines that either rely on nonanimal meth- ods or that do not incorporate such methods (or do not have them available). In the same way that research itself is not monolithic, patterns of ani- mal use and the use of nonanimal methods among research disciplines are not uniform. Ch. 5—The Use of Animals in Research s 709 CHAPTER 5 1. 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Chapter 6 Alternatives to Animal Use in Research Unless we get a handle 011 what is happening in the mammalian brain, there’s no way of knowing whether any of these [invertebrate] models is right or not. Richard F. Thompson Stanford University Science 85 6(4):33, 1985 Investigators often ask statisticians how many observations they should make (fortunately, usually before the study begins). To be answerable, this question needs fuller formulation. There is a resemblance to the question, How much money should I take when I go on vacation? Fuller information is needed there too. How long a vacation? Where? With whom? Three questions need to be answered before the sample size is determined. How variable are the data that will be collected? How precise an answer is needed? How much confidence should there be in the answer obtained? These questions can be well worth probing even if the question of sample size will foreseeably be answered by the size of the budget or the time available for the study. Sometimes a planned study is dropped because sample-size analysis shows that it has almost no chance of providing a useful answer under the constraints of time or budget that apply. Lincoln E. Moses Stanford University N. Engl. J. Med. 312:890-897, 1985 CONTENTS Page Continued, But Modified, Use of Animals in Biomedical Research . ............114 Reduction in the Number of Animals used . ...................,....,.,..114 Substituting One Species for Another . . . . . . . . . . . . . . . . . . . . . . . . . . ........116 Reduction of Pain or Experimental Insult . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .117 Use of Living Systems in Biomedical Research . ............................118 In Vitro Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..118 Invertebrates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....122 Micro-organisms ., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...........123 Plants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .....................123 Use of Nonliving Systems in Biomedical Research . ................,........124 Chemical and Physical Systems . . . . . . . . . . . . . . . . . . . . . . ......,..........,124 Epidemiology: Using Existing Databases . . . . . . . . . . . . . . . . . . . . . ............124 Computer Simulation in Biomedical Research . . . . . . . . . . . . . . . . . . . . . . ........124 Continued, But Modified, Use of Animals in Behavioral Research . ............126 Reduction in the Number of Animals Used . ............................,126 Substitution of Cold-Blooded for Warm-Blooded Vertebrates . ..............128 Reduction of Pain or Experimental Insult . . . . . . . . . . . . . . . . . . . . . . .........130 Use of Living Systems in Behavioral Research . . . . . . . . . . . . . . . .. .. ... ... ,...133 Invertebrates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....,.133 Plants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . , . . . . . . ....135 Use of Nonliving Systems in Behavioral Research . . . . . . . . . . . . . . . . . . . . . . . ,..136 Computer Simulation in Behavioral Research . . . . . . . . . . . . . . . . . . . . . . . . .. ....136 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....138 Chapter preferences. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..........139 List of Tables Table No. Page 6-1. Research Methods Involving Living Components. . . . . . . . . . . . . . . . . . . ., ., .113 6-2.Properties of In Vitro Culture Systems. . . . . . . . . . . . . . . . . . . . . . . . . . .. ....119 6-3. Some Examples of Computer Simulation of phenomena in Biomedical Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...125 6-4. Some Examples of Computer Simulation of Behavioral phenomena . .......137 List of Figures Figure No. Page 6-1. Apparatus for Remote Blood-Sampling via Chronic, Intravascular Catheter From Unrestrained Ferret . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ......,118 6-2, Schematic of Experimental Organ Perfusion . . . . . . . . . . . . . . . . . . . . . ......120 Chapter 6 Alternatives to Animal Use in Research Alternatives to animal use in biomedical and be- havioral research fall into four broad categories: G G G G continued, but modified, animal use, in- cluding a reduction in the number of animals used, improved experimental design and sta- tistical analyses of results, substitution of cold- blooded for warm-blooded vertebrates, substi- tution of laboratory mammals for domestic or companion mammals, and reduction of pain or experimental insult; use of living systems, including in vitro cul- tures (of cells, tissues, and organs; see table 6-l), embryos, invertebrates, micro-organ- isms, and plants; use of nonliving systems, such as chemical or physical systems; and computer simulation. In this chapter, various disciplines within bio- medical and behavioral research are surveyed in order to focus attention on the most promising areas for development of alternatives to animal methods. Areas not amenable to the implementa- tion of such alternatives are also identified. As noted in chapter 5, distinctions within and among the varied disciplines of biomedical and be- havioral research are artificial in one sense: Bound- aries among disciplines are often blurred, and broad areas of overlap exist. Yet the examination of discrete areas of research highlights the great variability among disciplines in the potential for using alternatives to animals. Using alternative methods in research holds sev- eral advantages from scientific, economic, and hu- mane perspectives, including: G G G G G G G G reduction in the number of animals used; reduction in animal pain, suffering, and ex- perimental insult; reduction in investigator-induced, artifactual physiological phenomena; savings in time, with the benefit of obtaining results more quickly; the ability to perform replicative protocols on a routine basis; reduction in the cost of research; a greater flexibility to alter conditions and vari- ables of the experimental protocol; reduction of error stemming from inter- individual variability; and the intrinsic potential of in vitro techniques to study cellular and molecular mechanisms. At the same time, these methods are fraught with inherent disadvantages, including: G G G reduced ability to study organismal growth processes; reduced ability to study cells, tissues, and or- gan systems acting in concert; reduced ability to study integrated biochem - ical and metabolic pathways; Table 6-1 .—Research Methods Involving Living Components Isolated perfused Isolated tissue or Isolated single Subcellular organs using: tissue sample using: cell using: constituents using: liver striated muscle fat cells nuclei muscle iris liver cells mitochondria heart trachea neurons microsomes lung bronchi glial cells Iysosomes adrenal gland lung striated muscle cells synaptosomes pituitary gland uterus smooth muscle cells cell membranes intestine intestine red blood cells muscle actin/myosin testis seminal vesicle leukocytes skin vas deferens platelets spleen bladder mast cells kidney spleen salivary gland fat pads liver slice SOURCE” Adapted from W. Paton, Man and Mouse: Anima/s in Medical Research (New York: Oxford University Press, 19S.4), 1 1 3 114 G Alternatives to Animal Use in Research, Testing, and Education reduced ability to study behavior; reduced ability to study the recovery of damaged tissue; reduced ability to study interaction between the organism and its environment; reduced ability to study idiosyncratic or species-specific responses; reduced ability to distinguish between male- and female-specific phenomena; and . a handicap to probing the unknown and phe- nomena not yet identified. This general listing of advantages and disadvan- tages provides a framework for examining the use of alternatives in specific disciplines of biomedi- cal and behavioral research. Many of these pros and cons are cited in this chapter’s detailed descrip- tion of alternatives. CONTINUED, BUT MODIFIED, USE OF ANIMALS IN BIOMEDICAL RESEARCH Animal use in biomedical research can be modi- fied in a number of ways, including strengthen- ing experimental design to use fewer animals, re- ducing the degree of experimental insult, and substituting one organism for another. In the case of substitution, cold-blooded vertebrates may supplant warm-blooded ones. Reduction in the Number of Animals Used Up to half the animals used in research protocols may be untreated, or control, animals. The impor- tance of using parallel, internally controlled de- signs for experimentation may be one of the first lessons learned by science students whose results are rejected for not providing comparable data from treatment groups of animals matched for size, age, sex, and dosage. Studies with investigator- initiated, internal controls support substantially stronger inferences than those without them. Common and Historical Controls Fewer animals may be used in an experiment by sharing a control group with other investiga- tors or by not using a concurrent control group. In both cases, all the physical and genetic charac- teristics of the treatment group(s) must be matched to those of the control group, and the conditions under which the data are collected must be as pre- cisely duplicated as possible. There are difficul- ties unique to each method. Investigators may en- counter constraints on their particular study when sharing controls. For example, sharing may be im- possible if one group needs to extend its studies beyond the time agreed for termination and au- topsy of the shared animals, or if the actions of one group adversely affect the other, as might hap- pen by the inadvertent spreading of a parasite or pathogen. In the case of historical controls, the difficulty rests in exactly duplicating earlier con- ditions. Use of such controls must be carefully doc- umented and justified (82). Animal Sharing Another way to use fewer animals is to share individual experimental animals or their tissues between research groups. Although this method may encounter the same types of difficulties de- scribed for the sharing of controls, it appears to be gaining in popularity among compatible groups, At the University of Virginia, investigators in en- docrinology (Department of Internal Medicine) and in the molecular genetics of heme synthesis (De- partment of Biology) use the pituitaries and livers of the same rats even though the two departments are on opposite sides of the campus (54). Research animals may also be shared among dif- ferent sites. This is especially practicable in the case of long-lived primates. As long as sequential protocols are not deemed inhumane or scientifi- cally conflicting, primates may be shipped from one research site to another. The Primate Research Institute (PRI) of New Mexico State University, for example, will loan chimpanzees and rhesus and cynomolgus monkeys to qualified U.S. scientists. PRI currently has 240 chimpanzees on campus, with another 150 animals on loan. Cjq. 6—Alernatives to Antirnal Use in Research Ž 115 Using animals maximally in a confined area is a mandatory part of experimental design in the research program of the National Aeronautics and Space Administration (NASA). Protocols typically call for investigators to combine projects and make efficient use of one small group of animals (125). Improved Experimental or Statistical Design “Every time a particle of statistical method “is properly used, fewer animals are employed than would otherwise have been necessary,” wrote Rus- sell and Burch some 27 years ago (174). Since then, progress has been made both in the number of statistical tools available and in the training of in- vestigators in the use of these tools. Yet training still lags behind the availability of tools. Insuffi- cient information for critical evaluation and inap- propriate statistical analyses appear frequently in the literature, particularly with investigators using the t-test in cases for which analysis of variance is the appropriate measure (82). An analysis of variance simultaneously tests two or more parameters of treatment groups for indi- cation of significant difference. When the test sta- tistic falls in the rejection region, the researcher can be reasonably sure that a real difference ex- ists between treatments. The t-test estimates the difference between the mean values of one param- eter of two treatments. It is a powerful measure of significance when the number of comparisons is small, but it is subject to an increasingly large potential for error as the number of parameters grows. Using multiple t-tests increases the risk of finding a significant difference between treatments where there is none. Such observations are not esoteric, since poor summarization and statistical usage may reflect poor experimental design, call- ing into question the results of an investigation and leading to otherwise unnecessary repetition. At least one group, the Harvard Study Group on Statistics in the Biomedical Sciences, is pursuing ways to improve statistical practice and report- ing (64). Serial sacrifice, crossover, and group sequen- tial testing are three experimental designs that can reduce animal use in laboratory research (82). In serial sacrifice, animals with induced effects are randomly selected for sacrifice and examination for the occurrence and progress of effects over time. Such studies, as in radiation oncology (22), have the dual advantage of cutting short the time some animals must spend in an affected state and providing information about changes within the animal other than those observed when it is al- lowed to die without further interference. The pri- mary disadvantage is that survival information is compromised; therefore, the resulting data can- not be compared with other studies in which sur- vival serves as an end point. A crossover design maybe appropriate for stud- ies in which short-term effects are expected. Each animal serves as its own control by first receiving either a drug or a placebo, and then receiving the reverse. Such a design can be highly useful in lab- oratory and clinical testing, but crossovers must be used judiciously. Should there be any unex- pected long-term effects, the entire test is invali- dated and would need to be repeated as two sepa- rate tests. In the group sequential design, treatment groups are compared with each other in stages. For ex- ample, if two groups are given the same dosage of two different drugs, experimentation at higher dosages is undertaken only if there is no statisti- cally significant difference between the responses of the two groups. The sooner a difference be- tween groups is observed, the fewer the number of trials run. Both crossover and group sequential designs have potential applications in anesthesiol- ogy, endocrinology, nutrition, pharmacology, ra- diology, teratology, and toxicology. A commonly mentioned method of reducing the number of animals used is smaller treatment groups. Yet within the biomedical research com- munity a frequently heard complaint is that too few animals to yield useful estimates are likely to be included in each treatment group, particularly in fields such as radiology (95). Problems of this nature generally grow out of the extreme economic pressures being applied to investigators to con- trol animal costs. Well-established techniques such as saturation analyses, particularly radioimmuno- assays, have radically reduced the number of ani- mals used for any one procedure, but they may have resulted in little or no reduction in overall 116 G Alternatives to Animal Use in Research, Testing, and Education use, since they have made previously difficult anal- yses more accessible to many more investigators. Substituting one Species for Another In some instances, laboratory mammals (e.g., ro- dents) or nonmammalian vertebrates can be used in place of companion mammals (e.g., dogs), domes- tic species (e.g., sheep), or primates (e.g., monkeys). As more information on the physiology, biochem- istry, and endocrinology of laboratory mammals and nonmammalian species accumulates and is demonstrated to be like or unlike that of humans, greater use can be made of laboratory species, which in turn can generate more information and reduce future needs for research. Comparative neuroscience is perhaps the most rapidly expand- ing field and is related to physiology, biochemis- try, pharmacology, developmental biology, and zoology (33). Some brain components have been found to be remarkably similar between vertebrate species (69). Economics plays a large part in the selection of how many and what kind of animals will be used in some research (see ch. 11). An investigator fol- lowing upon previous work will generally begin with the species already in use, changing only if money becomes scarcer or if a better model is clearly demonstrated. Investigators starting anew are likely to seek the advice of a facility veterinar- ian or of colleagues as to which species best fits their needs and to begin with the smallest accept- able animal. Still other researchers deliberately begin work with a novel animal model in order to create a new research niche. One of the principal reasons for the increased use of rodents in all areas of biomedical research has been the availability of genetically homogene- ous or pathogen-free strains. For some studies, however, a further degree of genetic definition is needed. These studies require that the research animal carry some specific genetic traits that are suited to the objectives of the research, Because of their high reproductive potential, rodents are ideal for this type of “custom designing” and ex- tensive use is being made of these animals in a va- riety of disciplines (109). Oncology and immunol- ogy are two of the more familiar areas of use (92). Pharmacological research using an ethanol-prefer- ring strain of rats has prepared the way for explo- ration of the genetics of alcoholism (212). Further, the male Lewis rat, an animal that rapidly acquires testicular lesions and antibodies to sperm after vasectomy, is a candidate for study of the rever- sal of vasectomy. This research could answer ques- tions of human concern in anatomy, physiology, immunology, endocrinology, and reconstructive surgery (100). Chickens and their embryos play an important role in developmental biology, endocrinology, histology, and zoology. Other current uses are in molecular biology, in which embryonic chicken brain tissue is being cultured to study the neural- cell adhesion molecule (193), and biochemistry, in which the embryonic chicken liver is being used to study the acquisition of hormone responsive- ness during embryogenesis (62). In cardiology, tur- keys with inherited Turkey Round Heart disease serve as models of cardiomyopathy (107), and tur- key erythrocytes are fused with amphibian eryth- rocytes to study receptors that mediate physio- logical functions in heart, smooth muscle, and other tissues (199). Frogs have long been used in anatomy, biochem- istry, developmental biology, physiology, and zool- ogy. They continue to be widely used in those dis- ciplines and, additionally, are currently being used by NASA in radiology studies (125). In dental re- search, frogs are used to assess digital transplants to augment tooth and jaw regeneration (101). The newt Triturus-able to regenerate its limbs, eye lens, tail, and spinal cord—is used in developmen- tal biology to explore mechanisms of organ regen- eration (90). Turtles are used in physiology to study, for example, retinal mechanisms subserving color vision. The cone cells of the turtle retina are espe- cially conducive to such research (161). Fish are used in research to a lesser degree than other vertebrates, considering that there are over 30,000 species and their care is relatively uncom- plicated. It has been suggested that fish would make excellent subjects for nutritional research, since many are known to show specific vitamin deficien- cy symptoms (210). Physiologists have used gold- fish to study the implications of myelin-sheath resistances in demyelinating diseases (73). Rain- bow trout embryos are being used in oncology re- Ch. 6—Alternatives to Animal Use in Research Ž 117 search (93). Further, there has been recent inter- est in a specialized feature of some piscine species: the electric organ. This tissue is exceptionally rich in a single class of cholinergic synapses. Biochem- ists, geneticists, and molecular biologists working with this material have determined that the struc- ture of the acetylcholine receptor protein is re- markably like a human’s (39). Reduction of Pain or Experimental Insult Until recently, the probability of a research ani- mal receiving the correct amount or type of anes- thetic depended largely on the inclination of indi- vidual investigators. They could accept information about anesthesia available from previous research or attempt to improve on it. In some cases where little information was available, guesswork was required. Now, the enhanced presence of facility veterinarians and animal care and use committees with oversight authority (see ch. 15) has resulted in experimental animals being recognized as veteri- nary patients entitled to protection from as much pain and distress as possible, while maintaining the integrity of research. Analgesics, anesthetics, and tranquilizers are the principle tools for the reduction of experimental pain and distress (see ch. 5). Terminal anesthesia and death has become the method of choice follow- ing major organ surgery on animals, even though it might be argued that observation of the healing process logically constitutes a part of surgical re- search (224). Where postsurgical study is consid- ered necessary, as in cardiology, intensive post- operative control of pain can be used in lieu of maintaining the animal under general anesthesia until death (24). Advances in Instrumentation New types of instruments are critical to a reduc- tion in experimental insult, as they can lead directly to the more refined or reduced use of live animals or living material. In the past decade, practically every piece of instrumentation in biomedical lab- oratories has been adapted to handle “micro” sam- ples or has been replaced by new microtechnol- ogy. Some examples of microinstrumentation include: G In reproductive physiology, a 1.0 microliter sample of rat epididymal fluid collected by micropuncture can be used to examine sperm motility, determine total protein, and deter- mine androgen-binding protein activity (207). G In biochemistry and molecular genetics, elec- trophy biological techniques are being used to explore the possibility of recording the open- ing and closing of single membrane channels, tiny pores controlling cellular function (105). G To study leukemias, blood diseases, and in- born errors in metabolism, a method for meas- uring the enzyme kinetics within a single white blood cell has been developed (134). G A device is available that will dispense a 1.0 microliter sample as 1)000 aliquots (1 nanoliter each) for use in biochemical enzyme research or for clinical samples such as cerebrospinal fluid from infants (97), The use of small samples for analysis by mass spectrometry (146) and by gas or liquid chroma- tography (86,208) exemplifies minimally invasive technology. Each year, an entire issue of Science magazine is devoted to trends in analytical instru- mentation (2,3). Continued developments in ana- lytical instrumentation, including noninvasive imaging techniques such as magnetic resonance imaging (MRI), will likely reduce the experimental insults faces by research animals. In vivo measurements using fiber optics now pro- vide miniaturized spectrophotometric analysis from within the ducts and blood vessels, deter- mine blood velocity, measure temperature changes, monitor intracranial and intracardiac pressure, measure fluorescent marker molecules in tumors, measure pH, and even determine glucose concen- tration (166). Fiber optics offer great promise: They can be inserted into vessels and ducts via small catheters with little discomfort and into the ab- dominal cavity using local anesthetics (a laparos- copy), and they can be used repeatedly within the same animal to obtain measurements without per- manent damage. Chronic intravascular catheters are used in a similar way to obtain repeated blood samples for hormone measurement from freely moving, undisturbed animals (see fig. 6-1) (189). Other minimally invasive techniques in animal research include immunoscintigraphy, amniocen- tesis, and use of the laser. In immunoscintigraphy, -. — 118 . Alternatives to Animal Use in Research, Testing, and Education Figure 6=1.–Apparatus for Remote Blood-Sampling via Chronic, Intravascular Catheter From Unrestrained Ferret - SOURCE: C.L. SIsk, Michigan State University. the production of target-specific monoclinal anti- bodies has improved the ability of external radio- imaging techniques to locate tumors and to iden- tify certain noncancerous diseases; radiolabeled antibodies attach to the target tissue and are then visualized (59). Amniocentesis is used for the early detection of genetic diseases, teratological events, and fetal distress, particularly in domestic species (67). A new application of the laser in oncology involves its ability to initiate a lethal photochemi- cal reaction in cancerous tissue during photoradi - ation therapy (41). Some apparently new noninvasive techniques are actually adapted, miniaturized, or computer- ized versions of older methods. One such exam- ple is a small, inflatable tail cuff used to measure blood pressure in a rat’s tail during hypertension studies (225). In another example, urine is used in some specialized methods: In physiological re- search, electrical impedance measures canine uri- nary output (1). Tandem mass spectrometry is being used for breath analysis to screen for diabetes, cirrhosis, renal disease, and ovulation. Many diseases remain to be examined, but there is potential for use of this technique in toxicology, nutrition, metabolic diseases, endocrinology, anesthesia, physiology, and pathology (133). A technique developed for the determination of the quality of agricultural crops (162) and the per- cent of fat in beef (135) uses amplified, digitized, computer-corrected diffuse reflectance spectro- photometry in the near-infrared region. It involves simply placing an appropriate sensor on the sur- face of the skin and it can be adapted for oncologi- cal, physiological, and nutritional research (102). Other increasingly popular noninvasive tech- niques include ultrasonography -which is used in cardiology to locate vessels (145), to determine blood-flow velocity (176), and to detect early ath- erosclerosis (108)-and magnetic resonance imag- ing, used to examine the energetic of skeletal mus- cle in gerontological research (201), to diagnose metabolic disorders (32), and to provide details of molecular structure and dynamics in liquids and solids (130). USE OF LIVING SYSTEMS IN BIOMEDICAL RESEARCH In Vitro Research In vitro biomedical research entails the mainte- nance of organs, tissues (or fragments of organs and tissues), and cells outside of the body. Depend- ing on the conditions of harvesting and preparing the living material for in vitro maintenance, the cells may be grown as a population of independ- ent cells (cell culture) or with the normal tissue or organ architecture preserved. In the former, the cells may be encouraged to proliferate, result- ing innumerous descendant cell populations suit- able for studies on growth, nutrition, cell division, and gene expression and regulation. Table 6-2, which summarizes the characteris- tics of in vitro systems, makes it clear that as orga- nization is disrupted or lost, the in vitro system has less and less of the kind of intercellular and intracellular interactions that characterize organs, Ch. 6—Alternatives to Animal Use in Research G 119 Table 6-2.-Properties of In Vitro Culture Systems Expression Genetic alteration Preparation and Level of tissue similar to by mutation Environmental consequences organization Reproducibility in vivo and/or selection control Intact system (no consequence). . . . + + + + + + + + + + + + + + Organ culture (remove influences of whole organism). . . . . . . . . . . . . . . . + + + + + + + + + to + + + + + + Tissue culture (remove influences of whole organisms). . . . . . . . . . . . . . . + + + + + + + to + + + + + + + Primary cell cultures (disrupt intercellular relationships) . . . . . . . 0 ++ + to + + + + + + + + Cell lines (intercellular relationships are reduced; cell proliferation is enhanced, at times with little control) . . . . . . . . . . . . . . . . . . . . . . . 0 ++++ + to + + + + to + + + + + + + + KEY: + + + + = High degree; + + + = Moderate degree; + + = Modest degree; + = Some degree; O = None. SOURCE: Adaoted from R.M. Nardone and L.A. Ouellette, “Scoge of ‘Alternatives’: Overview of the State of the Art, ” contract re~orl cweoared for the Office of Technoloav -. Ass&sment, US, Congress, July 1984. tissues, ’and cells in the body. Nevertheless, im- proved accessibility of added chemicals and the opportunity to achieve genetic homogeneity by cloning and genetic manipulation by selection and fusion are important trade-offs. Indeed, at times cell-to-cell interaction may interfere with an ex- perimental objective (156). The explosive growth of in vitro research dur- ing the 1960s and 1970s is illustrated by the fact that the index of Tissue Culture had 84 pages of entries in 1965, 207 pages in 1970, 566 pages in 1975, and 636 pages in 1980, when the publica- tion was discontinued because computerized in- formation retrieval was warranted. There is vir- tually no field of biomedical research that has not been affected by in vitro technology. In vitro mod- els for the study of cell senescence, atherosclero- sis, development, growth, and immune reactions are illustrative of the diversity of applications in biomedical research (156). The specific conditions that best support the maintenance, growth, or differentiation of each type of culture must be determined before any useful information can be garnered. Some of the general requirements of culture systems are com- binations of the proper gas atmosphere, humid- ity, temperature, pH, and nutrients. other culture systems may also have specific light, motion, pres- sure, and physical or chemical support require- ments, Under the proper conditions, many can be subculture for months or frozen in liquid nitro- gen for years without loss of their unique, differen- tiated properties. The ability to maintain many continuous cell lines has opened the floodgates of experimentation and made the new technologies accessible to all the disciplines of biomedical research. Other advan- tages include ease of transport from one labora- tory or country to another, the ability to culture both normal and abnormal tissue for comparison and research, the use of human cells to eliminate species variation, and the ability to expose cultures directly to exogenous molecules at specific concen- trations for precise time periods. Disadvantages include the changes in structure or function ob- served in some cultures, and the fact that isolated systems give isolated results that may bear little relation to results obtained from the integrated systems of whole animals. Organ Culture At some point in the history of research, inves- tigators have attempted with varying success to isolate and maintain every major and minor mam- malian organ, for a variety of purposes. In recent years, improved techniques, such as the availabil- ity of artificial blood media, have increased the probability of successful organ culture. Blood, or artificial blood media, can be pumped through the organ to sustain it (“perfusion”) (see fig. 6-2). Cur- rent applications of organ perfusion include the study of protein synthesis in lactating guinea pig mammary tissue (136) and the use of human pla- centas in toxicology studies, with additional po- tential for use in oncology and gerontology re- search (99). _ 120 G Alternatives to Animal Use in Research, Testing, and Education Figure 6=2—Schematic of Experimental Organ Perfusion PRESSURE- . ,A - OXYGENATOR SOURCE: C. Chubb, The University of Texas Health Science Center at Dallas. Organ Perfusion: Mouse Testis With Pipette Introduced Into Artery Photo credit: C. Chubb, The University of Texas Health Science Center at Dalias whole organs are not generally amenable to long- term in vitro culture or growth. The size and com- plexity of whole organs make it impossible for them to receive sufficient nourishment for normal func- tion without external support. Nevertheless, whole organs are fundamental to many types of anatomy, histology, and pathology because of their suitabil- ity for the examination of relationships between cells and tissues, and they can be sustained in cul- ture for hours or days. Cryostat sections (thin slices of frozen tissue cut with a microtome) through organs are maintained in vitro for oncology studies into organ-specific adhesion of metastatic tumor cells. This method closely reflects the in vivo event and therefore could eventually reduce the use of whole animals in a very active research area (159). Whole mammalian embryos, in addition to their obligatory use in the investigation of basic devel- opmental biology, have been cultured in vitro for other purposes. Protocols have included exami- nations of the effects of hormones and teratogens (42). Tissue Culture Many normal and pathological tissues from hu- mans and a variety of animal species can be suc- cessfully maintained and studied in culture. Indeed, the progress that has been achieved since 1907, when R.G. Harrison first maintained frog neural tissue outside of the body for weeks, has changed the field of tissue culture from an art into a science. Keeping cultures of anything other than bacteria or viruses alive for more than a few hours was problematic until the 1950s) when investigators began to gain a better understanding of the re- quirements of cells and the addition of antibiotics to culture systems. The viability of cultures was extended substantially by controlling bacterial con- tamination. In tissue culture, isolated pieces of a living organ- ism are maintained with their various cell types arranged as they were in the original organism and with their differentiated functions intact. Such cultures are both “better” and “worse” than cul- tures of a single cell type. They are better in that the effects of manipulation can be observed in a more natural environment and different cell types can interact as they would in vivo. They are worse in that they are much more difficult to maintain. Although tissue-culture experiments require the sacrifice of an animal, they can be viewed as alter- natives to animal use since numerous sections of adjoining tissue can be removed and compared. In this way, two or more treatments are adminis- tered to tissues, rather than to a number of indi- vidual animals. Ch. 6—Alternatives to Animal Use in Research G 121 Tissue culture is being successfully employed in many disciplines of biomedical research. In neu- rology, the use of embryonic rat mesencephalon tissues to examine the destruction of dopamine neurons has replaced the use of primates (154). Prior to this development, the monkey had served as the best model for the study of degenerative effects observed in humans. Adult rats, cats, and guinea pigs have been shown to be resistant to the destruction of dopamine neurons. Metabolic studies of an experimental antiarthritis agent have made use of the inside of hamster and rat intesti- nal walls (202). Physiological experiments exam- ined the dynamics of secretion with mouse epididymis (70). Cell Culture Although cell culture is not a new technique, developments and applications during the past dec- ade have come so rapidly as to create whole new research institutions and industries. Cell culture today touches every discipline of biomedical re- search, as well as clinical practice. The following illustrates the pervasiveness of this approach in biomedical research: G G G G G G G Eggs and sperm from many species have been used by endocrinologists, physiologists, and biochemists to study the mechanisms involved in fertilization and early development (58,157). A hamster ovary cell line and its mutants are being used to explore the biochemistry of a membrane-associated protein essential to all animal cell function (175). In oncology, human interferon derived from bacterial recombinant DNA induces a trans- formation of human white blood cells similar to that observed during infection, cancer, and rheumatoid arthritis. This change in white blood cells provides clues to the pathology of cancer (216). Steroid metabolism is being studied using cul- tured rat epididymal cells (29). A monkey kidney cell line was employed to demonstrate the metabolic effects of several general anesthetics (26). Geneticists are developing an in vitro method of studying heme gene expression (54). Surgical research into the use of cultured hu- man epitheliums for permanent coverage of large burn wounds has moved from the lab- oratory into clinical trials (76). In immunology, studies on antibody synthesis and response have been bolstered by the Nobel- prize-winning elucidation of monoclinal antibod- ies. In its initial steps, this technique consumes large numbers of animals, as the varying immune re- sponses of many mice are probed. Then, cloned cells from the spleen of one mouse can be exploited to produce valuable, highly specific antibodies. An- tibodies so produced can obviate the need for many rabbits, sheep, and even humans in the large-scale production of antibodies. Perhaps of even greater importance for research is the high quality of the antibody produced by monoclinal cells. A compre- hensive listing of current research being conducted with monoclinal antibodies from cloned cells is beyond the scope of this assessment. Some of the diagnostic potentials of monoclinal antibodies be- ing ‘ “ “ “ “ “ ‘ G G G G G G explored in biomedical research are: the characterization of malignant and benign tumors; the identification of autoimmune antibodies in rheumatoid arthritis, systemic lupus erythe- matosus, myasthenia gravis, and other auto- immune diseases; the identification and quantification of serum proteins, hormones, and their cell-surface receptors; the monitoring of therapeutic drugs and iden- tification of novel therapeutic drugs; the rapid diagnosis of bacterial, viral, fungal, and parasitic diseases; the monitoring and identification of lymphoid and hematopoietic cells in disease states; and pregnancy testing. Biologists have developed techniques for the con- trolled disruption of cells that can leave many or- ganelles intact or allow the harvesting of selected intracellular membranes. These fractions have proved to be invaluable in the search for informa- tion at the molecular level. For example, micro- somal membrane fractions from rat and human liver have been used in comparative anesthesia research (27). In endocrinology, human placental and ovarian microsomes were used to demonstrate inhibition of steroid hormone synthesis by plant chemicals (112). In dentistry, proteins purified from unerupted fetal buds were shown to be in- 38-750 0 - 86 - 5 —. 122 G Alternatives to Animal Use in Research, Testing, and Education hibitory toward seeded enamel growth in culture (56). In virology, infected cell nuclei isolated from a hamster cell line were used to study influenza virion RNA replication (16). In metabolic studies, the inhibition of chick-embryo-derived collagen fibril formation by glucose suggests a direct rela- tionship between excess glucose and poor wound healing observed in people with diabetes mellitus (126). One of the most unique uses of subcellular fractions involves the bringing together of mixed species systems in biochemical studies of protein transport across intercellular membranes. For ex- ample, researchers studying intracellular protein translocation used dog pancreatic microsomes, bo- vine pituitary and rabbit reticulocyte messenger RNA, bacterial nuclease, and a wheat germ cell- free system to elucidate the structure of the sig- nal recognition particle (213). Human Tissues and Cells Cultured human fetal lung cells have been found to be excellent hosts to support the developmental cycle of a protozoan parasite that causes severe, persistent, life-threatening diarrhea in immuno- deficient patients. There has been no effective ther- apy for this illness, so the in vitro system offers an opportunity to study the parasite’s behavior, development, and metabolism and provides a po- tential method for screening therapeutic agents (50). Virologist and oncologists have been very quick to take advantage of new human in vitro culture systems. For example, an embryonal carcinoma cell line from the stem cells of a teratocarcinoma is being used to study cytomegalovirus replication (83). Lysis of herpes simplex virus (HSV) type 2 is being investigated using human monocytes (117), and HSV latency is studied by using isolated neu- rons obtained from human fetuses (220). The use of postmortem material from humans has significance in many areas of biomedical re- search, but particularly in neurology. Investigators studying the unconventional slow-virus diseases use brain tissue from humans with Creutzfeldt- Jakob disease and from animals with scrapie (144). Postmortem material from schizophrenics has pro- vided evidence for two distinct categories of that disease (181), and temporal lobe structures from Alzheimer patients have revealed specific patho- logical cellular patterns in the brain hippocampal formation (104). Examples of the use of human tissue for investi- gations aimed at human treatment can today be drawn from every discipline of biomedical re- search. Advances in in-vitro culture methods are likely to increase this use further. Postmortem tis- sue use is likely to continue. Invertebrates Invertebrates represent over 90 percent of non- plant species on the earth. Although their body structure is much less similar to humans than is vertebrate body structure, invertebrate anatomy, physiology, and biochemistry offer avenues for new approaches that have been only partially explored. Caenorhabditis elegans, a 1 millimeter roundworm, is of intense interest to developmental biologists. As they have traced this nematode’s complete cell lineage, it offers an unprecedented opportunity for the study of individual living cells (38). Other terrestrial invertebrates are used in many disciplines of biomedical research. For example, flies, bees, earthworms, and leeches are involved in various aspects of anatomy, physiology, and bio- chemistry (5,33). Ants and bees are used in vision research (4). Fruit flies are well known for their participation in genetic studies. Age-related meta- bolic changes in other insects are being investi- gated for possible use in aging research (184). Marine invertebrates represent an important, largely untapped research resource. one commen- tator (190) has suggested that the lack of opportu- nity by medical scientists to learn marine biology and the failure of marine biologists to learn pathol- ogy have combined to leave marine species over- looked. A notable exception to the underuse of marine invertebrates is neurobiology. The coelen- terates, including hydra, corals, anemones, and jellyfish, have helped scientists understand primi- tive nervous system biochemistry. Lobsters and squid have contributed to knowledge of brain anat- omy and physiology, and the grazing snail and cray- fish have broadened understanding of cell biol- ogy (33). Ch. 6—Alternatives to Animal Use in Research G 123 Four advantages of using invertebrates in bio- medical research are: G G G G different phylogenetic levels of structural and functional specialization can be exploited (e.g., different types of circulatory systems, novel chemical compounds); invertebrates reproduce rapidly and produce numerous offspring; experiments can be exe- cuted in days and weeks instead of months and years; storage, upkeep, and maintenance are inex- pensive; and invertebrates are not prone to spreading dis - ease throughout a colony; The overwhelming disadvantage is the consid- erable phylogenetic distance between inverte- brates and humans. Micro-organisms From its origins within the medical disciplines of bacteriology, pathology, and virology, the study and use of microorganisms has branched out to influence practically every area of biomedical re- search, as these examples indicate: G G G G G G Salmonella typhimurium—bacteria used in mechanistic studies in genetics (124) as well as the Ames mutagenicity/carcinogenicity test (see ch. 8); Escherichia coli—bacteria used by develop- mental biologists to derive theories of gene control (90) and by molecular biologists in re- combinant DNA research (75); Streptococcus mutans—bacteria used in den- tal research on the metabolic activity of plaque (91); Bacillus subtilis (bacteria) spores, Artemina sa - lina (brine shrimp) eggs, and Sordaria fimil- cula (fungi) ascospores-all incorporated into NASA’s biostack (monolayer of biological test organisms sandwiched between thin foils of different types of nuclear track detectors) ra- diology experiments inside Spacelab I (31); Tetrahymena pyriformis—a ciliate protozoan being used to study the effects of anesthetics on metabolism (44); and a host of microscopic protozoans, metazoans, and rotifers used to investigate the physiol- ogy and biochemistry of photoreception and vision (4). The advantages of using micro-organisms in bio- medical research are fourfold: They reproduce rapidly at body temperature; rapid division (every 20 to 30 minutes) makes them useful for short- term studies; multigenerational studies can be per- formed in a short period of time; and they are in- expensive in terms of storage, upkeep, and main- tenance. The major disadvantage stems from the fact that these are unicellular organisms: As a con- sequence, the interaction of cells cannot be stud- ied (156). Plants One advantage of using organisms from the plant kingdom is that they lack anything resembling a nervous system. Presumably, plants do not feel pain; they appear to be good potential alternatives to animals. Plants, like micro-organisms, are rela- tively easy and inexpensive to propagate (156). Although there is some interest in the potential use of plant cells in toxicology and oncology re- search (191), the use of whole cells from plants is obstructed by their very rigid cell-wall struc- ture compared with the relatively fluid animal cell membrane. This prevents their use in many dis- ciplines where intimate cell surface contact or transmembrane communication is essential. Once removed from the cell, comparable organ- elles from plants and animals (including micro- organisms) are essentially indistinguishable in both appearance and function. For example, in studies having potentially broad applications in endocri- nology and immunology, yeasts have been found to contain active steroid hormone systems (118). Yeast is used in cell biology in studies of the im- port of proteins into mitochondria, organelles that are essentially the same whatever their source (129). An extensive literature in cell biology, genet- ics, molecular biology, and virology supports the use of subcellular fractions from plants and ani- mals, separately or together, for research into basic molecular mechanisms (213). 124 G Alternatives to Animal Use in Research, Testing, and Education USE OF NONLIVING SYSTEMS Chemical and Physical Systems Long before the advent of modern technology, researchers were constructing chemical models of certain phenomena that occur in living systems. There is a long and rich history in biochemistry, for example, of the application of nonliving sys- tems to experimentation (128,147). Enzyme biochemistry continues to be a principal area of application of nonliving methodology in biomedical research. Enzyme mechanisms maybe studied in a totally chemical system. Magnetic res- onance imaging is used to obtain from enzymes in solution detailed structural data and informa- tion about the mechanism of enzyme action. By combining MRI with cryoenzymology-the use of enzyme solutions held at subzero temperatures— enzymatic reactions can be slowed enough to study intermediate products that would ordinarily ex- ist for too short a time to be detected. Investiga- tions that had been restricted to in vivo manipula- tions can now be expanded into a far wider range in vitro (131). In dental research, a chemical system mimics the mechanics of the formation of dental caries. A two-chambered diffusion cell pairs an excess amount of specific protein crystals with a chemi- cal solution of artificial “plaque-saliva .“ Dissolution of the crystals can be studied under varying chem- ical conditions relevant to a better understanding of the caries process (40). In the field of membrane biophysics, the advent of synthetic membranes has proved a boon to re- search and stands as one of the premier examples of nonliving alternatives to animal use. Liposomes are synthetic vesicles made of protein and fatty molecules. Their structure can be dictated by the investigator, who can combine proteins and lipids of different types and in different ratios to yield COMPUTER SIMULATION Modern approaches to biomedical research de- scribe the functions of living systems at all organiza- tional levels by the language of science—mathe- matics. Knowledge is acquired by investigating IN BIOMEDICAL RESEARCH an artificial membrane. As with true biological membranes, the barriers formed by liposomes are selectively permeable. These artificial membranes are particularly useful in basic studies of the trans- port of molecules across membranes and of mem- brane damage (12). Except for the use of mannequins to simulate accident victims in the transportation industry and in trauma centers, the principal use of physical and mechanical systems today is in education (see ch. 9) rather than in biomedical research. How- ever, it is not inconceivable that future combina- tions of mechanical and electronic technology could provide biomedical researchers with artifi- cial research subjects capable of independent, un- anticipated responses. Epidemiology: Using Existing Databases The use of existing databases to gain new infor- mation and insights in biomedical research may be a major underused resource, if the paucity of published results is any criteria. One study that relied on such information concentrated on the relationship between 17 nutrients and the poten- tial for development of hypertension cardiovas- cular disease in more than 10,000 people from the database of the National Center for Health Statis- tics’ Health and Nutrition Examination Survey (HANES I) (137). The results proved to be highly controversial, with some of the criticism aimed at the use of the database (119). Too little information is currently available to evaluate fully the potential dimensions of the salu- tary use of epidemiologic databases in basic bio- medical research. The possibility exists that their enhanced use may constitute an important non- animal method. relationships among cells, tissues, fluids, organs, and organ systems. By the processes of trial and error and of hypothesis and testing, relationships begin to be understood and can be described by Ch. 6—Alternatives to Animal Use in Research G 125 mathematical expressions. These can range from simple, linear functions to various types of curved functions to multidimensional surface functions and may involve kinetic data expressed by differen- tial equations. Variables in these equations include physical terms, such as time, temperature, weight, energy, force, volume, and motion. Complex math- ematical relationships may be developed to express these cause-and-effect relationships more clearly. In some instances, a relatively simple relationship may be shown to exist, but this is unusual, since living systems are highly interactive and multi- dimensional in nature (48). A relationship that can be reasonably expressed in a mathematical equation maybe considered to be a candidate biological model. The limits within which the expression will hold determine the util- ity and validity of the model. If it is possible to change one or more parameters in the equation, and thereby obtain the same response or responses as found in live animal research, the model may be used to “simulate”a biological preparation. Simu- lation implies that an investigator can manipulate the parameters at will and observe the resultant effects on the model. Used in this way, computer simulation is a useful tool for research and espe- cially for suggesting new mechanisms or hypoth- eses for further study (48). At the subcellular level, information is usually gained by electron microscopic examination or by analytical methods for the sequencing of amino acids and nucleic acids. Such information tends to be of a descriptive or topological nature rather than numerical. Recent strides in genetic engineer- ing based on increased knowledge of DNA, RNA, and protein amino acid sequencing have required computers to store and match nucleic acid and amino acid sequences numbering in the millions (163). These capabilities are not equivalent to simu- lation, but they share with simulation a reliance on computers for storage and processing. At the level of one or a few cells, models are be- ing sought for computer simulation of sliding fila- ment systems—believed to be the basic movement of muscle fibers, cilia, and flagella (28). Modeling of the function of individual cone cells in the eye is under study at the National Institutes of Health (NIH) (167). Most efforts toward computer simulation of bio- logical systems are directed at higher levels of orga- nization, such as organs and organ systems. This bias is a consequence of the need to understand numerous feedback systems within living systems. Feedback systems are the basis for an organism’s ability to maintain a homeostatic, or steady, state, Feedback mechanisms involve several organs as well as communication via the bloodstream and nervous system. For a simulation to succeed, the system must be considered as a whole. In model- ing the cardiovascular system, for example, a simu- lation must take into account the heart, brain, lungs, and kidneys. In the 1980s, computer modeling of organ sys- tems is progressing on many fronts. The brief sam- pling of simulations listed in table 6-3 illustrates the variety of organ systems under study. One development in this field deserving particu- lar attention was the establishment by NIH’s Divi- sion of Research Resources in 1984 of the National Biomedical Simulation Resource, a computer fa- cility at Duke University that may be used onsite or over a telephone data network. Any project in which the results are free to be published in open scientific journals and where no profit is involved can apply to use the facility. Training sessions in- troduce biological scientists to the concepts of mod- eling, and special aid is provided in the develop- ment of simulation software (120). Projects under Table 6-3.-Some Examples of Computer Simulation of Phenomena in Biomedical Research Kidney function: Transport of electrolytes, nonelectrolytes, and water into and out of the kidney (142) Cardiac function: G Enzyme metabolism in cardiac muscle (214) G Cardiac pressure-flow-volume relationships (1 52) G Malfunctions of instrumented cardiovascular control systems (9) Lung function: G Respiratory mechanics (150) Sensory physiology: G Peripheral auditory system, and single auditory nerve fiber transmission of vibrations (180) Neurophysiology: G Impulse propagation along myelinated axons (73) Developmental biology: G Shape changes i n embryonic celIs that develop into mature organs (98) SOURCE: Office of Technology Assessment, 126 . Alternatives to Animal Use in Research, Testing, and Education way in 1985 involved research in cardiology, phys- Limitations on the utility of computer simula- iology, endocrinology, toxicology, and neurology. tions stem from the lack of knowledge of all possi- Specific simulations included: ble parameters that may play a role, however slight, G G G G G G G G G regulation of sodium, potassium, and calcium in heart muscle; electrolyte diffusion in heart muscle; propagation of activity in heart muscle; heart volume potentials; mathematical modeling of blood coagulation; regional dose responses in the human and ani- mal lung; ciliary motility; cochlear function in the inner ear; and a molecular model of ion transport in nerves in the melange of feedback-mechanisms that cons- titute living systems. Basic biomedical research at all levels, some of it involving live animals, will continue to provide the new knowledge required to improve existing simulations and develop models where no satisfactory one exists. The development of increasingly powerful computer programs de- pends on the use of animals in biomedical research. and muscles. CONTINUED, BUT MODIFIED, USE OF ANIMALS IN BEHAVIORAL RESEARCH As in biomedical research, the continued, but modified, use of animals in behavioral research encompasses reducing the number of animals used through changes in experimental design and sta- tistical analyses, substituting cold- for warm- blooded vertebrates, and lessening the degree of pain or experimental insult in general, and in pain research in particular. Compared with biomedi- cal research, behavioral research offers markedly fewer opportunities to substitute cold-for warm- blooded vertebrates and to use in vitro cultures, and it holds little chance of using nonliving systems. Reduction in the Number of Animals Used Improved Experimental Design and Statistical Analyses Individual animals vary in their behavior both between subjects and, in the case of one subject, over time. The goal of a behavioral experiment is to identify patterns that remain when these two sources of variability have been eliminated or taken into account. An investigator attempts to conclude that observed effects are due to the conditions be- ing manipulated in the experiment and not to ex- traneous factors. This decision usually rests on the outcome of statistical tests. Ensuring the validity of such tests or improving their design can mean that fewer experiments are needed. Enhanced sta- tistical rigor, however, may lead to increases or decreases in the number of animals required in a particular protocol. Statistical Power. —A statistical test’s sensitiv- ity in detecting experimental effects is termed its “power.” The most widely recognized method of increasing power and, hence, the sensitivity of an experiment is to use a large sample of subjects. Typically, the more variable the results, the more power is needed to detect an effect and, therefore, the greater the need for large samples. Although the magnitude of variability cannot be determined prior to an experiment, the amount of variability likely to be encountered can be estimated by con- ducting small, pilot studies or by examining previ- ous research in the same or related areas. Given an estimate of variability, statistical tables can be used to determine the sample size needed to at- tain certain levels of power (221). In certain instances, the methods of increasing power may reduce, not increase, the number of animals needed: G Choosing a lower level of statistical signifi- cance (i.e., the likelihood that the results were due to chance) increases power and reduces Ch. 6—Alternatives to Animal Use in Research “ 127 G G G G the number of subjects needed. However, this also increases the chances of concluding that the experimental procedure produced an ef- fect when in fact the effect was due to chance alone. By convention, researchers generally accept the probability of a chance effect of 5 percent or less as a statistically significant result. Greater precision in the conduct of an exper- iment may reduce variability and increase power. For example, highly precise behavioral measurements coupled with the elimination or control of extraneous variables would re- duce the need for large numbers of subjects (198). The use of different statistical analyses can increase the sensitivity and power of a proto- col (e.g., analysis of the data by parametric rather than nonparametric statistical tests) (198). Alterations in experimental designs can in- crease power. Factorial designs (where two or more treatments are manipulated concur- rently), for example, are more powerful and can be used instead of testing the effects of different treatments in separate experiments. Not only does the use of factorial designs in- crease power, it requires fewer untreated, control subjects than multiple concurrent studies do. It is important to note, however, that in areas that have not been heavily re- searched there are inherent dangers to the use of factorial designs. For example, there may be no observed effect of treatments given in combination, as one treatment cancels the effect of another. Without sufficient back- ground information on the effects of the treat- ments administered individually, this finding would be erroneously interpreted. Power is increased as the magnitude of the treatment effect is increased. ‘Treatment ef- fects can be maximized by choosing widely spaced levels of the treatment variables or by including conditions that are thought to max- imize the appearance of the phenomenon under study (113). Within-Subjects Design.–Many experiments on animal behavior are conducted using a between- subjects design, That is, different groups of ani- mals are given different treatments, and the per- formances of the different groups are compared. However, individuals also vary in their behavior. Depending on the degree of variability, large num- bers of subjects maybe needed in each group to obtain statistically significant results. Under cer- tain conditions, however, a within-subjects (or repeated measures) design can be used that re- quires only one group of animals instead of many. Under these conditions all members of the group serve in all treatment conditions. The advantage of this technique is that it minimizes variability by taking into account individual differences. The major drawback, however, is the possibility of con- taminating the data and nullifying the results: Treatments already received by a subject may in- fluence, and thereby confound, performance un- der subsequent treatments. Carry-over effects can be partially offset by counter-balancing, wherein the experimenter ensures an equal occurrence of each experimental treatment at each stage of the experiment; this balances any effect of prior test- ing equally overall treatment conditions (1 13). Al- though within-subjects designs are effective in re- ducing both variability and the number of subjects needed, the inherent danger of carry-over effects in many instances may invalidate the use of such designs. Random Block Design. -Randomized block de- sign consists of assigning subjects to groups based on evidence of their being similar to one another in one or more characteristics known to be related to the behavior under investigation. Two or more such blocks are formed and then each block is as- signed randomly to the treatment conditions. This design reduces variability by restricting the de- gree of individual differences within blocks, and thereby increases power (113). Although random- ized block designs are effective in lowering the number of animals needed in an experiment, they are not applicable to all areas of behavioral re- search. The technique requires substantial prior knowledge of the behavior being investigated and is therefore limited to intensively researched areas. Analysis of Covariance.—An analysis of covari- ance uses the same information as randomized block designs except that an estimate of variabil- ity is not needed beforehand. The covariance pro- 128 G Alternatives to Animal Use in Research, Testing, and Education cedure is applied to data after they are collected to adjust for chance differences among groups. The analysis increases power, and fewer animals may be needed to obtain statistically significant results (113). Single-Subject Design.—In some instances in- ferences can be made about populations from very small samples. This is common in psychophysical experiments in which there is a substantial prior body of evidence indicating that the behaviors un- der investigation do not vary appreciably within the population at large (e.g., visual sensitivity to light). Although such experiments can be con- ducted using just one subject, two or three are typically used to guard against the possibility of misleading results from an atypical subject (198). In other than psychophysical experiments, the general procedure in single-subject research con- sists of choosing a baseline (which involves meas- uring the frequency of occurrence of the behavior of interest), changing one treatment variable at a time, temporarily withdrawing the experimental treatment to assess its causal effects, and repeat- edly measuring the baseline behavior before and after each treatment. (More sophisticated experi- mental designs available for single-subject research are reviewed in ref. 96.) Single-subject designs are increasingly used in animal operant conditioning and human clinical research (187). Statistical anal- yses of these are reported infrequently due to the lack of many statistical techniques for handling such data, although using time-series analyses to test for changes over time is one acceptable method available (111), A limitation to studying a single subject is the uncertainty of the generality of the findings, a prob- lem commonly dealt with by replicating the experi- ment with different subjects (96). Thus the reduc- tion in animals used may be illusory. Inbred Strains.-One way of reducing variabil- ity (and hence increasing power) is to use highly homogeneous populations of subjects. Inbred strains of animals, produced as a result of 20 or more generations of brother-sister matings, rep- resent one approach, though it is usually much more expensive than using randomly bred animals. Inmost inbred strains all subjects are highly iden- tical genetically and genetically stable; they change only as a result of the slow accumulation of muta- tions. In contrast, outbred stocks of animals are genetically variable. They contain an unknown and uncontrollable degree of genetic variation that may obscure or mask experimental treatment effects. Inbred strains not only increase statistical power, they also reduce variability between experiments conducted indifferent laboratories or in the same laboratory at different times (68). It can be argued that experiments should rely on animals drawn from heterogeneous, outbred populations in order to get a broad genetic basis for results that can be extrapolated, for example, to heterogeneous human populations. Yet the differences between different inbred strains are usually greater than the differences between in- dividuals of an outbred stock. Greater generality, then, may be obtained by conducting experiments with two or more inbred strains (68). Sharing Animal&—A team approach to re- search questions across biomedical and behavioral research disciplines could reduce the number of animals needed for behavioral research (173). For example, researchers studying a behavioral phe- nomenon by noninvasive means could, at the ex- periment’s conclusion, give their animals to biolo- gists investigating the anatomy or physiology of that species. Likewise, scientists from different dis- ciplines could collaborate on research proposals: A psychologist maybe interested in studying preda- tor-prey relations, while a biologist wants to study endocrinological changes in response to stress; ef - fective collaboration could yield two different data sets from the same animals. Substitution of Cold-Blooded for Warm-Blooded Vertebrates The modified use of animals in biomedical re- search includes the replacement of mammals and birds with fish, amphibians, and reptiles. In be- havioral research, however, the often vast differ- ences between species are likely to make such sub- stitutions difficult. At the moment, researchers know more about why warm-blooded vertebrates cannot be replaced with cold-blooded ones, as this description of seven behavioral research dis- ciplines illustrates. Ch. 6—Alternatives to Anirnal Use in Research G 129 Aggression Aggressive interactions between members of the same species have been studied in a variety of fish and reptile species under both laboratory and field conditions. Considerable work has been done on intermale rivalry among sticklebacks (183) and cich- lid fish (11). Aggressive interactions among cold- blooded vertebrates are frequently stereotyped and species-specific. Among sticklebacks fish, for example, full-fledged attacks can be elicited by a model that is the same color but a different shape (30). In contrast, aggression in primates can em- body a variety of highly sophisticated introspec- tive social strategies such as deception, grudging, delayed retaliation, and reconciliation (77). Thus extrapolation among all vertebrates of the results of research into aggression is difficult. Animal Movements Migration and homing abilities have been inten- sively studied in several species of fish, particu- larly eels and Pacific salmon. Among American and European eels, for example, the eggs hatch in the Sargasso Sea, near Bermuda. The juvenile fish make a year-long migration toward the coasts of North America and Europe. On reaching sexual maturity in 7 to 15 years, the adult eels migrate back to the Sargasso Sea to breed (217), a move- ment primarily dependent on the use of chemical cues in the water (60). In contrast, birds use solar, stellar, and magnetic cues to navigate, and whales use the topography of the ocean floor and coast- line to remain on course for migratory purposes. Such dramatic differences in the way different spe- cies respond to and perceive the environment limit the use of cold-blooded vertebrates in modeling animal movements of warm-blooded ones. Communication Visual cues, such as changes in coloration, pos- ture, or body appearance, have been shown to be important determinants of social interaction among fish (3o), which, unlike most mammals, gen- erally have color vision. Fish also exhibit dramatic changes in appearance, such as flaring of the gill apertures, which are relatively rare among mam- mals. Auditory communication is marked by spe- cies differences, too. Communication among am- phibians and reptiles, primarily to attract a mate, consists of simple one- or two-note utterances. Vocalization in birds and mammals consists of a wide range and variety of sounds. Moreover, un- like cold-blooded species, many birds have to learn species-specific songs. Many rodents communicate by ultrasonic vocalizations (123) that have no ap- parent counterpart among cold-blooded species. Learning, Memory, and Problem Solving Learning has been studied in a variety of diverse species (179, 183), and many differences are mani- fested. Comparing learning in goldfish and turtles with that in rats yields both similar and distinguish- ing features. For example, rats show a decrement in performance when an accustomed reward is changed, while goldfish and turtles do not (23). The existence of so-called biological boundaries of learning (182), apparently shaped by unique eco- logical pressures, precludes most substitutions of one species for another in learning paradigms. Predator-Prey Relations Prey-catching behavior and predator avoidance have been studied in fish, frogs, and turtles (60, 103,203). The similarity across species in behaviors used by prey to avoid being caught suggests that when a general question about reactions to pre- dation (rather than the behavior of a given spe- cies) is of interest, cold-blooded vertebrates can substitute for warm-blooded ones (103). But there is growing evidence of neurochemical differences underlying predator avoidance behaviors even among birds and mammals (78). Predators exhibit marked differences across spe- cies. Frogs, for example, sit passively and wait for an insect to come within striking distance, while some carnivores have developed sophisticated hunting strategies that often embody elements of cooperation and may even culminate in sharing foods (30). Reproduction and Parental Care With some notable exceptions (e.g., the African Mouthbreeder fish), parental care of offspring is absent inmost cold-blooded vertebrates, since the eggs are typically abandoned shortly after fertili - 130 . Alternatives to Animal Use in Research, Testing, and Education zation. In contrast, all birds are subject to some type of parental care, and mammalian parent-off- spring relations become even more complicated. Species differences in external versus internal fer- tilization, seasonal breeding, courtship, pair- bonding, and nest-building preclude substitutions of one species for another in this research. Sensation and Perception The sensory and perceptual differences among species are vast. For example, many snakes’ pri- mary mode of prey identification is chemical cues transferred from the tongue to a structure at the roof of the mouth, called Jacobsen’s organ. Inges- tively naive baby snakes appear to have an innate preference for prey extracts that represent species- typical foods across a variety of different snakes. Each species shows unique attack profiles that ap- pear to be independent of maternal diet and not subject to modification by experience (e.g., baby snakes of a minnow eating species that are force- fed liver still show attack responses to minnow extracts but not to liver) (34,35). In contrast, baby rats seem predisposed to eat the same diet as their mother, and the flavor of the maternal milk serves as a medium for the trans- mission of cues that rat pups use to make their initial food choices. Manipulating maternal diet during lactation has produced corresponding changes in subsequent pup food preferences. Moreover, rat pups poisoned in association with a mother’s milk later avoid the types of food she had been eating (74). Reduction of Pain or Experimental Insult As noted earlier, a general anesthetic is prefer- able to a local one for surgical manipulations be- cause it suppresses both pain and fear (114). Pain- relieving drugs should be administered to animals after surgery whenever this would not interfere with the behavior under study, and animals should be carefully monitored so that any complications that develop may be treated (197). Transection of the spine or brain stem is recom- mended for surgical experiments when possible, because it renders the animal incapable of feeling pain (114). This technique has limited applicabil- ity in behavioral research, however, as postsurgi - cal behavioral assessment requires a relatively intact animal. Similarly, the nonrecovery experi- ments on completely anesthetized animals that were described earlier, in the biomedical research section, are rarely used in behavioral research, since most behaviors of interest do not occur when the animal is unconscious and behavioral testing is typically conducted postsurgically. Multiple sur- geries on the same animal are to be avoided when- ever possible, because painful consequences may be cumulative (197). The analysis of pain in behavioral research is complicated by recent theoretical and empirical developments suggesting that fear and pain acti- vate quite different kinds of behavior (25). Rather than being on a continuum, as might seem to be the case intuitively, data suggest that fear and pain are associated with fundamentally different mo- tivational systems. Fear activates species-specific defensive behaviors, such as freezing, flight, or fighting, that serve to minimize encounters with pain-producing stimuli (e.g., predators). Pain, on the other hand, appears conducive to the kinds of behaviors that provide for healing and recuper- ation (e .g., rest, grooming, licking the affected area, and sleep). A growing body of evidence shows that fear takes priority over pain, and that fear can ac- tually inhibit pain under some circumstances (pos- sibly through the release of endogenous opiates). For example, soldiers who are wounded in battle frequently continue fighting and feel no pain from their injuries until after they are removed from the front lines (211). Likewise, a deer wounded by hunters may flee the scene with defensive be- havior indistinguishable from that of uninjured animals, But once the deer is out of danger, pain- related recuperative behaviors predominate (25). Brain Manipulation In studies of the physiological bases of behavior, the recording of brain-wave patterns maybe sub- stituted for electrical stimulation whenever pos- sible, and brain areas may be stimulated instead of lesioning or ablating sections of the brain (121). These techniques, however, are not completely interchangeable. Recording neuronal firing as an Ch. 6—Alternatives to Animal Use in Research . 131 animal behaves allows correlational inferences to be made, but not causal ones. If the experimental goal is to determine a particular brain area that is responsible for a certain behavior, that area must be manipulated directly. Electrical stimulation of brain areas is useful in establishing causal rela- tionships, and the most definitive and reliable re- sults are obtained when stimulation is used in con- junction with lesioning or ablation (20). Drug Administration In research on the behavioral effects of experi- mental or currently available drugs, animals are injected either intraperitoneally (within the body cavity), intravenously, intramuscularly, or intra- cranially (within the skull, via an implanted can- nula). Depending on whether the drug must be given repeatedly, the injection procedure can be stressful and may cause discomfort. Within the last decade alternative administration methods have been developed that may replace the need for multiple injections in some chronic drug treatment studies. Capsules of porous rubber (Silastic@, produced by Dow-Corning) implanted beneath the skin release a drug slowly into the animal’s body, and stress produced by repeated injections is avoided. The method produces minimal discom- fort and is well tolerated by animals (63). Small, implantable minipumps are also available to de- liver drugs for days or weeks. The use of aerosols has also been suggested (174); although this would seem to hold promise for al- leviating the stress of injections, it has drawbacks. For example, animals may differ greatly in their inhalation rates, and dispersal of the drug into the air prevents adequate control of drug dosage. Food Deprivation It is important to distinguish between the differ- ent methods of depriving animals of food and the reasons for using any method. In most cases, ani- mals are deprived of food to motivate them to per- form various tasks or behaviors for food reward. The nature of the subject’s performance of such tasks—and not the food deprivation—is the object of study. Food deprivation is typically applied one of two ways: Animals are deprived of food for a stand- ard period of time (e.g., 24 hours) prior to testing or they are maintained at some percentage of their normal body weight (e.g., 80 percent) (43). Each procedure has advantages and disadvantages. Food deprivation for specified intervals of time is easy to implement, but it fails to take into account spe- cies differences in metabolic rates, For example, 24 hours of food deprivation for a mammal is less severe than it would be for a bird, while for a snake it would be inconsequential. Maintaining animals at a percentage of normal body weight avoids this problem, but it requires daily handling and the delay of the trial for long periods of time to stabi- lize body weights. When food deprivation is applied according to a standard time period in behavioral protocols, the most common interval is 24 hours (43). It is note- worthy that the feeding of domestic pets once a day parallels this laboratory protocol. When main- taining animals at some percentage of their normal body weight, behavioral protocols usually involve up to 20 percent weight loss (43). Experimental animals’ reduced food intake is associated in some instances with enhanced longevity (172). Several suggestions have been made to reduce, ameliorate, or eliminate food deprivation in be- havioral research: G G G G Water deprivation, sometimes used concur- rently with food deprivation, should be used to motivate behavior only if thirst or drink- ing is the object of study. Water deprivation affects an animal’s physical condition more severely than food deprivation does, because death by dehydration occurs much more rap- idly than death by starvation (121). The normal eating pattern of a species should be taken into account when deciding on the duration of food deprivation. For example, sparrows eat only during the light hours of the day; hamsters feed largely at night. In some cases, food deprivation might be avoided by using a highly preferred food as a reward (121). Food deprivation may also be avoided by tak- ing the experiment into the animal’s living quarters, so that it is required to perform for 132 Altematives to Animal Use in Research, Testing, and Education food as and when it wants to eat. In this way, any deprivation would be self-imposed as under natural conditions (121,151). This tech- nique has been used successfully in work on sensory-motor functioning in monkeys (168). Pain Research and the Use of Electric Shock The experience of pain is a highly adaptive ca- pacity. It prevents organisms from engaging in be- haviors that would otherwise prove maladaptive. For example, humans who are congenitally insen- sitive to pain become terribly scarred and muti- lated, often develop a sense of being invincible, and have short life expectancies (141)143,196). Per- haps because pain plays such an essential role in regulating the behavior of organisms, pain thresh- olds are surprisingly consistent across a great diversity of species (115). The discovery of en- dogenous opiates in earthworms (5) and recent findings with spiders (61) suggest that inverte- brates may also feel pain. Pain can be induced through mechanical, ther- mal, electrical, or chemical stimuli (127). Of the various stimuli used for research purposes, elec- tric shock at the levels normally used in experimen- tation is the only one that does not damage tissue. Most studies of pain in animals use what are called flinch-and-jump thresholds—an index of the min- imal amount of electric shock or heat needed to produce a reaction. Electric shock is used as a stimulus for research into the mechanism of pain for G G G several reasons: Electric shock is easily quantifiable. The pa- rameters of shock can be manipulated and specified with a high degree of precision over a wide range. Electric shock can be administered so as to have a discrete or gradual onset and offset. Electric shock of the type most often used (i.e., a brief current of 0.001 amperes, the equiva- lent of a tingling sensation in the finger) does not yield physical damage, bleeding, or tissue destruction. However, electric shock is a highly atypical stimu- lus (79). No contemporary terrestrial species ap- pears to have evolved under conditions of elec- tric shock. The question of whether data obtained this way are widely generalizable in mechanisms of pain remains unanswered. A survey of the 608 articles appearing from 1979 through 1983 in the American Psychological Asso- ciation journals that typically publish animal re- search (e.g., Journal of Comparative and Physio- logical Psychology and its successors Behavioral Neuroscience and Journal of Comparative Psychol- ogy) identified 10 percent of the studies as using electric shock. Four percent of the studies admin- istered inescapable shocks stronger than 0.001 am- peres. Most of the experiments with electric shock involved rodents; those with monkeys, dogs, and cats accounted for 0.5 percent of the total 608 arti- cles (43). Recommendations that have been made to re- duce pain or discomfort in animal experiments in- volving aversive stimulation include: G G G G G G The lowest possible level of electric shock should be used that will at the same time main- tain the behavior under study (52). However, this may reduce the statistical power and re- quire a large sample size. Animals should be given predictable rather than unpredictable shock and an opportunity to control its termination (52). Rats, for ex- ample, will choose to receive more shocks at greater intensity in order to receive a warn- ing cue prior to each shock delivery (10). If aversive stimulation must be used, alterna- tives to electric shock such as loud noise or bright lights should be considered (121). In developing models of chronic pain, the model should closely simulate a particular chronic pain syndrome in humans (e.g., arthri- tis or cancer). Otherwise, there is no justifi- cation for the procedure (114). Animals should have an opportunity to con- trol the intensity of the stimulus in chronic pain studies. While the objection to this might be that, given this option, the animal would “turn off” the pain stimulus, this might be cir- cumvented by giving a preferred food reward for keeping the stimulus “on” at a given level, as in experiments with electric shock titration techniques (114). A reward, such as a preferred food, should be used for the correct responses instead of a punisher, such as electric shock, for incor- rect response (121). Ch. 6—Alternatives to Animal Use in Research G 133 USE OF LIVING SYSTEMS In behavioral research, using living components derived from whole animals or living nonanimal systems as alternatives to animals could conceiva- bly involve embryos; cell, tissue, and organ cul- tures; invertebrates; and plants. The greatest po- tential in this area, however, appears to rest with the use of invertebrates. Several factors limit the use of embryos (used here to refer to the conceptus, embryo, and fetus prior to birth) as an alternative or complement to young or adult animals: G G G G G Some studies involving embryos may be con- ducted when the subject is very close to birth or hatching. The advanced developmental sta- tus of the organism at this point raises the same kinds of ethical considerations that would ap- ply to the use of postnatal animals (see ch. 4). In behavioral studies involving mammalian embryos, the mother is necessarily involved in most experimental manipulations performed on the embryo. As a consequence, embryo- logical manipulations on mammals cannot log- ically avoid the use of adult mammals. Behavioral studies using embryos may involve testing for effects later in adult life (e.g., be- havioral teratological studies). In these in- stances, embryos are not being used as alter- natives, since the procedures also require postnatal assessment. Only a limited number of behaviors can be studied in embryos, partly because of practi- cal problems associated with access to the embryo. Embryos live in a dramatically different envi- ronment than fully developed adult animals. This difference constrains the generalizabil - ity of behavioral data obtained from them. Cell, tissue, and organ cultures do not figure prominently in the equation of alternatives to ani- mal use in behavioral research. In isolation and in culture, cells, tissues, and organs exhibit few activities that fall among the disciplines of be- havioral research. A rare example of the use of cell culture in be- havioral research comes from studies of the bio- chemical basis of depression and manic mood BEHAVIORAL RESEARCH changes. Skin fibroblast cells obtained from hu- mans and maintained in culture for several months were assessed for their ability to bind a variety of pharmacologic agents. The cultured cells of pa- tients and relatives of patients with manic-depres- sive illness exhibited markedly different biochem- ical properties than the cultured cells of persons without a history of manic depression (155). One commentator characterized this as ‘(a step forward, applying to psychiatry the techniques of tissue sam- pling and cell culture that have been of great value in characterizing molecular abnormalities in nu- merous medical diseases” (192). Continued devel- opment of this line of research could reduce the use of animals in such investigations. Invertebrates Few behavioral studies use invertebrates as sub- jects (139). As a consequence, relatively little is known about invertebrate behavior. In behavioral research, invertebrates offer a fertile testing ground for any theory that claims to be broadly based across the phyla of the animal kingdom (138). Certain groups of invertebrates are promising sub- jects for behavioral research. The brains of octopuses and squid approach those of vertebrates in relative size and complex- ity (178). Visual discrimination learning has been studied extensively in the octopus. Octopuses can discriminate between pairs of geometric shapes that differ with respect to vertical, horizontal, and oblique orientations. The octopus and squid show learning performance on a par with mammals on such tasks as detour problems, reversal learning, delayed response, and delayed reinforcement (178,218). Among all the invertebrates, the only species with a neuroanatomy and learning ability compa- rable to vertebrates are the octopus and squid. Practical problems in obtaining, transporting, and housing these marine species have always pre- cluded their widespread use as alternatives in be- havioral research (178), However, recent advances in the laboratory culture of octopuses make them promising research candidates, although provid- ing live food (e.g., shrimp) on a consistent basis — 134 . Alternatives to Animal Use in Research, Testing, and Education remains a major logistical obstacle. The develop- ment of a dead or artificial food ration is currently a high priority in octopus culture (88). The same highly developed nervous system that makes the octopus and squid desirable replacements for ver- tebrates may cause some ethical objections to use of these invertebrates. In addition, their adapta- tion to a completely aquatic existence would also make tenuous any extrapolations to the behavior of terrestrial mammals. Starfish and sea urchins exhibit habituation— the waning of a response to stimuli, as a result of repeated elicitation of that response—and they can learn escape behaviors in response to a cue paired with aversive stimulation (46). Earthworms exhibit habituation (45), can learn to associate light with a food reward (66), and can learn to travel a maze to receive darkness and mois- ture as reinforcing stimuli (85). Flatworms are also of considerable interest, since they represent a bilateral body form, as do mammals. Flatworms exhibit a concentration of nervous tissue and sen- sory organs in the anterior, or head, portion of their bodies, and they have refined internal or- gan systems (47). Flatworms exhibit habituation, can be conditioned to avoid alight after it has been paired with shock, and can learn to approach an area for food reward. There are also claims that such learned events are remembered after these worms undergo regeneration, and that learning can be transferred from one animal to another by cannibalism (reviewed in ref. 47). Insects are valuable behavioral models in com- munication, navigation, learning and memory, and behavioral genetics. Ch. 6—Alternatives to Animal Use in Research G 135 G G G G Communication. Honeybees recruit others to a new food source through a dance per- formed at the hive that conveys distance and directional information (209). Many species of insects (e.g., moths and ants) communicate chemically by pheromones, which serve sex attractant, repellant, and/or trail-marking functions (reviewed in ref. 30). Other species, such as the cricket, communicate by songs produced by rubbing body parts together (19). Navigation. Honeybees have demonstrated extraordinary abilities to locate and return to a food source “mapped out” for them by other bees. They can also return to an artificial feed- ing source designed for experimental pur- poses to test their navigation abilities (37, 209)215). Learning and Memory. Habituation has been demonstrated in a variety of insect species (46). Honeybees also appear capable of more ad- vanced forms of learning, such as learning to associate a specific color with a food reward, and they can remember this association after a 2-week interval (219). Cockroaches can learn to leave their preferred dark retreats and stay in the light to avoid being shocked; ants have been trained to travel a maze to receive food rewards (85). Behavioral Genetics. Because of the relative ease with which their chromosomes and in- dividual genes can be identified, fruit flies have been used extensively to elucidate the genetic basis for a variety of behaviors (132). Habituation has been demonstrated in a variety of spiders (45), and spiders are capable of learn- ing and remembering the location of prey in their webs (85). They can also be trained to associate food dipped in quinine or sugar with different tones (46). Even though protozoa possess both plant- and animal-like characteristics and lack nervous tis- sue, some forms of learning have been demon- strated in these single celled organisms. Habitua- tion has been demonstrated in paramecia (45). Although the results generated much controversy (reviewed in ref. 45), one investigator claimed to have trained paramecia to enter a specific area of their water container in order to receive food reinforcement (81). It has also been reported that paramecia show spontaneous alternation in a T-maze, a phenomenon also observed in rodents (139). A recent study of learning ability in paramecia has demonstrated classical conditioning of an es- cape movement (94). This study also found that paramecia develop memory for the training event, since significantly fewer trials were needed 24 hours later to relearn the response. Data such as these challenge the widely held assumption that learning is a property of synaptic interactions be- tween nerve cells—absent in protozoa—and not of individual cells themselves. Plants From a behavioral perspective, plants differ from animals in two principal ways. First, plants lack the means of achieving rapid intra- and inter- organismal communication and coordination due to the absence of a nervous system. However, plants do regulate intra-organismal activities occur- ring at different sites through the use of hormones. Plants and animals thus share the basic principles of endocrine function. Second, plants differ from animals in that they are stationary. They must wait for energy to come to them, while most animals move about to obtain different sources of energy. Despite these differences, plants do show rudi- mentary forms of behavior (188). Plants can grow and move in response to light, and some plants have achieved the capacity for relatively rapid movement to exploit certain animals as prey (e.g., the venus fly trap). The mimosa plant, which can fold its leaves when touched, has been a subject of particular interest. Certain of its cells appear to generate primitive action potentials-electrical activity that may be analogous to neuronal func- tioning in animals (186). There have also been reports that the folding response of the mimosa plant shows habituation (6) and even some of the rudiments of classical conditioning (8). Although some claim evidence of feelings, emotions, and even thinking in plants based on polygraph record- ings (206), others contend these are artifactual (80). A number of plants defend themselves from predators via thorns, stickers, or toxic chemicals that produce sickness, irritation, or even death if touched or consumed. It has been demonstrated 136 Alternatives to Animal Use in Research, Testing, and Education that some plants, under attack by insects and These impressive features of the botanical world micro-organisms, develop highly sophisticated notwithstanding, it is unlikely that plants will make defenses involving the emission of antibiotic-like an important contribution to behavioral research. substances and chemicals that inhibit insect diges- The lack of a central nervous system, and in par- tive enzymes. Indeed, some plants can apparently ticular a brain, renders the plant an inappropri- communicate chemically with as-yet-unaffected ate model for use among the disciplines of be- neighboring plants to induce leaf-chemistry havioral research. changes in advance of infestation (reviewed in ref. 164). USE OF NONLIVING SYSTEMS IN BEHAVIORAL RESEARCH Inanimate chemical or physical systems are un- likely to prove useful in behavioral research, for reasons intrinsic to the nature of behavior. A dy- namic, emergent process, behavior functions to allow organisms to adapt to moment-to-moment changes in the environment. In a sense, all behavior ultimately functions to aid and abet survival and reproduction (50. Adaptation, survival, and repro- duction are not properties of nonliving systems. And behavior involves information processing and a continuous series of choices among an array of alternatives (140). Although chemical or physical systems may change in response to certain envi- ronmental stimuli, the nature of such changes does not involve decisionmaking or information-proc- essing. Behavior is a byproduct of interactions between sensory, neural, hormonal, genetic, and experien- tial factors. As such, it is influenced by the situa- tion at hand, the developmental history of the organism, and prior experience with similar and related situations. It appears inappropriate to im- bue inanimate chemical or physical systems with the capacity for experience. Devoid of such a ca- pacity for experience, nonliving systems are un- likely alternatives to using animals in behavioral research. Examples of the application of chemical or phys- ical systems to behavioral research are sparse. One involves the use of chemical reagents to mimic the properties of rhythmic behavioral phenomena in animals. Certain chemical reagents exhibit changes in state that oscillate periodically in a fashion sim- ilar to some biologically based rhythms. However, the chemical reactions themselves remain poorly understood (222). COMPUTER SIMULATION IN BEHAVIORAL RESEARCH A computer simulation is an operating model that depicts not only the state of a behavioral system at a particular point in time but also changes that occur in that system over time. Because dynamic processes are of quintessential importance in be- havioral research, computer simulation stands as a potentially useful tool for the behavioral scientist. In order to simulate a living system, a computer programmer must have information about that system. The more information at hand, the better the simulation (53). In the strictest sense, a com- pletely accurate simulation presupposes that every- thing that there is to know about the system in question is known. To construct a computer simu- lation that would fully replace the use of a live organism in behavioral research would require knowing everything about the behavior in ques- tion, which in turn would preclude the need for a computer simulation for research purposes. Yet, if computer simulation cannot fully replace living organisms, it can and does contribute to be- havioral research. Although the fundamental be- havioral qualities of adaptation, survival, and re- production do not pertain to computer programs, computer soft ware does, for example, embody in- formation processing and decisionmaking. Exam- Ch. 6—Alternatives to Animal Use in Research G 137 pies of recent attempts toward computer simula- tion in behavioral research are listed in table 6-4. Computer simulations are used in behavioral re- search in a number of ways. Statistical simulations, in particular, are increasingly frequent. For ex- ample, one computer program simulates random- choice behavior in mazes (194), and two programs simulate random movements of animals under various conditions (17,49). The output generated by these kinds of simulations is compared with animal-generated data to see if factors other than pure chance are influencing the animals’ behavior. Statistical simulations are also used to test hy- potheses that may not be subject to empirical con- firmation. One investigator used a computer simu- lation to test the proposition that “if enough monkeys were allowed to pound away at type- writers for enough time, all the great works of literature would result” (21). The larger objective Table 6-4.—Some Examples of Computer Simulation of Behavioral Phenomena Spacing mechanisms and animal movements: G Space use and movement patterns (17,49) G Movements of juvenile Atlantic herring (110) G Animal spacing (153) G Mosquito flight patterns (165) G Foraging of the honey eater bird (169) G Random choice in radial arm mazes (194) Learning, memory, and problem solving: Ž Classical conditioning (15,18) G Learning in neural systems (177) G Habituation (195) G Behavior in a psychoecological space (84) G Mechanisms for reducing inhibition (223) Sensation and perception: G Visual pattern analysis (13) G Landmark learning by bees (37) G Chemical recruitment in ants (106) Communication: G Bird song (55,185) G Animal vocalizations (57) Sensation and perception: Neuron models (122) G Neural basis for pain and touch (148) Body maintenance: G Food intake (14) Ž Control of drinking behavior (204) Reproduction and parental care: G Sexual behavior of the male rat (72,205) Ž Infanticide in Iangurs (89) G Evolution of reproductive synchrony (116) G Mating behavior of Spodoptera littoralis (200) SOURCE: Office of Technology Assessment. in this study was to determine if the extreme cases of human genius could be accounted for through chance processes. The simulation was based on an initial assump- tion that monkeys typing at random— or a com- puter simulation using random numbers–would generate huge volumes of nonsense. Statistical properties of the English language (e.g., the rela- tive frequencies of individual characters or se- quences of characters) were added to the simula- tion. As higher-order properties of English (i.e., the relative frequencies of three- and four-letter sequences) were incorporated into the algorithm, the rate of generation of intelligible words, phrases, and sentences increased. These results led to a hy- pothesis that genius could be simulated by a proc- ess of random choice with a weighting procedure, subject to a prior preparatory process in which an individual absorbs the necessary operational patterns that characterize the discipline. In studies such as this, it is not merely the output generated by the computer model that is of interest, but the simulation process as well. Computer simulations have considerable heuris- tic value (65): They may yield insight about the sys- tem or phenomenon being modeled (71) and, as a consequence, stimulate additional research. The value of computer simulation as a heuristic device has been summarized as follows (158): Simulation gives a means of exploring the plausi- bility of models in which theoretical sophistica- tion exceeds the state of the art in empirical test- ing. Simulations provide tools for empirically analyzing theories in order to better understand their implications and predictions. Simulations are a means of exploring interactions between components of complex models. They pose a prac- tical challenge to operationalize theoretical con- structs, which can lead to incidental discoveries about related processes. Finally, they engender a concern with issues of process control that con- tributes to the development of general principles with broad applications. Computer simulation holds promise for under- standing complex cognitive processes. For exam- ple, the computer is often considered analogous, at least in some ways, to the human brain (7)— both process large amounts of information, and their respective outcomes are a consequence of 138 Alternatives to Animal Use in Research, Testing, and Education multiple, relatively simple operations. On the other hand, there are major differences: G G G G computers have larger and many fewer com- ponents than the brain; computer operations occur with much greater speed than neural operations; computers operate through sequential proc- essing; and computers attend to all input, while the brain is selectively attentive. Such differences do not, however, preclude the use of computers as functional models (170). More- over, one advantage of computers is that they de- mand rigorous and exact description. And an in- vestigator need not invoke a variety of hypothetical or mentalistic variables (e g., hope, fear, desire, and intention) to describe their functioning (170. It is important to note that any predictions gen- erated by a computer simulation must be tested and verified using the system the computer was designed to replace (149). In this sense, the use of animals in behavioral research is likely to con- tinue in lockstep with the development of com- puter simulation software. One commentator summarized the use of com- puters as an alternative to animals in behavioral research in this way (114): At the present time and for the foreseeable fu- ture it seems clear that the computer will not be a feasible substitute for experiments on animals. The fundamental reason is that a computer can- not acquire data other than those that are gener- ated by carefully designed experimental studies in animals. What the computer does provide is a superb technique for processing vast amounts of data with great speed and accuracy and for presenting them in almost any manner the inves- tigator desires. To suggest that enough data are already available from previous work, so that from them programs can be generated and sub- jected to a variety of permutations that would lead to new insights, overlooks an important fact. In any animal experiment there are numerous vari- ables over which we have little control, and there are virtually always as many more about which we as yet know nothing but which may have very significant influences on the phenomenon under investigation. In real life, which after all is what matters in biologic research, these variables may be crucial and may give important clues to en- tirely unsuspected phenomena that are some- times far more important than the original sub- ject of the study. In a word, computers do not generate new concepts or acquire new data. They process data and permit the investigator to view it in more manageable or novel ways, and this may facilitate new hypotheses or insights. In summary, computer simulation can serve to facilitate behavioral research. The need for cer- tain protocols may be precluded, or protocols may be refocused by computer simulation before they commence. Modeling techniques using computer simulation lead to the refinement of experimental protocols to be conducted on animals (36). Yet as the preceding quote implies, in facilitating be- havioral research computer simulation may actu- ally increase, rather than decrease, the use of ani- mals because data can be analyzed more quickly and in much greater detail, leading to proportion- ately more hypotheses to investigate (87,160). SUMMARY AND CONCLUSIONS Animal use in research can be modified in a num- ber of ways, including strengthening experimental design to minimize the number of animals used, reducing the degree of experimental insult, and substituting one species for another. The outright replacement of animals with nonanimal methods in research is not at hand, and, because of the na- ture of biomedical and behavioral research, in many instances it is not likely to become feasible. Advances in instrumentation are critical to the more refined or reduced use of live animals or liv- ing material. In the past decade, practically every piece of instrumentation in biomedical laboratories has been adapted to handle ‘(micro” samples or has been replaced by new microtechnology. The use of small samples for analysis by mass spec- trometry and by gas or liquid chromatography leads to less invasive technology. Fiber optics, for Ch. 6—Alternatives to Animal Use in Research G 139 example, can be used to perform analyses inside ducts and blood vessels with little discomfort and no permanent damage to the animal. Continued developments in analytical instrumentation, in- cluding noninvasive imaging techniques such as magnetic resonance imaging, will likely ameliorate the degree of experimental insult faced by research animals. In vitro technology has affected virtually every field of biomedical research. This technology en- tails the maintenance of organs, tissues, and cells outside of the body and may affect research ani- mal use in two important ways. First, when or- gans, tissues, or cells are removed from animals and cultured, experiments may be conducted with fewer animals than would be necessary in whole- animal experiments and, of course, without pain. Cells from one animal, for example, may be divided among a dozen experimental cultures and a dozen control cultures, replacing 24 animals that might be used in a comparable whole-animal experiment. Second, when cells proliferate in culture, com- mercially available cell lines can completely elimi- nate animal use in some experiments. Such cell cultures are derived directly from preexisting cell cultures—not animals, Researchers have used, for example, a monkey kidney cell line to study the metabolic effects of general anesthetics. In vitro experiments are not equivalent to whole- animal experiments. In in-vitro systems, as orga- nization is disrupted or lost, the in vitro system has less and less of the kind of interactions that characterize cells in the body. This can be an advan- tage and a disadvantage. Interactions may cause extraneous phenomena that obscure the process under study. Conversely, the absence of interac- tions may produce results that are at variance with what actually occurs in the live animal. Conclu- sions drawn from in vitro studies must eventually 1. 2. CHAPTER 6 Abbey, J., Close, L., and Jacobs, M., “Pilot Study: Use of Electrical Impedance to Measure Urinary Volume in Canines, ’’Cmnmunic. Nurs. Res. 11:22, 1977. Abelson, P. H., “Analytical Instruments,” Science 226:249, 1984. be validated by comparison with results of whole- animal experiments. In some areas of biomedical research, inver- tebrates and micro-organisms can be used, and even plant parts can yield information about ani- mal systems. Structures within plant and animal cells, when removed from the cells, are essentially indistinguishable in both appearance and function. Cell biologists, for example, use yeast cells as a source of mitochondria—the energy-generating structures present in all plant and animal cells. 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E., “Asymp- totic Reversal Learning in Pigeons: Mechanisms for Reducing Inhibition,’)J. Exp. PsychoJ. (Animal Behavior Processes) 2:57-66, 1976. Wright, E. M., Jr., Vice-Chairman, Department of Comparative Medicine, University of Virginia, Charlottesville, VA, personal communication, Aug. 16, 1984. Zanberg, P., “Animal Models in Experimental Hypertension: Relevance to Drug Testing and Dis- covery,’’Handbook of Hypertension, Vol. 3: Phar- macology of Antihypertensive Drugs, P.A. van Zweiten (cd.) (Amsterdam: Elsevier Science Pub- lishers B. W., 1984). Chapter 7 The Use of Animals in Testing Distress caused by the Draize eye test is sometimes so acute that rabbits do scream out in pain. Close-Up Report Humane Society of the United States 1985 Laws should neither push the science where it is not yet ready to go nor hold the science to procedures that have been modified or replaced. Geraldine V. Cox Chemical Manufacturers Association March .20, 1985 CONTENTS Page Testing Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .l50 Designing a Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .150 Use of Standardized Test Methods and Guidelines . . . . . . . . . . . . . . . . . . . . ....152 Pharmacokinetics ..., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..., . . .153 Acute Toxicity Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..., . . . .153 Skin and Eye Irritation/Corrosion Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .154 Repeated-Dose Toxicity Tests.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1s4 Carcinogenicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ......................155 Developmental and Reproductive Toxicity . . . . . . . . . . . . . . . . . . . . ...........156 Neurotoxicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..............156 Mutagenicity . . . . . . . . . . . . . . . . . . . . ., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..157 Current Trends . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .157 The Role of Government in fasting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... ... ...157 Food and Drug Administration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .158 Environmental Protection Agency. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .161 Consumer Product Safety Commission. . . . . . . . . . . . . . . . . . . . . . . ...........164 Department of Labor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .164 Department of Transportation . . . . . . . . . . . . . . . . . . . . . ...................164 Department of Agriculture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..165 Centers for Disease Control ..+... . . . . . . . . . . . . . . . . . . . . . . ...............165 Federal Trade Commission. . ., . . . . . . . . . . . . . . . . . . . . . ...................165 State Uses of Animal Testing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. ...165 Pesticide Registrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...165 Identification and Classification of Toxic Substances . . . . . . . . . . . . . . . . . . . . . .166 Product Liability Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...........167 The Manufacturer’s Duty to Produce a Safe Product. . . . . . . . . . . . . . . . . . . . . .167 Methods of Testing Required, ,..., . . . . . . . . . . . . . . . . . . . . . ...............168 Summary and Conclusions , . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....168 Chapter preferences. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..., . . . . . . .. ...169 Table Table No. Page 7-1. Number of Animals Needed to Detect Carcinogenicity in 90 Percent of All Tests for a Statistical Significance of 0.05 . . . . . . . . . . . . . . . . . . . . . . . . . .156 Chapter 7 The Use of Animals in Testing Testing for the safety or efficacy of a substance or product accounts for a major use of animals as defined in this assessment, most of which are rats and mice (see ch. 3). Of these, probably the largest portion are used in developing drugs. A significant portion are also used to test other sub- stances—pesticides, industrial chemicals, and con- sumer products—to assess possible toxicity and to establish conditions under which they can be used safely. Research and testing have been differentiated for purposes of this assessment, but the bound- ary between them is not sharp. From the stand- point of developing alternatives, a key difference is that a particular test maybe performed for hun- dreds, perhaps thousands of substances and use hundreds or thousands of animals, whereas a given research method will be used on far fewer. As a corollary there are far more research procedures than testing procedures from which to choose, Fur- thermore, individual researchers are much more likely to develop their own methods than are those conducting testing. These differences make the task of developing alternatives more manageable for testing than for research. Testing for efficacy has some attributes of re- search and some of toxicity testing. A particular protocol may be used on a small number of sub- stances and is likely to be tailored either to the application or to the family of substances being tested. Experimenters testing for efficacy need to have a better understanding of the mechanisms by which a particular effect occurs than those test- ing for toxicity, primarily because efficacy test- ing is closely related to the physiological mecha- nisms that the new drug may affect, whereas toxic effects may be quite independent. Finally, an im- portant distinction of efficacy testing is that the animals used would ordinarily be diseased. Other kinds of tests include those for safety other than for toxicity, as in testing of diagnostic techniques or quality control tests in the manu- facture of medical devices. These have endpoints even more specific than those for toxicity, and are thus good candidates for the development of alter- natives (see ch. 8). Toxicity testing is the focus of this and the fol- lowing chapter for three reasons. First, this is an area of animal use in testing in which the govern- ment has great influence on nongovernmental ac- tivities. Second, these tests are used in a more rou- tine fashion than are tests for efficacy or general safety and therefore have a greater tendency to lag in the application of state-of-the-art technol- ogy. Third, toxicity tests include methods that have attracted the largest political attack. All substances can be toxic at some exposure level, even water. Conversely, even substances known to be highly toxic maybe harmless at low doses or under certain circumstances. Determin- ing the hazard to humans requires information about the potential hazard and the expected level of exposure, resulting in an estimate of the prob- ability that a substance will produce harm under certain conditions (8). This assessment of risk is a scientific endeavor, whereas the management of risk is a sociopolitical one (31,36). Although toxicity data on humans are invaluable in conducting risk assessments, they are usually unavailable. Some information comes from epi- demiologic studies or episodes of accidental hu- man exposure. Most often, however, testing on animals is used. An appropriate weight is given to the following factors on a case-by-case basis, considering the seriousness of the hazard and the kind of assumptions needed to estimate risks to humans: G G G G G the relationship between dose and response; the effects at the molecular, cellular, organ, organ system, and whole-organism levels; conflicting results between studies and pos- sible explanations for the conflicts; the effects of structurally similar substances on humans or animals; any known metabolic differences between hu- mans and the test species that could affect the toxic response; and statistical uncertainties and difficulties in ex- trapolating to a low dose (55). 149 150 G Alternatives to Anima/ use in Research, Testing, and Education TESTING Toxicology as a science began in the 16th cen- tury and has advanced with the growth of the chemical, pesticide, drug, and cosmetic industries. The concept of protecting the public from harm- ful effects of chemicals dates back to laws of an- cient civilizations that made it illegal to adulterate the food supply (25). The importance of toxicol- ogy to public health has received considerable at- tention in the United States since the 1930s. Pub- lic awareness of the value of toxicological testing has also been furthered by disasters such as Mina- mata disease (methyl mercury poisoning in Japan), the thalidomide tragedy, and, more recently, the development of cancer in those exposed to diethyl- stilbestrol (DES) in utero. Designing a Test There are two approaches to toxicology-mech- anistic and descriptive—and these affect the de- sign of experiments and the choice of biological end points to be measured. Mechanistic toxicol- ogy focuses on the chemical processes by which a toxic effect occurs and relies heavily on the tech- niques of physiology, biochemistry, and analyti- cal chemistry to monitor these processes. A sim- ple example of this approach would be a series of experiments showing that a certain substance is metabolized in the liver, that one of the by- products of metabolism happens to be a potent liver carcinogen, and that liver cancer typically follows administration of that substance. Mech- anistic tests are custom designed and are closely related to research. They can contribute greatly to the design and interpretation of descriptive tests, Mechanistic toxicology plays a major role in the development of methodologies that could replace whole-animal testing. Descriptive toxicology deals with phenomena above the molecular level and may rely heavily on the techniques of pathology, statistics, physi- ology, and pharmacology, e.g., the evaluation of changes in the appearance of an organ or its con- stituent cells, the presence of tumors, or signs of irritation. This approach does not necessarily re- quire an understanding of the mechanisms by which toxic effects occur, although if mechanis- tic information were available, it would be used. METHODS In terms of the test substance and species in the preceding hypothetical case, descriptive toxicol- ogy would show that a certain substance causes liver cancer in a particular species within a cer- tain time. It might also show the approximate rela- tionship between the substance dose and the inci- dence of the liver cancer. Regulatory schemes requiring testing most often rely on descriptive toxicology. Mechanistic toxicology provides an approach to extrapolation from one species to another based on known similarities and differences in physiol- ogy. The closer the test animal is biologically to humans or the greater the number of species in which the effect is detected, the more likely it will occur in humans as well. The reliability of extrap- olations from descriptive experiments is greatly enhanced when mechanistic information is also used. Similarly, the use of mechanistic informa- tion in the design of descriptive tests contributes greatly to the reasonableness of any later extrapola- tion to humans if human toxicity data are lacking. Most state-of-the-art toxicological tests require whole animals. Although in vitro alternatives are being developed (see ch. 8), different end points would be measured. For example, whole animals will probably continue to be needed to look for effects in previously unknown target organs, to evaluate effects that represent an interaction of multiple organ systems, to monitor metabolism and pharmacokinetics, or to evaluate healing or dimin- ished responsiveness to the toxic substance. Thus, whole-animal use is unlikely to stop entirely in the foreseeable future. Choice of Species and Strain In 1$104) the Food and Drug Administration (FDA) was still using human employees to test food pre- servatives (e.g., boric acid, salicylic acid, their de- rivatives, and formaldehyde) for toxicity (25). Use of animals remained limited until a few decades ago, when breeding technology provided large numbers of animals with carefully controlled genetic characteristics, thus allowing toxic effects to be more easily detected than had previously been the case. Animal use has grown with increas- Ch. 7—The Use of Anima/s in Testing Ž 151 ing demands by the public for safe and effective products. The most appropriate animals are ideally those that, for the substance being tested, predict the human response most accurately. There is no other animal wholly identical to humans in terms of toxic effects. The choice of animal is influenced by known similarity to humans for the organ system or mechanism of interest, as well as convenience of breeding or purchasing, familiarity with the spe- cies, existing data, lifespan, ease of handling under experimental conditions, cost of obtaining and maintaining, litter size, and gestation period. Ro- dents have been used extensively, as have rabbits, primates, and dogs. Rodents have been used in almost all carcino- genicity testing despite the fact that such tests are the most difficult to extrapolate to humans. Mice and rats have been used because their lifespan is short, they are small and easily handled, and they have a number of metabolic pathways and patho- logical responses similar to those of humans. Some specially developed strains are sufficiently suscep- tible to cancer that test groups can be small. These factors contribute greatly to the economic feasi- bility of conducting carcinogenicity testing with rodents. Extensive experience in using them, and in using particular strains, is often an important reason for continuing their use (55). A large amount of data are already available on spontaneous tumors at specific organ sites (l). Although rodents are routinely used for many kinds of tests, other animals maybe used for spe- cific reasons. For example, the rabbit is used for eye irritation tests because it has large, easily manipulated eyes and because its eyes have many characteristics found in human eyes (19). Hens have been shown to be a good model for delayed neurotoxic effects of organophosphorous com- pounds (12). Dose Levels and Route and Duration of Exposure The way in which exposure to a substance oc- curs can affect the kind and severity of toxic ef- fects. For example, if a chemical does not present a hazard when applied to skin because it is not absorbed, it may nonetheless be very toxic if taken orally. When the route of exposure does not affect the portion of the dose taken up or its distribution in the body, testing might be done in the manner most easily controlled. For other than the most preliminary tests to characterize toxicity, most would administer the substance by the same route as would occur in the course of accidental exposure or use by humans. Sometimes the palatability, volu- bility, stability, or volatility of a substance will de- termine which routes are feasible. Certain tests, such as the acute toxicity for a sin- gle exposure, are used as inexpensive screening tools for estimating the relative hazard presented by a substance. As discussed later in this chapter, the acute toxicity test known as the LD50 is used in classification schemes for the transportation or disposal of chemicals. Acute toxicity testing might also be used to determine the risks of one-time exposure, as might occur in an accident. Ordinar- ily, the duration of exposure in an animal study is greater (at least in proportion to the lifespan) than the exposure period for which data will be used in extrapolating the risks to humans. The dose levels administered depend on a vari- ety of factors. On the one hand, it is not possible to detect long-term effects if the dose is so large that many animals die before the end of the test. On the other hand, administered doses represent- ative of human exposure levels may not produce detectable effects with what may be considered a reasonable number of test animals. Generally, three dose levels are used; they are chosen so as to span the range of responses from a “no-observed- effect level” to fully observable toxic effects, For carcinogenicity and other long-term testing, the highest dose should be one that will produce measurable toxicity without significantly altering lifespan. Other levels may depend on whether the carcinogenicity is being looked for in combination with chronic toxicity (55). The lowest dose could be one for which there are no observed effects or it might be related to the level of estimated hu- man exposure (38). Another approach is to choose doses that will yield levels in the blood similar to those expected for humans. Although this is perhaps a more real- istic test, effects may be more difficult to detect. In addition, the criterion of similarity may require 152 “ Alternatives to Animal Use in Research, Testing, and Education more than one administration per day because metabolic rates and excretion rates tend to be faster in small animals than in humans (24). Statistical Considerations To obtain valid results, an experiment must be designed so that what is measured provides useful and sufficiently accurate information. Statistical methods allow a scientist to estimate the minimum number of test animals from which conclusions can be drawn to estimate the reliability of any con- clusions. Statistical analysis can help reduce the number of animals needed for a particular test procedure. To allow for the unexpected (including death, illness, or error), the number of animals used al- ways exceeds the minimum number needed to de- tect expected effects reliably. Determining that minimum number of animals is more difficult for longer tests, both because the passage of time makes the probability of something going wrong during the experiment increase, and because cer- tain problems are more likely to occur as the ani- mals age. Another factor affecting the number of animals needed is the variability in the sensitivity of indi- vidual animals to the substance involved. Thus, as few as 6 animals might be used for an eye irrita- tion test or 10 per dose level for an acute toxicity test. In carcinogenicity or teratogenicity testing, many of the animals maybe unaffected by the test substance, and 100 animals may be needed for each dose level. Most species experience some cancer and other diseases during their life. Any measurement of in- cidence as it relates to the dose given must be taken against this background incidence, which is gauged in an (untreated) control group. Control groups may also be important if a test substance is being carried in a particular vehicle needed to administer the test substance, such as in solution with another chemical, that is not itself being tested (vehicle con- trol group). The sensitivity of the test animals to a substance known to be toxic may also be meas- ured for comparison (a positive control group). Be- cause there are so many variables that can influ- ence a test, toxicologists consider it vital that the control and test groups be drawn from the same pool of animals and be tested concurrently. Any experiment suffers from experimental er- ror, of which there are three sources: the natural variation due to differences among test animals, the variation in experimental conditions, and error arising in measurement. Determining the amount of error is crucial to drawing reliable conclusions from experimental results, but it is also important to keep the error as low as possible by controlling conditions carefully. Differences among test ani- mals are controlled by using genetically similar and sufficiently large groups for each condition. Even minor environmental factors can influence toxic response (15,23). Sources of measurement error depend on the measurement technique and the equipment. Use of Standardized Test Methods and Guidelines Testing methodologies are standardized to con- trol experimental variables, thus allowing results to be easily compared. Methodologies may be- come standardized through round-robin testing in many labs, through publication and imitation, and through development by recognized organi- zations or agencies. Methodologies or guidelines are published by the Food and Drug Administra- tion, the Environmental Protection Agency (EPA), the Organization for Economic Cooperation and Development (OECD), the National Cancer Insti- tute, the American Society for Testing and Mate- rials, the American National Standards Institute, the British Standards Institute, the International Agency for Research on Cancer, and others (see app. A for information on FDA, EPA, and OECD guidelines). The most important reason to strive for com- patibility among guidelines is to avoid the need to repeat identical tests to satisfy particular require- ments of various governments and agencies. Com- patibility can also avoid nontariff trade barriers, Any government that would like to change its test- ing requirements to further the cause of animal welfare needs to consider the effects of its pol- icies on testing in other countries. Ch. 7—The Use of Animals in Testing G 153 Pharmacokinetics Pharmacokinetic studies provide information about the mechanisms of absorption, about a sub- stance’s distribution among the various body com- partments, and about metabolism and elimination. They facilitate the interpretation of results from other tests and their extrapolation to humans be- cause the distribution and elimination of a foreign substance will often explain its toxicity or lack thereof. Absorption of a substance into the body can oc- cur by a variety of routes. If exposure is by inhala- tion, absorption can occur in the lungs, in the path- ways leading to the lungs, and sometimes in the gastrointestinal tract. If exposure is by mouth, ab- sorption would occur as the substance passes through the gastrointestinal tract. What is not ab- sorbed is excreted in the feces, With dermal ex- posure, the substance must be absorbed through the skin. If exposure is via injection into a body cavity, the substance cannot be removed without the involvement of other parts of the body. Once a substance is absorbed, it maybe excreted unchanged. Excretion could be through the skin, in the urine, feces, semen, or breast milk, or, if it is volatile, in exhaled air. It might also be stored in tissues, organs, or body fluids, perhaps for the life of the organism. A substance might also be chemically modified until it can be excreted or until the body is unable to metabolize it any further. This metabolism normally takes place in the liver, the site where detoxification of substances takes place. A test substance or its metabolic products can react with the chemicals that make up the body, perhaps resulting in toxic effects. Pharmacokinetic studies are usually conducted through the sampling of body fluids, both those that are excreted (urine, saliva) and those that are not (blood, cerebrospinal fluid). Tissue samples are often taken, although normally not until the end of a study (4). Acute Toxicity Tests Acute toxicity testing is used to detect the toxic effects of single or multiple exposures occurring within 24 hours. These are frequently the first tests performed in determining the toxic characteris- tics of a substance and may serve as a basis for classification or labeling or for concerns about acci- dental exposure. The results are used to establish toxicity relative to other substances, to determine specific toxic effects, and to provide information on the mode of toxic action and the relationship between dose and adverse effects. Results may also help in designing long-term tests. One of the most common acute toxicity tests is the LD50 (from Lethal Dose for 50 percent), devel- oped in 1927 for comparing batches of dangerous drugs (52). The LD50 is calculated to be the dose, within statistically established confidence inter- vals, at which half the test animals can be expected to die upon exposure to a test substance. A sub- stance is administered once by the oral, dermal, or parenteral (injection into a vein or the body cavity) route or it is inhaled. The animals, usually rodents, are observed for 14 days and then sacri- ficed so that their organs and tissues can be evalu- ated for gross changes. Other measurements and observations can be added to increase the amount of information this test provides. A related procedure is the limit test. A high dose is given, often 5 g/kg body weight (54); if no ani- mals die, the test ends. This is based on the as- sumption that if an organism is not killed by an extremely large dose, it does not matter what dose it takes to actually cause death. Other tests using fewer animals have been devised and are receiv- ing growing acceptance (see ch. 8). Acute toxicity testing has its limitations, particu- larly because the end point is death. Death can come about in many ways and the mechanism is not conveyed in the numerical value of an LD50. In addition, the results may vary greatly both among and within species, with the animals’ sex, age, and diet, and with other test conditions. Acute toxicity testing, although not necessarily the clas- sic LD50 procedures, will continue to be of inter- est because there are many substances for which the toxic effects of acute exposure are quite differ- ent from those produced by chronic exposure (8). It may also continue to be used in selecting doses for long-term studies. Nonetheless, circumstances may be identified in which acute toxicity testing is not needed because other tests more relevant to the use should be performed. The Toxicity Com- mittee of the Fund for the Replacement of Ani- 38-750 0 - 86 - 6 154 . Alternatives to Animal Use in Research, Testing, and Education reals in Medical Experiments recommended that study of the consequences of not acquiring knowl- edge of acute toxicity of products be undertaken and that in the case of products such as drugs, LD~Otests should be replaced by acute toxicity tests that emphasize the nature of the effects observed (18). Skin and Eye Irritation/Corrosion Tests Irritation is the production of reversible tissue damage such as swelling, while corrosion is the production of irreversible tissue damage. Skin and eye irritation tests normally involve acute expo- sure. Repeated exposure can be used to test for allergic reactions, which involve the organism’s immune system, and cumulative effects. Skin irri- tation studies are used to initially characterize a substance’s toxicity and to develop precautionary information for situations in which human skin or eye exposure is possible. Although it is not yet possible to reliably predict the degree of irritation or corrosion a substance will cause, a considerable body of knowledge ex- ists. The factors that determine damaging effects to eyes or skin are: G intimacy and duration of contact, G physical properties that determine the amount of penetration, and G the reactivity of the substance with tissues (10). Intimacy is affected by both the ability of the sub- stance to spread over the surface (such as soaps or detergents) and its concentration. Penetration of the skin or other membranes is greatest in sub- stances with small molecular size and with abili- ties to mix with both water and oil. A substance that can react with proteins and enzymes in tis- sues is especially damaging if it can penetrate to the delicate structures of the eye (50). Skin irritation tests are usually conducted on rab- bits, guinea pigs, rats, and mice, although other mammals may also be used. The test substance is applied to a small area of skin from which the fur has been clipped or shaved and maybe held in place with a dressing. Using untreated skin of the same animal for comparison, the degree of red- ness or blistering is scored at intervals (e.g., 38,54). There are many similarities between the skin cells of humans and other mammals, but there are important differences as well. For example, there are structural differences that affect permeabil- ity (32). Animal models have been shown to be par- ticularly poor in the evaluation of mild irritants (27). The extrapolation of animal models is further complicated by large differences in the race, age, and skin condition of humans (21,26,58). The method most commonly used to evaluate eye irritation is the Draize test, which has remained largely unaltered since it was introduced more than 40 years ago (9). A single dose of a substance is applied to one eye of at least three adult rabbits. The other eye remains untreated. The degree of irritation or corrosion to the cornea, iris, and con- junctival is scored by comparison with standard pictures over a period of 3 days. The rabbits may be observed for 3 weeks to determine whether the effects are reversible. A substance shown to be highly corrosive to skin will be highly irritating to the eye and thus might not be tested. Similarly, a substance with a pH of 2 or less (strongly acid) or 11.5 or more (strongly alkaline) is assumed to be highly irritating or cor- rosive to skin or eye and need not be tested (38,54). The cornea tolerates substances with a pH rang- ing from 3 to 11 variably, with the severity of a reaction depending in large part on a substance’s ability to affect protein structure or function (17,35). Repeated-Dose Toxicity Tests Humans are often exposed repeatedly to a sub- stance and this does not necessarily cause the same effects as an acute, one-time exposure. Chronic toxicity effects differ from acute toxicity ones when the test substance or its metabolizes accumulate in the organism to a toxic level or when it causes irreversible toxic effects that accumulate with each administration (8). Rats are most frequently used, and testing in a second, nonrodent species, usu- ally a dog, is also common. Repeated or prolonged exposure to the test sub- stance is used in chronic, subchronic, and short- term toxicity tests. The term chronic generally Ch. 7—The Use of Animals in Testing G 155 refers to tests with exposure for at least 1 year or most of the lifetime of the test species. Sub- chronic usually refers to tests of intermediate dura- tion-3 to 6 months. Short-term repeated dose tox- icity tests last from 2 to 4 weeks. Some have suggested that there is little to be gained by exposures of more than 6 months dura- tion for chronic toxicity testing (18)34). One com- mentator has argued that studies of 3 to 6 months are easier to interpret because the complicating effects of aging are avoided (44)45). Another finds longer tests necessary for detecting effects that occur only late in life or for which cumulative tox- icity is an important consequence (42). Throughout repeated-dose testing, animals would be observed for general appearance, res- piratory problems, central and peripheral nerv- ous system function, coordination, and behavioral changes. During and following the course of ex- posure, observations are made of hematology (hematocrit, white cell count, platelet count, clot- ting factors), ophthalmology, electrolyte balance, carbohydrate metabolism, liver and kidney func- tion (as determined from concentrations of cer- tain substances in the blood), body weight, and the appearance of lesions. After the animals have been sacrificed, observations are made of body surfaces, orifices, cavities, and organs. Microscopic examinations are made of selected tissues and or- gans, of gross lesions, and of organs that changed in size. one technique used in repeated dose tox- icity testing to determine whether the toxic effects are reversible is to give a satellite group the high- est dose of the test substance and then give the animals time to recover before sacrificing them. Carcinogenicity Cancer is a major human health concern, strik- ing one out of four and killing one out of five Ameri- cans (53). Consequently, carcinogenicity is an im- portant animal test. Detecting human carcinogens presents special problems because a latency period of 20 years or more can occur. Animal testing, par- ticularly in rodents, is useful because the latency period for tumor formation is much shorter (1 to 2 years for rodents), thus allowing potential hu- man carcinogens to be detected during testing and before use, at which point they could become ma- jor public health problems. It is also much easier to control the animal environment than the hu- man environment, and therefore to investigate causal relationships. Although many human carcinogens were dis- covered without animal testing, several have been identified by first using such tests, e.g., DES, vinyl chloride, and bis(chloro-methyl) ether (55). Ani- mal use has its limitations; many substances cause cancer only in certain species. The known human carcinogens benzene and arsenic have never proved to be animal carcinogens. Hundreds of sub- stances have been identified as carcinogens in tests with one or more animal species but not in hu- mans, in part because of insufficient human epi- demiologic data and in part because some of them undoubtedly do not cause cancer in humans (41). Nonetheless, the use of animals in testing for car- cinogenicity is widely endorsed (55). Carcinogenicity testing is more costly and re- quires far more animals than other tests. Chronic toxicity testing may use about 160 rats and 32 dogs, whereas carcinogenicity testing would use about 400 rats and 400 mice. (In order to economize, car- cinogenicity testing and chronic toxicity testing are often combined. ) Cancer is easy to detect if tumors are visible, but it can only be detected in its early stages by microscopic examination of mul- tiple samples of 30 or more tissues and organs that may appear normal. Typically, 500,000 data points must be analyzed (41). These large numbers of animals and multiple data points are needed for statistical reasons. Can- cer has a high background incidence and large var- iations from animal to animal, making it difficult to establish that cancer was caused by the test sub- stance. The higher the incidence of spontaneous cancers, the more difficult it is to establish a link between cancer and the test substance. For ex- ample, if the background rate of cancer is 10 per- cent and the common criterion for statistical sig- nificance of 0.05 is used, the number of animals required to detect carcinogenicity in 90 percent of the tests is as shown in table 7-1. As can be seen, if a test substance causes cancer in 80 percent of the animals, 48 animals are needed to demonstrate carcinogenicity. If the incidence is only 15 percent, over 3,000 animals are needed. It has been sug- gested that the background incidence could be re- 156 “ Alternatives to Animal Use in Research, Testing, and Education Table 7.1 .—Number of Animals Needed to Detect Carcinogenicity in 90 Percent of All Tests for a Statistical Significance of 0.05 Number of animals Rate of incidence caused by (3 dose levels test substance (percent) plus control group) 80. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 60. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84 40. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184 20. . . . . . . . . . . . . . . . . . , . . . . . . . . . . 1,020 15. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3,304 SOURCE: Adapted from l.F.H. Purchase, “Carcinogenicity/’A nh-naLs artdA/ter. natives in Toxicity Testktg, M. Balls, R.J. Ridden, and A.N. Worden (eds,) (New York: Academic Press, 1983). duced, and the sensitivity of the method thereby improved, if animals were not kept under condi- tions that aggravate cancer (excessively nutritious diet, little exercise, and isolation) (42). Developmental and Reproductive Toxicity The effects of chemicals on human reproduc- tion are difficult to assess because of the complex- ity of the reproductive process and the many kinds of insults that can be inflicted before reproduc- tive maturity as well as during fetal development (8). Reproductive functions that can be harmed by foreign substances include the storage and maturation of the germ cells, fertility (including factors that affect sperm maturation and implan- tation of the fertilized egg), and the development of the fetus. Possible toxic effects to the fetus in- clude birth defects (teratogenicity), low birth weight, abnormal gestation time, and prenatal or postnatal death (7). There are a variety of experimental protocols by which these effects can be determined in ani- mals. Some involve more than one generation; others involve evaluation of a fetus before birth. Exposure to a substance can start before the fe- male ovulates or as late as some specific stage of fetal development. Exposure can be chronic or acute. The great variety of procedures available can lead to a certain amount of overlapping test- ing (2). Rats and rabbits are the most commonly used species. Mice and hamsters and other mammals are used as well. Three dose levels are normally used, the highest of which causes minimal toxicity in the adult female. Groups of about 20 pregnant females are typically used. In the OECD Testing Guidelines, if no teratogenic effects are observed at a dose of 1,000 mg/kg body weight, other dose levels are not necessary (38). Neurotoxicity Neurotoxicity (damage to the nervous system) is observed in acute and chronic testing, but the range of neurotoxic effects is so great and the signs so varied that special tests for damage to the nerv- ous system are sometimes warranted, Neurotoxic effects that tend to be associated with acute expo- sure are functional, sometimes reversible changes in the nervous system that might not involve struc- tural damage or degeneration. Most chronic neuro- toxic effects do involve structural changes or de- generation and are not readily reversible (6). The type of neurotoxic effect tends to depend on the size of the dose and the duration of exposure (46). There are many types of nerve cells, each per- forming special functions. Damage can occur to the functioning of the cell itself, to its connections to other nerve cells or to muscle cells, or to the supporting cells. Neurotoxicity can be manifested in the following ways: motor disorders such as weakness, lack of coordination, paralysis, tremor, convulsions, or slurred speech; sensory disorders such as numbness, pain, or auditory, olfactory, or visual deficits; disturbances of autonomic func- tion such as sweating, incontinence, vomiting, im- potence, or tear formation; increased state of excitability such as hyperactivity, irritability, or euphoria; impairment of short-or long-term mem- ory, disorientation, or confusion; sleep disorders; psychiatric disturbances; impaired temperature regulation; or alterations in appetite, or weight gain or loss (6). More than any other kind of toxicity test, neuro - toxicity does not lend itself to standard procedures or in vitro tests because the range of effects is so broad. There are considerable differences among species, and little standardization of tests across species has occurred. Neurotoxicity tests would typically follow acute or chronic toxicity ones in which neurotoxic effects had been observed or were suspected (6). Ch. 7—The Use of Animals in Testing G 157 Mutagenicity A mutation is a permanent change in a gene that is passed along to any descendants of the cell. Thus, mutations in germ cells will be passed along to off- spring. If recessive, the mutation will not be ob- served in the offspring but will become part of the gene pool from which future generations will draw. If the mutations dominant, it may be lethal to the developing fetus or it might affect the off- spring in a variety of ways, including impairing its fertility. If the damage is to a somatic cell, the mutation could lead to cancer or, in a developing fetus, birth defects. There are several nonanimal and in vitro tests based on mammalian or human cells that would be considered alternative mutagenicity tests (see ch. 8). There are several whole-animal tests as well. One is the dominant lethal assay, in which a male is exposed to the test substance and then mated with an untreated female. Part way through the pregnancy, the female is killed and the number and condition of the fetuses observed. Another is the heritable translocation assay, in which the male progeny of treated males are mated with un- treated females and the effect on fetuses deter- mined. The mutations transmissible to the next generation are of special interest because of their implications for the human gene pool (5). The in vivo sister chromatid exchange and mouse micronucleus tests rely on microscopic examina- tion of the chromosomes themselves after the test substance has been administered to the whole ani- mal. In vitro versions of these techniques also exist (see ch. 8). Changes can be observed using a micro- scope. Host-mediated assays are a hybrid of non- animal and whole-animal techniques in which the test substance and a micro-organism are adminis- tered to an animal and the effects on the micro- organism determined (5). Current Trends Many factors are likely to influence testing prac- tices in the near future. Public pressure to use alter- natives to whole animals, increasing costs of using animals, and improvements in toxicological meth- ods are likely to reduce the use of some tests, such as the LD50 and the Draize eye irritation tests. This pressure is also likely to result in changes in some existing tests in order to reduce animal suffering. These developments could bring about a review of current legal requirements for testing, perhaps reducing the amount of testing per chemical and the number of animals per test. Such a review, as well as advances in the state of the art, might better tailor testing to the substance being exam- ined and to the circumstances of human exposure. On the other hand, the number of substances be- ing tested could increase with greater regulatory or product liability requirements, with greater funding available for testing, or with less expen- sive tests available. Interpretation and extrapolation of test results to humans can be expected to improve as the mech- anisms of toxic responses are better understood. Increasing use of pharmacokinetics and mechanis- tic studies is likely to result in improved designs and better selection of tests. THE ROLE OF GOVERNMENT IN TESTING The Federal Government and each of the States are involved in testing in a variety of ways. Per- haps the most important are various explicit and implicit requirements for testing under existing statutes. Another area is the funding of research and development leading to new methods (see ch. 12). Yet another is the funding of toxicological test- ing, conducted primarily by the National Toxicol- ogy Program (NTP), supported largely by the Na- tional Institute of Environmental Health Sciences. This program, chartered in 1978, is a cooperative effort among agencies within the Department of Health and Human Services (see chs. 11 and 12). Four principal Federal agencies have a signifi- cant role in animal testing for regulatory purposes: FDA, EPA, the Consumer Product Safety Commis- sion (CPSC), and the Occupational Safety and Health Administration (OSHA). Other agencies whose reg- ulatory activities affect animal use include the 158 . Alternatives to Animal Use in Research, Testing, and Education Centers for Disease Control (CDC), the Department of Transportation (DOT), the Federal Trade Com- mission (FTC), and the US. Department of Agri- culture (USDA). Animal testing is also funded by the Department of Defense. Testing is covered by several types of statutes and regulations. Most common are laws that re- quire a product to be safe and effective. Given the state of currently accepted technology and prac- tice, such a statute implicitly (although not explic- itly) calls for animal testing. Such tests are rou- tinely expected as an indication of meeting the standard of product safety and effectiveness. A second stimulus for animal testing involves pre- market approval. Under this authority, testing with animals is explicitly required by regulations of the agency involved. Or, animal testing may be explic- itly required by statute, as in the case of the Fed- eral Hazardous Substances Act administered by CPSC. As a practical matter, it makes little differ- ence whether the tests involving animals occur under implicit or explicit statutory or regulatory authority: The procedures used are quite similar. The specific tests performed and the methodol- ogies used may be dictated by informal or formal requirements of the agency. These may take the form of promulgated regulations, published guide- lines, unpublished guidelines, or customary prac- tices. Some guidelines and the use of specific tests are accepted internationally (see app. A.) With these general principles in mind, this dis- cussion summarizes current Federal regulatory requirements relating to testing with animals (see also app. B). This review is not intended to evalu- ate the justification of such testing, only to describe its scope and magnitude. It is meant to provide sufficient background to permit an evaluation of the reasons testing is conducted and of the regu- latory needs that any alternatives to such testing must satisfy. Food and Drug Administration FDA is responsible for administering several stat- utes that regulate animal and human food, animal and human drugs, medical devices, cosmetics, color additives, and radiological products. This regulation takes place primarily under the 1938 Federal Food, Drug, and Cosmetic Act as amended (21 U.S.C. 301 et seq.) and the Public Health Serv- ice Act of 1944 (42 U.S.C. 200 et seq.). FDA evaluates each product on a case-by-case basis. The exact testing regime is determined by considering the type of product, the method of exposure, the amount and duration of intended use, and the potential hazards associated with the specific product. In support of its regulatory re- sponsibilities and to assure quality testing, FDA has issued standards for good laboratory practice (see ch. 13) and has developed guidelines and test- ing protocols. Although some special guidelines or testing protocols are established for specific products, most tests are the same as or similar to the toxicological tests used by other agencies. Ap- pendix A lists the types of tests used. The National Center for Toxicological Research (NCTR) in Jefferson, AR, and the National Toxicol- ogy Program are the research and testing arms of FDA. Although NCTR and NTP have no direct regulatory responsibilities, they provide informa- tion needed to evaluate the safety of chemicals. Research that involves the use of animals or alter- native methods includes studies of effects of low- dose, long-term exposure to chemicals; develop- ment of new methodology to investigate toxic ef- fects; study of biological mechanisms of toxicity; and investigation of methods for estimating human health risks using experimental laboratory data. The misbranding or adulteration of virtually any product regulated by FDA is prohibited. In addi- tion, testing is required both to substantiate label- ing claims and to demonstrate safety. These re- quirements should be assumed to apply to the substances and products discussed in this section unless otherwise stated. Food for Humans Under the law, a food additive is defined as a food substance that is not “generally recognized as safe” (as defined in the Federal Food, Drug, and Cosmetic Act) and that has not previously been approved as safe by FDA or USDA between 1938 and 1958. No such additive may be used until it has been subjected to extensive toxicity testing, a food additive petition has been submitted to FDA, and FDA has approved the additive as safe and promulgated a food additive regulation govern- ing its use. Ch. 7—The Use of Animals in Testing G 159 The safety of an additive is established by evalu- ating data from combinations of tests. The amount of testing that must be performed is determined by the amount of information already available and the degree of toxicological concern. Guidelines have been developed (Toxicological Principles for the Safety Assessment of Direct Food Additives and Color Additives Used in Food (56), known as the Red Book) that contain detailed procedures ade- quate to meet minimum requirements. However, manufacturers are permitted to modify the test- ing as they deem necessary as long as the data are equal to or better than what would be derived by using the guidelines. Food safety has been important to FDA since the early 1900s. However, the use of animals to test food additives was not begun until the passage of the 1954 Pesticide Chemical Amendments and the 1958 Food Additive Amendments. The most fa- mous amendment, sponsored by Delaney, required that any additive that induces cancer in animals or in humans be banned. Drugs for Humans FDA regulates all human drugs, including bio- logical ones. The 1938 amendments of the Federal Food, Drug, and Cosmetic Act require drug man- ufacturers to submit evidence to FDA that a new drug is safe prior to commercialization. Safety evaluations are primarily based on preclinical ani- mal testing and subsequent clinical testing in hu- mans. In 1962, amendments to the act required that the effectiveness of a new drug also be dem- onstrated, and this is accomplished through clini- cal testing. The requirements to use animals to test new hu- man drugs depend on the proposed scope of clini- cal investigation and on the drug’s anticipated use, Determining the best procedures for testing is com- plex because of the variation that exists in the use and activity of drugs. Testing must be tailored to each drug and specific requirements are deter- mined by considering the route of administration, the target population, the length of treatment, and the relationship of the drug to others already in use. In addition to the formal procedures required under the Good Laboratory Practices regulations (see ch. 13), guidelines are available to aid manu- facturers in designing test protocols. Manufac- turers commonly discuss their programs with FDA before and during testing, as well as afterward. Guidelines are available for tests required for drugs intended for oral, parenteral, dermal, inhala- tion, ophthalmic, vaginal, and rectal uses, and those used in combination. Duration of proposed human administration is a major factor for determining the particular animal test species, the number of animals, and the duration of the test. Biological products—any virus, therapeutic se- rum, toxin, antitoxin, vaccine, blood, blood com- ponent or derivative, or allergenic product used to prevent, treat, or cure human diseases or in- juries—are regulated under the Public Health Serv- ice Act and the Federal Food, Drug, and Cosmetic Act. As with drugs, before a new vaccine or aller- genic can be marketed, the manufacturer must provide test data to show that the product is safe and effective. FDA Center for Drugs and Biologics licenses the product and the manufacturing facil- ity. For some products, tests are performed on each batch to assure that standards of potency and safety are met prior to release. For most of these, requirements are specified in the Code of Federal Regulations. Food and Drugs for Animals Food for pets, food-producing animals, and any other animal is subject to the same basic regula- tory requirements as food for humans, with the addition of testing in the target species. The Federal regulation of animal drugs, medi- cated feeds, and feed additives began under the 1938 act. The 1968 Animal Drug Amendments con- solidated animal food and drug laws, keeping the 1962 standard for safety and effectiveness. The basic intent of these statutes and their resultant regulations is to avoid using substances that may leave harmful residues in animal products intended for human consumption, and to avoid harm to food-producing and other animals. FDA regulates all animal drugs except those derived from living matter (biologic), which are regulated by USDA. Animal drugs may not be mis- branded or adulterated. Testing is done to sub- stantiate labeling claims and to prove safety. A “new 160 G Alternatives to Animal Use in Research, Testing, and Education animal drug” is defined as one not “generally rec- ognized as safe” and effective. It must be tested to demonstrate both safety and effectiveness be- fore marketing is permitted. Medical Devices Extensive regulatory provisions relating to the safety and effectiveness of medical devices for hu- mans were enacted in 1976 (21 U.S.C. 321). For devices available before then, FDA may at anytime require that proof of safety and effectiveness be submitted. For post -1976 medical devices that are substantially equivalent to those for humans be- fore 1976, the same rule applies. But for those not substantially equivalent, testing must be under- taken to prove both safety and effectiveness, a pre- market approval application must be submitted to FDA, and FDA must approve the device as safe and effective before it maybe marketed. Because of the diversity of medical devices, the testing re- quired is tailored specifically to the product in- volved and there are relatively few guidelines. As the materials involved and methods of appli- cation are often unique, determining the safety of medical devices from the standpoint of toxicity presents special problems. Consequently, recom- mendations for specific tests are based on an evalu- ation of the following factors: G G G G G the population for which the device is in- tended, with special reference to the target group’s age and sex, and the benefit to be derived; the intended use of the device and its poten- tial to contact the body or, for leachable or absorbable materials, to be distributed in the body; the location of the device in the immediate vicinity of various organs that might be ad- versely affected by its presence; the size of the device and the amount of leachate potentially available to the body; and chemical or toxicological information suggest- ing the potential for adverse toxic effects, such as when a leachable substance belongs to a chemical family that contains compounds with known potential for these effects. Requirements for testing ophthalmic devices and products, color additives used in devices, and fe- male contraceptive devices are more standardized. For color additives used in devices, the same types of tests are recommended as for color additives used in foods. For female contraceptive devices, the requirements are the same as those used for contraceptive drugs. Medical devices for animals may not be mis- branded or adulterated either. Testing can involve animals and is undertaken to substantiate label- ing claims and safety. The law does not require premarket approval of such devices, however. Cosmetics Although the law prohibits misbranding or adul- teration of cosmetics, FDA has no statutory au- thority to require testing of cosmetics for safety (other than their color additives) before they are marketed. However, animal testing is commonly undertaken to substantiate labeling claims and, by regulation, FDA has stated that any cosmetic with an ingredient that has not been substantiated for safety or that itself has not been substantiated for safety in its final product form must bear a promi- nent label declaration that the safety of the prod- uct has not been determined. Color Additives The law requires that any color additive used in food or drugs for animals or humans, in medi- cal devices for humans, or in a cosmetic must be proved safe; must be the subject of a color addi- tive petition filed with FDA; and must be deter- mined by FDA to be safe before it is used (2 1 U.S.C. 321 et seq.). Color additives in use at the time of the enactment of this provision in 1960 have been placed on a provisional list and are subject to the same requirements for testing and approval as post-1960 color additives. Radiological Products The law authorizes FDA to regulate the emis- sion of radiation from electronic products through the establishment of performance standards and a program of research and other activities to min- imize human exposure. Testing on electronic prod- uct radiation is undertaken both in relation to pro- posed and promulgated performance standards and to determine other aspects of potential haz- ard for humans from such emissions. Ch. 7—The Use of Animals in Testing G 161 Environmental Protection Agency In fulfilling its statutory responsibilities, EPA uses toxicity data derived from animal testing in a variety of ways. EPA has the authority to require such data be submitted under laws it administers, but data are obtained through other means as well. They are submitted voluntarily by those who con- duct or sponsor testing and are obtained from the open literature, from other government agencies, through contracts and grants, and from EPA lab- oratories. This section describes the regulatory programs for which animal testing data are needed and the authorities under which existing data or testing can be required. (EPA’s testing guidelines are de- scribed in app. A.) Pesticides The Federal Insecticide, Fungicide, and Rodenti- cide Act (FIFRA) (Public Law 92-516, 7 U.S.C. 136 et seq.) is designed to protect human health and the environment from adverse effects of pesticides while allowing the benefits of their use. This is done by granting or denying registrations; approv- ing labeling; setting maximum residue levels on or in raw agricultural commodities; and establish- ing procedures for safe application, storage, and disposal. In registering the approximately 50,000 formulations of “pesticide products,” EPA uses comprehensive registration standards that include animal testing data, as well as physical properties, analytical methods, and descriptions of manufac- turing and use conditions. EPA also relies on animal toxicity data when it issues emergency exemptions, experimental-use permits, and temporary tolerances for experi- mental purposes in response to unexpected and temporary food or health emergencies. Emergency exemptions may be granted to State or Federal agencies for uses not included in the registration. Experimental-use permits allow large-scale test- ing of new pesticides or new uses of a registered pesticide. The Agency’s Data Requirements for pesticide Registration specify the kinds of material that must be submitted to EPA to support registration of each pesticide under Section 3 of FIFRA. EPA uses the information to determine the identity and com- position of pesticides and to evaluate their poten- tial adverse effects and environmental fate. Tests are either ‘(required” or “conditionally required” depending on such factors as the results of pre- liminary tests, whether the pesticide use is for a food crop, whether the use is experimental, where and how the pesticide is to be applied, and the fate of the pesticide residue. Certain tests are required for new products, and guidelines for conducting these tests have also been developed (40 CFR 158, 49 FR 42856). Many are conditionally required through “tiered testing,” whereby the results of the first tier of tests determine the need for addi- tional ones. Three tiers have been described. There is some flexibility in the application of these testing requirements, but EPA is to be con- sulted if test protocols other than those described are to be used. Additional flexibility in the testing requirements is available through EPA’s proce- dures for waivers and for minor uses (40 CFR 158). Virtually all data are submitted in the context of obtaining, maintaining, or renewing a registra- tion. Another requirement is that the registrant must submit any health or safety information that would be of interest to EPA regarding a registered pesticide. This includes the submission of ongoing or completed studies for pesticides subject to regis- tration standards, cancellation, or review; incidents involving adverse effects to human or nontarget organisms resulting from exposure; or incidents regarding lack of efficacy that could indirectly pose a hazard to human life. Industrial Chemicals The Toxic Substances Control Act (TSCA) (15 U.S.C. 2601) authorizes EPA to regulate chemical substances that present an “unreasonable risk” of injury to health or the environment and to require the reporting or development of data necessary for EPA to assess risks posed by a given substance. Toxicological testing data derived from animals form the basis for risk assessment and subsequent regulatory actions taken by EPA in implementing TSCA. If a chemical substance presents an unreason- able risk, EPA can regulate its manufacturing, proc- essing, distribution in commerce, use, or disposal. 162 G Alternatives to Animal Use in Research, Testing, and Education Such regulatory actions would be based on toxic- ity data and exposure data, as well as on data re- garding the beneficial uses of the substance. Reg- ulation can be in the form of prohibiting or limiting certain actions, requiring warnings or instructions for use, or requiring the submission or retention of certain records. If EPA has reason to believe that a substance presents an unreasonable risk but the agency lacks sufficient information to make such a finding, it can require reporting of existing toxicity or ex- posure data. EPA can also require that a substance be tested in animals for specific toxic effects. Under TSCA, EPA has authority to require test- ing of industrial chemicals if testing is needed to perform a risk assessment. To aid in identifying relevant chemical substances, TSCA authorized an interagency testing committee to make sugges- tions. EPA must consider these suggestions and either initiate rulemaking or publish reasons for not doing so. TSCA requires that 90 days before the manu- facture or import of a “new” chemical (a chemical not on the TSCA Inventory of Chemical Substances) can begin, a Premanufacture Notification must be submitted to EPA. The submitters must provide all information in their possession or control re- lated to health or environmental effects or to ex- posure. EPA can also require hazard or exposure information for substances already in commerce. Air The Clean Air Act (42 U.S.C. 7401 et seq.) requires the Federal and State Governments to take cer- tain actions to improve or maintain the quality of ambient air. Animal testing data support various activities under the act. EPA designates certain sub- stances as “criteria pollutants” and establishes na- tional standards for ambient air based on toxicity and other concerns. Under Section 112, EPA also designates certain very toxic pollutants as “haz- ardous” and establishes standards for their emis- sion or other control. For registrations of any fuel or fuel additive, the EPA Administrator may require the manufacturer to conduct tests to determine whether there are potential short- or long-term health effects. Tests may be for acute effects, chronic effects, immuno- toxicity, carcinogenicity, teratogenicity, or muta- genicity. Radiation EPA’s authority over radiation was delegated in the President’s Reorganization Plan of 1970 (35 FR 15623), under which EPA makes recommendations to other Federal agencies (the Nuclear Regulatory Commission, the Department of Energy, and OSHA) regarding acceptable levels of emissions for the byproducts of producing fuel-grade uranium and from other low-level wastes. Most of the data used to develop regulatory standards were gathered from humans inadvertently exposed to radiation, but data from animals are used for genetic and other effects, dose-response relationships, and me- tabolism. Water The Clean Water Act (33 U.S.C. 466) requires Federal and State efforts to restore and maintain the integrity of U.S. waters. Data needed to fulfill these requirements are obtained primarily from testing fish and other aquatic organisms. The 1977 amendments to the act listed toxic sub- stances that are commonly referred to as the 126 priority pollutants, primarily because of their toxic effects on humans and animals. These are con- trolled through nationally uniform limitations on the effluents containing them. Water Quality Cri- teria have also been promulgated for permissible ambient concentrations of these substances and are used to establish State water quality standards. Other toxic chemicals will also be regulated under the Clean Water Act. The Clean Water Act calls for National Water Quality Criteria to be derived. The complete data set is developed by conducting a series of acute and long-term bioassays using organisms from at least eight different families. Acute tests are re- quired on a salmonid, another family belonging to the class Osteichthyes (bony fish), and another representative of the phylum Chordata. The long- term tests required are chronic tests with one spe- cies of fish and a bioconcentration test with one aquatic species, Ch. 7—The Use of Animals in Testing . 163 In 1982, EPA published a Water Quality Stand- ards Handbook that provides guidance for develop- ing site-specific water quality criteria that reflect local environmental conditions based on toxicity testing in fish. The Safe Drinking Water Act (42 U.S.C. 300) is designed to protect public drinking water supplies through minimum national standards that are im- plemented by the States. Under this act, EPA also regulates the underground injection of fluids and other imminent or substantial hazards to drink- ing water. In addition, health advisories are pre- pared on specific problems. Primary drinking water regulations are devel- oped for certain contaminants that may have ad- verse effects on human health, Maximum contami- nant levels are established or health advisories published using mammalian testing data. EPA’s authority over groundwater is based on a number of the laws that the agency administers. The management of groundwater is a joint Fed- eral and State responsibility, but EPA provides tech- nical assistance to State agencies and prepares advisories dealing with common problems that en- danger groundwater. To some extent, these sup- port activities rely on toxicity data. Because groundwater is the source of drinking water for about half the U.S. population, the iden- tification and characterization of groundwater problems is an important part of the drinking water program. Over 700 synthetic organic chemicals have been identified in various drinking water sup- plies. Some epidemiologic evidence is available, and more is being collected to help characterize the toxicity of these contaminants, but animal testing data are mainly used. Solid Waste The Resource Conservation and Recovery Act (RCRA) (Public Law 94-580, 48 U.S.C. 6901) pro- tects public health and the environment by con- trolling the disposal of solid waste and by regulat- ing the management and handling of hazardous waste materials. EPA is authorized to develop reg- ulations governing the generation, transportation, treatment, storage, and disposal of hazardous wastes. These regulations, in addition to State laws on waste, are enforced by the States. Animal testing is used to identify hazardous wastes. Toxicity is one of the criteria. RCRA regu- lations list chemicals that have been determined to be hazardous and processes that are presumed to generate hazardous waste. Analytical proce- dures for determining the contents of waste are also described, as are criteria for determining whether the contents are toxic or otherwise haz- ardous. When information does not exist for cer- tain wastes, EPA must develop it. RCRA does not require those who generate hazardous waste to test the toxicity of the waste. Because RCRA deals with solid waste, the pre- dominant health problems arise from the leach- ing of waste from disposal sites. EPA is in the process of selecting and validating tests for char- acterizing waste. These will look for acute and chronic effects on aquatic animals, primarily fat- head minnows. Partial or full life-cycle bioassays and fish bioaccumulation tests will also be required. The potential hazards to humans are character- ized with several mutagenicity tests. Data from tests with humans and animals are used under RCRA to develop “acceptable daily in- take” levels that are regulated under the act. Be- cause of the nature of exposure to these wastes, data from short-term and dermal tests are not used. Superfund The Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) (42 U.S.C. 9601), known as Superfund, authorizes the Federal Government to clean up or otherwise re- spond to the release of hazardous substances or other pollutants that may endanger public wel- fare. The most significant activity under CERCLA, from the standpoint of animal testing, is the desig- nation of hazardous substances. Substances des- ignated as hazardous under certain sections of other laws (TSCA, the Clean Air Act, the Clean Water Act, and RCRA) are also considered haz- ardous under CERCLA, and the EPA Administra- tor is to designate specific amounts of hazardous substances to be “reportable quantities, ” based in part on toxicity data. One activity under CERCLA that diminishes the need for animal testing (because it assembles data on humans) is the compilation of a Toxic Substances — 164 G Alternatives to Animal Use in Research, Testing, and Education and Disease Registry under the Department of Health and Human Services. This registry will track persons exposed to hazardous substances, along with the medical testing and evaluation that fol- lows the exposure. Consumer Product Safety Commission The CPSC administers the Consumer Product Safety Act (I5 U.S.C. 401 et seq.), the Federal Haz- ardous Substances Act (15 U.S.C. 1261 et seq.), the Poison Prevention packaging Act (15 U.S.C. 1471 et seq.), and the Flammable Fabrics Act (15 U.S.C. 1191 et seq.). The Consumer product Safety Act empowers CPSC to prevent unreasonable risks of injury from consumer products. Included are both the risk of acute and chronic toxicity and the risk of physical injury. Under this statute, industry regularly con- ducts animal toxicity testing to determine the safety of consumer products. The Federal Hazardous Substances Act provides for the regulation of hazardous substances used in or around the household. These are defined as any substance or mixture that is toxic, corrosive, flammable, or combustible, that is an irritant or a strong sensitizer, or that generates pressure through decomposition, heat, or other means, if such substance may cause substantial personal in- jury or illness during customary or reasonably foreseeable handling or use. Unlike its usual meth- od of letting a regulatory agency or the manufac- turer determine what kind of testing is needed to determine safety, in this act Congress defines a “highly toxic” substance in terms of the results of the LD50 test and requires certain labeling when the LD50 is less than 50 mg/kg body weight, 2 mg/l of air inhaled for an hour or less, or 200 mg/lcg of dermal exposure for 24 hours or less. Although the act does not literally require that these tests be done, a manufacturer cannot know whether they are in compliance with the act unless they perform the tests. CPSC has issued regulations re- garding testing requirements needed to determine whether a substance is a skin or an eye irritant (16 CFR 1500). The Flammable Fabrics Act authorized regula- tion of wearing apparel and fabrics that are flam- mable. Industry regularly conducts animal test- ing to determine the toxicity of substances applied to fabric in order to reduce or eliminate flamma- bility. Department of Labor The Occupational Safety and Health Act of 1970 (29 U.S.C. 651 et seq.) requires the National Insti- tute for Occupational Safety and Health (NIOSH) to conduct health hazard evaluations of the work- place (see section on Centers for Disease Control). A goal of the act is that no employee suffer diminished health as a result of conditions in the workplace. To this end, employers have a duty to communicate safety information about substances present in the workplace through labels, material safety data sheets, and training. Most safety test- ing is done with animals. Under the Federal Mine Safety and Health Act of 1977 (30 U.S.C. 801 et seq.), employers must determine whether substances found or used in mines are potentially toxic at the concentrations at which they occur. Department of Transportation The Hazardous Materials Transportation Act (49 U.S.C. 1801 et seq.) requires that any materials shipped in interstate commerce be properly la- beled and contained in a manner reflecting the degree of hazard present. DOT requires that acute toxicity studies be carried out on substances not already classified or for which toxic effects to hu- mans or test animals are not already known. A substance would be treated as a class B poison (and thus as presenting a health hazard during trans- portation) if its administration to 10 or more rats at a single dose of a specified amount (orally, der- mally, or by inhalation) killed at least half the ani- mals within 48 hours. Analogous authority exists for the U.S. Coast Guard under the Dangerous Cargo Act (46 U.S.C. 179) and the Ports and Water- ways Safety Act (33 U.S.C. 1221 et seq.). Ch. 7—The Use of Animals in Testing G 165 Department of Agriculture USDA administers the Virus-Serum-Toxin Act of 1913 (21 U.S,C. 151 et seq.), under which it licenses animal biologics. The regulatory require- ments are similar to those administered by FDA for other animal drugs. Animal testing is under- taken to substantiate labeling claims for animal drugs and to prove their safety. The testing re- quired by USDA for proof of safety and effective- ness of these animal biological drugs is extensive. Under a series of statutes, USDA exercises close inspection authority over the processing of meat, poultry, and eggs for human consumption. These statutes prohibit any misbranding or adulteration, Testing is required to substantiate labeling claims. Although most safety issues are handled by FDA, testing may also on occasion be required by USDA to demonstrate safety under particular conditions. USDA administers a number of statutes designed to control and eradicate disease in plants and ani- mals. This authority extends from research through to control of interstate and foreign transportation. Substantial testing is undertaken by USDA in pur- suing these statutory mandates. Centers for Disease Control The Public Health Service Act (42 U.S.C. 201 et seq.) authorizes CDC to take appropriate action to prevent the spread of communicable disease. Pursuant to this authority, CDC regulates any agent that could cause such illnesses. CDC uses animal data to determine the agents that should be reg- ulated. Under the authority of the Occupational Safety and Health Act of 1970, the National Institute for Occupational Safety and Health, a component of CDC, develops and periodically revises recommen- dations for limits of exposure to potentially haz- ardous substances or conditions in the workplace. When morbidity cannot be explained on the basis of current toxicological knowledge, NIOSH must design toxicological investigations to discover the cause. Such occupational hazard assessments are based on data on humans and animals collected by NIOSH. Federal Trade Commission The Federal Trade Commission Act (15 U.S.C. 41 et seq.) prohibits any advertisement that is mis- leading in a material respect. FTC has adopted the position that an advertiser must have adequate sub- stantiation for any claims relating to safety or ef- fectiveness. Thus, manufacturers and distributors regularly test their products, using data on humans and animals to substantiate their claims. STATE USES OF ANIMAL TESTING DATA States engage in a variety of regulatory activi- Pesticide Registrations ties that rely directly or indirectly on animal test- ing data. One of the most important longstanding All States are required to register pesticides un - uses is the registration of pesticides. Air, water, der Section 24 of FIFRA. Most States have 5,000 and waste have also been the subject of State leg- to 10)000 pesticides registered and grant 5 to 10 islation in recent years. State laws often use ani- emergency exemptions per year. As part of the mal testing data for the identification and classifi- registration process, States receive animal testing cation of substances for control. Several States have data for evaluation. Much of the time, the infor- also enacted right -to-know laws that may give peo- mation is required only in summary form, unless ple greater access to testing data, although such the State specifically requests the raw data. The legislation does not necessarily affect the amount data are usually obtained directly from the regis- of testing done. trant to avoid possible delays or confidentiality 166 G Alternatives to Animal Use in Research, Testing, and Education problems. Although States generally rely on EPA’s assessment of data for registration purposes, they regularly review it for emergency exemptions and special local needs (22). California and Florida have the largest pesticide programs. These States also have the authority to require additional testing (e.g., field testing locally). In addition, California also recently passed a law giving its Director of Food and Agriculture the au- thority to require data for which EPA has granted a waiver or exemption (e.g., experimental-use per- mits). California law also requires that data gaps for 200 pesticides be filled and that the first re- port of an injury to a worker exposed to a pesti- cide be reported to the Health Department (Cali- fornia Food and Agriculture Code, Div. 7, ch. 2). Identification and Classification of Toxic Substances Identification and classification of substances is an important function in most environmental laws. Such activities take place under each Federal envi- ronmental statute. Coordination among offices in EPA or with other agencies is common. State agen- cies also coordinate these activities with their Fed- eral counterparts. Sometimes, Federal law or regulations are sim- ply adopted by a State and remodified. For exam- ple, certain provisions of the New York and Florida regulations governing hazardous wastes incor- porate, by reference, EPA regulations appearing at 40 CFR 261 and its Appendices (New York Com- pilation of Rules and Regulations, Title 6, ch. 366). These regulations list hazardous waste and their constituents, provide analytical procedures to de- termine the composition of a waste so that it can be classified, and provide for variances from these regulations that may be granted by EPA’s Admin- istrator. Much more common are statutes that in- corporate Federal laws and regulations and that add other requirements or combine Federal re- quirements in new ways. The Wisconsin Pollution Discharge Elimination Law (Wisconsin Statutes Annotated, ch. 147) adopts EPA effluent limitations, effluent standards, and prohibitions. In addition to substances already regulated by EPA, Wisconsin effluent limitations apply to all toxic pollutants “referred to in table 1 of committee print number 95-30 of the Commit- tee on Public Works and Transportation of the U.S. House of Representatives.” Additional pollutants are to be identified under Section 147.07 of the Wisconsin law. The Colorado Hazardous Waste Management Regulations (Code of Colorado Regulations, Title 5, ch. 1007) adopt EPA toxicity provisions of 40 CFR 261 but include “any other substance which has been found to be fatal to humans at low doses, or in the absence of human data, has an oral LD50 in the rat of 50 mg/kg or less, an inhalation LC50 (lethal concentration) in the rat of 2 mg/l or less, or a dermal LD50 in the rabbit of 200 mg/kg or less .“ The Texas Water Quality Acts (Texas Water Code, Title 2, chs. 5, 26, 30, 313) use several Fed- eral laws to classify a substance as hazardous: CERCLA; the Water Pollution Control Act; the Solid Waste Disposal Act; the Clean Air Act; and TSCA. If it is hazardous under any one of these laws, it is hazardous for purposes of Texas law. Under Oregon Hazardous Waste Management Regulations (Oregon Administrative Rules, ch. 340, div. 62, 63), a substance is considered toxic if it is a pesticide or pesticide manufacturing residue and has one of the following properties: G oral toxicity in a 14-day test with an LD50 less than 500 mg/kg, . inhalation toxicity over 1 hour with an LC50 less than 2 mg/l gas or 200 mg/m3 dust or mist, G dermal toxicity over 14 days with an LD50 less than 200 mg/kg, or G aquatic toxicity over 96 hours at an LC5o less than 250 mg/l. It would also be considered toxic if it contains a car- cinogen identified by OSHA at 29 CFR 1910.93(C). Washington Dangerous Waste Regulations (Wash- ington Administrative Code, Title 173, ch. 303) re- quire the polluter to use EPA toxicity information, EPA’s Spill Table, NIOSH’s Registry of Toxic Effects of Chemicals (see ch. 10), and any other reason- ably available sources to determine if a pollutant is toxic. Carcinogens are identified by an Interna- tional Agency for Research on Cancer finding that a substance is a positive or suspected human or animal carcinogen. Additional criteria are provided Ch. 7—The Use of Animals in Testing G 167 in the Toxic Category Table, which contains five categories of hazards based on an LC50 test for fish, an oral LD50 for rats, an inhalation LC50 for rats, and a dermal LD50 for rabbits. Some State laws do not explicitly provide for har- monization with Federal requirements regarding the identification and classification of toxic sub- stances. Under the California Air Pollution Laws (California Air Pollution Control Laws, 1979 Edi- tion), the California Air Resources Board and the State Department of Health Service are to prepare recommendations for substances to be regulated and to consider all relevant data. State officials may PRODUCT LIABILITY Toxicological testing and research play an im- portant role in the law of product liability. Manu- facturers are responsible for knowing what dan- gers their products may present and must pay for any damages these products cause. Animals are used to discover possible dangers, and courts may award damages to a party whose injuries could have been prevented with additional testing or re- search (see ch. 11). This discussion of product liability law focuses primarily on drugs because animal use plays such an important role in determining safety. Drugs are also an interesting case study because they are re- viewed for safety and effectiveness by the Food and Drug Administration before they are mar- keted, and yet satisfying FDA’s testing require- ments does not necessarily fulfill the manufac- turer’s duty to test. The Manufacturer’s Duty to Produce a Safe Product In general, a manufacturer has a duty to pro- duce a safe product with appropriate warnings and instructions. This is based on an individual’s responsibility to exercise care to avoid unreason- able risks of harm to others. The duty extends to all persons who might foreseeably be injured by the product manufactured. Under the Uniform Commercial Code—a law governing commercial transactions involving goods, which varies only request information on any substance under evalu- ation, although they do not have the authority to require testing. However, any person who wishes the board to review one of its determinations must specify additional evidence that is to be consid- ered. Similarly, the California Hazardous Waste Control Act (California Health and Safety Code, Div. 20, chs. 6.5, 1039; California Administrative Code, Title 22, div. 4, ch. 30) directs the California Department of Health Services to prepare lists of hazardous waste and extremely hazardous waste and to develop regulations for their management. CONSIDERATIONS slightly from State to State—failure to produce a safe product results in liability for the manufac- turer for the damages thereby caused. Generally, an injured plaintiff must prove that the drug in question was unreasonably danger- ous, that the defect existed at the time the drug left the manufacturer’s control, that the consumer was injured or suffered damages from the use of the drug, and that the defect in the drug was the proximate cause of the injury (13,37). product liability law inmost jurisdictions follows the “strict liability” standard—that is, no matter how careful a manufacturer is, it is liable for inju- ries caused by its products. Some jurisdictions only hold the manufacturer to a high standard of care, and many that do have strict liability standards also have exceptions. One exception is for drugs that are necessary but that cannot be made safe. Some have a high risk of harmful side effects but treat conditions that are even more harmful if left untreated, such as rabies (57). (Conversely, when the advantages a product offers are small, such as where vaccines were combined instead of using multiple injections, the manufacturer is more likely to be held liable (51).) Another exception is for products for which no developed skill or foresight could have avoided the harm (14). Even though a toxic effect might not have been tested for using existing methods with animals, a manufacturer must not ignore in- 168 G Alternatives to Animal Use in Research, Testing, and Education juries its product may cause after marketing (13). Similarly, if a new test becomes available, the man- ufacturer may be required to use it (14,29,47). Methods of Testing Required A manufacturer must normally use the safest and most effective testing method available. Thus, when monkeys provided the only reliable means for testing polio vaccine, they had to be used to test individual batches of drugs, despite the diffi- culty and expense of obtaining them (20). Although no cases could be found pertaining to drugs, this standard might not apply when testing is imprac- tical in relation to the risk of harm (30,48). Testing must reflect conditions of actual use as closely as possible. Thus, where the drug DES was to be used on pregnant women, the manufacturer should have tested pregnant animals and was held SUMMARY AND The most widespread kind of testing with ani- mals is conducted for the elucidation of toxicity from drugs, chemicals, and so forth. Toxicology has advanced with the growth of the synthetic chemical industries and the use of chemicals in consumer products. Toxicological testing is used in the assessment of hazards and the management of health risks to humans. The use of animals for such testing did not become common until a few decades ago; it now accounts for several million animals per year. Many toxicological tests are standardized to aid in the comparison of results and because they have been shown to be acceptable tools for measuring certain phenomena. Most of the standard tests are descriptive in that they indicate an end result but do not necessarily elucidate the processes leading to it. Knowledge of the mechanisms by which a toxic effect occurs allows much greater reliabil- ity in extrapolation to humans. The design of a test involves many trade-offs. The choice of species is affected by its physiologi- cal similarity to humans, its cost and availability, and the amount of data for other substances avail- liable for cancer in offspring (3). Several smokers have tried to recover from cigarette manufactur- ers. They have been denied recovery to date be- cause when they started smoking, the risk of can- cer had not been demonstrated (28,40)43). A judge or jury would normally decide whether testing was adequate, but if there was a failure to comply with regulatory requirements, this would normally prove insufficient testing (16)33,39). How- ever, compliance with such requirements would not prove that testing was adequate (14). In addition to examining what tests were done, the judge or jury might look at the adequacy of the test protocols themselves. For example, the in- jured plaintiff might argue that the number of test animals was not large enough to determine if a risk was presented (11) or that the conditions un- der which the drug was tested did not represent actual use conditions (49,51). CONCLUSIONS able for comparison. The route of exposure, dura- tion of exposure, and size of doses are affected by the possible nature and extent of exposure in humans, by the dose needed to produce a meas- urable toxic effect, and by convenience. Expected variability in the toxic response governs the num- bers of animals used. Commonly used tests include the following: G G G G G acute toxicity—a single dose at high enough concentrations to produce toxic effects or death, often used to screen substances for rela- tive toxicity; eye and skin irritation—usually a single ex- posure, generally used to develop warnings for handling and predict accidental exposure toxicity; repeated-dose chronic toxicity—repeated exposure for periods ranging from 2 weeks to more than a year, used to determine the possible effects of long-term human exposure; carcinogenicity—repeated exposure for most of lifespan, used to detect possible hu- man carcinogens; developmental and reproductive toxicity– Ch. 7—The Use of Animals in Testing G 169 a variety of exposures to determine the pOS - sible production of infertility, miscarriages, and birth defects; . Ž neurotoxicity--a variety of doses and routes to determine toxic effects to nerves, with toxic end points such as behavioral changes, lack of coordination, or learning disabilities; and . mutagenicity—a variety of methods for de- termining if genetic material of germ or so- matic cells has been changed. To aid in the design of tests and in the extrapola- tion of results to humans, studies are sometimes done to determine the mechanisms by which tox- icity occurs or to characterize the processes by which the test substance enters, is handled, and leaves the body. The Federal Government has considerable im- pact on testing practices through a variety of laws and regulations. Sometimes testing is required for premarket approval; more often, it is implied by requirements for safe and effective products. In only a handful of instances, such as the Federal Hazardous Substances Act administered by the Consumer Product Safety Commission and the Hazardous Materials Transportation Act admin- istered by the Department of Transportation, do Federal statutes explicitly require animal testing. The four agencies with the largest roles are the Food and Drug Administration, the Environmental Protection Agency, the Consumer Product Safety Commission, and the occupational Safety and Health Administration. FDA uses animal testing data in the approval of food additives, drugs, bio- logics, medical devices, and color additives for hu- mans and animals. EPA and State Governments use such test results in the registration of pesti- cides and the regulation of industrial chemicals, as well as in the protection of water and air and in the regulation of waste disposal. CPSC relies on animal data in identifying and regulating risks to consumers, while OSHA indirectly uses them in requiring employers to maintain a safe workplace. Testing also plays an important role in the liabil- ity of a manufacturer for unsafe products. In most States, a manufacturer is responsible for any in- juries arising from use of its products, regardless of how much testing was done. Exceptions may be made where suitable tests do not exist or the product is known to present risks but those risks are preferable to the harm that would occur with- out the product, as in the case of rabies vaccine. Despite the problems of extrapolating to humans and other shortcomings of animal testing tech- niques, the use of animals in testing is an integral part of the Nation’s attempt to protect human health. Ideally, as the practice of toxicology ad- vances, there will be less emphasis on numerical values in certain tests and more consideration of the mechanisms by which toxic effects occur. CHAPTER 7 REFERENCES 1. Altman, P.L. (cd.), Padmlogyof Laboratory Mice and Rats (New York: Pergamon Press, 1985). 2. Berry, C. L., “Reproductive Toxicity, ” Animals and Alternatives in Toxicity Testing, M. Balls, R.J. Rid- den, and A.N. Worden (eds.) (New York: Academic Press, 1983). 3. Bichler v. Eli Lilly and Company, 436 N. Y.S.2d 625 (1981). 4. Bridges, J. W., Chasseaud, L. F., Cohen, G. M., et al., “Application of Pharmacokinetics, ” Animals and Alternatives in Toxicity Testing, M. Balls, R.J. Rid- den, and A.N. Worden (eds.) (New York: Academic Press, 1983). 5. Brusick, D., Principles of Genetic Toxicology (New York: Plenum Press, 1980). 6. Dewar, A. J., “Neurotoxicity, ’’Animals and Alterna - tives in Toxicity Testing, M. Balls, R.J. Ridden, and A.N. Worden (eds.) 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M., “Assessment of Mild Irritants, ’’Prin - ciples of Cosmetics for the Dermatologist, P. Frost and S.N. Horwitz (eds. ) (St. Louis, MO: C.V. Mosby, 1982). 28. Lartique v. R.J. Reynolds Tobacco Company, 317 F.2d 19 (1963). 29. Leibowitz v. ortho Pharmaceutical Co., 224 Pa. Su - per, 418, 307 A.2d 449 (1973). 30. Livesley v. Continental A40tors Corporation, 331 Mich. 434, 49 N.W.2d 365 (1951). 31. Lowrance, W .W., Of Acceptable Risk—Science and 32, 33, 34. 35. 36. 37. 38. 39. 404 41. 42. 43. Determination of Safety (Los Altos, CA: William Kaufmann, Inc., 1976). Marks, R., “Testing for Cutaneous Toxicity)” Ani- mals and Alternatives in Toxicity Testing, M. Balls, R.J, Ridden, and A.N. Worden (eds.) (New York: Aca- demic Press, 1983). McComish v. DeSoi, 42 N.J. 274,200 A.2d 116 (1964). McNamara, B. P., “Concepts in Health Evaluation of Commercial and Industrial Chemicals, ” New Con- cepts in Safety Evaluation, M .A. Mehlman, R .E. Sha- piro, and H. Blumenthal (eds.) (New York: John Wiley & Sons, 1976). Morton Grant, W., and Kern, H. L., “Cations and the Cornea,” Am. J. Ophthalmo]. 42:167-181, 1956. National Academy of Sciences, Toxicity Testing Strategies to Determine Needs and Priorities (Wash- ington, DC: National Academy Press, 1984). Nicklaus v. Hughes Tbo] Company, 417 F.2d 983 (8th Cir. 1969). Organization for Economic Cooperation and Devel- opment, Guidelines for Testing of Chemicals, and addenda (Paris: 1981). Orthopedic Equipment Company v. Eutsler, 276 F.2d 455 (4th Cir. 1960). Pritchard v. Liggett and Meyers Tobacco Company, 295 F.2d 292 (3rd Cir. 1961). Purchase, I. F. H., ‘( Carcinogenicity,” Animals and Alternatives in Toxicity Testing, M. Balls, R.J. Rid- den, and A.N. Worden (eds.) (New York: Academic Press, 1983). Roe, F.J .C., “Carcinogenicity Testing, ’’Animals and Alternatives in Toxicity Testing, M. Balls, R.J. Rid- den, and A.N. Worden (eds.) (New York: Academic Press, 1983). Ross v. Phi]lip Morris and Company, 328 F.2d 3 (8th Cir. 1964). 44. Salsburg, D. S,, “The Effects of Lifetime Feeding Studies on Patterns of Senile Lesions in Mice and Rats,” Drugs Chem, Tox. 3:1-33, 1980. 45, Salsburg, D. S., “Statistics and Toxicology: An Over- view ,“ Scientific Considerations in Monitoring and Evaluating Toxicological Research, E.J. Gralla (cd.) (Washington, DC: Hemisphere, 1981). Ch. 7—The Use of Animals in Testing s 171 46. Schaumburg, H. H., and Spencer, P., “Central and Peripheral Nervous System Degeneration Produced by Pure n-Hexane: An Experimental Study, ” Brain 99:183.192, 1976. 47. Schenebeck L’. Sterling Drug, Inc., 423 F.2d 919 (8th Cir. 1970). 48. Sieracki v. Seas Shipping Compan&v, 57 F. Supp. 724 (E. D. Pa. 1944). 49. S[romsodt v. Parke-Davis and Companvv, 257 F. Supp. 991 (D.N.D. 1966), affirmed, 411 F.2d 1390 (8th Cir. 1969). 50. Swranston, D. W., “Eye Irritancy Testing, ” Animals and Ahernatilres in Toxicitkv Testing, M. Balls, R.J. Ridden, and A.N. Worden (eds. ) (New York: Aca- demic Press, 1983). 51. Tinnerholm v. Parke-Davis and Companuv, 411 F.2d 48 (2nd Cir. 1969). 52. Tre\~an, J. W., ‘(The Error of Determination of Tox- icity, ” PI*OC. R. Soc. Lend. (Bio/.) 101:483 (1927). 53. U.S. Congress, Office of Technology Assessment, Assessment of Technologies for Determining C’an - cer Rjsks From the Environment,OTA-H-138 (Wash- ington, DC: U.S. Government Printing Office, June 1981). 54. U.S. Environmental Protection Agency, “Acute Ex- posure: Oral Toxicity ,“ Offi”ce of Toxic Substances Health and Environmental Effects Test Guidelines (Washington, DC: updated october 1984). 55. U.S. Executive Office of the President, Office of Science and Technology Policy, “Chemical Carcino- gens: A Review of the Science and Its Associated Principles, ” Mar. 14, 1985. 56. U.S. Food and Drug Administration, Toxicological PrjncjpJes for the Safetuv Assessment of Direct Food Additiies and Color Additit’es Used in Food (Wash- ington, DC: Bureau of Foods, 1982.) 57. U.S. v. An Artjcle of Drug–Bacto Unidisk, 394 U.S. 784, 792 (1968). 58. Weigland, D. A., Haygood, C., and Gay]or, J. R., “Cell Layer and Density of Negro and Caucasian Stratum Corneum, ” .J. invest. Derznato). 62:563-568, 1974. Chapter 8 Alternatives to Animal Use in Testing Queen: I will try the forces Of these compounds on such creatures as We count not worth the hanging, but none human . . . Cornelius: Your Highness Shall from this practice but make hard your heart. Shakespeare, Cymbeline Act I, Scene VI The experimental means to be used for safety evaluations is left open to suggestion. As unorthodox as this might sound, leaving such means open for consideration is the best solution. Safety evaluations should not be based on standard, specified series of tests. They are best approached by first raising all pertinent safety questions and then searching for the experimental means to provide the best answers. Under such circumstances, even the standard LD test might on occasion be the best experimental means to resolve outstanding satfety questions. Constantine Zervos Food and Drug Administration Safety Evacuation and Regulation of Chemicals 2, D. Homburger (cd.) (Base]: Karger, 1985) —.— CONTENTS P a g e ‘ Continued, But Modified, Use of Animals in Testing . . . . . . . . . . . . . . . . . . . . . ..175 Avoiding Duplicative Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 176 Reducing Pain and Distress . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177 Use of Living Systems in Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177 In Vitro Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177 Nonanimal Organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .179 Use of Nonliving Systems in Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180 Chemical Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180 Mathematical and Computer Models . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180 Epidemiologic Data on Humans . . . . . . . . . . . . . . . . . . . . . . . . . . ..:...... .181 The LD50Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...181 Using Fewer Animals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .182 The Limit Test and Other Refinements . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..182 In Vitro and Nonanimal Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .182 Skin and Eye Irritation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ]83 In Vitro Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .183 Chick Embryo . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183 Repeated-Dose Toxicity Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184 Hepatotoxicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ......185 Neurotoxicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185 Mutagenicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185 Microorganism Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....186 In Vitro Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186 Tests Using Insects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186 Carcinogenicity ...,.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..187 The Ames Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .187 Use of the Ames Test in a Battery of tests . . . . . . . . . . . . . . . . . . . . . . . . . .188 Current Trends . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...188 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190 Chapter preferences.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Table Table No. Page 8-1. The Response of Known Human Carcinogens to Rodent Carcinogenicity and Bacterial Mutagenicity Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..188 Figure Figure No. Page 8-1. Chronological Sequence of Chick Embryo Chorioallantoic Membrane Assay.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..184 Chapter 8 Alternatives to Animal Use in Testing Alternatives to using animals in testing serve the same purposes that using whole animals does— protecting and improving human health and com- fort. The technologies on which alternatives are based result primarily from biomedical and bio- chemical research. Several of them are reviewed in this chapter, though they are discussed in greater detail in chapter 6. Some alternatives that might eventually replace the tests covered in chap- ter 7 are also described here. Notable progress in the move to alternatives has been achieved in certain areas (78). For example, biochemical tests to diagnose pregnancy have re- placed those using rabbits, and the Limulus ame- bocyte lysate test, which relies on the coagulation of a small amount of blood from a horseshoe crab, has replaced rabbits in testing for the presence of bacterial endotoxins that would cause fever (25,117). Many companies have modified the widely used LD5O test to use fewer animals (22) and have otherwise refined the methods used to test for tox- icity (100). Mammalian cell culture assays are used extensively in industrial laboratories for safety test- ing of medical devices (52,53) and pharmaceutical CONTINUED, BUT MODIFIED, It has been suggested that many more animals are used for testing than are needed (90) and that changes in experimental design or improved meth- ods of data analysis could substantially reduce the number of animals used. Each experiment has unique requirements (see ch. 7), and the ways in which the number of animals might be reduced will vary accordingly. Many of the methods discussed in chapter 6 for the modified use of animals in research are also applicable to testing, such as gathering more data from each animal or improving the analysis of re- sults by using random block design or covariance analysis. In random block design, animals with a particular characteristic, such as litter mates or animals of a certain size, are randomly assigned to different groups to balance whatever effect substances (1,84) and as immune response assays (97,98). The development of alternatives to animals in testing has accelerated in recent years with the establishment of programs having development and implementation of alternatives as their goal (see ch. 12). However, the barriers to adoption of these tests are more than the technical barrier of developing and validating anew technology. Test- ing is an integral part of many regulatory schemes and product liability law, and validation ultimately rests on acceptance by the scientific, regulatory, and legal communities. Public concern over animal to be increasing in tandem for product and drug safety use in testing appears with public concern . Ironically, the pub- lic’s-increasing concern for safety could lead to more testing. Yet it also provides an incentive to develop new techniques, particularly those that promise to be cheaper and faster than current whole-animal methods. A further irony is that de- veloping alternatives, as well as validating them, sometimes requires animal use. USE OF ANIMALS IN TESTING these variables might have. If the groups being dis- tributed are sufficiently large, the results can also be analyzed to determine the effect of the mask- ing variable (47). Covariance can be used to ana- lyze results when some of the experimental varia- bles are uncontrolled but known, thus estimating their effect on the results. As in research, the number of animals needed as controls can be reduced by using the same group as a control for several simultaneous experiments. A laboratory’s ability to do this will be limited by its size and the amount of lead time available to allow testing to be coordinated. Another difficulty is that environmental conditions must be exactly the same and the tests must start and finish at ex- actly the same times. The reduction in animal use that simultaneous experiments brings about is 175 176 G Alternatives to Animal Use in Research, Testing, and Education modest because the control group should be larger if it is being used in several simultaneous experi- ments (34), The use of historical data for control groups is constrained by the difficulty of exactly duplicat- ing the conditions of a study. However, the size of the groups and other controlled variables can be better planned if historical data are used to dis- cover the background incidence of specific tumors or other diseases before testing begins. This use of historic controls has been recognized by the National Cancer Institute, the world Health Orga- nization, the Canadian Government, and the now- defunct Interagency Regulatory Liaison Group (104). The Federation of American Societies for Experimental Biology has developed a data book containing such information based on the Labora- tory Animal Data Bank (see ch. 10) (2). Avoiding Duplicative Testing Animal use in testing can and has been reduced by industry and others through improved commu- nication and cooperation in the planning and exe- cution of testing, thereby avoiding unintentional duplication. Trade groups such as the Chemical Manufacturers Association, the Pharmaceutical Manufacturers Association, and the Soap and De- tergent Association play important roles in this co- ordination. The sharing of data after testing has occurred is often done for pesticides (see chs. 10 and 11). And in 1978, the Food and Drug Administration implemented a policy of permitting approval of new drug applications solely on the basis of pub- lished scientific papers (113). The possibility of an unintentional repetition of an experiment is also avoided through the work of organizations such as the Chemical Industry Institute of Toxicology (CIIT) (Research Triangle Park, NC). Using contri- butions from member companies, CIIT conducts toxicological tests and distributes the results widely. Governments contribute greatly to information sharing, which allows duplicative testing to be avoided, by providing both access to test results and information about their own planned and on- going tests. The International Agency for Research on Cancer makes it easy for duplicative carcinoge- nicity testing to be avoided by informing testing facilities and governments about planned and on- going testing, Federal and international databases and publications also contain information about planned tests and those under way (see ch. 10). Reducing Pain and Distress As with research, testing can be modified to re- duce animal pain or distress in two ways: by pro- viding relief with drugs or by changing the proce- dures so that less pain or distress is produced (see ch. 6). A third alternative might be to use a less sensitive species, but there is no method by which relative distress among species can be discerned. Relief from pain and distress is accomplished through analgesics, anesthetics, tranquilizers, or sedatives and modification of the test itself. Few pain-relieving drugs have been developed and marketed for animals. Little information is available on recommended doses (122) or on the likely effect on test results. Thus, before pain re- lief could be incorporated into a test, it would be necessary to determine the needed dose and the effect on the toxic response, thus using additional animals as well as subjecting them to pain. Several small changes that do not interfere with the experimental design can be made by an inves- tigator. Small needles can be substituted for large. Animals can be comforted by petting. Social ani- mals can be caged in groups, although there are often reasons that multiple housing cannot be used. Smaller doses can be used and tests can be ended at the earliest feasible time. Sometimes, smaller doses will actually result in increased sensitivity of the test (38). Making such changes sometimes depends on the attitude and expertise of individ- ual researchers rather than the contents of test- ing guidelines, which may not be sufficiently detailed. Ch. 8—Alternative to Animal Use in Testing . 177 USE OF LIVING SYSTEMS IN TESTING As detailed in chapter 6, two kinds of living sys- tems can reduce whole-animal use—in vitro sys- tems based on animal or human components (cell, tissue, and organ cultures) and systems based on organisms not considered animals for purposes of this report (micro-organisms and invertebrates). (Some people consider both of these in vitro system s.) In Vitro Systems Cells, tissues, and organs can be kept alive out- side a living organism and used for testing. Al- though animals are still required as a source for these in vitro systems, the animal would experi- ence distress for a much shorter time, and per- haps less distress overall, than occurs with whole- animal testing because it would be killed before any experimental manipulations were carried out. Occasionally, different cells, tissues, or organs from the same animals can be used for different inves- tigations. In addition, many fewer animals would be required for a given test, in part because varia- bility in the toxic response is smaller than it is with whole-animal tests and in part because one ani- mal can be used for multiple data points, further reducing variability. The fact that human tissues sometimes can be used confers an additional ad- vantage because the need for extrapolation from animal data is obviated. These isolated components also have disadvan- tages. They are usually unable to produce the com- plete physiologic responses of a whole organism. The components often become undifferentiated and lose their ability to perform their special func- tions when isolated from the organism, particu- larly when the sample is broken up into its con- stituent cells, and even more so when the cells replicate. Another disadvantage is that the effect of the route of exposure, a variable that can have profound effects on test results, is often impossi- ble to determine. There are many measures of damage to differen- tiated or undifferentiated cells—the rate of repro- duction, the rate of synthesis of certain substances, Microscopic View of Cell Culture From Rabbit Corneal Epitheliums Photo credit: Kwan Y. Chan, University of Washington 178 G Alternatives to Animal Use in Research, Testing, and Education changes in membrane permeability, and damage to some part of the cell structure. Those functions having to do with viability and growth are most frequently measured because they require an in- tegration of many physiologic events within the cell, are sensitive, and lend themselves to automa- tion (73). Quantifiable tests are preferred over subjective ones, and a wide variety of quantitative approaches are available to measure irritation, including the release of prostaglandins (35); the production of enzymes (46), proteins (57), antigens, antibodies, or hormones (73); and the migration of certain white blood cells (macrophages) to the area of ir- ritation (12,101). Irritation can also be measured by the extent to which cells exfoliate from the sur- face of the tissue. The extent of damage can be determined by counting cells and by examining the nuclei (1O2). Another indicator of irritation, the integrity of cell membranes, can be monitored through the uptake of nutrients through the cell wall. Where the nutrient uptake is active (that is, when the cell is required to expend energy for transport), uptake can also be used to indicate changes in metabolism (86,102). Liver cells have been the subject of considerable research, in part because they play such an im- portant role in an organism’s removal of toxic sub- stances and in part because they retain most of their special functions when cultured. The re- sponse of liver ceils to toxic substances may be measured in many ways: the use of sugar as an indication of metabolic activity; the production of proteins or other substances that have been cor- related with toxicity; uptake of amino acids as an indication of protein synthesis; changes in appear- ance that parallel those observed in livers of whole animals (106); and morphological changes and re- ductions in viability (75). Other promising tech- niques in this rapidly expanding field include cul- turing: G G G G beating heart cells to detect the effect of cer- tain vapors on irregularities in heartbeat (68); rabbit kidney tubules to detect substances that can cause acute renal failure, and rat vaginal tissue to test vaginal irritancy of contracep- tives (27); various kinds of cells to test for biocompati- bility of implants (15,52,53); and nerve cells to test for the synthesis of neuro- Dispensing Apparatus for Deiivery of Cuiture Medium to Ceiis Within a Piastic Cuiture Piate Photo credit: The Johns Hopkins University Ch. 8—Alternative to Animal Use in Testing G 179 transmitter chemicals, the formation of syn- apses, and the conduction of impulses (7). Although tissue and organ cultures may approx- imate more closely the physiology of the human or whole-animal model, they are more difficult to manipulate than cell cultures (see ch. 6). Sophisti- cated equipment must be used to monitor and con- trol the environment and to perfuse the sample with nutrients. Where the sample is more than a few cell layers thick, uniform delivery of the test substance, nutrients, and oxygen is difficult, as is the removal of waste products. Cell differentia- tion can usually be maintained in tissue and or- gan cultures, albeit with some difficulty (50). Human placentas have proved quite useful in testing the ability of a drug to cross the placenta from mother to fetus. There are certain logistical problems with this method, however. The placenta must be transferred to the perfusion apparatus within 5 minutes after it is eliminated from the uterus, and it is only useful for about 3 hours af- terward (77). Nonanimal Organisms There are a variety of nonanimal organisms that can replace some animals in testing, ranging from plants to single-celled organisms to invertebrates. All of these can respond to certain noxious stimuli, and some may experience pain. However, many commentators believe that they do not experience pain or suffering in the same way that animals do, particularly in those cases where there is no brain or neural tissue (90). The use of such organisms, which has never been controlled under any Fed- eral or State law, is regarded as a replacement for animals in this report. Micro-organisms In recent years, increased emphasis has been placed on the use of bacteria and fungi to meas- ure certain genotoxic effects. A major advantage of these organisms is that they can be cultivated much more easily and quickly than most animal or human cells. Their genetic makeup is simple compared with that of animals and humans and the fact that a great deal is known about it facili- tates their use, particularly in toxicological re- search leading to new methods (74). A change in genetic material is relatively easy to detect and characterize. Fungal systems have been shown to be especially useful in mutagenicity testing and seem to be more sensitive than bacteria (126), per- haps at the expense of falsely indicating a hazard. Other species that have proved useful include slime molds, algae, and protozoa (74). Protozoa, although rather primitive overall, fre- quently have specialized functions that mimic those of humans. For example, the cilia of protozoa re- spond to smoke or phenols as do the cilia in the human bronchial tube (5). Various protozoans have been used in toxicity testing of cigarette smoke. protozoans are currently being evaluated for use in screening tests for carcinogenesis, mutagene- sis, and reproductive toxicity (93). Invertebrates Invertebrates have made major contributions in biomedical research because certain aspects of their physiology are sufficiently similar to that of mammals (74). Although models for toxicity test- ing require greater similarity to animals or more thorough characterization of differences than models for research, invertebrates offer exciting possibilities. Of the invertebrates, insects offer the greatest selection of models, there being over 2 million spe- cies from which to choose (74). Among them, the fruit fly, Drosophila rnelanogaster, is the best un- derstood. procedures have been developed for de- tecting mutagenicity (18), as well as teratogenic- ity (11) and reproductive toxicity (93). The sea urchin has long been a favored test organism for basic reproductive research (74). Con- sequently, the mechanisms and procedures of testing this invertebrate can easily be developed and performed. The sea urchin model for fertili- zation and development can be used in screening for reproductive toxicity, teratogenicity, and muta- genicity. Nematodes, annelids, and mollusks are also used for alternative mutagenesis testing re- gimes and, additionally, mollusks are used in the area of reproductive toxicology. Sponges, mollusks, crustaceans, and echinoderms are being used in metabolism studies, as understanding metabolize formation in nonmammalian species can lend in- sight to interspecies variation (93). 180 G Alternatives to Animal Use in Research, Testing, and Education USE OF NONLIVING SYSTEMS IN TESTING Animal use can sometimes be avoided altogether with nonliving biochemical or physiochemical sys- tems, although most such systems currently re- quire animal derived components. Computer simu- lation can also be used when there are sufficient data available for substances related to the one of interest and when the mechanisms of toxicity are at least partially understood. Chemical Systems Whole animals have been replaced with analyti- cal chemistry for tests involving detection of a sub- stance or measurement of potency or concentra- tion, such as for vaccines, anticancer drugs, and vitamins (10). However, toxicity testing in nonliv- ing systems is quite limited at this time. Recently developed methods of detection or measurement are based on the selective binding that occurs between a particular substance and the antibodies to it. In an assay for botulism toxin (which traditionally required up to 200 mice), an- tibodies obtained from rabbits are modified so that the binding of the toxin can be detected easily. The rabbits are initially injected with a small, harm- less dose of the botulism toxin. Small amounts of blood are then removed from the rabbits at regu- lar intervals. In 4 weeks, a rabbit can produce enough antibody, with little discomfort, to perform tests that would otherwise require thousands of mice (32). Chemical systems that test for toxicity are based on determining whether a substance undergoes a specific reaction. For example, it is well known that carbon monoxide binds to hemoglobin in the blood, thus greatly reducing the blood’s ability to carry oxygen. The extent to which a substance would displace oxygen in hemoglobin can be a measure of its ability to produce asphyxiation. Sub- stances can also be tested in isolation for their ef- fects on enzymes crucial to certain bodily functions. An important limit of chemical systems is that they do not indicate the extent to which an organ- ism can recover from or prevent these reactions. For example, a substance that binds strongly to hemoglobin may not be a problem because it is not absorbed. A substance will not have a signifi- cant effect on an enzyme of interest if it is excreted before it has an effect. Physiochemical systems have some ability to determine whether a substance will be absorbed and what will happen to it. The tendency of a sub- stance to accumulate in a biological system can be roughly estimated by the relative proportions that dissolve in equal volumes of water and the organic solvent octanol (34,55). Artificial skin made with filter paper and fats is being tried as a means of mimicking absorption of cosmetics and drugs (45). Reactivity and other toxicity-related proper- ties can be deduced from chemical structure alone (109). Mathematical and Computer Models Advances in computer technology during the past 20 years have contributed to the development of sophisticated mathematical models of quantita- tive structure activity relationships (QSAR). These models are used to predict biological responses on the basis of physical and chemical properties, structure, and available toxicological data. The limi- tations of such models are due in part to a lack of understanding of the mechanisms by which toxic effects occur. In applying QSAR, the biological effects of chem- icals are expressed in quantitative terms. These effects can be correlated with physiochemical properties, composition, and/or structure. Fre- quently used properties include an affinity for fats versus water (octanol/water partition coefficient), the presence of certain reactive groups, the size and shape of molecules, and the way reactive frag- ments are linked together. The simplest extrapolation is for a series of closely related chemicals. The several character- istics they have in common need not be incorpo- rated into the model as variables. This type of analysis has been performed for several hundred families of chemicals and has established that rela- tionships within a series are fairly predictable (64). Another approach, more broadly applicable, is to examine the contributions of various portions of a molecule. In more elaborate computer pro- Ch. 8—Alternative to Animal Use in Testing . 181 grams, it is possible to identify likely reactions and cascading physiological events in various species, techniques first developed for pharmacology (54). A similar approach is the use of multitiered clas- sification schemes that use large databases to draw semiempirical conclusions (36). Epidemiologic Data on Humans Perhaps the most useful alternative to animal testing is epidemiologic studies on humans. Such studies were used to detect carcinogenicity in hu- mans as early as the 18th century (49,85,87). The most well known study detected scrotal cancer in chimney sweeps (85). A more recent example in which epidemiologic evidence was used to de- tect a human carcinogen was the finding that vi- nyl chloride causes a rare liver cancer in humans (26). A major disadvantage of epidemiologic studies is that considerable human exposure can take place before a toxic effect is detectable, particularly in the case of diseases that take many years to de- velop. Another disadvantage is that they can be quite expensive to conduct. Privacy must also be considered (112), preventing many data that would be useful from being collected or analyzed. Epidemiologic studies may be divided into three general types: experimental, descriptive, and ob- servational. Experimental epidemiology is the hu- man equivalent of animal testing—providing or withholding a substance to determine its toxic or beneficial effects. Such studies are greatly limited by ethical and legal considerations, as well as the difficulties involved in securing the cooperation of a large number of people. Descriptive epidemiology analyzes data on the distribution and extent of health problems or other conditions in various populations, trying to find correlations among characteristics such as diet, air quality, and occupation. Such comparisons are frequently done between countries or smaller geo- graphic regions, as is the case for cancer statistics collected and analyzed by the National Cancer In- stitute (9). observational epidemiology uses data derived from individuals or small groups. Data would be evaluated statistically to determine the strength of the association between the variable of interest and the disease. In cohort studies, a well-charac- terized and homogeneous group is studied over time. In case-control studies, a control group is selected retrospectively based on variables thought to be relevant to the effect, Both methods rely on an accurate prediction of the variables that are important and are subject to various selection biases (62)112). THE LD50 TEST The LD5O testis one of the most widely used tox- icity tests, and the development of alternatives to it is regarded by many as a high priority. As de- scribed in chapter 7, this acute toxicity test meas- ures the amount of a substance needed to kill half the population of the test species. The LD5O is used as a rough indicator of the acute toxicity of a chemical, The LD5O is useful for testing biological thera- peutics, although there remain few such sub- stances for which the LD5O is the only available means of standardization (13)90). Other applica- tions, perhaps not so well justified (90), are deter- mining doses for other toxicological tests and set - ting regulatory priorities. There has been political pressure to abolish the LD5O and it has been criticized by many toxicolo- gists on scientific grounds. It has poor reproduci- bility and the results are difficult to extrapolate to humans because there are so many mechanisms by which death could occur (70,90,125). Despite the many criticisms of the LD5O, most toxicologists agree that acute toxicity information has valid uses, and that measurements of lethality also are important. Nevertheless, the precision with which the LD5O is measured is often unjustified for several reasons. First, most applications of the information do not require precision. Second, even if the information were precise for a given spe- cies, the LD5O varies so much from species to spe- 182 Ž Alternatives to Animal Use in Research, Testing, and Education cies that extrapolation to humans is only rough. Third, the LD50 of a given substance varies signifi- cantly from laboratory to laboratory, and even in the same laboratory. Various regulatory classification schemes make distinctions between levels of toxicity (“highly toxic” versus “toxic, ” versus “moderately toxic, ” versus “nontoxic”). The LD50 for two neighboring levels typically differs by a factor of 4 to 10. Yet, the reproducibility of test results does not justify even these distinctions. A recent study, though not nec- essarily typical, indicates the magnitude of the problem. A series of LD5O tests were performed in 60 European laboratories for five substances on one species. The LD5O for one substance ranged from 46 mg/kg body weight to 522 mg/kg, possi- bly ranging over three toxicity levels in some clas- sification schemes. Although the variations were not this large for the four other chemicals tested, the smallest variation was 350 to 1,280 mg/kg. Each test was done with 50 or more animals so that the results would be precise (61). Using Fewer Animals The standard LD5O requires at least three groups of 10 animals or more each. An alternative proce- dure for determining the Approximate Lethal Dose (ALD) was developed as early as the 1940s (29), in which individual animals are administered doses that increase by 50 percent over the previous dose. Depending on the initial dose level, the total num- ber of animals needed is usually 4 to 10. Because the test substance might not be cleared between doses or because there maybe cumulative effects, the ALD can be lower than the LD50, perhaps by 70 percent, though more typically by less than 20 percent (29). Many other acute toxicity tests that require fewer animals than the LD50 have been developed (14, 17,33,61,69,71,94,105,107). Most require that the doses increase sequentially, thereby allowing the experiment to stop when a certain limit is reached. Thus, fewer animals die in the conduct of a test, but its duration could increase from 2 weeks to a month or more. Although many investigators are moving to less precise LD5O tests, no generally accepted alternative seems to have emerged. The Limit Test and Other Refinements If a substance is not lethal at high doses, its pre- cise LD5O is not very important. In the limit test (80), a small number of animals is given a single oral dose, e.g., 5 g/kg body weight. If no animals die and no major ill effects occur, no further test- ing is needed. However, this limit is so high that this approach may have little practical value in re- ducing animal use (24). Rather than determining the dose that is lethal, studies can also be done to detect toxic effects at doses that are not lethal. As with the LD5O, increas- ing doses can be administered to a small number of animals, perhaps stopping when some limit is reached. This approach can be further refined so that animals that are in distress could be sacrificed without affecting the outcome of the test (14). In Vitro and Nonanimal Methods Cell toxicity—changes in cell function or death of cells-can sometimes be used to detect acute toxicity. However, cell toxicity cannot be expected to function as a replacement for the LD5O because lethality can occur by so many mechanisms that are supercellular. Cell toxicity is particularly use- ful in comparing members of chemical families, such as alcohols and alkaloids (79). At present, mathematical modeling has limita- tions, although it may have some utility in range- finding and in screening substances for testing (109). Modeling of acute toxicity fails to meet one of the criteria suggested by a working party on quantitative structure activity relationships, namely that the mechanism by which the response occurs should involve a common rate determining step (88). Nonetheless, in a large study involving thou- sands of substances, a computer program was de- veloped that predicted LD5O values within a fac- tor of 2.5 for 50 percent of the substances and within a factor of 6 for 80 percent. Considering Ch. 8—Alternative to Animal Use in Testing Ž 183 the reproducibility of the test itself, this might be satisfactory for some purposes, and it certainly warrants further investigation. Furthermore, many of the larger deviations in this study, upon fur- ther examination, were found to involve report- ing errors. This program relied on a multi-tiered classification scheme based on chemical structure (36). SKIN AND EYE IRRITATION The widely used Draize eye irritation test and, to a somewhat lesser extent, the skin irritation test have been criticized because of the amount of pain inflicted and because they are unsatisfactory mod- els for human irritation (91,95). First, the rabbit eye has structural differences, such as a thinner cornea and differing tearing apparatus (103), and animal skin is much less sensitive and discriminat- ing than human skin (56,63). Second, both of these tests are sensitive to too many variables, making reproducibility poor (83,118). As with most tests, the number of animals used can sometimes be reduced. Several refinements have also been proposed. For example, screening tests based on pH or skin irritancy might also serve as alternatives to eye irritancy tests in limited cir- cumstances, although preliminary studies indicate that this approach is frequently misleading (119). Other refinements involve local anesthetics (51,65, 110), applying smaller (43) or more dilute (120) doses, and testing whole eyes in vitro (20). The lat- ter method has particular appeal when cow eyes are used because they are so readily available from slaughterhouses. In the case of smaller doses, a recent comparison with over 500 accidental human exposures showed that doses smaller than those now in use yielded results more predictive of the human response while causing less severe irrita- tion (38). Skin and eye irritation are similar in many re- spects. Thus, even though little work has been done to develop alternatives to skin irritation tests, the many approaches just summarized for eye irrita- tion may eventually be applied to skin testing as well (91). In Vitro Tests Several in vitro alternatives have been examined, and it appears to some commentators that no sin- gle alternative will be adequate, but that a battery of in vitro tests might be a useful replacement (67). Several types of cell cultures have been used in developing an in vitro test for eye irritation. The cells used are rabbit and human corneal cells (72), mouse and hamster fibroblasts, human hepatoma cells, and mouse macrophages (96). A variety of effects have been used as surrogates for eye irritation, such as the rate of uptake of uri- dine as an indication of cell functioning and re- covery, visible changes in cell structure, decreases in the concentration of cell protein (96), and re- lease of plasminogen activator from the injured cells (21). Some techniques appear promising, par- ticularly in their ability to rank substances based on irritancy, Rapid progress is being made in the development of techniques, but none can be con- sidered validated at this time (91). To date, little work has been done on in vitro replacements for skin irritancy testing. However, the growth of skin in tissue culture is of interest for treating burn victims, and it is expected that culture techniques currently being developed for that purpose can be used in testing methods. In addition, it has also been suggested that suitable specimens can be obtained from cadavers and surgery and from judicious use of human volun- teers (63). Chick Embryo One test system receiving considerable attention is the fertilized chicken egg. A part of the eggshell is removed and the test substance applied to the chorioallantoic membrane surrounding the devel- oping embryo (see fig. 8-l). This test has the po- tential for assessing both eye and skin irritancy. The chorioallantoic membrane of the chick em- bryo is a complete tissue, including arteries, capil- 184 G Alternatives to Animal Use in Research, Testing, and Education Figure 8-1.—Chronological Sequence of Chick laries, and veins, and is technically easy to study. Embryo Chorioallantoic Membrane Assay An embryonic membrane tested after 14 days of incubation responds to injury with a complete in- flammatory reaction, a process similar to that in- Day O duced in the conjunctival tissue of the rabbit eye. The embryonic membrane can show a variety of signs of irritation and has capabilities for recov- ery (59,60). / Assessment of toxicity is made and the embryo G G G Day 14 size, contours and surface, color, retraction of surrounding chorioallantoic membrane, spokewheel pattern of vessels, G overall grade of severity, and G necrosis (confirmed microscopically). Although this is, strictly speaking, an in vivo test, the chorioallantoic membrane does not have nerve cells, and thus it is unlikely that the organism ex- periences any discomfort. In addition, fertile eggs are inexpensive and do not require elaborate ani- mal room facilities. Day O. Fertile eggs are incubated at 37” C. Day 3. The shell is penetrated in two places: A window is cut at the top, and 1.5 to 2 milliliters of albumin is removed with a needle and discarded. The chorioallantoic membrane forms on the floor of the air space, on top of the embryo. The window is taped. Day 14. A test sample is placed on the embryonic membrane and contained within a plastic ring. Day 17. The chorioallantoic membrane is evaluated for its response to the test substance, and the embryo is discarded. SOURCE: J. Leighton, J. Nassauer, and R. Tchao, “The Chick Embryo in Toxicol- ogy: An Alternative to the Rabbit Eye,” Food Cherry. Tox/co/. 23:293-298. Copyright 19S5, Pergamon Press, Ltd. REPEATED-DOSE TOXICITY TESTS Repeated-dose toxicity testing involves the re- peated-dose testing, the long-term effects of peated application of a substance to a biological repeated, sublethal exposure to a substance are assay system and subsequent measurement of of interest, rather than acute, lethal effects. Cell many different effects of the substance. In re- cultures may be useful adjuncts for suspected tar- Ch. 8—Alternative to Animal Use in Testing G 185 Chick Embryo Chorioallantoic Membrane Assay Photo credit: Joseph Leighton, Medical College of Pennsylvania Typical react ion seen 3 days after certain concentrations of household products have been placed on the 14-day- old chorioallantoic membrane. The thin white plastic ring has an internal diameter of 10 millimeters. The area of injury within the ring is well defined with a distinct edge. All of the cells in the injured area are degenerating or dead. The severity of this positive lesion is quantified by measuring its diameter. get organs or tissues, but they are not a replace- ment for whole-animal testing. The most promising alternatives in the near future involve modifica- tions of animal use (for example, by combining tests), and the use of screening tests and computer simulation for improved experimental design. The screening tests with the greatest promise are for hepatotoxicity and neurotoxicity. Hepatotoxicity Several in vitro alternatives for hepatotoxicity have been developed, including perfused liver (108), liver cell suspensions (39), and liver cell cul- tures (39)44). Liver perfusions can only be main- tained for a few hours, and with some difficulty. Cell cultures can retain the special functions of liver cells with specially prepared culture media (76,81). However, the cells are viable for only a limited period of time and do not replicate in a reproducible manner. Although these techniques have been used to study mechanisms of liver tox- icity, only limited attention has been given to their use in screening or as alternatives (91). Neurotoxicity The development of alternatives for neurotox- icity is more difficult than for hepatotoxicity. The nervous system is the most complex organ in the body, both in terms of structure and its function. Because many neurotoxins affect only one kind of cell, a battery of in vitro tests would probably be required to replace whole-animal testing–if anything could. Substances can also affect vari- ous areas differently, partly because of distribu- tion factors, For example, very few substances are able to enter the brain because of the ‘(blood-brain barrier.” Thus, pharmacokinetic studies will con- tinue to be very important. Some in vitro tests (41) and tests using inverte- brates (8) seem useful, at least for screening. As yet, however, the primary use of in vitro tech- niques has been the elucidation of mechanisms of known toxic effects (31). Many toxic effects to neural tissue have been correlated with concen- trations of specific chemicals in or around the cells, thus offering the means for developing in vitro tests (31). MUTAGENICITY Mutation, the change in the DNA sequence of is passed from the mutated cell to its descendants. genes, is a mechanism by which toxic effects may Mutation can lead to cell death or the gain or loss be initiated. If the DNA replicates, the mutation of certain functions. When it occurs in germ cells, 38-750 0 - 86 - 7 186 G Alternatives to Animal Use in Research, Testing, and Education the gene pool is affected, even if the mutation is not expressed in the progeny. The mutations that occur in somatic cells that are of greatest concern are those that lead to cancer (18). Recent advances in the techniques of cell biology have led to an increase in the types and sophisti- cation of mutagenicity tests available. Mutations can be detected by analyzing DNA or its fragments or by observing changes in the size, shape, or num- ber of the chromosomes (which contain DNA), as well as by observing changes in a whole organism (34). Mutation can also be detected by measuring the amount of DNA repair. Micro-organism Tests The most commonly used test for mutagenicity is the Ames test for “reverse mutation” in Salno- nella typhimurium (3). Mutagenicity is detected by exposing an already mutated strain to poten- tial mutagens. If the mutation is reversed, the bac- teria regain their ability to produce the amino acid histidine and will proliferate in a histidine-deficient culture medium. The Ames test, as well as most other mutagenic- ity tests involving micro-organisms, does not avoid animal use entirely, To determine whether the meta- bolic products of a substance might be mutagenic even if the substance itself is not, liver prepara- tions from rats or other rodents are used to pro- duce at least some of the likely metabolic products. Microorganism systems may fail to detector may overpredict mutagenic changes that could occur in whole animals or humans. For example, the sys- tem provided for metabolism may not be capable of reproducing conditions in vivo, or in the case of screening for carcinogenicity, mutation may not be the initiating event. on the other hand, such systems may indicate mutagenicity when the DNA repair system of mammals would reverse the mu- tation. . Other bacterial tests have been developed using S. typhimurium, Escherichia coli, and Bacillus sub- tilis. These systems do not seem to offer any par- ticular advantage over the Ames test, although thorough evaluation is hampered by lack of a com- parable database of results (28). Tests have also been developed for molds (30)) fungi (16), and yeasts (18,82). In Vitro Tests In vitro mutagenicity tests maybe done with cul- tured mammalian cells that are exposed to toxic substances, although many mammalian in vitro tests also have an in vivo variant. Such tests typi- cally measure acquired resistance or lost resistance to the effects of the toxic substance. Most com- monly used are a mouse lymphoma ceil line or ham- ster ovary cells, but almost any well-characterized cell can be used. ovary cells are often used be- cause, as germ cells, they have half the number of chromosomes to be evaluated (18). A test known as the specific locus test can be done with Chinese hamster ovary cells. They are exposed to a test substance and their response to the normally lethal 8-azaguanine or 6-thioguanine in cell culture determined. The cell’s ability to sur- vive, requiring the ability to metabolize the 8- azaguanine or 6-thioguanine, is an indication of the occurrence of mutation as a result of exposure to the test substance. This test can also be done with mouse lymphoma cells exposed to 5-bromo- deoxyuridine or trifluorothymidine (23). The sister chromatid exchange test relies on the fact that certain substances will cause DNA break- age and reunion. This damage can be observed by staining the original chromosomes so that any segments exchanged during replication can be ob- served. Commonly used cells include human lym- phocyte cells and rodent and human fibroblasts (37). Both the specific locus test and the sister chro- matid exchange can also be performed as in vivo procedures (see ch. 7). Although the cells are usually derived from ani- mals, there is a considerable net savings in animal lives when in vitro mutagenicity tests are per- formed. For example, the rat mast cell assay can be used to screen severe irritants, and one rat can supply enough tissue to replace the use of 48 ani- mals in in-vivo procedures (103). Tests Using Insects The most widely used insect for genetic studies is the fruit fly, Drosophila melanogaster (114, 115). The fruit fly has well-characterized genetics and is similar to mammals in many key reactions, A variety of end points can be detected. The most common, and probably most sensitive, test is the Ch. 8—Alternative to Animal Use in Testing 187 sex-linked recessive lethal assay (18). Treated males ured include the loss, gain, or breakage of chro- are mated with untreated females, and the progeny mosomes detected by examining germ cells. With are mated to each other. The number and charac- the availability of mutant strains, the measurement teristics of the male progeny are evaluated to de- of reverse mutations can be a valuable tool. Eye termine if lethal mutations (that is, mutations that color is a popular method of following genetic ef- prevent viability) have occurred. fects in the fruit fly (18). Other tests involving fruit flies also exist or are likely to be developed. End points that can be meas - CARCINOGENICITY Many assays meant to replace carcinogenicity testing are designed to detect the initiation of can- cer rather than the formation of tumors. First, de- tecting initiation is faster and easier than detect- ing cancer. Second, although not all initiation leads to cancer, certain kinds are considered reliable surrogates for the disease. A major problem with evaluating the predictive- ness of alternatives to whole animals for carcinoge- nicity testing is that very few human carcinogens have been positively identified. Most substances treated as human carcinogens, although docu- mented to be known animal carcinogens, must be viewed as probable or suspected human carcino- gens. The development of alternatives is somewhat hampered by a lack of epidemiologic data on hu- mans. Various molecular and physiochemical prop- erties of substances have been correlated to car- cinogenicity. Some structure-activity models de- veloped for families of chemicals have predicted the carcinogenic properties for 75 to 97 percent of them. The chemicals modeled include polycyclic aromatic hydrocarbons (123), nitrosamines (89,99, 121), and aromatic amines (124). The Ames Test Because mutation is often the first step in car- cinogenesis, the Ames test has been suggested as a possible screen or replacement for carcinoge- nicity testing. It has been evaluated for this pur- pose, both alone and as one in a battery of tests. Alone, it is less predictive than whole-animal tests. In a battery, it has been shown to be about as pre- dictive as animal testing for certain families of chemicals and substantially less predictive for others for the substances tested. Table 8-1 shows the predictiveness of mouse and rat bioassays and the Ames test for some known human carcinogens. The Ames test has been performed thousands of times in over 2,000 laboratories throughout the world and has provided results on over 1,000 chemical substances since it was developed less than two decades ago. Portions of this large body of analytical data have been reviewed in over a dozen evaluation studies with the intent of deter- mining the test ability to predict carcinogenicity (6,19,66). These evaluations show that the percent- age of human carcinogens that are also mutagens (mutagenic carcinogens) ranges from 50 to 93 per- cent and is most likely about 80 percent (48). About 20 percent of the human carcinogens were not mutagens (nonmutagenic carcinogens) in the Ames test, and it is believed that cancer associated with these carcinogens is initiated by a mechanism other than mutation. A critical analysis of several studies (19) identi- fied several sources of variation. These include methods of chemical selection, sample coding, use of a high proportion of chemicals known to work well or poorly with Ames testing, and differences in metabolic activation during the test procedure. The conclusion was that a reasonably careful ap- plication of the Ames technique to a nonbiased group of chemicals would be expected to yield a predictive accuracy of approximately 80 percent for mouse and rat carcinogens. The Ames test tends to be positive for a large proportion (about 40 percent) of substances that have not been identified as carcinogens in rodent bioassays. It should be noted, however, that these 188 G Alternatives to Animal Use in Research, Testing, and Education Table 8.1.—The Response of Known Human Carcinogens to Rodent Carcinogenicity and Bacterial Mutagenicity Assays Rat Mouse Ames Chemical bioassay bioassay test 4-Aminobiphenyl . . . . . . . . . . . . . . . . . . . . . . . . . Arsenic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Asbestos . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Benzene. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Benzidine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Bis(chloromethyl)ether . . . . . . . . . . . . . . . . . . . . Chromium; some chromium compounds . . . . . Cyclophosphamide . . . . . . . . . . . . . . . . . . . . . . . Diethylstilbestrol . . . . . . . . . . . . . . . . . . . . . . . . . Melphalan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mustard gas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-Naphthylamine . . . . . . . . . . . . . . . . . . . . . . . . . Soot, tars . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Vinyl chloride.. . . . . . . . . . . . . . . . . . . . . . . . . . . + — + — + + + + + + n.d. — — + + — + + + + — + + + + + + + + — — + + + + — + + + + + KEY:+ = Positive results (carcinogenic to rodents or mutagenic to bacteria) – = Negative results (not carcinogenic ornot mutagenic) n.d. = No data. SOURCES: From H. Bartsch, L. Tomatis, and C. Malaveille, ’’Mutagenicity and Carcinogenicityof Environmental Chemicals:’ Regu/ Tox/co/. Pharmacoi. 2:94-105, 1982;D. Brusick, devaluation of Chronic Rodent Bioaasays andAmes Assay Tests as Accurate Modeis for Predicting Human Carcinogens,” Application of Bioiogicai Markets to Carcinogen Testing, H. Milman and S. Sell (ads.) (New York: Plenum Press, 1963); B.D. Goldstein, C.A. Snyder, S. Laskin et al., “Myelogenous Leukemia in Rodents Inhaling Benzene,” TcJx/coi. f.ett. 13:169-173, 1962; and J.V. Soderman (cd.), Handbook of identified Carcinogens and Noncarcinogens, Vols. / and Ii (Boca Raton, FL: CRC Press, 1982), substances have not been shown to be noncarcino- genic, and many authorities maintain that the in- formation is insufficient to make any statement about the proportion of noncarcinogens that are also nonmutagens in the Ames test (4,116). Use of the Ames Test in a Battery of Tests The predictive value of the Ames test, or other mutagenicity tests, can be improved by combin- ing it with additional short-term assays to form a test battery. Although no US. regulatory agency has yet recommended a specific combination, most authorities recommend that an appropriate bat- tery should include information from a minimum of three types of tests: G G G gene mutation (Ames test, mouse Iymphoma test); chromosomal mutation (in vivo Chinese ham- ster ovary cell cytogenetics); and DNA damage (sister chromatid exchange, — unscheduled DNA repair). At least one test should include a mammalian in vitro cell, tissue, or organ culture assay (4). In a recent study, 18 Ames tests averaged 66 per- cent “accuracy” (number of chemicals correctly identified/number of chemicals tested). Compara- tive results from six batteries of short-term tests that included the Ames test increased the accuracy to 82 to 90 percent (58,111). CURRENT TRENDS As long as toxicological data continue to be re- thermore, there are several impediments to devel- quired by regulators and by the courts to protect opment and implementation: human health, animal testing will continue for the foreseeable future. Even major progress in the de- . A large number of scientists have been trained velopment and implementation of alternatives will to solve health problems and to invent new not necessarily eliminate whole-animal tests. Fur- products using animal models. Ch. 8—Alternative to Animal Use in Testing G 189 G G G Regulatory schemes, product liability law, and patent law also incorporate notions of animal models. A large body of animal testing information al- ready exists that is useful in interpreting new testing data. There are substantial costs and delays associ- ated with the development and adoption of alternatives. One study indicated that it takes about 20 years for an in vitro test to be devel- oped, validated, adopted, and implemented (92). At the same time, there are several factors facilitating the development and implementation of alternatives: G G G G Rapid progress is being made in techniques for culturing mammalian cells and organs, in instruments for detecting and quantifying cellular and molecular changes, and in the understanding of the cellular and molecular processes underlying toxicity. Improved un- derstanding is leading to the ability to predict long-term effects and carcinogenicity from short-term biochemical and morphological changes. As such advances are made, the research lab- oratories that have developed the expertise are often willing to apply it to the develop- ment of new testing methods, and can do so efficiently (42). Organizations such as The Johns Hopkins Cen- ter for Alternatives to Animal Testing and the Rockefeller University laboratory have been set up to facilitate and coordinate research on alternatives (see ch. 12). Many organizations have been established to pressure those who conduct animal testing or use data based on it to adopt alternatives or conduct research that will lead to alter- natives. Strategies to speed the development and adop- tion of alternatives will depend on the needs and resources of the organization involved. The fol- lowing recommendations encompass a variety of perspectives. They were promulgated by the Tox- icity Committee of the Fund for the Replacement of Animals in Medical Experiments, which met from 1979 through 1982 (40). Some involve re- assessment of testing needs and priorities; others involve technical strategies thought to be likely to lead to better methods, both in testing and in evalu- ating results: G G G G G G G G G G G G G Provide a mechanism for reviewing the need for a given test. Investigate the consequences of not requir- ing or possessing testing data other than what already exists. Particular attention should be given to widely used tests such as the LD5O and skin and eye irritation tests with a view toward eliminating unnecessary requirements. Encourage flexible use of testing guidelines and frequent reappraisal of them in light of new knowledge. Strive for broader-based international har- monization and mutual recognition of data from other countries so that duplicative test- ing can be avoided. Encourage detailed publication of all testing results, particularly for costly or painful tests or those requiring many animals. Investigate the possibility of time limits on the confidentiality of test results. Make greater use of studies on absorption, distribution, biotransformation, and excretion in humans, as well as in test animals, to select the most relevant exposure conditions, to aid in extrapolation of results, and to improve the reliability of test results. Perform preliminary studies before undertak- ing long-term studies so that results can be as useful as possible. Make greater use of the structural and con- formational computer models used in devel- oping drugs for the prediction of toxicity. Standardize screening tests based on in vitro and nonanimal tests, both to promote efficient use of testing resources and to evaluate the predictiveness of these tests. Try to predict toxic reactions before testing, both as a means for improving prediction tech- niques and to avoid testing highly irritating substances, particularly in the eye, if possible. Conduct research on the mechanisms by which toxic effects occur to facilitate the develop- ment of new testing methods. Develop more accurate, reproducible instru- mentation for measuring toxic effects, avoid- 190 G Alternatives to Animal Use in Research, Testing, and Education G G G G G G G G G ing subjective measurements and reducing measurement errors. Make greater use of depositories in standard- izing cell lines or strains of micro-organisms used for testing. Study the relationship between physiochemi- cal properties and pharmacokinetic proper- ties, as well as between physiochemical and toxicologic properties. Develop techniques for detecting nonmuta- genic carcinogens. Develop systematic methods for objectively evaluating new techniques. Conduct postmarketing surveillance for ad- verse effects, noting any discrepancies with test results from animals. Substitute very specific tests for the LD5O and other general toxicity tests, particularly for substances having specialized uses, such as drugs. Use skin irritation testing as a rough screen- ing tool for eye irritation. Attempt to describe specific effects in eye ir- ritation studies, rather than reporting only the magnitude of the response. Investigate specific effects such as neurotox- icity to the extent possible when conducting general toxicity tests. G G G G G G G G Search for cell lines that retain their special functions upon replication and develop tech- niques for culturing them. Evaluate the statistical precision needed in various circumstances with a view toward using the smallest number of animals likely to be adequate. Use statistics to maximize the utility of results. Techniques such as blocking, covariance anal- ysis, and factorial design should be used rou- tinely. Improve standards of care and diet to reduce background effects. Take care that those conducting tests are qual- ified to do so, including having been trained in humane handling of animals. Combine tests wherever possible and keep them as short as possible, compatible with the nature of the test, Place greater emphasis on “no observed effect levels” than on lethal doses when they have greater predictive value. Use more than one species only to answer spe- cific questions, and not for general safety as- sessments. SUMMARY AND CONCLUSIONS There has been a small but significant shift away from whole-animal testing to in vitro and non- animal techniques in recent years, partly as a re- sult of advances in biological techniques and partly in response to political and economic pressures. Many new methods are being developed for com- monly used tests. Most of these are not yet vali- dated, but they already have potential uses for screening substances for the animal testing they may eventually replace. There are several kinds of alternatives. The first entails the continued, but modified, use of ani- mals-changes in experimental design or data anal- ysis so that fewer animals are needed or changes in protocols to reduce pain or distress. Living tis- sues, organs, and cells derived from humans or animals can sometimes be used instead of whole animals. These systems require a larger investment of time and money to develop than do modifica- tions of whole-animal techniques, but their advan- tages may also be greater. They are usually faster and often cheaper than the corresponding whole- animal test, and they have scientific advantages as well. However, they almost always are less predictive than whole-animal tests and often fail to provide reliable dose-response data, informa- tion that is critical in estimating potential toxicity to humans. Data, both anecdotal and epidemiologic, on toxic effects in inadvertently exposed humans are some- times useful. However, these data are often con- founded by lifestyle and exposure to other toxic Ch. 8—Alternative to Animal Use in Testing Ž 191 factors. Another drawback is that human exposure can be great if there are long delays between ex- posure and observable effects. The LD5O, probably the most common and most criticized toxicity test, is well suited to the limited use for which it was first developed. The biggest obstacle to limiting or eliminating use of the LD5O is institutional: Many regulatory schemes rely on it for classifying substances. The most promising alternatives in the short term are testing sequences that require fewer animals. Cell culture techniques and computer modeling show some promise, but they have limited value at this time. Another common and widely criticized test is the Draize eye irritation test. Several promising in vitro alternatives have been developed with cell cultures. Another technique uses the outer (chorio- allantoic) membrane of a 14-day-old chicken em- bryo. This technique, although it uses a whole ani- mal embryo, is thought to involve no pain because the membrane has no nerves. These alternatives may also apply to skin irritation. Alternatives to carcinogenicity testing and re- peated dose toxicity testing are of special interest, in part because the potential savings in testing costs and time are quite large, and in part because these tests require large numbers of animals. The most promising replacements are batteries of tests in- volving cell cultures and living, nonanimal organ- isms. Mutagenicity testing uses many in vitro or nonanimal protocols. Mutagenicity is of particu- lar interest because mutation can be the first event in other kinds of toxicity, including carcinogenic- ity, and because it can permanently affect the hu- man gene pool. The most well known nonanimal mutagenicity assay is the Ames test. When it is com- bined with other tests, the Ames shows promise as an alternative to carcinogenicity testing, but it is not yet validated for this use. In general, the development of alternatives is being facilitated by the rapid development of bio- logical techniques, which are being applied to the search for- alternatives in many different labora- tories. Major contributions to the coordination of these developments in the United States are being made by Rockefeller University and The Johns Hopkins Center for Alternatives to Animal Testing. The implementation of alternatives is hindered by various forms of institutional inertia, such as regulatory schemes (see ch. 7), product liability law (see ch. 7), and general resistance to change. Important impediments are the large body of ex- isting information —derived from animals—that is relied on for the interpretation of new data and the lack of sufficient information to support the use of alternatives. CHAPTER 8 REFERENCES 1. 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Deichmann, W .B., and Leblanc, T.J., “Determina- tion of the Approximate Lethal Dose With Six Ani- mals)” J. Ind. Hyg. Toxicol. 25:415-417, 1943. De Serres, F.J., and Mailing, H. V., “Measurement of Recessive Lethal Damage Over the Entire Ge- nome and Two Specific Loci of the ad-3 Region of Neurospora crassa With a Two Component Het - erokaryon, ” Chemical Mutagens, Principles and Methods for Their Detection, Vol. H, A. Hollaender (cd.) (New York: Plenum Press, 1971). Dewar, A.J., “Neurotoxicity,” Animals and Aher- natives in Toxicity Testing, M. Balk, R J. Riddell, and A.N. Worden (eds.) (New York: Academic Press, 1983). Dezfulian, M., and Barlett, J. G., “Selective Isola- tion and Rapid Identification of Clostridium botu - linum; Type A and Type B by Toxin Detection,” J. Clin. Microbiol. 2:231-233, 1985. Dixon, W. J., and Mood, A. M., “A Method of Ob- taining and Analyzing Sensitivity Data,’)J. Am. Stat. AsSOC. 43:109-126, 1948. Doull, J., Klassen, C. D., and Amdur, M.O. (eds.) Casserett and DouWs Toxicology: The Basic Science of Poisons (New York: Macmillan Publishing Co., 2d cd., 1980). 35. Eakins, K. E., “Prostaglandins and the Eye,’ ’Prosta - 36 37 glandins:Physiological, Pharmacolo#”caland Path- dog”cal Aspects, S.M.M. Karim (cd.) (Baltimore, MD: University Park Press, 1976). Enslein, K., Lander, T. R., Tomb, M.E., et al., Bench- rnark Papers in Toxicology: Vol. 1, A Predictive Model for Estimating Rat Oral LD~O Values (Prince- ton, NJ: Scientific Publishers, Inc., 1983). Evans, H.J., and O’Riordan, M. L., “Human Periph- eral Blood Lymphocytes for the Analysis of Chro- mosome Aberration in Mutagen Tests)” Handbook of Mutagenicity Test Procedures, B.J. Kilbey, M. Legator, W. Nichols, et al. (eds.) (Amsterdam: El- sevier/North Holland, 1977). 38. Freeberg, F. E., Griffith, J. F., Bruce, R. D., et al., “Correlation of Animal Test Method With Human Experience for Household Products,”J. Toxicol. - Cut. Ocular Toxicol. 1:53-64, 1984. 39. Fry, J. R., and Bridges, J. W., “The Metabolism of Xenobiotics in Cell Suspensions and Cell Cultures,” Progress in DrugMetabolism, Vol. 2, J.W. Bridges Ch. 8—Alternative to Animal Use in Testing G 193 and L.F. Chasseand (eds.) (New York: John Wiley & Sons, 1977). 40. Fund for the Replacement of Animals in Medical Experiments, “Report of the FRAME Toxicity Com- mittee, ” Animals and Alternatives in Toxicity Test- ing, M. Balls, R.J. Ridden, and A. N’. Worden (eds.) (New York: Academic Press, 1983). 41. Goldberg, A. M., “Mechanisms of Neurotoxicity as Studied in Tissue Culture Systems,” Toxicology 17:201-208, 1980. 42. Goldberg, A. M., Director, The Johns Hopkins Cen- ter for Alternatives to Animal Testing, Baltimore, MD, persona] communication, 1985. 43. Griffith, J. F., Nixon, G. A., Bruce, R. D., et al., “Dose- Response Studies With Chemical Irritants in the Albino Rabbit Eye as a Basis for Selecting optimum Testing Conditions for Predicting Hazard to the Human Eye,” Toxicol. Appl. Pharmaco). 55:501-13, 1980. 44. Grisham, J. W., “Use of Hepatic Cell Cultures to De- tect and Evaluate the Mechanisms of Action of Toxic Chemicals, ’’lnt. Rev. Exp. Pathol. 20:123-210, 1979. 45. Guy, R. H., and Fleming, R., “Transport Across a Phospholipid Barrier, ” J. Colloid Interface Sci. 83:130-137, 1981. 46. Hassid, A., and Levine, L., “Induction of Fatty Acid Cyclooxygenase Activity in Canine Kidney Cells (MDCK) by Benzo(a) Pyrene,” J. Biol. Chem. 252: 6591-6593, 1!377. 47. Healey, G. F., “Statistical Contributions to Experi- mental Design, ’’Animals and Alternatives in Tox- ici(.y Testing, M. Balls, R.J. Ridden, and A.N. Wor- den (eds.) (New York: Academic Press, 1983). 48. Hertzfeld, H. R., and Myers, T. D., “Alternatives to Animal Use in Testing and Experimentation: Eco- nomic and Policy Considerations, ” contract report prepared for the Office of Technology Assessment, U.S. Congress, January 1985. 49. Hill, J., Cautions Against the Immoderate Use of Snuff (London: Baldwin and Jackson, 1761). 50. Huot, R., Fodart, J., Nardone, R., et al., “Differen- tial Modulation of Human Chorionic Gonadotropin Secretion by Epidermal Growth Factor in Normal and Malignant Placental Cultures, ” J. Clin. EzIdo- crinol. Metab, 53:1059-1063, 1981. 51. Johnson, A. W., ‘(Use of Small Dosage and Corneal Anesthetic for Eye Testing In Vivo)’’Proceedings of the CTFA Ocular Safety Testing Workshop: In Vivo and In Vitro Approaches (Washington, DC: Cosmetic, Toiletry, and Fragrance Association, 1980). 52. Johnson, H.J., Northup, S.J., Seagraves, P. A., et al., “Biocompatibility Test Procedures for Polymer Evaluation In Vitro: I. Comparative Test System Sensitivity,” J. Biomed. Mater. Res. 17:571-586, 1983. 53. Johnson, H.J., Northup, S.J., Seagraves, P. A., et al., “Biocompatibility Test procedures for Materials Evaluation In Vitro: II. Quantitative Methods of Toxicity Assessment, ” J. Biomed. Mater. Res. 19:489-508, 1985. 54. Kaufmann, J. J., Koski, W. S., Hariharan, P. C., et al., “Theoretical and Quantum Prediction of Toxic Effects,” Drug Metab. Rev. 15:527-556, 1984. 55. King, L. A., and Moffatt, A. C., “Hypnotics and Seda- tives: An Index of Fatal Toxicity, ” Lancet 11:387- 78, 1981. 56. Kligman, A.M., “Assessment of Mild Irritants, ’’Prin - ciples of Cosmetics for the Dermatologist, P. Frost and S.N. Horwitz (eds. ) (St Louis, MO: C.V. Mosby, 1982). 57. Knowles, B. B., Howe, C. C., and Aden, D. P., “Hu- man Hepatocellular Carcinoma Cell Lines Secrete the Major Plasma Proteins and Hepatitis B Surface Antigen)” Science 209:97-99, 1980. 58. Lave, L., Omenn, G., Hefferman, K., et al., ‘(Model for selecting Short Term Test of Carcinogenicity, ” J. Am. CO1l. Toxicol. 2:125-130, 1983. 59. Leighton, J., Nassauer, J., and Tchao, R., “The Chick Embryo in Toxicology: An Alternative to the Rab- bit Eye,” Food Chem. Toxicol. 23:293-298, 1985. 60. Leighton, J., Nassauer, J., Tchao, R., et al., “Devel- opment of a Procedure Using the Chick Egg as an Alternative to the Draize Rabbit Test, ” Product Safety Evaluation, A.M. Goldberg (cd.) (New York: Mary Ann Liebert, Inc., 1983). 61. Lorke, D., “How Can We Save Animals in Toxicity Testing,” Progress Without Pain (Lord Dowding Fund, National Anti-Vivisectionist Society, Ltd., London) 22: 1984. 62. MacMahon, B., and Pugh, T. F., Epidemiology: Prin- ciples and Methods (Boston, MA: Little, Brown & co., 1970). 63. Marks, R., “Testing for Cutaneous Toxicity, ” Ani- mals and Alternatives in Toxicity Testing, M. Balls, R.J. Ridden, and A.N. Worden (eds,) (New York: Academic Press, 1983). 64. Martin, Y, C., QuantitativeDru gDesign :A Critical Introduction (New York: Marcel Dekker, Inc., 1978). 65. Maurice, D., “Pain and Acute Toxicity Testing in the Eye, ” Alternatives to the Draize EwVe Test, A. Goldberg (cd.) (New York: Mary Ann Liebert, Inc., 1985). 66. McCann, J., Choi, E., Yamasaki, E., et al., “Detec- tion of Carcinogens as Mutagens in the Salmonella/ Microsome Tests: Assay of 300 Chemicals)” Proc. Natl. Acad. Sci. USA 72:5135-5139, 1975. 67. McCulley, J. P., “Chairman’s Summary,” Alterna - 194 G Alternatives to Animal Use in Research, Testing, and Education tives to the DraizeEye Test: A]ternativeMethodism Toxicology, Vol. 3, A. Goldberg (cd.) (New York: Mary Ann Liebert, Inc., 1985). 68. Miletich, D.J., Khan, A., Albrecht, R. F., et al., “Use of Heart Cell Cultures as a Tool for the Evaluation of Halothan Arrhythmia,” Toxicol. Appf. Z%arrnacol. 70:181-187, 1983. 69. Molingengo, L., “The Curve Doses v. Survival Time in the Evaluation of Acute Toxicity, ” J. Pharm. Pharmacol. 31:343-344, 1979. 70. Morrison, J.K, Quinton, R. M., and Reinert, H., “The Purpose and Value of LDSO Determinations, ’’Mod- ern Trends in Toxicology, Vol. I, E. Boyland and R. Goulding (eds.) (London: Butterworths, 1968). 71. Muller, H., and Kley, H. P., “Retrospective Study of the Reliability of an Approximate LD~O Deter- mined With a Small Number of Animals,” Arch. Toxicol. 51:189-196, 1982. 72. Nardone, R. M., and Bradlaw, J., “Toxicity Testing With In Vitro Systems: 1. Ocular Tissue Culture,” J. Toxicol.— alncludes frogs, sheep, and Pi9e0ns. SOURCE: Association of American Medical Colleges, Use of Animals in Undergraduate and Graduate Medical Education (Washington, DC: 1985). 206 . Alternatives to Animal Use in Research, Testing, and Education Dogs and pigs are used to teach techniques for incubation (establishing an emergency airway) and the installation of intravenous/intra-arterial cath- eters, In the AAMC survey, one anesthesia depart- ment used dogs to teach insertion of Swan-Ganz catheters into the right chamber of the heart, a common procedure in cardiac intensive care units. Two otolaryngology departments used dogs to teach the musculature and innervation of the tra- chea and oropharynx to ear, nose, and throat resi- dents. One obstetrics and gynecology department used dogs as models to teach exposure and isola- tion of the Fallopian tubes from the nearby ure- ters, and three pediatrics departments use young cats as models for instruction in incubation of pre- mature newborn babies. All of the techniques taught in these graduate medical programs must be learned to achieve competence in the desired specialty. In those programs that do not use ani- mals, the techniques are mastered through experi- ence with human patients during surgery (2). The AAMC survey found no relation between a medical school’s level of research expenditures (high, medium, or low) and its use of animals in education. The medium expenditure schools used the most animals in education, perhaps because in the more research-intensive schools there is a greater opportunity for students to observe ani- mal surgery in the course of participation in faculty research and less need to include such experience in the curriculum. Most of the schools surveyed expressed regret that they were not able to use Instruction in Incubation of Premature Newborn Babies, Using a Young Cat as a Model Redrawn by: Office of Technology Assessment. animals to a greater extent in student instruction, often citing cost as a factor limiting instruction with live animals (2). National estimates of the numbers of animals used in medical education (see table 9-5) were cal- culated based on the assumptions that the 16 schools surveyed are typical of the 127 accredited schools in the United States. The mean number of animals of each species used in the sample schools was accepted as the best estimate of the mean for all schools, and an extrapolation was made to 127 schools (2). Rats and dogs are the principal species used in medical education, accounting for about 70 per- cent of the estimated 36,700 animals used annu- ally. These figures are very rough—the potential error inherent in the estimates ranges from 22 and 25 percent for rats and dogs to 100 percent for pigs and hamsters. The great uncertainty stems from variability among the 16 institutions in the sample. One school used 10 primates, for example, while another used 4, and 14 schools used none at all. Use of dogs and cats was more general; less uncertainty is associated with the national esti- mates of those species’ use (2). It is unlikely that any of the 127 medical schools in the United States train physicians without using any live animals. This is neither surprising nor alarming, particularly in light of the fact that the ultimate recipients of medical attention-humans— are not available for many of the types of educa- Table 9-5.—Estimated Animal Use in Medical Education in the United States, 1983.84 Kind of animal Number useda Rat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Dog . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mouse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rabbit . . . . . . . . . . . . . . . . . . . . ... , . . . . . . . . Cat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Hamster . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pig . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Primate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Guinea pig , . . . . . . . . . . . . . . . . . . . . . . . . . . . Other b . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14,000 12,000 3,000 1,700 800 200 130 70 4,000 Total. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36,700 aEst/mate /s b-On an extrapolation of a survey of 16 Seiected mSdiC@ schools evenly distributed by geographic region (Northeast, Midwest, South, or West), ownership (public or private), and research expenditures (low, medium, or high). bfncludes frogs, sheep, and Pi9eons. SOURCE: Association of American Medical Colleges, Use of Anima/s in Under- graduate and Graduate Medical Education (Washington, DC: 1965). Ch. 9—Animal Use in Education and the Alternatives G 207 tional exercises that medical students routinely must perform. It should be noted, however, that it is possible for a student to complete medical school without using animals. Veterinary Education Being admitted to the profession of veterinary medicine, I solemnly swear to use my scientific knowledge and skills for the benefit of society through the protection of animal health, the re- lief of animal suffering, the conservation of live- stock resources, the promotion of public health, and the advancement of medical knowledge. I will practice my profession conscientiously, with dignity, and in keeping with the principles of veterinary medical ethics. I accept as a lifelong obligation the continual improvements of my professional knowledge and competence, The Veterinarian’s Oath American Veterinary Medical Association Twenty-seven accredited veterinary schools in the United States educate and train veterinary scientists and veterinarians in the basic biomedi- cal sciences and comparative animal health. An OTA survey of the 27 schools indicated that every veterinary school uses animals in its curriculum. As in medical education, the question of the use of animals in veterinary education is a matter of degree and practice. Veterinary students—unlike medical students— train on models identical to their prospective pa- tients. Animals are used in laboratory exercises and demonstrations, and students have the addi- tional opportunity to interact with clinical cases owned by their schools as well as those brought in by clients. Privately owned pets, domestic live- stock, and zoo animals all serve as resources for the clinical education of veterinary students, Most animal use occurs in the third year of the curriculum, when surgical training takes place, using principally dogs and sheep. In earlier basic science courses, anatomy involves dissection of cadavers with live animals present in the lab for comparison, and physiology exercises involve the observation of live animals. The fourth year of veterinary studies is largely clinical apprenticeship. With cooperation from the Association of Amer- ican Veterinary Medical Colleges, OTA conducted a census of animal use in veterinary education in the 27 accredited veterinary schools in the United States for the school year 1983-84. The survey counted only those animals that began an exer- cise alive and either died or were subjected to euthanasia during the course of the laboratory ses- sion or demonstration. Cadavers or animals sub- jected to euthanasia prior to educational use were not counted, and clinical patients were not counted. Of 16,655 animals used in 1983-84, half (8,020) were dogs. Mice, rats, and birds accounted for the bulk of the remaining animals (see table 9-6). No primates were killed during or after educational exercises in veterinary schools. Laboratory-Animal Training Technicians with specialized training in public health and animal care are needed at all levels by public health organizations, research institutions, pharmaceutical manufacturers, and universities. During the 1970s, several 2-year training programs were developed in response to an increasing need for personnel formally qualified to assist in pri- Table 9.6.—Animals Used in Veterinary Education in the United States, 1983-84 Kind of animal Number useda Dog . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8,020 Mouse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2,180 Rat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2,083 Bird . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1,323 Reptile. ., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 433 Sheep . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 423 Cat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 414 Horse. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 378 Rabbit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195 Goat. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194 Pig . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 Guinea pig . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112 cow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111 Hamster . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 Other . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 578 Total. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16,655 aThis census of all 27 U.S. veterinary schools does not include privately owned or pet animals used for clinical demonstrations, animals purchased as cadavers, or those subjected to euthanasia prior to the laboratory exercise. It krcfudes only those animals that began the course alive and then either died or were subjected to euthanasia during the course of the laboratory session. bincludes fish, frogs, and exotic SPecies. SOURCE: Office of Technology Assessment, 208 Alternatives to Animal Use in Research, Testing, and Education vate veterinary practices, biological laboratories, animal research, food inspection, and other areas requiring expertise in both science and animal care and use. Graduates of these programs are generally re- ferred to as animal technicians. The terminology may vary slightly among different schools or with individual State laws and regulations. Many em- ployees of animal care and research and testing facilities have received training on the job, in sec- ondary schools, or at less than the 2-year college level. These individuals are commonly referred to as animal attendants, animal caretakers, or ani- mal health assistants. Two other types of animal- support personnel are laboratory-animal techni- cians (whose training has been oriented primarily toward laboratory animals) and animal technolo- gists (who have had training in a 4-year bacca- laureate degree program). Most accredited animal technician cover 2 academic years of college-level programs study and lead to an Associate in Applied Science degree or its equivalent. The core curriculum usually in- cludes animal husbandry, animal care and man- agement, animal diseases and nursing, anesthetic monitoring and nursing, ethics and jurisprudence, veterinary anatomy and physiology, medical ter- minology, animal nutrition and feeding, necropsy techniques, radiography, veterinary urinalysis, veterinary parasitology, and animal microbiology and sanitation (l). Many States require animal technicians to be reg- istered or certified. The Laboratory Animal Tech- nician Certification Board sponsored by the Amer- ican Association of Laboratory Animal Science provides examinations and registry for technicians who are eligible and employed in laboratory-animal facilities. In addition to increasing interest in laboratory- animal technician degree programs, a number of graduating veterinary students have begun to seek additional training and certification in laboratory- animal medicine. To date, about 700 full-time veter- inarians are certified in this field nationwide (see ch. 15). As more’ laboratory-animal technicians are trained and as the number of veterinarians spe- cializing in laboratory-animal medicine increases, the resulting base of skills and knowledge will likely improve animal care in the laboratory. THE ALTERNATIVES Finding alternatives to the use of animals in edu- cation is a complex challenge. Alternatives must satisfy the demands of science education, teach- ing both the scientific method and the fundamen- tal skills and techniques necessary to carry out scientific investigation. Yet science education does more—as it trains aspiring students, it establishes a framework of values and molds attitudes that will long influence their work. Therefore, exposure to alternatives, particularly the concepts under- lying animal use and alternative methods, strongly influences the paths investigators choose to fol- low in the future. Viewed from this perspective, the acceptance (or rejection) of a specific alterna- tive method in education assumes an importance that is, in fact, secondary to the impact it may have on the development of a student’s overall attitude toward animal use in research, testing, or edu- cation. Implementing alternative technologies and meth- ods in education does not necessarily mean banish- ing animals from the classroom or laboratory. As in research (see ch. 6) and testing (see ch. 8), cer- tain techniques are available that allow for the con- tinued, but modified, use of animals, the use of living systems, the use of nonliving systems, and the use of computers. In education, computer simu- lation stands as a particularly promising alter- native. Continued, But Modified, Use of Animals in Education Demonstrations In contrast to animal experimentation in re- search and testing, animal use in the educational laboratory is unlikely to result in novel findings. Ch. 9—Animal Use in Education and the Alternatives . 209 In education, a traditional laboratory exercise with a well-known outcome is usually repeated by a new student or group of students. The process of self- discovery and training is generally of greater im- portance than the specific data being collected. Under these circumstances, live demonstrations can often provide experiences that combine the best of direct student participation in animal lab- oratories with a reduction in animal use. Such ex- ercises, when carried out by practiced professional instructors, avoid clumsy errors that students may make at the expense of laboratory animals. They also provide a convenient intermediate in the con- stant tension between active student participation in the laboratory and the limitations imposed by large class sizes. In a variation on the laboratory demonstration, students may work together in groups on a single animal, again using fewer animals than if each stu- dent worked alone on a single animal. Exercises based on animal cells, tissues, or organs may be coordinated such that the minimum number of animals required can be sacrificed. Or animals may be subjected to multiple procedures, although if these involve sequential survival surgeries the advantage of reducing the number of animals used stands in conflict with the undesirability of repeated insults imposed on a surviving animal. Noninvasive Procedures observation can give rise to an appreciation of the diversity of the animal kingdom in general and important principles of physiology and behavior in particular. A sense of responsibility and an understanding of the life processes of animals are also conveyed when animals are maintained for observation in the laboratory. Areas for study in which animals can be used in a noninvasive man- ner include: Ž simple Mendelian genetics (e.g., the inheri- tance of coat color in successive generations of small rodents); G reproductive behavior (e.g., behavioral recep- tivity of a female during estrus); . normal physiological processes of maturity, aging, and death (e.g., the relationship be- tween aging and body weight); G disease processes (e.g., the incidence of spon- taneous tumor growth in a population); Ž biological rhythms (e.g., nocturnal and diur- nal feeding and drinking patterns); and G social interactions (e.g., territoriality and dom- inance relationships among males). Reduction in Pain Reduction in pain and distress may be accom- plished with the use of anesthetics, analgesics, and tranquilizers. In education, this is of primary im- portance in surgical training, when animals are anesthetized, operated on, and then subjected to euthanasia. Principles of pain and pain relief— common to research, testing, and education—are discussed in chapters 5, 6, and 8. Substitution of Species The substitution of nonmammalian for mam- malian species, of cold-blooded vertebrates for warm-blooded ones, or of nonpet species for com- panion animals is occasionally possible in educa- tion. Swine have replaced dogs in one surgical teaching and research laboratory (20). The pigs were especially successful replacements in a basic operative surgery course offered as an elective t o medical students. The principal advantages cited were closely shared anatomic and physiologic char- acteristics with humans, better health than dogs, and economic factors. Use of Other Living Systems in Education Invertebrates The use of invertebrates as an alternative is al- ready widespread in primary and secondary schools. Most laboratory manuals include common exer- cises that teach biological principles and introduce students to the scientific method of inquiry using organisms such as hydra, planaria (flatworms), an- nelids (earthworms), mollusks, and a variety of ar- thropods (e.g., insects and crustaceans). The use of invertebrates at the college and graduate levels may also increase as more is known about them. These deceptively simple systems are valuable re- sources for the laboratory investigation of sophis- ticated biological principles. 210 G Alternatives to Animal Use in Research, Testing, and Education In Vitro Methods Like the use of invertebrates, in vitro manipula- tion and maintenance of animal components such as cells, tissues, or organs (see chs. 6 and 8) can illustrate many biological principles. The incorpo- ration of in vitro techniques into students’ educa- tion and training also bears potential for shaping their later attitudes about the utility of in vitro methods. The stimulus provided by in vitro lab- oratory exercises can therefore ultimately alter the general course of research and testing. One noteworthy endeavor in training research- ers in in-vitro methods is the program of the Cen- ter for Advanced Training in Cell and Molecular Biology at Catholic University of America in Wash- ington, DC. With funding from the American Fund for Alternatives to Animal Research, the American Anti-Vivisection Society, and the Albert Schweit- zer Fellowship, the Center offers courses to stu- dents interested in the biomedical sciences and to professional researchers. In 1985, its third year of existence, the Center offered: G Basic Cell and Tissue Culture, G In Vitro Toxicology: Principles and Methods, G Tissue Culture Technology in Neuroscience Research, and G An Introduction to Tissue Culture and In Vitro Toxicology. The first three courses were attended by techni- cians and Ph.D. and M.D. researchers. The last course was specifically designed for high school seniors and college freshmen (9). Activities of this nature are useful in that they enable professionals and, particularly, beginning students to become acquainted with and proficient in in-vitro meth- odologies and to comprehend the possibilities as well as the limitations of alternative methods. The debate about whether or not the training of medical and veterinary students requires ani- mals has spawned development of an alternative technique in microsurgery training. The most prominent use of microsurgery is for reconnect- ing arteries and veins, for example in restoring circulation to severed fingers. To reproduce vas- cular circulation for microsurgical training, a Brit- ish plastic surgeon connected human placentas to a pump and an artificial blood supply, thereby simulating a heartbeat and typical blood pressures. Because the placenta contains blood vessels of widely ranging diameters, a single placenta can provide material for a substantial amount of prac- tice (14). At present, the human placenta cannot fully sub- stitute for living animals. One of the problems is that the placenta contains an anti-blood-clotting agent or mechanism that is not understood and cannot be controlled. Clotting therefore does not occur in placental vessels. since learning how to avoid clotting during repair is a critical aspect of training, and since students training on placental tissue cannot detect their errors that cause clot- ting, the existing system is not fully adequate in microsurgical training (14). Use of Nonliving Systems in Education Audiovisual presentations bring the abstract prose of lecture and text one step closer to the biological reality of living organisms. Films and videotapes can demonstrate principles and proto- cols performed with live animals, while sparing additional animals. They may also present experi- ments and situations that cannot be performed live in the average classroom setting. As replace- ments for animals, however, they lack the living dimension; most cannot behave interactively. Re- cently developed computerized videodisks offer an opportunity for student interaction with an au- diovisual program. When audiovisual aids are used in concert with animals, they may enhance the value of live ani- mals used in the laboratory. Students may learn a technique from a taped demonstration, for ex- ample, and then build on that experience as they perform the actual laboratory exercise in vivo. Medical education substitutes audiovisual tech- niques for animals in several cases. This has less to do with educational philosophy than with fac- tors external to the particular laboratory exercise, Those factors include the costs of animals and the facilities required to perform quality experiments, large medical school classes, lack of faculty time, and competition within a tightly packed curriculum. Animal cadavers (e.g., frogs, sharks, cats, and fetal pigs) are currently used at all levels of educa- Ch. 9—Animal Use in Education and the Alternatives . 211 Use of the Human Placenta for Training in Microvascular Surgery Human placenta perfused under dissecting microscope. Phcto credits: Pau/l LG Townsend, Consultant Plastic Surgeon, Frenchay Hospita/, Brlstol Sutures and valves implanted in vessels of human placenta. tion as models for dissection. Commercially pre- pared specimens are often used in junior high and high school education; medical and veterinary schools are more likely to prepare their own speci- mens. In some situations, cadavers may provide adequate replacements where living animals were once used. Computer Simulation in Education Computer simulation offers a variety of alter- natives for studying animal and human biology at all levels of education, and the field is evolving quickly as experience grows and computer tech- nology advances. Although at this time popular expectations for computer simulation still out- distance actual performance, the options that simu- lations present to educators can be expected to increase. Educational computer simulations fall into two categories: computer models of biologi- cal events and interactive simulations of biologi- cal experiments. Computer simulations of biological events-pri- marily mathematical models of physiological and cellular phenomena—present in quantitative form phenomena that might be difficult or impossible to study in animals or humans. By altering param- eters within the programs and noting results, stu- dents learn principles of biology from an ersatz animal system, the computer program. For exam- ple, a dog’s circulatory functions are converted to a series of mathematical equations, which are programmed into a computer. As students change individual values or groups of values, the program resolves the various equations and reports values that mimic the effects of altering those parame- ters of the circulatory system in a living dog. Fig- ure 9-1 depicts a portion of such a simulation. An array of computer models of physiological processes are used in undergraduate and gradu- ate laboratory exercises. The range of physiological simulations includes simulations of blood chemis- try, cardiovascular physiology, the digestive sys- tem, the musculoskeletal system, respiratory phys- iology, and renal physiology. Computer simulations currently used in physiology laboratory exercises include: G HUMAN: a comprehensive physiological mod- el (3), 212 G Alternatives to Animal Use in Research, Testing, and Educatkm Overview of a computer simulation of the complete cardiovascular system, showing student-controlled variables such as heart rate (HR%), total active blood volume (BV%), and total peripheral resistance (TPR%). SOURCE: N.S. Peterson and K.B. Campbell, “Teaching Cardiovascular Integrations With Computer Laboratories,” Physlo/oglst 26(3):159-169, 1965. G G G G G G G G pH regulation and carbon dioxide (24), pulsatile hemodynamics in the aorta (5), determinants of cardiac output (16), effects of medically important drugs on the circulatory system (25), simulation of the digestion of a meal (25), responses of organisms to exposure to high and low temperatures (25), influence of hormones on muscle cells (25), and renal excretory response to volume and os- molarity changes (12). Computer simulation of a particularly sophisticated laboratory exercise—for example, one that is too difficult for beginning veterinary students to per- form-can enable students to carry out laboratory exercises they otherwise would not have had (13). Table 9-7 summarizes the advantages, disadvan- tages, and barriers to substituting computer mod- els of biological systems for animals in education. Some characteristics apply to one type of computer application more than another. Viewed as a whole, the descriptors of computer simulations listed in table 9-7 illustrate the potential as well as the limi- Table 9-7.—Advantages, Disadvantages, and Barriers to Using Computer Simulations in Education Advantages: Quality of teaching material: G G G G G Simplification. Some biological events that are too complicated or not accessible to human study by vivisec- tion or dissection are better approached through com- puter simulation. Quantitative ski//s. Physical mechanisms and mathemat- ical variables that underlie biological events are em- phasized. Emphasis. Student attention is shifted from techniques to concepts, supporting lecture and textbook material. Reliability. Strong consistency from experiment to ex- periment. Response time. Simulations yield immediate results. Cost and efficiency: G Long-range cost reduction. Following initial purchase of computer hardware, computer laboratory costs are often lower than relatively high animal laboratory costs. G Speed and coordination. Increased teaching efficiency through expeditious testing, drills, and tutorials. . Laboratory availability. Increased access for students to laboratories. Disadvantages: G G G G G Bio/ogica/ complexity. Computers cannot be programmed to simulate many integrative interactions between inter- nal organs. Missed experiences. In the view of some teachers, stu- dents should have experience with living tissue. Bioogical variability. Computers do not accurately por- tray the large degree of uncertainty that arises from bi- ological variability, whereas comparisons of animals do present this concept. Publication of results. Developers of computer simula- tions sometimes find publication of their work in the usual scientific journals difficult since some simulations require ponderous documentation; in cases where pub- lications are intrinsic to tenure and other faculty deci- sions, computer modelers may be discriminated against, Student attitudes. In some cases, dubious student out- look on computer replacement of animals undermines teaching of concepts. In other cases, simulations may unintentionally train students (e.g., medical students) to ignore the behavior and appearance of patients and to place unwarranted importance on data from instruments. Barriers: G G G G Incompatibility. Hardware components and software sys- tems often are not interchangeable; this is especially true of graphic simulations. Computer /imitations. Some complex digital computer programs are not fully realistic because they must ap- proximate biological processes that are continuous and simultaneous by using a series of discrete steps. The only way to make such a computer approximation more realistic is to reduce the time the computer takes be- tween steps. This may require more sophisticated hardware. Tradition. Widespread lack of training in mathematics modeling leads many talented people to write textbooks rather than computer models. Proprietary considerations. Many of those who are de- veloping programs or catalogs of programs for commer- cial purposes will only disseminate useful information about computer simulations if they are paid, restricting applications. SOURCE: Office of Technology Assessment Ch. 9—Animal Use in Education and the Alternatives G 213 tations of this alternative in a variety of teaching situations. Several of the disadvantages listed in table 9-7 underscore the small likelihood that com- puters can completely replace animals in the class- room. Those who are developing computer simu- lations are among the most vocal in maintaining that this technique is not the optimal method in every teaching situation; in some cases, they say, animals serve the lesson better (8,13,15,26). In addition to providing models of biological ex- periments, computer programs serve in the class- room and laboratory as reusable training devices to teach specific skills, just as airline pilots train in flight simulators. These simulations are based on graphic presentation of the experiments and involve interaction between the program and trainee. An interactive videodisk program, for example, enables students to simulate dissections using pho- tographic images stored on the disks, rather than animals. Production of such a videodisk can cost from $60,000 to $200,000 for a 30-minute program and involves thousands of still photographs, com- puter overlay, and touch screen interaction. The sales price of such a videodisk can range from $1,000 to $5,000. The most sophisticated types of videodisk pro- grams have not achieved widespread use, largely because of economic factors. Apart from steep ini- tial production costs, the hardware supporting videodisk use is expensive. Computer-linked mannequins and robots cur- rently provide the most sophisticated simulations. Resusci-Dog, developed at the New York State Col- lege of Veterinary Medicine at Cornell University in Ithaca, NY, is a canine cardiopulmonary resus- citation training mannequin, the equivalent of the human dummies used in training paramedical tech- nicians. Constructed of plastic, Resusci-Dog can simulate a femoral artery pulse, and pressure can be applied to its rib cage for cardiac massage or cardiopulmonary resuscitation. The first micro- processor-laden canine simulator cost $7,000; the second $700. Resusci-Dog has replaced about 100 dogs per year in veterinary classes at the New York school (19). Despite the widespread enthusiasm for the po- tential of computer models and interactive simu- lations in the life sciences, three general problems Scenes From Interactive Videodisk Laboratory Exercise —Canine Hemorrhagic Shock Photo credits: Charles E. Branch and Gregg Greanoff, Auburn University These photographs were taken from the monitor screen of a video program on blood flow and hemorrhagic shock in use at the Auburn University School of Veterinary Medicine. The interactive video simulated experiment depicts actual experiments conducted by experts. Several treatments are videotaped and students then simulate performing the experiment, testing different treatments and dealing with the results as if they were actually performing the study. Top: Anesthetized dog in experimental setup. Bottom: Response of dog’s pupil to light. confront computer-based education in the mid- 1980s (7,8): G The rapid advance of computer technology has resulted in many–frequently incompat- ible—machines in competition for the same market. This has limited the transportability of existing computer-based education mate- rials. Users of different systems cannot eas- 214 . Alternatives to Animal Use in Research, Testing, and Education Canine Cardiopulmonary Resuscitation Simuiator (Resusci-Dog) in Use Photo credit: Charles R. Short, New York State College of Veterinary Meidiine, Cornell University ily share or exchange materials. As a result, there is a serious problem of duplication of effort, with individuals and institutions devel- G G oping similar teaching programs. Although ideas are clearly portable, actual computer programs may not be, and the avenues for effective dissemination of programs remain limited. The resources available to support research and development in computer-based educa- tion are too limited. Few institutions have com- mitted funds for such activity, and much cur- rent work is supported by departmental or individual resources. Many new computer- based education materials are developed by individuals on their own time out of personal interest. There is virtually no external fund- ing available to support advances in this field. In the long run, the most serious problem may well be the lack of professional academic re- wards for faculty members working in this area. Promotion, tenure, and salary increments are awarded predominantly for productivity in the research laboratory, not for efforts to develop innovative teaching techniques and materials, with essentially no external grant support for computer-based education activ- ities and with few refereed high-quality jour- nals in which to publish, two of the measures by which rewards are apportioned are not available to developers of novel educational soft ware. This is a particular problem for jun- ior faculty members, who often must devote their major efforts to climbing the academic ladder. Computer-based education seemingly fails to meet the perception of an academi- cally valid and creditable enterprise. SUMMARY AND CONCLUSIONS In elementary school, student exposure to ani- Taken together, the approximately 53,000 ani- mals in the classroom generally takes the form of reals used in accredited medical and veterinary exercises in humane awareness. Later, involve- schools for education and training make up less ment in science becomes more active and the role than one-half of 1 percent of the estimated 17 mil- of the animal as a tool of science is explored. As lion to 22 million animals used annually in the students advance to and through college, animal United States for research, testing, and education. use often becomes more invasive during instruc- (No data are available on the number of animals tion in laboratory techniques. At the highest levels, used in primary, secondary, and college educa- especially in professional and research training, tion.) Yet the development of students’ attitudes students are expected to attain levels of skill that toward animals during the classroom years over- may be difficult to reach without the use of animals. shadows in importance the actual quantity of ani- — Ch. 9—Animal Use in Education and the Alternatives . 215 mal subjects used in education. Each phase of pri- mary and secondary education appears to offer an opportunity for shaping students’ attitudes toward animals. Grades 8 through 11 seem to be the most appropriate times for influencing the de- velopment of attitudes toward animals. Alternatives applicable to different levels of schooling vary with the educational goals of each level. whereas classroom demonstrations or non- invasive observation could be appropriate in pri- mary and secondary education to teach the scien- tific method and aspects of biology, a nonliving system is inadequate to teach surgical technique and manual dexterity to medical and veterinary students. Computer models of biological phenom- ena and interactive simulations of biological ex- periments are especially promising alternatives to animal use, even in sophisticated laboratory phys- iology exercises. Interactive videodisk programs— although expensive and not currently widely avail- able-offer particularly realistic training simu- lations. CHAPTER 9 REFERENCES 1. American Veterinary Medical Association, Your Ca - reer in Animal Technology (Washington, DC: Jan- uary 1981). 2. Association of American Medical Colleges, Use of Animals in Undergraduate and Graduate Medical Education (Washington, DC: 1985). 3. Coleman, T. G., and Randall, J. E., “HUMAN: A Com- prehensive Physiological Model,” Physiologist 26: 15-21, 1983. 4. International Science and Engineering Fair, Rules of the 35th International Science and Engineering Fair (Washington, DC: Science Service, Inc., 1984). 5. Katz, S., Hollingsworth, R. G., Blackburn, J. G., et al., “Computer Simulation in the Physiolo~ Stu- dent Laboratory,” Physiologist 21:41-44, 1978. 6. Kellert, S. R., “Attitudes Toward Animals: Age- Related Development Among Children, ’’.J. Environ. Educ. 16:29-39, 1985. 7. Michael, J. A., “Computer-Simulated Physiology Experiments: Where Are We Coming From and Where Might We Go?” Physiologist 27:434-436, 1984. 8. Michael, J. A., Associate Professor, Department of Physiology, Rush-Presbyterian-St. Luke’s Medical Center, Chicago, IL, personal communication, Mar. 4, 1985. 9. Nardone, R. M., Director, Center for Advanced Training in Cell and Molecular Biology, Department of Biology, Catholic University of America, Wash- ington, DC, personal communication, Sept. 4, 1985. 10. National Science Teachers Association, Code of Practice on Animals in Schools (Washington, DC: 1981). 11. Oelsner, G., Boeckx, W., Verhoeven, H., et al., “The Effect of Training in Microsurgery, ’’Am. .J. Obstet. Gynecoi. 152:1054-1058, 1985. 12. Packer, J. S., and Packer, J. E., “A Teaching Aid for Physiologists—Simulation of Kidney Foundation, ” The Physiology Teacher 6:15, 1977. 13. Peterson, N. S., Professor, Department of Veteri- nary and Comparative Anatomy, Pharmacology, and Physiology, College of Veterinary Medicine, Washington State University, Pullman, WA, per- sonal communication, Aug. 23, 1985. 14. Progress Without Pain (Lord Dowding Fund, Na- tional Anti-Vivisection Society, Ltd., London), “De- velopment of a Dynamic Model Using the Human Placenta for Microvascular Research and Practice,” 23:6-10, 1985. 15. Randall, J. E., Professor, Medical Sciences Program, Indiana University School of Medicine, Blooming- ton, IN, personal communication, Apr. 25, 1984. 16. Rothe, C. F., “A Computer Model of the Cardiovas- cular System for Effective Learning, ” Physiologist 22:29-33, 1979. 17. Rowan, A. N., Of Mice, Models, & Men: A Critical Evaluation of Animal Research (Albany, NY: State University of New York Press, 1984). 18. Scientists Center for Animal Welfare (Bethesda, MD), “Co]lege Courses on Ethics and AnimaIs, ” Newsletter 5(2):3-6, 1983. 19. Short, C. E., Chief, Department of Anesthesiology, New York State College of Veterinary Medicine, Cornell University, Ithaca, NY, personal commu- nication, March 1984. 20. Swindle, M. M., “Swine as Replacements for Dogs in the Surgical Teaching and Research Laboratory,” Lab. Anim. Sci. 34:383-385, 1984. 21. Tarp, J., “Toward Scientific Literacy for All Our Students,” The Science Teacher 45:38-39, 1978. 22. U.K. Home Office, Scientific Procedures on Living Animals, Command 9521 (London: Her Majesty’s Stationery Office, 1985). 23. U.S. Department of Health and Human Services, 216 “ Alternatives to Animal Use in Research, Testing, and Education Public Health Service, National Institutes of Health, 25. Walker, J. R., “Computer Simulation of Animal Sys- National Survey of Labora tory AnimalFacilities and terns in the Medical School Laboratory,” A]terna - Resources, NIH Pub. No. 80-2091 (Bethesda, MD, tives to Laboratory Animals (U. K.) 11:47-54, 1983. 1980). 26. Walker, J. R., Assistant Director, Integrated Func- 24, Veale, J. L., “Microcomputer Program for Teach- tional Laboratory, University of Texas Medical ing pH Regulation and COZ Transport, ” Fed. Proc. Branch at Galveston, TX, personal communication, 43:1103, 1984. Feb. 22, 1984. Chapter 10 Information Resources and Computer Systems One of the biggest harriers to using available information is that most people do not know how to use existing resources or what systems are available for use. John S. Wassom Oak Ridge National Laboratory March 1985 The best computer programs evolve into large creations. It is rare !v possible to imagine a very large computer activity at the outset and build it as such. Charles S. Tidball The George Washington University Medical Center March 4, 1985 CONTENTS P+!@ Sources of Research and Testing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .219 Primary Literature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ., . . . . . . . . . . . . .219 Secondary Literature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..., . . . ~ ..219 Unpublished Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...........219 Information Centers. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..220 The Availability of Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .......221 Journal Publication Policies. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...221 Federal Laws Affecting Unpublished Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ~ ...221 Barriers to Using Available Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..222 Data Quality and Comparability. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...222 International Barriers to Sharing Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..223 Retrieving Research and Testing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ......223 Abstracting and Citation Services ~ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...........223 Retrieving Unpublished Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...224 Computer Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...........228 Advantages of Computers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .228 Toxicology Data Bank . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ............229 Registry of Toxic Effects of Chemical Substances . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ,229 On-Line Literature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .231 Laboratory Animal Data Bank . . . . . . . . . . . . . . . . . . . . . . . . . . . ~ . . . . . . . . . . . . . . . . . . . . ..233 Building Phase, 1975-80 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...................233 Public Accessibility, 1980-81 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... ... ..235 Reasons for the Failure of LADB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .............235 Lessons Learned From LADB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...236 Expanding the LADB Concept: A Computerized Registry of Research and Testing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ~ . .................237 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .............238 Chapter 10 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ~ . . . . .239 List of Tables Table No. Page 1()-1 Growth and Publication Frequency of Literature Related to Genetic Toxicology, Carcinogenicity, Mutagenicity’, and Teratogenicity . . . . . . . . .220 10-2. Examples of Databases Available for Searches of Literature Involving Animal Research and Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .230 10-3. Examples of On-line Databases of the National Library of Medicine. . . . . ..231 List of Figures Figure No. P a g e 10-1. A Scientific Abstract and Corresponding Index Entry in BIOSIS . ................225 IO-2. Sample Bibliographic Entries in Biological Abstracs/RRM . . . . . . . . . . . . . . . . . . . . ..226 10-3. Promotional Material From Commercial Supplier of Full Texts of’ Scientific Publications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .227 10-4. A Typical Substance Entry in the Registry of Toxic Effects of Chemical Substances (RTECS) . . . . . . . . . . . . . . . . . . . . . ~ . . . . . . . . . . . . . . . . . . . . .. ..232 10-5. A Representative Page of the Eight-Page Data Input Form for the Laboratory Animal Data Bank . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..234 Chapter 10 Information Resources and Computer Systems Earlier chapters have described the quantity and creases information exchange reduces the need variety of data generated by using animals in re- of other investigators to perform the same exper- search, testing, and education. To assess fully the iments. The pivotal role computers can play in that alternatives to animal use in these areas, there- process has recently become an important topic fore, it is important to consider how the data are for consideration and is examined in this chapter. shared once they are generated. Anything that in- SOURCES OF RESEARCH AND TESTING DATA Primary Literature One of the most important ways to make data publicly available is through the “primary litera- ture” in which they are published for the first time and in greatest detail. A significant form of this is the scientific journal, the most up-to-date and ubiquitous of the published sources available. Jour- nal articles that are reviewed by knowledgeable peers before they are accepted for publication are considered especially reliable. Most normally con- tain a description of the methodology of the ex- periment, the results obtained, the conclusions drawn by the author or authors, and references to and discussions of related published and un- published information. Other primary sources are published reports (e.g., of Government-sponsored research), proceed- ings of technical meetings, or similar collections of articles. As a rule, reports and proceedings are not as widely available as journal articles. They may or may not have been peer-reviewed, Secondary Literature Secondary sources contain information drawn solely from other published material. The most common forms are books, reviews, and reports. (A book that contains original material would not be considered a secondary source.) Handbooks are a useful secondary source for numerical data and for citations to the primary literature in which they were first published. Because secondary sources draw from primary sources, the information they report can be somewhat dated, as there is a time- lag ranging from months to years between the pub- lication of a primary source and that of any sec- ondary sources that rely on it. Many reviews and reports are prepared to meet the specific needs of various organizations. Gov- ernment agencies, such as the Food and Drug Administration and the Environmental Protection Agency (EPA), prepare reports to support regula- tory activities. Research institutions, such as the National Institutes of Health and the Chemical Industry Institute of Toxicology (CIIT), prepare reports to announce the results of a particular study. Other organizations, such as the Chemical Manufacturers Association and the World Health Organization, prepare reports to further their programs. Unpublished Information Unpublished information about recent, planned, and ongoing research and testing can be of even greater interest than older, published information. The timelag between submission or acceptance of data for publication and their actual publica- tion is often a handicap to those waiting to learn of experimental results. Time lost while waiting to obtain another investigator’s published research results can cost a laboratory its claim to priority in obtaining research results. In testing, proprie- tary interests create pressure to obtain informa- tion as quickly as possible, One of the oldest sources of unpublished infor- mation is networking-that is, the use of personal 219 220 G Alternatives to Animal Use in Research, Testing, and Education contacts. Networking is affected by the economic factors discussed in chapter 11, such as the pro- prietary value of testing data and the incentives to make it public. Membership in scientific and professional societies and attendance at profes- sional meetings facilitates this form of informa- tion exchange. Recent test results are often pre- sented at meetings of professional societies, and valuable information about work in progress is exchanged by participants. Unpublished data may not be written in report form, which makes it difficult to share the infor- mation. Although the data are stored in some kind of organized fashion, the way one person organizes information may not be useful to someone else. Thus, even if it is possible to determine that un- published useful research or testing data do exist, it is often difficult to share them. In addition to unpublished material, a separate category of information that is fairly inaccessible includes many Government reports, research insti- tute reports, and obscure journals. This informa- tion falls into a grey area—’’published” in a literal sense, but not in a practical one. Information Centers Because of the large volume of published and unpublished information that is generated, spe- cial services called “information centers” have been set up to collect, organize, and disseminate it. An information center, to be comprehensive, must have a fairly narrow scope. These centers are a good vehicle for sharing unpublished information, although they do not have the resources to seek it out. The most well known information center with holdings of research and testing data is the Inter- national Agency for Research on Cancer, in Lyons, France. The United Nations maintains several col- lections of published and unpublished data on chemicals potentially of international interest, e.g., through the International Program on Chemical Safety and the International Registry for Poten- tially Toxic Chemicals in Geneva, Switzerland. These agencies have a much broader scope than a typical information center, although they carry out many of the same functions. The Oak Ridge National Laboratory in Oak Ridge, TN, has indi- vidual information centers for environmental car- cinogens, teratogens, and mutagens. Statistics on the volume and rate of growth of publications in the areas for which Oak Ridge has holdings are given in table 10-1. Table 10.1 .—Growth and Publication Frequency of Literature Reiated to Genetic Toxicology, Carcinogenicity, and Teratogenicity Papers Publication sources published Increase in papers providing Subject per yeara published per yearb information Genetic toxicology. . . . . . . . . . . . . 4,000-5,000 200-300 3,400 In vivo animal carcinogenicity studies . . . . . . . . . . . . . . . . . . . . . 1,500-2,000 50-100 1,000 In vitro cell transformation studies . . . . . . . . . . . . . . . . . . . . . 400-500 25-50 500 Teratogenicity. . . . . . . . . . . . . . . . . 2,000-2,800 100-150 3,500 aFi9ureS Would & substantially greater if augmented with unpublished or Inaccessible published materiai. bNumbers shown are projected increases based on trends cataloged from the literature fOr the period 1979-M. clncludes journais, books, symposium proceedings, 90VWr7mt3nt reports, and abstracts. SOURCE:J.S. Wassom, Director, Environmental Mutagen, Carcinogen, and Teratogen Information Program, Oak Ridge Nation- al Laboratory, Oak Ridge, TN, personal communication, November 1985. Ch. 10—information Resources and Computer Systems Ž 221 THE AVAILABILITY One of the most important incentives to pub- lish, both for people and for organizations, is to establish a professional reputation. Although “pub- lish or perish” is an enduring part of academic tra- dition, in nonacademic research and testing sec- tors there is often little incentive to publish. As a rule, industry is more concerned with the pro- tection of proprietary information and the con- servation of financial resources than with pub- lishing. Federal organizations are likewise more inter- ested in carrying out missions required by law than in the publication of research and testing data (un- less that is their mission). As a result, many agen- cies’ reports are never sent to the National Tech- nical Information Service (NTIS) for distribution and cataloging, or too little time is spent indexing them in a fashion that facilitates easy retrieval of the information. Journal publication Policies Because of the importance of journals as a source of testing data, their publication policies are cru- cial to the effective exchange of information. Some journal policies (e.g., the limitations on the length of an article and the amount of detail it contains) are related to high printing and distribution costs. others, such as an unwillingness to publish results that have already been disclosed publicly, are a result of the stiff competition that exists among journals. One of the most frustrating publication policies from the standpoint of avoiding duplicative re- search and testing is that most journals (and there- fore secondary sources) rarely publish negative results. It is natural that people would be more interested in knowing, for example, which chem- icals have been found to be hazardous than which chemicals have not. As a consequence, a certain number of experimental protocols are repeated because the negative results of earlier experiments were not published. This policy is not likely to change without dramatic alterations in the stance of journal publishers, the policies of professional societies, and, indeed, the tradition of scholarly publication in academia. One notable exception OF INFORMATION to this is the journal Mutation Research, which in 1977 made it a policy to also publish negative results. Federal Laws Affecting Unpublished Data one method available to the Federal Government for collecting testing data is to require them, ei- ther through registration requirements such as those under the Federal Insecticide, Fungicide, and Rodenticide Act (Public Law 92-516, as amended by Public Laws 94-140 and 95-396), or through reporting rules such as those promulgated under the Toxic Substances Control Act (TSCA) (Public Law 94-469). Section 8(d) of TSCA requires manu- facturers and processors to submit citations or cop- ies of health and safety studies they have spon- sored, or about which they are aware, for specified chemicals. As of June 1984, EPA had received over 6,000 such submissions, about half of which were health-effects studies. For the specified chemicals, when regulatory notices were published in the Fed- eral Register, about one-quarter of the citations were to data received under Section 8(d) (10). Some unpublished data are given to Government agencies voluntarily, either through personal con- tacts or in response to publicity that the govern- ment is working on a particular problem. Much of the data concern adverse effects, but some con- cern negative results as well. Unlike most countries, the United States has a policy of making information held by the Gov- ernment as available as possible, consistent with protecting its proprietary value. Key laws in implementing this policy are the Administrative Procedures Act (Public Law 79-404, as amended by Public Law 89-554), which encompasses the Freedom of Information Act (Public Law 90-23, as amended by Public Laws 93-502, 94-409, and 95-454). This act makes all information held by the executive branch of the Federal Government avail- able to anyone who asks for it, unless the infor- mation is specifically exempted or is protected under another law. The person requesting the in- formation is frequently required to pay search and duplication costs, but the burden is on the Govern- ment to show why information should be withheld. 222 G Alternatives to Animal Use in Research, Testing, and Education Under these laws, the public also has access to collections of published and unpublished nonpro- prietary data gathered to support administrative actions such as rulemaking. This ‘(public docket” contains all reports, literature, memos, letters, and other information considered in taking the action. Once information has been obtained by the Fed- eral Government, it may be shared within and among Government agencies. Often such sharing is very informal, and with informality comes un- predictability and oversights. Various committees have been set up to facilitate intragovernmental networking, such as the Interagency Regulatory Liaison Group of the late 1970s, the Interagency Risk Management Council, and the Interagency Toxic Substances Data Committee. These efforts increase the amount of information available to solve particular problems. They also reduce du- plicative information requests made of industry. In 1980, an interagency Toxic Substances Strat- egy Committee examined the sharing of informa- tion, focusing principally on the data held by Fed- eral agencies (20). The Committee noted there were then more than 200 independent data systems, mostly incompatible. Barriers to sharing informa- tion included diverse methods of identifying chem- icals and differing reliability and review of the data- bases. The Committee noted that coordination of Federal agencies’ chemical data systems could re- duce duplication of information gathering, mini- mize delay, and, to some extent, decrease uncer- tainties in decisionmaking. The benefits of such coordination would likely extend beyond the Fed- eral Government to State and local governments, industry, labor, public interest groups, academic institutions, international organizations, and for- eign governments. BARRIERS TO USING AVAILABLE INFORMATION Data Quality and Comparability Before data are to be used, the user must be con- fident of their quality. This judgment is based on a variety of facts and inferences. People will fre- quently take into account the professional repu- tation of the investigator or the investigator’s in- dustrial, academic, or professional affiliation or organization. If the person has no reputation, good or bad, many scientists will not rely on that inves- tigator’s data. This phenomenon is most acute with investigations carried out in foreign countries and published overseas (14). Further, many scientists will not (and perhaps should not) trust results that have not been peer-reviewed. Lastly, some orga- nizations tend not to trust any data that they have not generated. It is important to assess the quality of data. Thus, even though numerical databases are convenient because they contain data in summary form, often there is no way to determine from the informa- tion contained there how reliable the data are (un- less they were peer-reviewed before being put into the system). This problem has been addressed by the National Bureau of Standards (NBS), the Chem- ical Manufacturers’ Association, and others, A workshop held in 1982 (16) recommended that computerized databases (discussed at length later in this chapter) include the following “data qual- ity indicators” that would allow the user to deter- mine reliability for specific needs: G the method(s) used to obtain the data, G the extent to which the data have been eval- uated, G the source of the data, and G some indication of the accuracy of the data. An important part of evaluating data is compar- ing them with data obtained using similar meth- ods—that is, validating the data. In deciding, for example, to rely on a particular test protocol, it is necessary to be confident not only that the test is a useful model of the effect of interest, but also that the results can be trusted, even though they are unexpected. For many investigators, valida- tion involves repeating at least a portion of an ex- perimental protocol in their own laboratories. They might also compare the results with those generated by other procedures with which they are more familiar. Ch. 10—information Resources and Computer Systems . 223 International Barriers to Sharing Information Animal research and testing is conducted in many countries (as described in ch. 16). The im- portance of communicating scientific information among nations has been recognized in the United Nations, in the Organization for Economic Coop- eration and Development (OECD), and in regional and bilateral forums. Although much has been done to facilitate this, many barriers must still be overcome. International communications cost more and take longer than domestic communications. More- over, there are fewer international personal ac- quaintances on whom to rely for information than there are on a national level. Communication prob- lems are exacerbated by institutional differences. It is difficult for industry-to-industry communi- cations to occur, for example, when one industry is privately owned and another is government- owned, because governments typically deal through diplomatic channels. Political animosities hinder information ex- change. Defense-related information is affected the most, but all information sharing must suffer in such a climate. Even political differences cause problems in sharing information. It is difficult for agencies within the U.S. Government to obtain in- formation from governments that have close work- ing relationships with their industries, such as Ja- pan, particularly when any information received would be subject to Freedom of Information Act requests in the United States. Language differences are a large problem, both in the use of written materials and in personal com- munications. Translation and interpreting are ex- pensive, particularly in the United States, where the number of people who speak more than one language has been decreasing. English translation costs for the four principal languages of science (French, German, Russian, and Japanese) range from $40 to $88 Per thousand words. An estimated $4 billion to $5 billion would be required to trans- late the current foreign-language holdings of the National Library of Medicine (NLM), for example, with an ongoing yearly translation cost of $150 million (9). Duplicative translations are avoided through the clearinghouse effort of the John Crerar Library in Chicago, IL. Translations donated by a variety of sources on a broad spectrum of topics are made available to others. Common protocols can also facilitate the inter- national exchange of, for example, testing data. OECD members decided in 1981 that health-effects data generated according to OECD test guidelines should be mutuallv acceptable in all member coun- tries, regardless of where the testing was done (see app. A) (17). Although this decision has not been fully implemented, OECD test guidelines are readily available and are receiving considerable use. RETRIEVING RESEARCH AND TESTING DATA The ways data are obtained and the amount sought are functions of the resources available for searching, how the data are to be used, the likeli- hood that the information exists at all, and how reliable the information is likely to be. Many meth- ods for finding information are available, and most of them overlap to some extent. Abstracting and Citation Services In research and testing, several hundred thou- sand scientific articles in thousands of journals are published each year in the primary literature (6). Abstracting and indexing services and biblio- graphic services play a vital role in making these accessible to those who need them. (An index based on references cited, or citations, permits the user to follow the literature into the future to locate pertinent articles. For example, a user with a 1981 article in hand who is seeking related, more re- cent publications can consult a citation index to identify 1985 publications that referenced the 1981 article.) Because animals are used for a variety of research purposes (see chs. 5 and 6), however, and because testing is interdisciplinary (see chs. 7 and 8), information may be indexed in the fields of chemistry, biology, pharmacology, medicine, and so on. 224 G Alternatives to Animal Use in Research, Testing, and Education Abstracting and indexing services and biblio- graphic services have existed since the 17th cen- tury and have grown in number and size as pub- lished literature has expanded. The first major services for scientific information were published by professional societies (e.g., Chemical Abstracts). Some were sponsored by the Federal Government (e.g., Air Pollution Abstracts and AGRICOLA) or by commercial enterprises (e.g., Current Contents and Environmental Abstracts) (8). Some, such as the Chemical Information System, originated in Government and were later converted into com- mercial enterprises (12). The largest abstracting and indexing service for biological and biomedical research is BIOSIS, the Biosciences Information Service. In 1985, its cov- erage extended to 440,000 items from over 9 )000 sources worldwide. The file accumulated to date contains over 6 million items, the largest biologi- cal file in the English language. Items covered in- clude abstracts and citations for journal articles and other serial publications, and citations to reports, reviews, and scientific meetings (6). A typical abstract of a journal article and an il- lustration of how it is indexed by BIOSIS appear in figure 10-1. Information like this is contained in the semimonthly publication Biological Ab- stracts. Another publication, Biological Abstracts/ RRM, contains bibliographic entries for research reports, reviews, meetings, and books (see fig.lo- 2). BIOSIS also offers several computer-based serv- ices that provide citations tailored to the custom- er’s information needs. All of these resources are regularly used by scientists. As the figures illus- trate, however, it is often difficult to tell from a title, or even from an abstract, whether a particu- lar article would satisfy a reader’s needs. Once a citation has been obtained, it is easy to acquire the full text of a research report. Most libraries have the necessary services available, or the inquirer can write to the author and ask for a reprint. In addition, some commercial vendors offer to supply by mail the full text of virtually any article (see fig. 10-3). A recent comparison of databases for literature on 10 pesticides illustrates the problem of over- lap (15). Eight databases had to be searched in or- der to get 90 percent of all data relevant to a par- ticular regulatory decision. The share of citations produced by these databases that were not rele- vant ranged from 11 to 27 percent. Used together, the four most consistently relevant databases— TOXLINE, CAB Abstracts, BIOSIS, and Chemical Abstracts—produced 25 to 91 percent of all rele- vant citations, with an average of 69 percent. These statistics illustrate the fragmentation that may accompany a literature search. Although the number of databases that need to be searched may be small for some fields, questions of an interdis- ciplinary nature require substantial resources for a complete literature search. Retrieving Unpublished Information Citation services are available for some unpub- lished data and testing in progress. Federal data- bases and publications include the Bioassay Status Report and Tox-Tips of the National Toxicology Program (NTP), the EPA Chemical Activity Status Report, the Current Research Database of the Na- tional Institute for Occupational Safety and Health, NTIS’s Federal Research in Progress, and the Smith- sonian Science Information Exchange (no longer active). There are also many small databases used to keep track of specialized data, such as informa- tion used in the implementation of a specific law. Similar citation services to unpublished data or ongoing testing exist on an international level. The International Agency for Research on Cancer, which has substantial U.S. support, coordinates the sharing of information about current carcino- genicity testing in laboratories around the world and publishes an information bulletin, Survey of Chemicals Being Tested for Carcinogenic Activity. The International Program on Chemical Safety of the United Nations Environment Program (UNEP) is establishing a database for Chemicals Currently Being Tested for Toxicological Effects. This data- base is designed for long-term or otherwise expen- sive studies other than those on carcinogenicity. Participants in both programs include govern- ments, industry, academia, and research institutes. In addition, Infoterra, a service of UNEP, publishes a directory through which experts in numerous subject areas can be located. Assistance is also pro- vided by national representatives. The U.N.’s In- ternational Registry of Potentially Toxic Chemi- Ch. 10—information Resources and Computer Systems G 225 Figure IO-l.—A Scientific Abstract and Corresponding Index Entry in BIOSIS ABSTRACT FORMAT TOXICOLOGY- Major Headlng E N V I R O N M E N T A L A N D I N D U S T R I A L ~ Subheadlng Authors / \ Author Roforsnco Address N u m b e r ~ 23330. CARSONS, JOANNE N and JOHN O. GOULDEN (Arch= Oceanogr. Inst., Phila., Pa. 19103, USA.) The effects of chlorine Article pollutlon on growth and respiration rates of larval lobsters ~ T i t l e (Homarus americanus). BIOL RES 11(12): 1433-1438. 1985. The - length, dry weight and standard respiration rate of Iarval lobsters (H. americanus) were measured following 20 days Immersion In coastal waters surrounding a power plant. Significantly lower increases in dry weight (P<. 05) and significant reductions in stan- dard respiration rates (P<. 01) were measured in exposed organisms when compared to control organisms. Water samples taken from the immersion site contained high concentrations of free Cl. BIOSIS’ INDEXING SYSTEM AUTHOR INDEX (Personal or Corporate Names) Author Index NAME REF. NO. NAME REF. NO. CARSONS J N 23330+ ELL A W 26787 (Personal or Corporate Names) CASEY N 29606 FINEMAN C 26884 DAVIES R 24001 GOULDEN J O 23330 BIOSYSTEMATIC INDEX (Taxonomic Categories) Biosystematic index ARTHROPODA. . . . . . . . . . . . . . HIGHER TAXONOMIC CATEGORY Crustacea. . . . . . . . . . . . . . . . . . . Malacostraca. . . . . . . . . . . . . I LOWER TAXONOMIC CATEGORIES (Broad Taxonomic Categories) Environmental and Industrial Toxicology. . . . . . . . . . . . . . MAJOR CONCEPT 23330 23572 25352 . . . . . . . . . . . . . REFERENCE NUMBERS GENERIC INDEX (Genus-species Names) Generic index GENUS-SPECIES MAJOR CONCEPT REF. NO. (Genus-species Names) HOMARUS-AMERICANUS TOXIC INDUS 23330WILDLIFE AQU 24063 MICROCERUS- BERONI CRUSTAC SYST 19145’s SUBJECT INDEX (Specific Words) A l p h a b e t i c P O S I T I O N Subject index SUBJECT CONTEXT KEYWORD REF. NO. GHT/ THE EFFECTS OF CHLORINE POLLUTION ON GROWTH 23330 (Specif ic Words) TOBACTER/ EFFECT OF SUBSTITUTION ON THE 26575LORINE POLLUTION ON GROWTH AND RESPIRATION RATES 2 3 3 3 0 GERMINATION RADICAL BARRIER TEMPERATURE 27304 SOURCE: The 1985 8/0S/S Information Catalog (Philadelphia, PA’ Biosciences Information Service, 19S5). 226 Ž Altematives to Anirnal Use in Research, Testing, and Education Figure 10=2. -Sampie Bibliographic Entries in Blological Abstracts/RRM EXAMPLES OF BIBLIOGRAPHIC ENTRIES IN BA/RRM: CONTENT SUMMARY FORMAT SOURCE: The 1985 EUOS/S Information Catalog (Philadelphia, PA: Biosciences Information Service, 1985). 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Alternatives to Animal Use in Research, Testing, and Education cals sometimes refers information requests among per month would be referred to some combina- member countries through its national corre- tion of government agencies, industry, academia, spondents. and research institutes that might have unpub- . lished data relevant to the request, The mecha-A recent U.S.-led project of the OECD, generally nisms for referring&quests on the national levelreferred to as “Switchboard,” has also addressed the problems of obtaining information from other and the enlistment of various organizations, eitheras requesters or responders, is the responsibility countries. Unpublished information may be re- quested through the Switchboard system for use of Switchboard’s national focal point. This project in risk assessments or to otherwise protect health and the environment. A pilot system is to be run in which two requests per participating country will begin on a small scale and will be monitored. If appropriate, it could be expanded (18). COMPUTER Computers have two applications as an alterna- tive to using animals in research, testing, and edu- cation. First, they can be used to model or simu- late biological, chemical, and physical systems. In this way, a computer could be used as a direct replacement for some number of animals used in laboratories. This form of computer use is dis- cussed in chapters 6,8, and 9. Second, computers are used to disseminate information that has been generated from prior use of animals in research and testing, thus avoiding the needless repetition of a procedure by other scientists. It is this role of computers as information disseminators that is discussed in the rest of this chapter. Advantages of Computers Biological testing (see ch. 7) can be described as the repetitive use of a standard biological test situ- ation, or protocol, employing different chemicals or different test parameters (e.g., species or bio- logical end points). Because the protocols in test- ing are more stereotyped and less varied than those in research, biological testing is more amenable than research to the institution of a computerized data retrieval system. In fact, testing emerged in the 1970s as the first discipline in which such a system was developed. If a comprehensive, computerized registry of bio- logical research or testing data were established, certain benefits might accrue. These benefits are predicated on the inclusion in the computerized registry of both control and experimental data, and of both positive and negative results. (Data SYSTEMS obtained from testing fall into two broad catego- ries: those derived from untreated (control) sub- jects, and those from treated (experimental) sub- jects. Data obtained from treated subjects may either show an effect from the treatment (“posi- tive results”) or no effect (’(negative results’’).) Fur- thermore, the advantages of such a registry de- pend on the acceptance by working scientists of the data contained in it—acceptance that seems possible only with the imprimatur of peer review of the data. The anticipated benefits of a computer- based registry of research or testing data include: G G Decreased Use of Animals in Research or Testing. In some instances, an investigator would locate the exact data desired, possibly from a previously unpublished source, thus avoiding unintentional duplication of animal research or testing. Baseline data could per- mit the selection of a dose, a route of adminis- tration, or a strain of animal without the need for new animal experiments to establish these factors. Efficiencies could also include the use of fewer doses on smaller numbers of animals. Conceivably, the number of animals required for control groups could be reduced, although many experimental protocols require the use of concomitant control subjects, rather than of data from a pool of control subjects, in or- der to achieve statistical significance. A Check for Genetic Drift. Certain experi- mental results can change over a span of many generations due to subtle, progressive changes in the underlying genetic constitution of the strain of animals ((’genetic drift”). The regis- Ch. 10—information Resources and Computer Systems G 229 G try would provide baseline data within speci- fied time frames of measurement, and make it easy to check for the possibility of genetic drift. New Perspectives on Old Data. By perform- ing statistical comparisons across data sets and identifying relationships not already obvious, unforeseen relations could be established without animal experimentation. The scientific community makes use of a num- ber of computerized literature retrieval services to obtain bibliographic citations and abstracts to the published literature. Most abstracting and in- dexing services started as publications, but most are now available on-line as well. Others, such as AGRICOLA, are only available on-line. Many handbooks and other numerical databases are also available on-line. Several numerical data- bases are sponsored by the Federal Government. The most comprehensive, the recently terminated Laboratory Animal Data Bank, is reviewed in de- tail in the next section. Two current systems, the Toxicology Data Bank and the Registry of Toxic Effects of Chemical Substances, are discussed in some detail here. Table 10-2 lists a number of data- bases available for searches of the research and testing literature. Table 10-3 lists some widely used databases of the NLM. Toxicology Data Bank The Toxicology Data Bank (TDB) was made public by NLM in 1978. It is designed to address some of the needs of the testing and regulatory com- munities for toxicity information. TDB is organized by individual chemicals or substances, now totaling more than 4,000. Its fixed format includes: G G G data on the production and use of each chemical; a description of the physical properties of each chemical; and the results of pharmacological and biochemi- cal experiments, and information on toxico- logical testing. TDB is based on conventional published sources and does not include unpublished data. Thus, base- line data on control animals, which might be used in place of a control group, could not be included because so little has been published. The most valuable feature of TDB is the fact that all the data it contains are peer-reviewed. As a con- sequence, its data summaries are acceptable to most users (5). (Another database containing only peer-reviewed data is the Environmental Protec - tion Agency’s Gene-Tox.) Registry of Toxic Effects Chemical Substances o f The Registry of Toxic Effects of Chemical Sub- stances (RTECS) has been published annually since 1971 by the National Institute for Occupational Safety and Health, under Section 20(a)(6) of the Occupational Safety and Health Act of 1970 (Pub- lic Law 91-596). RTECS is a compendium, extracted from the scientific literature, of known toxic and biological effects of chemical substances. RTECS does not evaluate the data it cites, leaving that responsibility to the reader. An example of the in- formation contained in a typical substance entry in RTECS is given in figure 10-4. By congressional mandate, those data that indi- cate a toxic effect of a chemical are to be included in RTECS; those that show no toxicity are to be excluded. Thus, RTECS does not include negative results. Moreover, a chemical might not be in- cluded in the registry for a variety of reasons, in- cluding the following: G G G The test results could not be cited because the protocol of the study did not meet the RTECS selection criteria. The substance has not yet been tested or the results have not yet been published. The substance has been tested and the results published, but the information has not yet been entered into the RTECS file. The exclusion of negative results from RTECS and its incompleteness for these other reasons may lead to the repetition of toxicity testing of essen- tially nontoxic substances. The production of RTECS costs approximately $500,000 per year. The current quarterly update includes a total of 68,000 compounds, and it con- tinues to grow steadily toward the estimated 100,000 unique substances for which toxicity data may be available. If RTECS were expanded to include all results of whole-animal toxicity testing, including 230 . Alternatives to Animal Use in Research, Testing, and Education Table 10.2.—Examples of Databases Avaiiabie for Searches of Literature Involving Animal Research and Testing Database Description First year covered AGRICOLA Worldwide journal and monograph literature on agriculture and related subjects; from 1970 the National Agricultural Library AQUACULTURE AQUALINE ASFA (Aquatic Sciences and Fisheries Abstracts) BIOSIS Previews CA Search Comprehensive Dissertation Abstracts Conference Papers Index CRIS (Current Research Information System) Enviroline Environmental Bibliography Excerpta Medica INSPEC I PA (International Pharmaceutical Abstracts) IRL Life Sciences Collection ISI/BIOMED ISI/COMPUMATH ISI/lSTP&B LISA (Library Science Abstracts) Microcomputer Index NIMH Oceanic Abstracts Pollution Abstracts Population Bibliography Psychological Abstracts SCISEARCH Growth requirements, engineering, and economics of marine, brackish, and freshwater organisms; from National oceanic and Atmospheric Administration Abstracts from world literature on water, waste water, and aquatic environments; from Water Research Centre, Stevenage, U.K. Life sciences of seas and inland waterways plus legal, political, and social implica- tions of aquatic life; from UNESCO International coverage of life science research; from Biological Abstracts International coverage of chemical sciences; from Chemical Abstract Service Author, title, and subject guide to nearly all American dissertations since 1861 and many from foreign countries; abstracts added beginning in July 1981; from Xerox Univer- sity Microfilms Records of scientific and technical papers presented at major regional, national, and international meetings each year; from Data Courier, Inc. Research in agricultural sciences; from U.S. Department of Agriculture’s State Research Service International coverage of biology, chemistry, economics, geology, law, management, planning, political science, and technology of environmental issues; from Environment Information Center, Inc. Atmospheric studies, energy, general human ecology, land resources, nutrition and health, and water resources; from Environmental Studies Institute Worldwide citations and abstracts from 3,500 biomedical journals; from Excerpta Medica Coverage of literature in computers, electrotechnology, and physics; from the Amer- ican Institute of Electrical Engineers Literature on drug development and use of drugs; from the American Society of Hos- pital Pharmacy Worldwide coverage of life sciences including conferences; from Information Retrieval, Ltd. Index of 1,400 biomedical journals; from the Institute of Scientific Information Covers literature in computer science, mathematics, statistics, operations research, and related areas; from the Institute for Scientific Information Computerized version of Scientific and Technical Proceedings and Books. Covers 3,000 proceedings and 1,500 books annually; from the Institute for Scientific Information international coverage of library and information science literature; from Learned in- formation, Ltd. Subject and abstract guide to 21 microcomputer journals; form Microcomputer infor- mation Services Mental health literature from 950 journals, symposia, government reports, and other sources; from the National Institute of Mental Health International literature on geology, governmental and legal aspects of marine resources, marine biology, marine pollution, meteorology, and oceanography; from Data Courier, Inc. Literature on the sources and control of environmental pollution; from Data Courier, Inc. international coverage of population research: abortion, demography, family planning, fertility studies, and migration; from Carolina Population Center, University of North Carolina Worldwide coverage of literature in psychology and related social-behavioral litera- ture; from the American Psychological Association International literature of sciences and technology; from the institute for Scientific Information 1970 1974 1978 1969 1967 1861 1973 1974 1971 1973 1974 1969 1970 1978 1979 1976 1978 1969 1981 1969 1964 1970 1966 1967 1974 Ch. 10—Information Resources and Computer Systems Ž 231 Table 10-2.—Examples of Databases Availablea for Searches of Literature Involving Animal Research and Testing (Continued) First year Database Description covered SOCIAL SCISEARCH Worldwide coverage of social and behavioral sciences literature; from the Institute 1972 for Scientific Information TELEGEN Covers literature on biotechnology and genetic engineering in 7,000 sources includ- 1973 ing conference and symposia papers, government studies, periodicals, and the popu- lar press; from Environment Information Center, Inc. Zoological Record Covers zoological literature from 6,000 journals; from Biosciences Information Serv- 1978 ice and the Zoological Society of London aThe~~ d~~aba~~~ are available bY tel~~h~n~ ~On”~Ctl~n to one or more of the following: Lockheed Information System DIALOG, SyStem Development Corp ‘S ORBIT, and Bibliographic Retrieval Service, Inc. SOURCE Adapted from R,V. Smith, Graduate Research (Philadelphia, PA: ISI Press, 19S4). Table 10-3.—Examples of On-Line Databases of the National Library of Medicine Number of records Name Description (average length) Type of record MEDLINE 1966-present. Bibliographic citations and abstracts from 3,300,000 Bibliographic primary biomedical literature (1,250 char.) TOXLINE 1965-present. Abstracts from primary toxicological literature 1,400,000 Bibliographic (1,050 char.) CHEMLINE Dictionary to chemicals contained in TOXLINE and other MED- 500,000 Chemical compound LARS data bases (275 char.) RTECS Brief summaries of toxicity results from primary literature 68,000 Chemical compound (1,000 char.) TDB Detailed chemical, pharmacological, and toxicological data and 4,000 Chemical compound extracts from monographs and handbooks (17,000 char.) SOURCE: Office of Technology Assessment negative results, its size would be increased by an estimated 10 to 15 percent (11). RTECS is avail- able in hard copy (19), on microfiche, on magnetic tape, and on-line from both the MEDLINE service of NLM and the Chemical Information System, a joint resource of several Federal agencies that is managed by EPA. On-Line Literature The research community makes use of a num- ber of computerized literature retrieval services to obtain bibliographic citations and abstracts from primary literature. Among these, for example, is NLM’s MEDLINE database, a bibliographic file now exceeding 3,300)000 entries. In the private sector, Biosciences Information Services prepares hun- dreds of thousands of abstracts each year, pro- viding access to essentially the entire published biological research literature. However, the re- search community is not presently served by a computerized database that includes comprehen- sive descriptions both of experimental protocols and of the resulting data. Movement toward on-line delivery of the full text of scientific publications has begun in the private sector. For example, Mead Data Central (Dayton, OH) offers MEDIS, a medical literature database. In 1985, the MEDIS service included about 70 pub- lications, with some stored journal articles going back to 1980. MEDIS includes the full text of the Journal of the American Medical Association (since 1982), Archives of Internal Medicine, and some textbooks and newsletters. In 1984, Bibliographic Retrieval Services (Latham, NY) joined with pub- lisher W.B. Saunders Company to offer the full text of the New England Journa/ of A4edicine and several other journals on-line. A serious limitation to any current full-text literature retrieval system is the inability to retrieve graphs, photographs, and other images (7). 232 G Alternatives to Animal Use in Research, Testing, and Education Figure 1O-4.—A Typical Substance Entry in the Registry of Toxic Effects of Chemical Substances (RTECS) Q4 76 MOLFM: C. Re-S-Te “ TSL (06 S.) - A M I H B C AMIHBC PNASA6 10,61,64 10,61,64 72,513.75 MUREAV4,53,67 TXAPA9 23,288,72 GISAAA 42(1),32,77 TJADAB 19.41A,79 NTIS*” AD-900-000 TOXID9 1,125,81 29ZUA8 -,183,80 JTEHD6 - [SUPPL.2;,69,77 BJCAAI 16,275,62 AIHAAP 23,95.62 MarJV# 26 Apr 76 FCTXAV 17(3),357.79 WQCHM” 2,-.74 IARC 20,151,80 PLMJAP 6(1),160,75 DTLVS* 4,368,80 DTLVS” 4,358.80 FEREAC 39,23540,74 FEREAC 41,57018,76 NTIS** STATUS. SELECTED BY NTP FOR CARCINOGENESIS BIOASSAY AS OF SEPT 1982 STATUS: NTP SECOND ANNUAL REPORT ON CARCINOGENS, 1981 STATUS: NIOSH MANUAL OF ANALYTICAL METHOOS, VOL 3 S255 STATUS: NIOSH CURRENT INTELLIGENCE BULLETIN 41, 1980 STATUS: REPORTED IN EPA TSCA INVENTORY, 1962 STATUS: EPA TSCA 8(a) PRELIMINARY ASSESSMENT INFORMATION FINAL RULE FEREAC 47,26992,82 Ch. 10—Information Resources and Computer Systems G 233 Key to Figure 10-4 A. RTECS accession number, a sequence number assigned to each substance in the Registry. 1. 2. 3. 4. 5. 6. 8.: 9. 10. 11. 12. 13. 14. 15. 16. 17. Substance name. Date when substance entry was last revised. American Chemical Society’s Chemical Abstracts Service unique identification number for the substance. Molecular weight of the substance, Molecular or elemental formula of the substance. Synonyms, common names, trade names, and other Skin and eye irritation data. Mutation data. Reproductive effects data. Tumor-causing data. Toxicity data. chemical names for the substance. Acronyms for the references from which the data and other citations were abstracted. Aquatic toxicity rating. Reviews of the substance. Standards and regulations for the substance promulgated by a Federal agency. A Criteria Document supporting a recommended standard has been published by NIOSH. Status information about the substance from NIOSH, EPA, and the National Toxicology Program. SOURCE: US. Department of Health and Human Services, National Institute for Occupational Safety and Health, Registry of Toxic Effects of Ctremica/ Substances, R.L, Tatken and R,J. Lewis, Sr. (eds.) (Cincinnati, OH: DHHS (N IOSH) Pub. No. 83-107, 1983). LABORATORY ANIMAL DATA BANK The Laboratory Animal Data Bank (LADB) is a computerized set of records of baseline data of physiological, histological, and other biological properties of mammalian species (largely rodents) used in research and testing. The data contained in LADB were derived from both research and test- ing, and are relevant to both areas of animal use. Although LADB exists today only as an archival reference, and is no longer publicly available on- line, it is of great historical interest in a considera- tion of computer-based information resources. In 1970-73, as the carcinogenesis bioassay pro- gram of the National Cancer Institute (NCI) was developed, NCI’s Division of Cancer Cause and Pre- vention anticipated needing better access to base- line data for experimental animals. In 1973-74, NLM helped formulate the concepts leading to LADB. The major contributor of funding for LADB was NCI. Data for LADB were derived from published and unpublished reports. Only control, or baseline, data from groups of animals were included. The data were collected and entered into LADB via a standard, eight-page form (reproduced in ref. 2) that surveyed 306 variables, including: G G G G G G name and manufacturer of the animals’ feed, vaccinations given to the animals, organs or tissues routinely examined at autopsy, blood variables that were analyzed, detergent used in washing cages, and source of the animals. The first page of that form s reproduced in fig- ure 10-5. Building Phase, 1975-so Battelle Laboratories (Columbus, OH) was awarded an NLM contract in 1975, after a competitive pro- curement, and began detailed design activities in 1975-76. Methods for obtaining data were devel- oped, and the data file was designed to permit inter- active access, or time-sharing, by users. Sufficient data were entered to permit initial study by NLM staff in 1976, and in the following year 13 outside users were allowed to test the system. In June 1976, NLM requested the Institute of Lab- oratory Animal Resources (ILAR) of the National Academy of Sciences to provide advice on scien- tific and technical aspects of LADB. A Committee on Laboratory Animal Data was formed by ILAR 234 G Alternatives to Animal Use in Research, Testing, and Education Figure 1O-5.—A Representative Page of the Eight-Page Data Input Form for the Laboratory Animal Data Bank Animal Group Environment and Husbandry Conditions [ FOR INTERNAL LADB USE ONLV LADB Animal Group Number + Related Animal Group Numbers Carnivores Now go to qtitlori 3s 1 Now(pJLoquowbon35 Iv G Includes orn~s of Application Under the CrueltLv to Anjmals Act, 18T6 (London: 1971). 69. U.K. Home Office, Advisory Committee on the ACi - ministration of the Cruelty to Animals Act, 1876, Report on the LD,O Test to the Secretaty of State (London, 1979). 70. U.K. Home Office, Scientific Procedures on LiI’ing Anhnafs Command 9521 (London: Her Majesty’s Sta- tionery Office, 1985). 71. Voetz, D., Federal Ministry of Food, Agriculture, and Forestry, Federal Republic of Germany, Bonn, West Germany, personal communication, No\rem - ber 1984. 72. t$[eibel, E. R., “Man’s Relation to Experimental Ani - 73 74 reals. Modern Times—The Present Situation in S\\~it- zerland,” at the Second CFN Symposium, The Ethics of Anjmal Experimentation, Stockholm, S\veden, Aug. 12-14, 1985. weihe, w., “problems of Alternatives to Aninlal EX - pediments, ” Fortschritte der Medizin 100:2162- 2168, 1982. Weihe, w., “Regulation of Animal Experimenta- tion—The International Experience: S\iitzerland,” presented at the Second CFN Symposium, The Ethics of AnjmaI L’xperimentatjon, Stockholm, Sweden, Aug. 12-14, 1985. 75. Yokoyama, A., Handbook on E,~pe[’iI1leI]talAnjI1]als (Tokyo: Yokendo, 1983). 380 “ Alternatives to Animal Use in Research, Testing, and Education 76. Zbinden, G., and Flury-Roversi, M., “Significance 77. Zobrist, S., Attache for Science and Technology, Em- of the LD~O Test for the Toxicological Evaluation bassy of Switzerland, Washington, DC, personal of Chemical Substances, ” Arch. Toxicol. 47:77-99, communications, October 1984 and December 1981. 1985. Appendixes Appendix A Testing Guidelines Testing Guidelines Testing guidelines are developed for a variety of rea- sons: to allow results of various test substances or spe- cies to be easily compared, to encourage the use of cer- tain protocols so that testing need not be repeated, and to facilitate the work of those who design and carry out tests. Many organizations have developed testing guidelines. Three such compilations have been selected for discussion. FDA Guidelines Involving Whole Animal Testing To the extent possible, the Food and Drug Adminis- tration (FDA) makes its animal testing guidelines con- sistent throughout the agency and consistent with those of other agencies and organizations. However, special uses of products require special testing, and guidance is available from agency staff to help manufacturers meet those requirements. In this table, tests that gen- erally can be considered common or standard toxico- logical tests usually used throughout the agency are grouped together. Those that are more specific for evaluation of the safety of certain products are identi- fied with the FDA Center responsible for regulating that product. I. Agency-wide A. General Toxicity 1. Acute oral—rodent, nonrodent 2. Acute dermal—rodent, nonrodent 3. Acute inhalation—rodent 4. Subchronic oral—rodent, nonrodent 5. Chronic oral—rodent, nonrodent 6. Carcinogenicity —rodent 7. Combined chronic/carcinogenicity —rodent B. Specific Effects 1. Dermal sensitization—guinea pig 2. Dermal irritation—rabbit 3. Eye irritation—rabbit 4. Teratogenicity—rodent, rabbit 5. Reproduction—rodent 6. Absorption, distribution, metabolism, elimination—rodent, nonrodent 7. Neural-behavioral—rodent, rabbit H. Center-oriented A. Human Drugs 1. Subchronic inhalation—rodent, nonrodent 2. Subchronic dermal—rodent, nonrodent 3. Vaginal and rectal administration—rodent, nonrodent 4. Immunotoxicity—rodent B. Food Additives/Color Additives I. Immunotoxicity—rodent 2. Protein quality—rodent 3. Vitamin D assay—rodent C. Biologics 1. All biologics administered by injection a. Safety—guinea pigs, mice b. pyrogenicity–rabbits 2. Vaccines a. Safety—mice, suckling mice, chimpanzees, monkeys, guinea pigs, rabbits b. Potency–guinea pigs, mice, monkeys c. Hypersensitivity-guinea pigs d. Toxicity—mice 3. Antitoxins a. Potency—guinea pigs, mice 4. Toxins a. Potency—mice 5. Toxoids a. Potency—mice 6. Immune globulins a. Potency—guinea pigs 7. Tuberculin a. Safety—guinea pigs b. Potency—mice D. Devices 1. Corneal metabolism—rabbit 2. Biomaterial implant—rabbit, primate, cat 3. U.S.P. intracutaneous—rabbit E. Cosmetics 1. Primary skin irritation and corrosivity - rabbit 2. Phototoxicity—nude mouse, rabbit, guinea pig F. New Veterinary Drugs 1. 2. 3. 4, 5. 6. 7. 8. Safety, efficacy—target species Drug tolerance—target species Reproduction studies—target species Tissue irritation—target species Combination drug—target species Drug disposition—target species Route of administration—target species Intramammary infusion-dairy cows, goats 383 384 G Alternatives to Animal Use in Research, Testing, and Education OECD Guidelines Involving Whole Animal Testing The Organization for Economic Cooperation and De- velopment (OECD) guidelines have wide acceptance in the United States and abroad because of the Mutual Acceptance of Data Decision (l). Under the terms of this decision, member countries of OECD must accept data generated in other countries if done so according to these guidelines. Animal tests contained in the guide- lines are listed below. 1. Effects on Biotic Systems 202 Daphnia, acute immobilization test and re- production test 203 Fish, acute toxicity test 204 Fish, prolonged toxicity test: 14 day study 205 Avian dietary toxicity test 206 Avian reproduction test 2. Degradation and Accumulation 305A Bioaccumulation: Test 305B Bioaccumulation: 305C Bioaccumulation: Bioconcentration 305D Bioaccumulation: 305E Bioaccumulation: 3. Health Effects Sequential Static Fish Semi-static Fish Test Test for the Degree of in Fish Static Fish Test Flow-through Fish Test Short-Term Toxicology 401 Acute oral toxicity 402 Acute dermal toxicity 403 Acute inhalation toxicity 404 Acute dermal irritation/corrosion 405 Acute eye irritation/corrosion 406 Skin sensitization 407 Repeated dose oral toxicity—rodent: 14/28 day 408 Subchronic oral toxicity–rodent: 90 day 409 Subchronic oral toxicity—nonrodent: 90 day 410 Repeated dose dermal toxicity: 14/28 day 411 Subchronic dermal toxicity: 90 day 412 Repeated dose inhalation toxicity: 14/28 day 413 Subchronic inhalation toxicity: 90 day 414 Teratogenicity 415 One-generation reproduction toxicity study 416 Two-generation reproduction toxicity study 417 Toxicokinetics 418 Acute delayed neurotoxicity of organophosphorous substances 419 Subchronic delayed neurotoxicity of organophosphorous substances: 90 day Long-Term Toxicology 451 Carcinogenicity studies 452 Chronic toxicity studies 453 Combined chronic toxicity/carcinogenicity studies Genetic Toxicology 474 Genetic toxicity: micronucleus test 475 In vivo mammalian bone marrow cytogenetic test—chromosomal analysis 478 Rodent dominant lethal test Pesticide Assessment Guidelines Involving Whole-Animal Testing The Office of Pesticide Programs of the Environmen- tal Protection Agency (EPA) has developed guidelines for testing required under the Federal Insecticide, Fun- gicide, and Rodenticide Act. These Pesticide Assessment Guidelines contain standards for conducting acceptable tests, guidelines for the evaluation and reporting of data, guidelines as to when additional testing might be re- quired, and examples of acceptable protocols (2). Simi- lar guidelines have been developed by EPA’s Office of Toxic Substances (OTS) for testing required under the Toxic Substances Control Act (3). Subdivision E: Hazard Evaluation: Wildlife and Aquatic Organisms Series 70: General Information and Requirements Series 71: Avian and Mammalian Testing 71-1 Avian single-dose oral LD5O test 71-2 Avian dietary LC5O test 71-3 Wild mammal toxicity test 71-4 Avian reproduction test 71-5 Simulated and actual field tests for mammals and birds Series 72: Aquatic Organism Testing 72-1 Acute toxicity test for freshwater fish 72-2 Acute toxicity test for freshwater aquatic invertebrates 72-3 Acute toxicity test for estuarine and marine organisms 72-4 Fish early life-stage and aquatic invertebrate life-cycle studies 72-5 Life-cycle tests of fish 72-6 Aquatic organism accumulation tests 72-7 Simulated or actual field testing for aquatic organisms Subdivision F: Hazard Evaluation: Humans and Domestic Animals Series 80: Overview, Definition, and Genera] Requirements Series 81: Acute Toxicity and Irritation Studies 81-1 Acute oral toxicity study App. A—Testing Guidelines G 385 81-2 Acute dermal toxicity study 81-3 Acute inhalation toxicity study 81-4 Primary eye irritation study 81-5 Primary dermal irritation study 81-6 Dermal sensitization study 81-7 Acute delayed neurotoxicity of Series 82-1 82-2 82-3 82-4 82-5 Series 83-1 83-2 83-3 83-4 83-5 Series 84-1 84-2 Series 85-1 85-2 organophosphorous substances 82: Subchronic Testing Subchronic oral toxicity (rodent and nonrodent): 90 day study Repeated dose dermal toxicity: 21 day study Subchronic dermal toxicity: 90 day study Subchronic inhalation toxicity: 90 day study Subchronic neurotoxicity: 90 day study 83: Chronic and Long-Term Studies Chronic toxicity studies Oncogenicity studies Teratogenicity study Reproductive and fertility effects Combined chronic toxicity/oncogenicity studies 84: Mutagenicity Purpose and general recommendations for mutagenicity testing Mutagenicity tests (described in very general terms, with reference to the OTS guidelines) 85: Special Studies Metabolism study Domestic animal safety testing Subdivision G: Product Performance Series 95: Efficacy of Invertebrate Control Agents 95-1 Genera] considerations 95-8 Livestock, poultry, fur and wool-bearing ani- mal treatments 95-9 Treatments to control pests of humans and pets Series 96: Efficacy of Vertebrate Control Agents 96-1 96-2 96-3 96-4 96-5 96-6 96-7 96-8 96-9 General considerations Fish control agents Aquatic amphibian control agents Terrestrial amphibian and reptilian control agents Avian toxicants Avian repellents Avian frightening agents Mole toxicants Bat toxicants and repellents 96-10 Commensal rodenticides 96-11 Rodenticides in orchards 96-12 Rodenticides on farm and rangelands 96-13 Rodent fumigants 96-14 Rodent repellents on tree seeds 96-15 Rodent repellents on cables 96-16 Rodent reproductive inhibitors 96-17 Mammalian predacides 96-18 Domestic dog and cat repellents 96-19 Browsing animal repellents 96-30 Methods and protocols Subdivision M: Biorational Pesticides (This subdivision duplicates many of the provisions of other subdivisions, and is therefore not described in detail. ) Series 150: Overview, Definitions, and General Provisions Series 152: Toxicology Guidelines Subseries 152A: Toxicology Guidelines Subseries 152B: Toxicology Guidelines for Microbial Pest Control Agents Series 154: Nontarget Organism Hazard Guidelines Subseries 154A: Nontarget Organism Hazard Guidelines for Biochemical Agents Subseries 154B: Nontarget Organism Hazard Guidelines for Microbial Agents Series 157: Experimental Use Permit Guidelines Subdivision N: Environmental Fate Series 165: Accumulation Studies 165-4 Laboratory Studies of Pesticide Accumulation in Fish 165-5 Field Accumulation Studies of Aquatic Nontarget Organisms Appendix A References 1. Organization for Economic Cooperation and Development, Guidelines for Testing of Chemicals, and addenda (Paris: 1981). 2. US. Environmental Protection Agency, Pesticide Assess- ment Guidelines (Springfield, VA: National Technical In- formation Service, 1984). 3. U.S. Environmental Protection Agency, Office of Toxic Sub- stances Health and Environmental Effects Test Guidelines (Washington, DC: update October 1984). Appendix B Regulation of Animal Use within Federal Departments and Agencies Six Federal departments and four Federal agencies conduct animal experimentation within Federal facil- ities, or “intramurally. ” Of those, only the Departments of Commerce and Transportation, which use few ani- mals, have no specific guidelines. A seventh Federal department, the Department of Energy (DOE), conducts no intramural animal experimentation, but has a pol- icy on animal experimentation for its extramural con- tracted work. The other entities all have some type of policy for intramural use of animals. Effective December 1986, each Federal research fa- cility will be required to establish an animal care and use committee with composition and function as de- scribed in the 1985 amendments to the Animal Welfare Act (see ch. 13). Each Federal committee will report to the head of the Federal entity conducting the animal experimentation. Several generalizations can be drawn about the guide- lines of the Federal entities conducting intramural ani- mal experimentation. Most policies on proper animal care and treatment include: G G G G G G G adherence to the Animal Welfare Act and to the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (NIH) (26) as well as the Public Health Service (PHS) Policy on Hu- mane Care and Use of Laboratory Animals by Awardee Institutions (see app. C); an animal care and use committee with at least three members (the attending veterinarian and two scientists within the agency); an attending veterinarian responsible for maintain- ing the proper animal care standards; some prior review of protocols and animal species use, usually accomplished by an animal care and use committee; no real mechanism for enforcement of the policy, with the primary responsibility for maintaining the proper standards and adhering to agency guide- lines lying with the individual investigator; a minimal number of site inspections and no real oversight mechanism; and a policy calling for using as few animals as possi- ble and encouraging the use of alternative meth- ods wherever feasible. Some agency policies are noteworthy for additional provisions intended to promote high standards of ani- mal care and use: NIH requires all animal research committees to in- clude one member sensitive to bioethical issues and G G G G not employed in the same NIH bureau, institute, or division. This person must be a Federal Govern- ment employee and so may or may not be a lay- person. These committees have explicit responsi- bilities and a detailed administrative structure in which to carry out duties. The Ames Research Center of the National Aero- nautics and Space Administration (NASA) has dem- onstrated the successful participation of lay com- mittee members in the consideration of animal welfare issues: 40 percent of the committee are laypeople, a format set up at NASA’s instigation. Since 1971, the Veterans’ Administration (VA) has required that all facilities using animals seek and ob- tain accreditation by the American Association for Accreditation of Laboratory Animal Care (AAALAC). The VA has a contract with AAALAC covering all its research facilities, thus prohibiting failure of accreditation of any constituent facility solely for financial reasons. The Department of Energy also requires the facilities of its extramural contractors to be AAALAC-accredited. The Department of Defense (DOD) has a policy and committee distinct from its general animal policy to ensure proper care and use of nonhuman primates. The policies at the Food and Drug Administration (FDA) and the Department of the Interior give a great deal of flexibility to the research centers to allow specific policies tailored to the needs and de- mands of each animal facility. Although this may have many advantages, it may make the mainte- nance and monitoring of a standard of care through- out the agency difficult. Department of Agriculture Regulation of animal use in research within the U.S. Department of Agriculture (USDA) involves adherence to the Animal Welfare Act and to the NIH Guide (26). Much of the animal research performed by USDA in- volves farm animals, which are largely excluded from these policies. The system of compliance involves peri- odic checking of intramural research facilities. For ex- tramural research, no enforcement occurs; hence the system is largely voluntary and self-regulating (15). Department of Defense The general policy on animal use in all Department of Defense programs is contained in DOD Directive No. 386 App. B—Regulation of Animal Use Within Federal Departments and Agencies G 387 3216.1, issued by the Deputy Secretary of Defense in 1982. This statement sets policy on the humane treat- ment and appropriate care of animals used in research and the responsibilities of different DOD personnel to carry out the directive. In general, it follows the Ani- mal Welfare Act and the NIH Guide, along with attempt- ing to incorporate alternatives to animal use in the form of replacement, reduction, and refinement. Other, spe- cial policies treat the general use of nonhuman primates and prohibit the use of dogs, cats, and nonhuman pri- mates for developing nuclear weapons. The directive also requires that all proposals or designs for animal experiments undergo appropriate animal welfare re- view to confirm: “I) the need to perform the experi- ment or demonstration; 2) the adequacy of the design of the experiment or demonstration; and 3) compliance with established policy on the use of animals” (20). Army Regulation 70-18 (a Joint Service regulation) implements the directive’s policies uniformly for all DOD components. The authority for enforcing this reg- ulation is conferred to the Secretary of the Army, who is required to develop and issue, in consultation with the other DOD components, regulations implementing the directive. Army Regulation 70-18 states that all DOD facilities using animals should seek AAALAC accredi- tation. Also, it sets up a long chain of responsibilities for establishing and policing animal welfare policies. The regulation states that the Under Secretary of De- fense for Research and Engineering will:” 1) issue pol- icies and procedural guidance under DOD directive 3216.1, 2) allocate nonhuman primate resources, and 3) designate a veterinarian as the DOD representative to IRAC [Interagency Research Animal Committee]” (21). The Surgeon General of each DOD component involved in animal research must supervise animal use and im- plement this regulation in each component, establish a joint working group to identify and conserve non- human primate resources, and establish and provide representatives to a joint technical working group that periodically reviews the care and use of animals in DOD programs. Finally, the local commander of a facility must ensure that: all programs involving animals conform to the guidelines cited in Army Regulation 70-18; local animal care and use, procurement, and trans- portation policies and procedures comply with the regulation; animals used or intended to be used will experi- ence no unnecessary pain, suffering, or stress, and their use will meet valid DOD requirements; alternatives to animal species will be used if they produce scientifically satisfactory results; and dogs, cats, or nonhuman primates are not used in research conducted to develop nuclear, biological, or chemical weapons (21). Thus, the powers and responsibilities for carrying out DOD animal welfare policies are decentralized. DOD does not do any inspections of its facilities. The facil- ities are required to submit annual reports to USDA under the regulations implementing the Animal Wel- fare Act. The Army regulation builds the institutional review structure around the local animal care committee. Lo- cal commanders must form a committee to oversee the care and use of animals in their facilities. The committ- tee must have at least three members, including at least one person not involved in the proposed project and one veterinarian. The committee reviews: 1) all aspects of animal care to ensure that established policies, stand- ards, and regulations are complied with; and 2) all research protocols and proposals for proper animal wel- fare policies and good animal experimentation stand- ards. Sufficient information to do this animal care and treatment review must be presented with all research proposals. In addition, proposals that involve experi- mentation on nonhuman primates are reviewed sepa- rately by the proper DOD component office (21). As with other departments in the Federal Govern- ment, DOD contracts with outside investigators for some of its research. The DOD extramural animal re- search policy requires that the same standards outlined in Army Regulation 70-18 be followed by contractors in order to receive DOD funds. Assurance is obtained by written statements from the recipient animal care committee or other responsible official. An assurance is also required that the proposal or protocol has been reviewed and approved by the local animal care and use committee or by the attending veterinarian (21). Enforcement of these policies for extramural research is more difficult than the intramural policy, since in- vestigators and administrators are not directly respon- sible to the military line of command. In addition to DOD-wide policies issued by the Office of the Secretary, a recommendation is pending in the Army Medical Research and Development Command that an Advisory Committee on Animal Welfare be ap- pointed, including non-DOD representatives, to meet periodically about concerns related to the use of ani- mals for research and training purposes (7). In 1983, the Air Force commissioned an outside re- view panel to study animal use in its Aerospace Medi- cal Division. The panel looked at Brooks Air Force Base (San Antonio, TX) and Wright -Patterson Air Force Base (Dayton, OH), which together account for 95 percent of the service’s animal use. The panel found the cur- rent policy in place to be satisfactory and was (17): ... impressed with the thoroughness and genuine con- cern of all those involved to ensure that appropriate measures are taken to effect proper care and use of animals. Furthermore, there was a clear emphasis on selection of alternatives to animal use where feasible. Excellent progress was shown in the use of simulation models for a variety of radiation and toxicological studies. 388 G Alternatives to Animal Use in Research, Testing, and Education The panel did note that the system of care and treat- ment policies was too informal and based on the cur- rent personnel; it was unconvinced the system would remain in place if staff were transferred. The Aero- space Medical Division of the Air Force drafted a Sup- plement to Army Regulation 70-18 to implement some of the review panel’s recommendations. The most sub- stantial change deals with the animal care and use com- mittee membership (21): The local commander will appoint at least one lay person from the local community who has no direct Department of Defense connections to serve as a mem- ber of the Committee. This lay member should not be a veterinarian or research scientist who works with animals; however, a background in sciences would be helpful. The Committee may have permanent or ad hoc membership. Its specific purpose is to review all pro- tocols, experimental designs, or lesson plans that in- volve the use of animals and assure compliance with [DOD policy]. The Air Force Supplement to Army Regulation 70-18 also requires that each organization using animals sub- mit not just the Annual Report of Research Facility of USDA’s Animal and Plant Health Inspection Service (APHIS), but also an Annual Animal Use Report, listing all species used, the inventory at the beginning of the year, additions and losses to the facility, the ending in- ventory, the utilization of the animals, the different ex- perimental situations, and the projected use of animals for the next fiscal year (18). Department of Energy The Department of Energy has no intramural re- search facilities and so contracts for all its research. The division involved with animals is the Office of Health and Environmental Research (OHER); programs involving research with animals represented less than 15 percent of OHER’s total research budget for fiscal year 1985 (5), Proposals for OHER-funded research are subjected to outside peer review for scientific merit. An OHER research committee from the Office’s four divi- sions has final approval before funding a research proposal. The OHER policy for animal use by its extramural contractors places the prime responsibility for the main- tenance of animal facilities and for animal care on the contractor. OHER contract research facilities are bound by law to comply with the Animal Welfare Act and its regulatory policies, and OHER personnel maintain close liaison to assure such compliance. In addition, the IRAC principles are part of the OHER policy statement, along with the requirement to maintain AAALAC accredita- tion (5), To enforce these policies, one OHER staff member has responsibility for monitoring animal research pro- grams for compliance. This staff member must main- tain contact with the research facilities to assure ac- creditation and to affirm, at least yearly, that it is being maintained. Site visits with at least one noncontract veterinarian who is an expert in laboratory-animal care may be conducted to evaluate the care and treatment of experimental animals (5,6). Department of Health and Human Services Food and Drug Administration The Food and Drug Administration has recently played a major role in attempting to address animal welfare issues. In 1983, the agency took two steps in this direction by sponsoring an acute studies workshop and by establishing an Agency Steering Committee on Animal Welfare Issues. The workshop helped clarify FDA’s position on its need for toxicity data, especially from the LD50 test. The points emerging from the workshop were that: FDA had no regulations mandating use of the LD5O test; the requirement by Federal agencies for LD5O data from regulated parties was much less than per- ceived by the public; government and industry agreed that there are bet- ter determinants of acute toxicity than the LD5O test and that they supported developing valid alter- natives to the use of animals for testing chemicals; U.S. Government agencies are cooperating with other countries through organizations like the Organization for Economic Cooperation and De- velopment; and improvements in the way animals are used for tox - icity testing can and should be made administra- tively rather than through legislation (1,22). The steering committee, which in part grew out of the acute studies workshop, found several FDA refer- ences to the LD5O that could be misinterpreted as re- quirements to perform the test, and one involving three antitumor antibiotics where the requirement still ex- isted (in contrast to the workshop findings). Its 1984 report states that, in all these instances (except for the antitumor antibiotics), regulations and guidelines are being rewritten to resolve any misunderstandings. They will then reflect the position of FDA that “the use of this test should be avoided except for those rare sit- uations where no alternative exists. ” In the case of the antitumor antibiotics, FDA is considering eliminating the requirement (23). Addressing five specific considerations, all part of its investigation of agency testing guidelines and practices App. B—Regulation of Animal Use Within Federal Departments and Agencies G 389 to answer questions raised at the acute studies work- shop, the steering committee concluded that: G G G G G FDA practices and procedures are designed to ob- tain the maximum amount of data from the mini- mum number of animals; despite general references to the use of LD5O tests, FDA has no requirements for LD5O data obtained by using the classical, statistically precise test, ex- cept for batch release toxicity tests of three anti- tumor antibiotics; there are many alternative tests being studied and developed throughout FDA; practices and procedures for assuring humane care and treatment of animals are agency-wide; and FDA has a number of regular channels of commu- nication to industry, consumers, and the private sector in general and efforts to improve commu- nication channels will continue (23). The steering committee recommended workshops on acute toxicity studies throughout the agency, on the use of in vitro alternatives by various centers, and on agency and PHS practices and procedures for the care and handling of animals. The recommendations also called for the establishment of an agency-wide animal welfare committee (23). FDA is now setting up two in- house workshops to address the first two topics (l). Furthermore, it has established a Research Animal Council to see that the recommendations of the report are carried out, to consider animal research issues at FDA in a broad context, and to serve as an oversight committee for individual FDA centers. FDA’s Research Animal Council began meeting quarterly in 1984 and will report to the Commissioner; its membership in- cludes one representative from each of the centers within FDA (3). FDA policies on humane animal care and treatment require compliance with the Animal Welfare Act as well as with other standards for humane care and use of animals. The steering committee report found that all centers have acceptable procedures, but that they var- ied in specific details. The centers conduct different amounts of research and testing; some have more for- mal procedures than others and stronger veterinary staff capabilities. Accreditation by AAALAC is sought on a voluntary basis, and two of FDA animal facilities, the National Center for Toxicological Research (NCTR) and the Center for Drugs and Biologics of the Office of Biologics Research and Review, are fully accredited (23). The policies and procedures in place at the National Center for Toxicological Research (Jefferson, AR) are a good example of FDA system for addressing animal welfare issues, since NCTR is the primary animal re- search facility within FDA (24): The policy of NCTR management is to use labora- tory animals under practical and reasonable conditions of humane treatment, in carefully planned experiments with in vitro methodologies balanced against minimally required test species numbers in in vivo bioassays, and via procedures set forth in national standards and guidelines. The Director of NCTR has primary responsibility for assuring compliance with the policy but delegates some aspects of that control. The duties of the Senior Scien- tists in NCTR’s Office of Research include technical over- view of animal use, strain selection, genetic quality con- trol, state-of -the-art reviews, and recommendations for adopting new concepts in animal care and control. The Director of the Division of Animal Husbandry is respon- sible for breeding-colony operations, animal produc- tion and laboratory-animal care in NCTR’s various hold- ing areas, and quarantine procedures (25). The animal care committee has adopted an “Animal Use Form for Experimental Protocols” and requires every investiga- tor using animals to provide the committee with detailed information for evaluation (23). Finally, the Director has set up ad hoc committees of in-house personnel to evaluate specific areas of animal care, such as change in feed for the facility (1,24). The FDA policy on extramural research requires ad- herence by awardee institutions to the PHS policy and procedures (23): This includes (1) having in place a program of animal care which meets federal and Department standards, (2) providing, through AAALAC accreditation or de- fined self-assessment procedures, assurance of institu- tional conformance, and (3) maintaining an animal research committee to provide oversight of the insti- tution’s animal program, facilities and associated activities. National Institutes of Health NIH has a specific animal care and use program for intramural research and for research within NIH- controlled space (25). The NIH policy requires individ- ual investigators to adhere to the NIH Guide. In addi- tion, each bureau, institute, or division (BID) is encour- aged to pursue accreditation of its animal facilities by either AAALAC or any other NIH-approved accrediting body (at present AAALAC is the sole body) and to re- port its accreditation status each year to the Deputy Director, who ensures compliance with the policy by each BID. The NIH policy delegates responsibility to five differ- ent authorities, including two types of committees. The 390 . Alternatives to Animal Use in Research, Testing, and Education first is the local BID Animal Research Committee (ARC). This committee must have at least five Federal Govern- ment employees; the BID Scientific Director is respon- sible for annual appointments of the chairperson and members and for carrying out the committee’s recom- mendations. Included among the five ARC members must be the attending veterinarian on the BID staff, a tenured investigator representing laboratories and divisions that use animals, and “a person who is sensi- tive to bioethical issues, does not possess an advanced degree in one of the life sciences, and is an employee from outside that BID” (26). The NIH policy gives the BID ARCS many specific responsibilities beyond the general duties of many such committees. As with other local animal care commit- tees, each ARC is required to make recommendations on animal care matters to its Scientific Director and to review proposals and protocols for humane stand- ards of animal care. It is also supposed to advise indi- viduals on the BID’s policies and oversee their imple- mentation within the facility, The major specific duties of the ARC are: G G G G G to hold quarterly meetings at which a majority of the ARC members are present; to maintain a file of all minutes, memorandums, waivers, and project review documents; to perform site visits of each facility within the BID at least annually to assess compliance, and to sub- mit written reports on these inspections to the Scientific Director; to develop a plan for attaining accreditation of the animal facilities or for pursuing accreditation standards; and to prepare an annual report for the NIH Deputy Director for Intramural Research addressing prob- lems and accomplishments related to attaining ac- creditation. Individual investigators are responsible for submit- ting appropriate information needed for ARC review of a proposal, advising the ARC chairperson of any sig- nificant deviations from procedures described in the most recent project review, and ensuring that all per- sonnel working directly with animals have been trained in the proper care and use of that species. Thus, the system puts much of the burden for proper animal care during an experiment on each investigator. The second authority set up by the NIH intramural policy was the NIH Animal Research Committee (NIHARC). Committee members are appointed annu- ally by the Deputy Director for Intramural Research and must include a veterinarian, the chairperson from each BID ARC, and a nonaffiliated member. NIHARC holds quarterly meetings, advises the Deputy Director on animal care and use at NIH, discusses issues referred from the BID ARCS, develops and coordinates training programs for NIH employees on animal care and use, and prepares NIH’s Annual Report of Research Facility for USDA. Department of the Interior The Department of the Interior does more than 95 percent of its research in-house. All research and de- velopment facilities must comply with both the Ani- mal Welfare Act and with the Department Research and Development Policy Procedures Handbook (27), which calls for an approved animal welfare plan. The National Wildlife Health Laboratory (NWHL) must pro- vide assistance upon request in the development, im- plementation, and maintenance of each program. Due to the diversity of the research programs and the uniqueness of the species involved, each facility is al- lowed to develop an animal welfare plan peculiar to its own needs as long as it is approved by NWHL. Each division plan must discuss: G G G G G G G persons responsible for compliance; reporting and recordkeeping procedures for ani- mals used; all components of the Animal Welfare Act and the Department animal health and husbandry stand- ards that cannot be complied with, due either to the general design of anticipated studies or the unique natural requirements of the species in- volved; quarantine procedures for exotic species; personnel health monitoring and disease preven- tion programs; a schedule for periodic onsite evaluations by the NWHL Veterinary Medical Officer; and procedures for handling carcasses following un- expected mortalities (27). The NWHL Veterinary Medical Officer oversees en- forcement of these policies. Consumer Product Safety Commission The Consumer product Safety Commission (CPSC), as part of its mission to enforce the labeling require- ments of the Federal Hazardous Substances Act (FHSA) (see ch. 7), conducts its own oral acute toxicity studies to determine the toxic potential of regulated substances. If the demand for testing exceeds the capacity of the CPSC’s Health Sciences Laboratory Division, the agency contracts with FDA’s NCTR (13). App. B—Regulation of Animal Use Within Federal Departments and Agencies G 391 In addition to requiring its own personnel, contract- ing agencies, and regulated parties to observe the re- quirements of the Animal Welfare Act and the NIH Guide in performing required safety tests on animals, CPSC has published an Animal Testing Policy, “which is intended to reduce the number of animals tested to determine hazards associated with household products and to reduce any pain that might be associated with such testing” (49 FR 22522). The policy states that CPSC itself and manufacturers of substances covered by the FHSA “should wherever possible utilize existing alter- natives to conducting animal testing [including] prior human experience, literature sources which record prior animal testing or limited human tests, and expert opinion. ” Citing the provision in FHSA regulations that gives preference to studies based on humans over those with animals, the policy states that CPSC “resorts to animal testing only when the other information sources have been exhausted. ” It also states that: G G G “limit” tests for acute toxicity studies, rather than the “classic” LD5O, are performed when necessary, requiring fewer animals; eye irritancy testing is not performed if the test substance is a known skin irritant; and agency-required Draize (eye irritation) tests are modified to eliminate the need for restraining test rabbits, allowing them full mobility and access to food and water (49 FR 22522). Environmental Protection Agency The guidelines and policies that the Environmental Protection Agency (EPA) follows governing humane treatment and appropriate veterinary care for labora- tory animals involve AAALAC accreditation for its two major laboratories, adherence to the NIH Guide, and adherence to the Animal Welfare Act. In addition, EPA has an intra-agency committee that oversees animal re- search issues. There is no separate policy for extramural research; NIH Guide principles and requisites are en- forced in such cases by a signed statement from the investigator that the proper animal care is being ob- served (16). The EPA facility at Research Triangle Park, NC, has an animal care committee that oversees and carries out an institutional review of animal care and welfare is- sues. The committee is composed of representatives of the different research divisions within that facility along with the attending veterinarian. Its 8 to 10 mem- bers, who meet approximately once a month and keep records of their proceedings, are responsible for ani- mal care issues only, and do not conduct scientific re- views of research proposals, Scientific review is done separately before proposals reach the committee. The overall responsibilities for the committee are to: G oversee the functioning of the animal care facility, G plan improvements for the facility and carry them out , set policy for humane treatment of animals, G set policy for sharing facility resources, G address any day-to-day animal care problems brought to its attention, and G review proposals for appropriate animal use and care (2). In addition, the committee can recommend experi- mental changes to improve animal care and treatment and has the authority to interrupt or terminate an ex- periment if it finds any instances of inhumane treat- ment or inappropriate care of the animals, a step that has been taken at least once since the committee was established (2). The committee does not monitor experiments while in progress or handle the day-to-day activities of the animal care facility. These powers are delegated to the attending veterinarian (who is under contract with EPA to work at the facility 3 days a week) and a staff of ap- proximately 20 (2). National Aeronautics and Space Administration The overall National Aeronautics and Space Admin- istration policy on animal research is based on the Ani- mal Welfare Act, the NIH Guide, and the IRAC princi- ples. All NASA facilities, all users of NASA facilities, aircraft, or spacecraft, and all NASA contractors using animals are subject to this policy. The overriding phi- losophy of the policy is based on three principles: G G G Animals will be used only to answer valid ques- tions that improve the health, welfare, or general medical and scientific knowledge of humans. Experimental animals must not be subject to avoid- able discomfort or distress. Experiments requiring the use of invasive proce- dures without benefit of anesthetic agents demand strong justification and attention to possible alter- natives (12). Although the NASA policy exists today as only a pro- posed NASA Management Instruction (NMI), it is already being implemented. For example, the NMI establishes an Animal Care and Use Committee (ACUC) in each facility with animals (12); the committee includes a research veterinarian, a biomedical scientist, a non- scientist, and a person not affiliated with NASA. It is responsible for overseeing the animal care facility, establishing specific guidelines, reviewing proposals, and making recommendations for approval or dis- 392 Alternatives to Animal Use in Research, Testing, and Education approval of funding (9). The committee must ask the following questions for each experiment (12): G Will the minimum possible number of animals be used? G Is the use of animals necessary in this experiment? G Are provisions for care of these animals adequate? Different compliance with these policies is needed for intramural versus extramural research. For NASA facilities, ACUC reports are required to be sent to the Director of the Life Sciences Division at NASA head- quarters reviewing facility procedures. AAALAC accred- itation is required for all NASA installations. Currently all facilities are moving toward AAALAC accreditation but have not yet obtained it (12). For extramural re- search, the institution must submit a written assurance that its animal care policies are equivalent to the NASA policy. (AAALAC accreditation is one way of showing compliance. ) Noncompliance will result in termination of the research by the ACUC and possibly sanctions after review by the Director of the Life Sciences Divi- sion (19). The Ames Research Center (Moffett Field, CA), NASA’s primary center for nonhuman research, illustrates the implementation of NASA policy. The Ames Research Center has established the Animal Users Guide for Ames-sponsored laboratory experiments using animals. This guide sets up two entities to ensure that all legal requirements are met: The animal care facility is re- sponsible for housing and maintaining the animals prop- erly, and the animal care and use committee must mon- itor all animal care and experimentation progress at the center. In addition, the guide states (28): EVERY RESEARCH SCIENTIST AND ALL RESEARCH PERSONNEL, CONTRACTORS, AND GRANTEES ARE RESPONSIBLE FOR OBSERVING THE LEGAL REQUIRE- MENTS CONCERNING LABORATORY ANIMALS. The Ames committee reports to the center’s Direc- tor of Life Sciences and is responsible for: G reviewing the use of animals in proposed and on- going experiments; G reviewing all animal experimentation performed by contractors or grantees; G serving as an advisory committee on all questions of animal care and use, and as a forum for resolv- ing differences that may occur; and G reviewing animal-related inventions and devices (28). At present, the Ames committee has 10 members—4 non-NASA, non-life-sciences laypersons; 1 veterinarian; 1 scientist-veterinarian; 1 engineer; 2 scientists; and 1 science manager. In addition, 2 veterinarians accred- ited in Laboratory Animal Medicine are advisors. The lay members include an attorney, a professor of relig- ion (ethics), the chairman of the Department of Educa- tion at a local college, and the public relations director of the Santa Clara Valley Humane Society. This is one of the few such committees in the country with a 40 percent lay membership. According to the Acting Di- rector of Life Sciences at Ames Research Center, “the out -of -house members have contributed materially to the [committee].” Two of the lay members head sub- committees that are reviewing and updating the Ani- mal Users Guide and committee charter and develop- ing an animal user’s orientation program (14). National Science Foundation A summary of the animal care requirements of the National Science Foundation (NSF) is found in Section 713 of the NSF Grant Policy Manual (30) and included in the NSF document “Grant General Conditions, ” that is sent to each grantee when an award is made. Any grantee performing research on warm-blooded animals must comply with the Animal Welfare Act and its reg- ulations and follow the NIH Guide. NSF has no formal inspection system to check on compliance with these policies, as that is judged to be the responsibility of USDA/APHIS (8). The result is a voluntary adherence system by NSF grantees, Beginning in 1986, NSF imposed two new require- ments on grant applicants and grantees who perform research on vertebrate animals: G Each proposal must be reviewed by an institutional animal care and use committee. G Each proposal must be accompanied by a statement from the grantee that assures the grantee’s com- pliance with the PHS policy. Grant proposals submitted to NSF thus face three sep- arate reviews-one by the grantee’s institutional com- mittee, one by outside reviewers, and one by NSF staff. Although these are primarily scientific in nature, reviewers are asked to comment on animal welfare is- sues. If a proposal involves the use of animals, suffi- cient information must be provided to allow evalua- tion of the appropriateness of experimental protocols with respect to the choice of species, the number of animals to be used, and any necessary exposure of ani- mals to discomfort, pain, or injury (29). With this infor- mation, the reviewers are asked to (29): . . . comment if you have any concerns regarding the violation of animal welfare laws or guidelines, the ex- posure of animals to unnecessary pain or mistreatment, or the use of excessive numbers of animals. If the spe- cies being used is not the one most appropriate, or if alternative or adjunct methods could be used to elimi- nate or reduce the need for animal experimentation, please comment. Veterans’ Administration The Veterans’ Administration is unique in its policies governing humane treatment and appropriate veteri- — . App. B—Regulation of Animal Use Within Federal Departments and Agencies Ž 393 nary care for laboratory animals because it has required all its facilities using animals to seek and obtain AAALAC accreditation (see ch. 15). This policy was originated in 1971, and 81 out of 174 VA facilities (as of Apr. 1, 198.5) had some level of AAALAC accreditation. Not all VA constituents apply for accreditation, since some do not engage in animal research. In fact, the VA has a contract with AAALAC covering all its research facil- ities that prohibits failure of accreditation of any con- stituent facility solely for financial reasons (10). In addition to requiring adherence to the PHS policy, the VA has a lengthy research review process with a strong committee structure. At the local research fa- cility, each research and development committee has a subcommittee for animal studies that oversees all such research. The membership varies, though it includes at least one member of the research and development committee, a Veterinary Medical Officer (VA employee), and two to four investigators who are involved in studies using animals. Thus, there are no laypersons or persons not affiliated with the research facility on the subcommittee. Except for the veterinarian, who serves indefinitely, members serve 3-year terms (31). The subcommittee has three primary functions: G G G to approve the use or uses made of animal sub- jects in all research studies as they relate to animal welfare laws, regulations, and policies; to review all animal studies for need, adequacy, and availability of essential animal research facil- ity support; for the appropriateness, quality, and availability of the animal models; for the humane- ness and appropriateness of procedures and con- ditions surrounding animal subjects before and throughout the study; and to evaluate, at least annually, the animal research facility and recommend appropriate actions to cor- rect deficiencies noted (11). Proposals are reviewed again at a regional VA office by two committees, first for veterinary medical review (appropriate use and care of animals) and then for scien- tific merit (10). The animal welfare review is done by a Veterinary Medical Panel of specialists chosen for their experience, knowledge, and research in labora- tory-animal science and medicine. This panel attempts “to assure that proposals include sound, acceptable ani- mal medicine and husbandry practices in animal re- search facilities that are operated in conformance with all pertinent animal welfare laws, regulations, and pol- icies” (11). Specifically, the panel conducts reviews: G to ascertain the description of the animal model; G to ascertain the biological and medical definition of the animal model; to ascertain the environmental and experimental- animal-related factors; G to determine if there is evidence of adequate ex- perience with the proposed technology of manipu- lations, monitoring, or measuring; G to determine if use of intact animals is required or if animal parts could be obtained from or shared with other investigators who have scientifically compatible studies; G to determine if painful procedures are involved and whether these can be avoided or if their con- trol has been satisfactorily planned; and G to relate the budget of the experiment to the ani- mal costs and to the animal maintenance needs (11). In 1984, the VA required that all research proposals have an appendix with a detailed discussion of animal protocols, the number of animals to be used, and why the specific choice of organism was made. This appen- dix is signed by three people from the local facility– the researcher, the animal committee chairperson, and the research and development chairperson—to guar- antee that the procedures are carried out. The enforcement of the VA’s animal research pol- icies rests with the committee structure and is over- seen by the Chief Veterinary Medical Officer for the VA, whose duties include making sure all Federal and State animal research laws are observed and that the individual facilities have the funds to continue to re- main AAALAC-accredited. In addition, the VA began in fiscal year 1984 strict enforcement of the comple- tion of the Annual Reports of Animal Research Facil- ities for APHIS by every VA facility, whether the facil- ity used animals in research the preceding year or not (lo). At the local VA facilities, the attending veterinarian has authority for veterinary medical matters. This per- son must monitor the housing, general treatment, and care of the experimental animals while the experiment is in progress as often as needed. If inhumane treat- ment or inappropriate care is found, the veterinarian and animal subcommittee do not have the authority to interrupt or terminate an experiment. The subcom- mittee would make a recommendation to the research and development committee and to the Associate Chief of Staff for Research and Development, who may make a decision or a recommendation to the Director (4). This means there is some enforcement of the proper ani- mal care standards at each local VA facility on a day-to- day basis. Appendix B References 1. Borsetti, A., Staff Scientist, Office of Science Coordina- tion, Food and Drug Administration, U.S. Department of Health and Human Services, Rockvil]e, MD, personal com- munications, October and November 1984 and March 1985. 2. Chernoff, N,, U.S. Environmental Protection Agency, Re- search Triangle Park, NC, personal communication, 1984. 394 G Alternatives to Animal Use in Research, Testing, and Education 3. Crawford, L., Director, Bureau of Veterinary Medicine, Food and Drug Administration, US. Department of Health and Human Services, Rockville, MD, personal communi- cation, November 1984. 4. Ditzler, J., Chief Medical Director, Department of Medi- cine and Surgery, Veterans’ Administration, Washington, DC, persona] communication, Jan. 17, 1985. 5. Edington, C., Associate Director, Office of Health and Envi- ronmental Research, Office of Energy Research, U.S. De- partment of Energy, Washington, DC, personal commu- nication, Nov. 16, 1984. 6. Edington, C., Procedures for Selection of Animal Research Projects for Funding Through OHER/DOE (Washington, DC: Department of Energy, Jan. 24, 1985). 7. Kainz, R,, Office of the Commander, U.S. Army Medical Research and Development Command, Ft. Detrick, MD, personal communication, September 1984. 8. Kingsbury, D,, Assistant Director, National Science Foun- dation, Washington, DC, personal communication, Nov. 2 , 1 9 8 4 . 9. Lewis, C. S., “NASA’s Use of Animals in Research,” pre- pared for the Life Sciences Division, National Aeronau- tics and Space Administration, Washington, DC, Sept. 28, 1983. 10. Middleton, C., Chief Veterinary Medical Officer, Veterans’ Administration, Washington, DC, personal communica- tion, Oct.3, 1984. 11, Moreland, A., “Animal Research Protocol Review Within the Veterans’ Administration,” mimeo, Gainesville, FL, 1984. 12. Nicogossian, A., Director, Life Sciences Division, National Aeronautics and Space Administration, Washington, DC, personal communication, Oct. 19, 1984. 13. Porter, W., Health Sciences Laboratory Division, Con- sumer Product Safety Commission, Washington, DC, per- sonal communication, Nov. 19, 1984. 14. Sharp, J., Acting Director of Life Sciences, Ames Research Center, National Aeronautics and Space Administration, letter to A. Nicogossian, Director, Life Sciences Division, NASA, on Animal Care and Use Committee, Moffett Field, CA, Sept. 12, 1984. 15. Stewart, W., Senior Veterinarian, Animal and Plant Health Inspection Service, U.S. Department of Agriculture, Hyattsville, MD, personal communication, November 1984. 16. Ulvedal, F., Acting Director, Water and Toxic Substances Health Effects Research Division, U.S. Environmental Pro- tection Agency, Washington, DC, personal communica- tions, September and October 1984. 17. US. Department of Defense, Air Force, Aerospace Medi- cal Division Animal [Jse Review Panel Meetings (Wash- ington, DC: May 1984). 18. U.S. Department of Defense, Air Force, Aerospace Medi- cal Division, ‘(Animals in DOD Research and Training AMD Supplement 1“ (draft), Brooks Air Force Base, TX, 1985. 19, U.S. Department of Defense, Assistant Secretary of De- fense for Health Affairs, Memorandum to the Secretaries of the Uniformed Services, President of the Uniformed Services University of the Health Sciences, and Directors of Defense Agencies (Washington, DC: Jan. 4, 1984). 20. U.S. Department of Defense, The Use of Animals in DOD Programs, DOD Instruction 3216.1 (Washington, DC: Feb. 1, 1982). 21. U.S. Department of Defense, The Use of Animals m DOD Programs, Army Regulation 70-18 (Washington, DC: June 1, 1984). 22. U.S. Department of Health and Human Services, Food and Drug Administration, Office of Science Coordination, Fi- nal Report of Acute Studies Workshop (Washington, DC: Nov. 9, 1983). 23. U.S. Department of Health and Human Services, Food and Drug Administration, Final Report to the Commissioner, FDA Agency Steering Committee on Animal Welfare Zs- sues (Rockville, MD: Aug. 15, 1984). 24. U.S. Department of Health and Human Services, Food and Drug Administration, National Center for Toxicological Research, iWTR Quality Assurance Program (Jefferson, AR: May 1983). 25. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, Animal Care and Use in the Intramural Program, NIH Policy 3040-2 (Bethesda, MD: Dec. 30, 1983). 26, U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, Guide for the Care and Use of Laboratory Animals, NIH Pub. No. 85-23 (Bethesda, MD: National Institutes of Health, 1985). 27. U.S. Department of the Interior, Research and Develop- ment PolicyRocedures Handbook (Washington, DC: July 26, 1984). 28. US. National Aeronautics and Space Administration, Ames Research Center, Animal Users Guide AHB 7180-1 (Mof - fett Field, CA: June 1982). 29. U.S. National Science Foundation, Office of the Assistant Director for Biological, Behavioral, and Social Sciences, ATSFADDBS circular NO. 1.3 (Washington, DC: June 15, 1982). 30. US. National Science Foundation,lWFG rant Policy A4an - ual, Section 713 (Washington, DC: Apr. 15, 1984). 31. U.S. Veterans’ Administration, Department of Medicine and Surgery, Research and Development in Medicine Gen- eral (Washington, DC: Apr. 27, 1982). Appendix C Public Health Service Policy on Humane Care and Use of Laboratory Animals by Awardee Institutions The following is reprinted from U.S. Department of Health and Human Services, Public Health Service, Na- tional Institutes of Health, “Laboratory Animal Wel- fare, ” NIH Guide for Grants and Contracts 14(8), June 2.5, 1985. Introduction It is the policy of the Public Health Service (PHS) to require institutions to establish and maintain proper measures to ensure the appropriate care and use of all animals involved in research, research training and biological testing activities (hereinafter referred to as activities) supported by the PHS. The PHS endorses the “U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research and Training” developed by the Interagency Research Animal Committee (IRAC). This policy is intended to implement and supplement those Principles. Applicability This policy is applicable to all PHS-approved activities involving animals, whether the activities are performed at an awardee institution or any other institution and conducted in the United States, the Commonwealth of Puerto Rico, or any territory or possession of the United States. Institutions in foreign countries receiv- ing PHS support for activities involving animals shall comply with this policy, or provide evidence to the PHS that acceptable standards for humane care and use of the animals in PHS-supported activities will be met, No PHS support for an activity involving animals will be provided to an individual unless that individ- ual is affiliated with or sponsored by an institution which can and does assume responsibility for compli- ance with this policy for PHS-supported activities, or unless the individual makes other arrangements with the PHS. This policy does not affect applicable state or local laws or regulations which impose more strin- gent standards for the care and use of laboratory ani- mals. All institutions are required to comply, as appli- cable, with the Animal Welfare Act, and other Federal statutes and regulations relating to animals. A. B. c. D. E. F. G. H. Definitions Animal Any live, vertebrate animal used or intended for use in research, research training, experimenta- tion or biological testing or for related purposes. Animal Facility Any and all buildings, rooms, areas, enclosures, or vehicles, including satellite facilities, used for animal confinement, transport, maintenance, breed- ing or experiments inclusive of surgical manipula- tion. A satellite facility is any containment outside of a core facility or centrally designated or man- aged area in which animals are housed for more than 24 hours. Animal Welfare Act Public Law 89-544, 1966, as amended, (P.L. 91- 579 and P.L. 94-279) 7 U.S.C. 2131 et seq. Imple- menting regulations are published in the Code of Federal Regulations (CFR), Title 9, Subchapter A, Parts 1, 2, 3 and 4, and are administered by the U.S. Department of Agriculture. Animal Welfare Assurance or Assurance The documentation from an awardee or a pro- spective awardee institution assuring institutional compliance with this policy. Guide Guide for the Care and Use of Laboratory Ani- mals, DHEW, NIH Pub. No, 78-23, 1978 edition or succeeding revised editions. Institution Any public or private organization, business, or agency (including components of Federal, state and local governments). Institutional Official An individual who has the authority to sign the institution’s Assurance, making a commitment on behalf of the institution that the requirements of this policy will be met. Public Health Service The Public Health Service includes the Alcohol, Drug Abuse, and Mental Health Administration, the Centers for Disease Control, the Food and Drug Administration, the Health Resources and Services Administration, and the National Institutes of Health. 395 396 . Alternatives to Animal use in Research, Testing, and Education I. Quorum A majority of the members of the Institutional Animal Care and Use Committee. Implementation by Awardee Institutions A. Animal Welfare Assurance No activity involving animals will be supported by the PHS until the institution conducting the ac- tivity has provided a written Assurance acceptable to the PHS, setting forth compliance with this pol- icy for PHS-supported activities. Assurances shall be submitted to the Office for Protection from Re- search Risks (OPRR), Office of the Director, Na- tional Institutes of Health, 9000 Rockville Pike, Building 31, Room 4B09, Bethesda, Maryland 20205. The Assurance shall be typed on the institution’s letterhead and signed by an institutional official. OPRR will provide the applicant institution with necessary instructions and an example of an ac- ceptable Assurance. All Assurances submitted to the PHS in accordance with this policy will be evaluated by OPRR to determine the adequacy of the institution’s proposed program for the care and use of animals in PHS-supported activities. On the basis of this evaluation OPRR may approve or dis- approve the Assurance, or negotiate an approva- ble Assurance with the institution. Approval of an Assurance will be for a specified period of time (no longer than five years) after which time the insti- tution must submit a new Assurance to OPRR. OPRR may limit the period during which any par- ticular approved Assurance shall remain effective or otherwise condition, restrict, or withdraw ap- proval. Without an applicable PHS approved Assur- ance no PHS-supported activity involving animals at the institution will be permitted to continue. 1. Institutional Program for Animal Care and Use The Assurance shall fully describe the insti- tution’s program for the care and use of animals in PHS-supported activities. The PHS requires in- stitutions to use the Guide for the Care and Use of Laboratory Animals (Guide) as a basis for de- veloping and implementing an institutional pro- gram for activities involving animals. The pro- gram description must include the following: a, a list of every branch and major component of the institution, as well as a list of every branch and major component of any other in- stitution which is to be included under the As- surance; b. the lines of authority and responsibility for ad- ministering the program and ensuring com- pliance with this policy; c. the qualifications, authority and responsibil- ity of the veterinarian(s) who will participate in the program; d. the membership list of the Institutional Ani- mal Care and Use Committee (IACUC) es- tablished in accordance with the require- ments set forth in IV.A.3.; e. the procedures which the IACUC will follow to fulfill the requirements set forth in IV. B.; f. the health program for personnel who work in laboratory animal facilities or have frequent contact with animals; g. the gross square footage of each animal facil- ity (including satellite facilities), the species housed therein and the average daily inven- tory, by species, of animals in each facility; and h. any other pertinent information requested by OPRR. 2. Institutional Status Each institution must assure that its program and facilities are in one of the following cate- gories: Category l—Accredited by the American Asso- ciation for the Accreditation of Laboratory Animal Care (AAALAC). All of the institution’s programs and facilities (including satellite fa- cilities) for activities involving animals have been evaluated and accredited by AAALAC, or other accrediting body recognized by PHS.2 G Category 2—Evaluated by the Institution. All of the institution’s programs and facilities (in- cluding satellite facilities) for activities involv- ing animals have been evaluated by the IACUC and will be reevaluated by the IACUC at least once each year. The IACUC shall use the Guide for the Care and Use of Laboratory Animals as a basis for evaluating the institution’s pro- gram and facilities. A report of the IACUC evaluation shall be submitted to the institu- tional official and updated on an annual ba- sis.3 The initial report shall be submitted to IThe name Institutional Animal Care and Use Committee ([ AC UC) M used in this policv is intended as a generic term for a committee whose function is to ensure that the care and use of animals in PHS-supported activities is appropriate and humane in accordance with this policy. Howe\,er, each in. stitution may identify the committee by whatever name it chooses. Member- ship and responsibilities of the IACLJC are set forth in IL’.A,3 and It’ B. ‘As of the issuance date of this policy the only accrediting body recognized by PHS is the American Association for Accreditation of Laboratory)’ .Animal Care (AAA1.AC). 3The IACUC may, at its discretion, determine the best means of conduct- ing an evaluation of the institution’s programs and facilities. The IACUC may invite ad hoc consultants to conduct or assist in conducting the evaluation. However, the 1.4CL1C remains responsible for the evaluation and report. App. C—Public Health Service Policy on Humane Care and Use of Laboratory Animals by Awardee Institutions G 397 OPRR with the Assurance. Annual reports of the IACUC evaluation shall be maintained by the institution and made available to OPRR upon request. The report must contain a description of the nature and extent of the in- stitution’s adherence to the Guide and this pol- icy.4 The report must identify specifically any departures from provisions of the Guide and this policy, and state the reasons for each departure, If program or facility deficiencies are noted, the report must contain a reason- able and specific plan and schedule for cor- recting each deficiency. The report must dis- tinguish significant deficiencies from minor deficiencies. A significant deficiency is one which, in the judgment of the IACUC and the institutional official, is or may be a threat to the health or safety of the animals. Failure of the IACUC to conduct an annual evaluation and submit the required report to the institu- tional official may result in PHS withdrawal of its approval of the Assurance. 3. Institutional Animal Care and Use Committee (IACUC) a. Each institution shall appoint an Institutional Animal Care and Use Committee (IACUC), qualified through the experience and exper- tise of its members to oversee the institution’s animal program, facilities and procedures. b. The Assurance must include the names, posi- tion titles and credentials of the IACUC chair- person and the members. The committee shall consist of not less than five members, and shall include at least: (1) one Doctor of Veterinary Medicine, with training or experience in laboratory animal science and medicine, who has direct or delegated program responsibility for activ- ities involving animals at the institution; (2) one practicing scientist experienced in re- search involving animals; (3) One member whose primary concerns are in a nonscientific area (for example, ethi- cist, lawyer, member of the clergy); and (4) one individual who is not affiliated with the institution in any way other than as a mem- ber of the IACUC, and is not a member of the immediate family of a person who is af- filiated with the institution. c. An individual who meets the requirements of more than one of the categories detailed in —.———— 41f some of the institution’s facilities are accredited b}r AAAI,AC or other accrediting body recognized by PHS, the report should” identify those facil- ities and need not contain any further information about e~,aluation of those facilities IV. A.3.b. (1)-(4) may fulfill more than one re- quirement, However, no committee may con- sist of less than five members, B. Functions of the Institutional Animal Care and Use Committee As an agent of the institution the IACUC shall, with respect to PHS-supported activities: 1. review at least annually the institution’s program for humane care and use of animals; 2. inspect at least annually all of the institution’s animal facilities, including satellite facilities; 3. review concerns involving the care and use of animals at the institution; 4. make recommendations to the institutional offi- cial regarding any aspect of the institution’s ani- mal program, facilities or personnel training; 5. review and approve, require modifications in (to 6 7. secure approval) or withhold approval of those sections of PHS applications or proposals related to the care and use of animals as specified in IV.C.; review and approve, require modifications in (to secure approval), or withhold approval of pro- posed significant changes regarding the use of animals in ongoing activities; and be authorized to suspend an activity involving animals in accord with specifications set forth in IV.C.5. C. Review of PHS Applications and Proposals I. In order to approve applications and proposals or proposed significant changes in ongoing activ- ities, the IACUC shall conduct a review of those sections related to the care and use of animals and determine that the proposed activities are in accord with this policy. In making this deter- mination, the IACUC shall confirm that the activ- ity will be conducted in accord with the Animal Welfare Act insofar as it applies to the activity, and that the activity is consistent with the Guide unless acceptable justification for a departure is presented. Further, the IACUC shall determine that the activity conforms with the institution’s Assurance and meets the following requirements: a. Procedures with animals will avoid or mini- mize discomfort, distress and pain to the ani- mals, consistent with sound research design. b. Procedures that may cause more than momen- tary or slight pain or distress to the animals will be performed with appropriate sedation, analgesia, or anesthesia, unless the procedure is justified for scientific reasons in writing by the investigator. c. Animals that would otherwise experience se- vere or chronic pain or distress that cannot be relieved will be painlessly sacrificed at the 398 G Alternatives to Animal Use in Research, Testing, and Education end of the procedure or, if appropriate, dur- ing the procedure. d. The living conditions of animals will be appro- priate for their species and contribute to their health and comfort. The housing, feeding and nonmedical care of the animals will be di- rected by a veterinarian or other scientist trained and experienced in the proper care, handling and use of the species being main- tained or studied. e. Medical care for animals will be available and provided as necessary by a qualified veteri- narian. f. Personnel conducting procedures on the spe- cies being maintained or studied will be appro- priately qualified and trained in those pro- cedures. g. Methods of euthanasia used will be consistent with the recommendations of the American Veterinary Medical Association (AVMA) Panel on Euthanasia,s unless a deviation is justified for scientific reasons in writing by the inves- tigator. 2. Prior to the review, each IACUC member shall be provided with a list of applications and pro- posals to be reviewed, Those sections of appli- cations and proposals that relate to the care and use of animals shall be available to all IACUC members, and any member of the IACUC may upon request obtain full committee review of those sections, If full committee review is not re- quested, at least one member of the IACUC, des- ignated by the chairperson and qualified to con- duct the review, shall review those sections and have the authority to approve, require modifi- cations in (to secure approval) or request full committee review of those sections. If full com- mittee review is requested, approval of those sec- tions may be granted only after review at a con- vened meeting of a quorum of the IACUC and with the approval vote of a majority of the quo- rum present. No member may participate in the IACUC review or approval of an application or proposal in which the member has a conflicting interest (e.g., is personally involved in the project), except to provide information requested by the IACUC; nor may a member who has a conflicting interest contribute to the constitution of a quorum. 3. The IACUC may invite consultants to assist in the review of complex issues. Consultants may not ‘Journal of the American Veterinary Medical Association (JAVMA), 1978, Vol. 173, No. 1, pp. 59-72, or succeeding revised editions. approve or withhold approval of an application or proposal or vote with the IACUC. 4. The IACUC shall notify investigators and the in- stitution in writing of its decision to approve or withhold approval of those sections of applica- tions or proposals related to the care and use of animals, or of modifications required to se- cure IACUC approval. If the IACUC decides to withhold approval of an application or proposal, it shall include in its written notification a state- ment of the reasons for its decision and give the investigator an opportunity to respond in per- son or in writing. 5. The IACUC shall conduct continuing review of applications and proposals covered by this pol- icy at appropriate intervals as determined by the IACUC, but not less than once every three years. 6. The IACUC may suspend an activity that it pre- viously approved if it determines that the activ- ity is not being conducted in accordance with applicable provisions of the Animal Welfare Act, the Guide, the institution’s Assurance, or IV. C.l.a.-g. The IACUC may suspend an activity only after review of the matter at a convened meeting of a quorum of the IACUC and with the suspension vote of a majority of the quorum present. 7. If the IACUC suspends an activity involving ani- mals, the institutional official in consultation with the IACUC shall review the reasons for suspen- sion, take appropriate corrective action and re- port that action with a full explanation to OPRR. 8. Applications and proposals that have been ap- proved by the IACUC maybe subject to further appropriate review and approval by officials of the institution. However, those officials may not approve those sections of an application or pro- posal related to the care and use of animals if they have not been approved by the IACUC. D. Information Required in Applications and Propos- als Submitted to PHS 1. All Institutions Applications and proposals submitted to PHS that involve the care and use of animals shall contain the following information: a. identification of the species and approximate number of animals to be used; b. rationale for involving animals, and for the appropriateness of the species and numbers to be used; c. a complete description of the proposed use of the animals; d. assurance that discomfort and injury to ani- mals will be limited to that which is unavoid- App. C—Public Health Service Policy on Humane Care and Use of Laboratory Animals by Awardee Institutions . 399 able in the conduct of scientifically valuable research, and that analgesic, anesthetic, and tranquilizing drugs will be used where indi- cated and appropriate to minimize discomfort and pain to animals; and e. a description of any euthanasia method to he used. 2. Institutions That Have an Approved Assurance Applications or proposals covered by this pol- icy from institutions which have an approved As- surance on file with OPRR shall include verifi- cation of approval by the IACUC of those sections related to the care and use of animals, With the authorization of PHS, such verification may be filed at a time not to exceed 60 days after sub- mission of applications or proposals.6 If verification of IACUC approval is submitted subsequent to the submission of the application or proposal, the verification shall state the modifications, if any, required by the IACUC. The verification shall be signed by an individual authorized by the institution, but need not be signed b-y the institutional official who signed the Assurance. 3. Institutions That Do Not Have an Approved As- surance Applications and proposals involving animals from institutions that do not have an approved Assurance on file with OPRR shall contain a dec- laration that the institution will establish an IA- CUC and submit an Assurance upon request by OPRR. After OPRR has requested the Assurance, the institution shall establish an IACUC as re- quired by IV.A.3. and the IACUC shall review those sections of the application or proposal as required by IV.c. The institution shall then sub- mit to OPRR the Assurance and verification of IACUC approval. The verification shall state the modifications, if any, required by the IACUC. The verification shall be signed by an individual authorized by the institution, but need not be signed by the institutional official who signed the Assurance. E. Recordkeeping 1. The awardee institution shall maintain: a. an Assurance approved by the PHS; b. minutes of IACUC meetings, including records of attendance, activities of the committee, and committee deliberations; 6(lnt Il further notwe PHS hereh\, authorizes all institutions with approked Assuraores to file \ ~r]ftration of 1,4(;{ 1(’ appro~ al either along with the ap. plicatwn or proposal or within f-XI da}s of submission of the application or proposal From time to time P}iS w ill rem aluate this hl,]nket authorization An}’ deriwn to withdra}~ this authorization will take place on]? after ample Opporfllnlt}’ IS pr”ol ided for romment by the Put)]ic c. d, e. records of applications, proposals and pro- posed significant changes in the care and use of animals and whether IACUC approval was given or withheld; records of any IACUC reports and recommen- dations as forwarded to the institutional offi- cial; and records of accrediting body determinations. 2. All records shall be maintained for at least three years; records that relate directly to applications, proposals, and proposed significant changes in ongoing activities reviewed and approved by the IACUC shall be maintained for the duration of the activity and for an additional three years af- ter the completion of the activity. All records shall be accessible for inspection and copying by authorized OPRR or other PHS representatives at reasonable times and in a reasonable manner. F. Reporting Requirements 1. On or before each anniversary of approval of its Assurance, the institution shall report in writ- ing to OPRR: a. any change in the institution’s program or fa- cilities which would place the institution in a different category than specified in its Assur- ance (see IV.A.2.); b. any change in the description of the institu- tion’s program for animal care and use as re- quired by IV. A.l.a.-h.; c. any changes in IACUC membership; and d. if the institution’s program and facilities are in Category 2 (see IV.A.2.), verification that the IACUC has conducted an annual evaluation of the institution’s program and facilities and sub- mitted the evaluation to the institutional official. 2. Institutions that have no changes to report as specified in IV.F. 1.a.-c. shall submit a letter to OPRR stating that there are no changes. 3. Institutions shall provide OPRR promptly with a full explanation of the circumstances and ac- tions taken with respect to: a. any serious or continuing noncompliance with this policy; b. any serious deviation from the provisions of the Guide; or c. any suspension of an activity by the IACUC. Implementation by PHS A. Responsibilities of OPRR OPRR is responsible for the general administra- tion and coordination of this policy and will: I. request and negotiate, approve or disapprove, . 400 G Alternatives to Animal Use in Research, Testing, and Education and, as necessary, withdraw approval of As- surances; 2. distribute to executive secretaries of initial re- view and technical evaluation groups, and to PHS awarding units, lists of institutions that have an approved Assurance; 3. advise awarding units and awardee institutions concerning the implementation of this policy; 4. evaluate allegations of noncompliance with this policy; 5. have the authority to review and approve or dis- approve waivers to this policy (see V.D.); and 6. conduct site visits to selected institutions. B. Responsibilities of PHS Awarding Units PHS awarding units may not make an award for an activity involving animals unless the institution submitting the application or proposal is on the list of institutions that have an approved Assurance on file with OPRR, and the institution has provided verification of approval by the IACUC of those sec- tions of the application or proposal related to the care and use of animals in PHS-supported activi- ties. If an institution is not listed, the awarding unit will ask OPRR to negotiate an Assurance with the institution before an award is made. No award shall be made until the Assurance has been submitted by the institution, approved by OPRR, and the in- stitution has provided verification of approval by the IACUC of those sections of the application or proposal related to the care and use of animals in PHS-supported activities. C. Conduct of Special Reviews/Site Visits Each awardee institution is subject to review at anytime by PHS staff and advisors, which may in- clude a site visit, in order to assess the adequacy of the institution’s compliance with this policy. D. Waiver Institutions may request a waiver of a provision or provisions of this policy by submitting a request to OPRR. No waiver will be granted unless suffi- cient justification is provided and the waiver is ap- proved in writing by OPRR. Appendix Laboratory-Animal Facilities Fully Accredited by the American Association for Accreditation of Laboratory Animal Care As of April 1, 1985, there were 483 facilities listed as fully accredited by the American Association for Accreditation of Laboratory Animal Care (AAALAC) (New Lenox, IL). Institutions are categorized as univer- sities, medical schools, combined facilities for health sciences, veterinary schools, dental schools, colleges of pharmacy, colleges of biological science, colleges of arts, colleges of engineering, Veterans’ Administration medical centers, pharmaceutical manufacturers, gov- ernment laboratories, commercial laboratories, hos- pitals, nonprofit research laboratories, or laboratory animal breeders. The following list of AAALAC-accred- ited facilities numbers 538, as some institutions are listed in more than one category. (Facilities receiving accreditation since April 1, 1985, are not listed.) Universities (Programs serving an entire campus) Alabama: University of Alabama, University University of Alabama at Birmingham and the Veterans’ Administration Medical Center, Birmingham Arkansas: University of Arkansas at Little Rock, Little Rock California: University of California-Davis, Davis University of California-San Diego, San Diego University of California at Los Angeles, Los Angeles University of Southern California, Los Angeles Georgia: Medical College of Georgia, Augusta Illinois: University of Illinois at the Medical Center and the Veterans’ Administration Medical Center, Chicago Kansas: University of Kansas-Lawrence, Lawrence Massachusetts: Massachusetts Institute of Technology, Cambridge Michigan: University of Michigan, Ann Arbor, Dearborn, and Flint Wayne State University, Detroit Missouri: University of Missouri-Kansas City, Kansas City Montana: University of Montana, Missoula Nebraska: University of Nebraska at Omaha, Omaha New York: St. John’s University, Jamaica State University of New York at Buffalo, Buffalo Rockefeller University, New York North Carolina: Duke University, Durham Oklahoma: Oral Roberts University, Tulsa Rhode Island: Brown University, Providence South Carolina: University of South Carolina, Columbia Tennessee: Oak Ridge Associated Universities, Oak Ridge Vanderbilt University, Nashville Utah: University of Utah, Salt Lake City Virginia: Virginia Commonwealth University, Richmond University of Virginia, Charlottesville Washington: University of Washington, Seattle Universities (Programs serving only a portion of a campus) California: Divisions of Animal Resources, University of California, Berkeley Georgia: Yerkes Regional Primate Research Center, Emory University, Atlanta Ohio: Laboratory Animal Center, The Ohio State University, Columbus 401 402 Alternatives to Animal Use in Research, Testing, and Education Medical Schools Arizona:, Arizona Medical Center, University of Arizona, Tucson Arkansas: Medical Center, University of Arkansas, Little Rock California: California College of Medicine, University of California, Irvine School of Medicine, University of California at Los Angeles, Los Angeles School of Medicine, University of California at San Diego, San Diego Charles R. Drew Postgraduate Medical School, Los Angeles School of Medicine, Loma Linda University, Loma Linda Colorado: Medical School, University of Colorado, Denver Connecticut: School of Medicine, University of Connecticut Health Center, Farmington School of Medicine, Yale University, New Haven District of Columbia: School of Medicine, Georgetown University College of Medicine, Howard University Florida: College of Medicine, University of Florida, Gainesville Medical Center, University of South Florida, Tampa Illinois: Chicago College of Osteopathic Medicine, Chicago The Chicago Medical School/University of Health Services, North Chicago College of Medicine-Rockford, University of Illinois, Rockford Stritch School of Medicine, Loyola University, Maywood School of Medicine, Southern Illinois University, Springfield Iowa: College of Osteopathic Medicine and Surgery, Des Moines Kentucky: Medical Center, University of Kentucky, Lexington School of Medicine, University of Louisville, Louisville Louisiana: School of Medicine, Tulane University, New Orleans Delta Regional Primate Research Center, Tulane University, Covington Maryland: School of Medicine, University of Maryland at Baltimore, Baltimore School of Medicine, Uniformed Services University of the Health Sciences, Bethesda Massachusetts: Medical School, Harvard University, Boston Medical Center, University of Massachusetts, Worcester School of Medicine, Tufts-New England Medical Center, Boston Michigan: Medical Center, University of Michigan, Ann Arbor Medical School, Wayne State University, Detroit Minnesota: School of Medicine, University of Minnesota- Duluth, Duluth Medical School, University of Minnesota, Minneapolis Missouri: The University of Health Sciences, Kansas City Kirksville College of Osteopathic Medicine, Kirksville School of Medicine, University of Missouri, Columbia Nebraska: College of Medicine, University of Nebraska, Omaha New Hampshire: Dartmouth Medical School, Hanover New Jersey: Medical School, College of Medicine and Dentistry of New Jersey, Newark New Mexico: School of Medicine, University of New Mexico, Albuquerque New York: Albany Medical College of Union University, Albany Albert Einstein College of Medicine, Bronx College of Physicians and Surgeons, Columbia University, New York Medical College, Cornell University, New York Mount Sinai Medical Center, New York Downstate Medical Center, State University of New York, Brooklyn App. D—Laboratory-Animal Facilities Fully Accredited by AAALAC . 403 School of Medicine, University of Rochester, Rochester North Carolina: School of Medicine, University of North Carolina, Chapel Hill Bowman Gray School of Medicine, Wake Forest College, Winston-Salem Ohio: Medical College of Ohio, Toledo College of Medicine, University of Cincinnati, Cincinnati College of Medicine, Northeastern Ohio Universities, Rootstown Department of Animal Laboratories, Hospitals, and College of Medicine, The Ohio State University, Columbia Oregon: Oregon Health Sciences University, Portland Pennsylvania: School of Medicine, Hahnemann University, Philadelphia Milton S. Hershey Medical Center, Pennsylvania State University, Hershey Jefferson Medical College, Thomas Jefferson University, Philadelphia School of Medicine, University of Pittsburgh, Pittsburgh Puerto Rico: Medical Sciences Campus, University of Puerto Rico, San Juan South Carolina: School of Medicine, University of South Carolina, Columbia Tennessee: Meharry Medical College, Nashville Texas: Texas College of Osteopathic Medicine, North Texas State University, Ft. Worth Medical School, University of Texas at Houston, Houston Utah: College of Medicine, University of Utah, Salt Lake City Vermont: College of Medicine, University of Vermont, Burlington Virginia: Medical College of Virginia, Virginia Commonwealth University, Richmond Medical Center, University of Virginia, Charlottesville Eastern Virginia Medical School, Norfolk Washington: School of Medicine, University of Washington, Seattle Wisconsin: Medical College of Wisconsin, Milwaukee Medical School, University of Wisconsin, Madison Combined Facilities for Health Sciences Connecticut: University of Connecticut Health Center, Farmington District of Columbia: Georgetown University Medical Center School of Medicine and Health Sciences, George Washington University Florida: University of Florida, J. Hillis Miller Health Center and the Veterans’ Administration Medical Center, Gainesville Illinois: Life Sciences Vivarium, Southern Illinois University, Carbondale Indiana: Lobund Laboratory, University of Notre Dame, Notre Dame Kansas: University of Kansas-Lawrence, Lawrence Louisiana: Medical Center, Louisiana State University, New Orleans Maryland: Johns Hopkins Medical Institutions, Baltimore Massachusetts: Boston University School of Medicine and Graduate School of Dentistry, Boston Harvard University Medical School, Dental School, School of Public Health, Animal Research Center, and the New England Regional Primate Research Center, Southboro Tufts-New England Medical Center, Boston Minnesota: Health Sciences, University of Minnesota, Minneapolis Missouri: John M. Dalton Research Center, Graduate School, University of Missouri, Columbia New Jersey: College of Medicine and Dentistry of New Jersey, Newark New York: Health Sciences Center, State University of New York at Stony Brook, Stony Brook School of Medicine and Dentistry, University of Rochester, Rochester North Carolina: School of Medicine and School of Dentistry, University of North Carolina, Chapel Hill — 404 . Altematives to Animal Use in Research, Testing, and Education Oklahoma: University of Oklahoma Health Sciences Center at Oklahoma City, Oklahoma City Texas: University of Texas Health Science Center, San Antonio Virginia: School of Basic Sciences, Virginia Commonwealth University, Richmond West Virginia: West Virginia University Medical Center, Morgantown Wisconsin: University of Wisconsin, Madison Graduate School, University of Wisconsin, Madison Veterinary Schools California: School of Veterinary Medicine, University of California-Davis, Davis Florida: College of Veterinary Medicine, University of Florida, Gainesville Louisiana: School of Veterinary Medicine, Louisiana State University, Baton Rouge Massachusetts: School of Veterinary Medicine, Tufts-New England Medical Center, Boston New York: New York State College of Veterinary Medicine, Cornell University, Ithaca Tennessee: College of Veterinary Medicine, University of Tennessee, Knoxville Wisconsin: School of Veterinary Medicine, University of Wisconsin, Madison Dental Schools California: School of Dentistry, University of California at Los Angeles, Los Angeles Connecticut: Dental School, University of Connecticut Health Center, Farmington District of Columbia: School of Dentistry, Georgetown University Florida: College of Dentistry, University of Florida, Gainesville Illinois: Dental School, University of Illinois at the Medical Center, Chicago School of Dentistry, Maywood Indiana: School of Dentistry, Indianapolis Maryland: School of Dentistry, Baltimore Massachusetts: School of Dentistry, Loyola University, Indiana University, University of Maryland, Harvard University, Boston School of Dental Medicine, Tufts-New England Medical Center, Boston Michigan: School of Dentistry, University of Michigan, Ann Arbor Minnesota: School of Dentistry, University of Minnesota, Minneapolis New Jersey: School of Dentistry, Fairleigh Dickinson University, Hackensack Dental School, College of Medicine and Dentistry of New Jersey, Newark New York: School of Dentistry, University of Rochester, Rochester North Carolina: School of Dentistry, University of North Carolina, Chapel Hill Ohio: College of Dentistry, The Ohio State University, Columbus Oregon: School of Dentistry, Oregon Health Sciences University, Portland Washington: School of Dentistry, University of Washington, Seattle Colleges of pharmacy Florida: College of Pharmacy, University of Florida, Gainesville Indiana: School of Pharmacy and Pharmaceutical Sciences, Purdue University, Lafayette Kansas: School of Pharmacy, University of Kansas- Lawrence, Lawrence — App. D—Laboratory-Animal Facilities Fully Accredited by AAALAC 405 Massachusetts: Massachusetts College of Pharmacy and Allied Health Sciences, Boston Michigan: College of Pharmacy, University of Michigan, Ann Arbor Minnesota: College of pharmacy, University of Minnesota, Minneapolis Nebraska: School of Pharmacy, Omaha New Mexico: College of Pharmacy, Albuquerque Ohio: College of Pharmacy, Columbus South Carolina: College of Pharmacy, Carolina, Columbia Virginia: University of Nebraska, University of New Mexico, The Ohio State University, University of South School of Pharmacy, Virginia Commonwealth University, Richmond Washington: School of Pharmacy, University of Washington, Seattle College of Pharmacy, Washington State University, Pullman Wisconsin: School of Pharmacy, University of Wisconsin, Madison Colleges of Biological Science California: College of Biological Sciences and Scripps Oceanography, University of California-San Diego, San Diego Pomona College, Claremont South Carolina: College of Humanities and Social Sciences, University of South Carolina, Columbia College of Science and Mathematics, University of South Carolina, Columbia Utah: College of Science, University of Utah, Salt Lake City College of Social and Behavioral Science, University of Utah, Salt Lake City Washington: College of Biological and Laboratory Animal Resource Center, Washington State University, Pullman Colleges of Arts Alabama: College of Arts and Sciences, University of Alabama, University Tennessee: College of Liberal Arts, University of Tennessee, Knoxville Virginia: School of Arts and Sciences, Virginia Commonwealth University, Richmond College of Arts and Science, Virginia, Charlottesville Washington: College of Arts and Science, Washington, Seattle College of Arts, Washington Pullman College of Engineering New York: University of University of State University, Biomedical Engineering Laboratory, Rensselaer Polytechnic Institute, Troy Veterans’ Administration Medical Centers Alabama: Birmingham Arizona: Tucson Phoenix Arkansas: Little Rock North Little Rock California: Fresno Loma Linda Long Beach Martinez San Diego San Francisco Sepulveda West Los Angeles Colorado: Denver Connecticut: West Haven Delaware: Wilmington District of Columbia: Washington Florida: Gainesville 406. Alternatives to Animal Use in Research, Testing, and Education Tampa Miami Lake City Bay Pines Georgia: Decatur Augusta Illinois: Chicago (2) North Chicago Hines Indiana: Indianapolis Iowa: Iowa City Des Moines Kentucky: Lexington Louisiana: New Orleans Shreveport Maryland: Perry Point Baltimore Massachusetts: Bedford Boston Brockton West Roxbury Michigan: Allen Park Ann Arbor Minnesota: Minneapolis Mississippi: Jackson Missouri: Kansas City Columbia St. Louis Nebraska: Omaha New Mexico: Albuquerque New Jersey: East Orange New York: Albany Brooklyn Buffalo Castle Point New York Northport Syracuse North Carolina: Asheville Durham Ohio: Cleveland Cincinnati Dayton Oklahoma: Oklahoma City Oregon: Portland Pennsylvania: Coatesville Philadelphia Puerto Rico: San Juan South Carolina: Charleston Tennessee: Memphis Nashville Texas: Dallas Houston San Antonio Utah: Salt Lake City Virginia: Richmond Washington: Seattle Tacoma West Virginia: Huntington Wisconsin: Madison Wood Vermont: White River Junction pharmaceutical Manufacturers California: Hyland Division, Travenol Laboratories, Glendale Quidel, La Jolla Connecticut: Boehringer Ingelheim Ltd., Ridgefield Miles Laboratories, Inc., West Haven Medical Research Laboratory, Charles Pfizer & Co., Inc., Groton Delaware: Stuart Pharmaceuticals, Division of ICI Americas, Inc., Wilmington App. D—Laboratory-Animal Facilities Fully Accredited by AAALAC G 407 Illinois: Abbott Laboratories, North Chicago American Critical Care, American Hospital Supply Corporation, McGaw Park Division of Biological Research, G.D. Searle & Co., Chicago Travenol Laboratories, Inc., Morton Grove Indiana: Bristol-Myers Company, Evansville Eli Lilly and Company, Indianapolis and Greenfield Miles Laboratories, Inc., Elkhart Michigan: The Upjohn Company, Kalamazoo Warner-Lambert/Parke-Davis, Ann Arbor and Detroit Minnesota: Riker/3M, St. Paul Missouri: Mallinckrodt, Inc., St. Louis Marion Laboratories, Inc., Kansas City Mississippi: Travenol Laboratories, Inc., Cleveland New Jersey: Berlex Laboratories, Inc., Cedar Knolls Biological Research Division, Bristol-Myers Products, Inc., Hillside Pharmaceuticals Division, Ciba-Geigy, Inc., Summit Hoechst-Roussel Pharmaceuticals, Inc., Somerville Ethicon Research Foundation, Somerville Hoffman-La Roche, Inc., Nutley Johnson & Johnson Baby Products Company, Skillman Johnson & Johnson Research Foundation, New Brunswick Merck Institute for Therapeutic Research, Merck Sharp & Dohme Research Laboratories, Rahway Merck Sharp & Dohme Research Laboratories, Branchburg Farm, Somerville Ortho Pharmaceutical Corporation, Raritan Sandoz Inc., East Hanover Biological Research Division, Schering Corp., Bloomfield Toxicology and Pathology Division, Schering Corp., Lafayette Squibb Institute for Medical Research, E.R. Squibb & Sons, Inc., Princeton and New Brunswick Wallace Laboratories, Carter Wallace, Inc., Cranbury New York: American Cyanamid Company, Lederle Laboratories, Pearl River Bristol-Myers Company, Buffalo Norwich-Eaton Pharmaceuticals, Norwich Pennwalt Corporation, Rochester Revlon Health Care Group, Tuckahoe North Carolina: Becton Dickinson and Company Research Center, Research Triangle Park and Durham Wellcome Research Laboratories, Burroughs- Wellcome, Co., Research Triangle Park Ohio: Merrell Research Center, Merrell Dow Pharmaceuticals, Cincinnati Pennsylvania: McNeil Pharmaceutical, Inc., Spring House Merck Institute for Therapeutic Research, Merck Sharp & Dohme Research Laboratories, West Point Veterinary Services and Veterinary Pathology, Merck Sharp & Dohme, West Point William H. Rorer, Inc., Fort Washington Research and Development Divison- Pharmaceuticals, Smith Kline & French Laboratories, Philadelphia Wyeth Laboratories, Radnor Virginia: A.H. Robins Research Laboratories, A.H. Robins, Co., Richmond France: Searle Recherche et Developpement, G.D. Searle and Company, Valbonne Government Laboratories Alabama: U.S Army Aeromedical Research Laboratory, Fort Rucker Arizona: National Center for Toxicological Research, Jefferson California: Health Protection Systems/Laboratory Services Program, California Department of Health, Berkeley Letterman Army Institute of Research, Animal Resources Division, Presidio of San Francisco, San Francisco Colorado: Fitzsimons Army Medical Center, Aurora Connecticut: Naval Submarine Medical Research Laboratory, Naval Submarine Medical Center, Groton 408 . Altematives to Animal Use in Research, Testing, and Education District of Columbia: Armed Forces Institute of Pathology, Washington Florida: Naval Aerospace Medical Research Laboratory, Naval Aerospace Medical Institute, Pensacola John F. Kennedy Space Center, John F. Kennedy Space Center Georgia: Centers for Disease Control, Atlanta Hawaii: Tripler Army Medical Center, Honolulu Illinois: Argonne National Laboratory, Argonne Naval Dental Research Institute, Great Lakes Naval Base, Great Lakes Iowa: National Animal Disease Center, U.S. Department of Agriculture, Ames Louisiana: Naval Biodynamics Laboratory, New Orleans Maryland: Bureau of Biologics, Food and Drug Administration, Bethesda Frederick Cancer Research Facility, National Cancer Institute, Frederick Medical Laboratory Veterinary, Medicine Service, Department of Pathology, Ft. Meade Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore Veterinary Resources Branch, National Institutes of Health, Bethesda U.S. Army Environmental Hygiene Agency, Edgewood U.S. Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick Massachusetts: Human Nutrition Research Center at Tufts University, Boston U.S. Army Research Institute of Environmental Medicine, Natick Mississippi: USAF Medical Center Keesler, Keesler Air Force Base Montana: National Institute of Allergy and Infectious Disease, National Institutes of Health, Hamilton New Mexico: Los Alamos National Laboratory, University of California, Los Alamos New York: Brookhaven National Laboratory, Upton Food and Drug Research Laboratories, Inc., Waverly North Carolina: National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park United States Environmental Protection Agency, Research Triangle Park Ohio: National Institute for Occupational Safety and Health, Cincinnati United States Environmental Protection Agency, Cincinnati USAF 6570th Aerospace Medical Research Laboratories, Wright-Patterson AFB Oklahoma: Civil Aeromedical Institute, FAA Aeronautical Center, Oklahoma City Oregon: Oregon Regional Primate Research Center, Beaverton Pennsylvania: Naval Air Development Center, Warminster Tennessee: Oak Ridge National Laboratory, Biology Division, Oak Ridge Texas: William Beaumont Army Medical Center (U.S. Army), El Paso Brooke Army Medical Center, Department of Pathology and Laboratory Services, Fort Sam Houston Texas Research Institute of Mental Sciences, Houston USAF School of Aerospace Medicine, Brooks AFB Washington: Madigan Army Medical Center, Tacoma Commercial Laboratories Arizona: Armour Research Center, Armour-Dial Company, Scottsdale Arkansas: INTOX Laboratories, Inc., Little Rock California: American Pharmaseal Laboratories, Irwindale Bio-Devices Laboratories, Inc., Orange Chevron Environmental Health Center, Richmond App. D—Laboratory-Animal Facilities Fully Accredited by AAALAC G 409 Edwards Laboratories, Santa Ana McGaw Laboratories, Irvine Science Applications, Inc., La Jolla North American Science Associates of California, Irvine Shell Development Company, Modesto Stauffer Chemical Company, Mountain View Connecticut: United States Surgical Corporation, Norwalk Stauffer Chemical Company, Farmington Delaware: Haskell Laboratory for Toxicology and Industrial Medicine, Newark Florida: Life Sciences, Inc., St. Petersburg Sherwood Medical Laboratories, Inc., DeLand Georgia: American McGaw, Milledgeville Illinois: American Biogenics Corporation, Decatur Kendall Company Health Research Center, Barrington Quaker Oats Company, Barrington Kansas: Mobay Chemical Corporation, Stilwell BAVET Division of hliles Laboratories, Inc., Shawnee Missions Maryland: BioCon, Inc., Rockville Biotech Research Laboratories, Inc., Rockville Borriston Laboratories, Inc., Temple Hills Gillette Capital Corporation, Rockville Laboratory Animal Services, Inc., Rockville Litton Bionetics, Inc., Bethesda Microbiological Associates, Bethesda Tegeris Laboratories, Inc., Laurel Massachusetts: Arthur D. Little, Inc., Cambridge Bioassay Systems Corporation, Woburn Biotek, Inc., Woburn EG & G Mason Research Institute, Inc., Worcester SISA Laboratories Inc., Cambridge Michigan: Toxicology Research Laboratory, The Dow Chemical Company, Midland Dow Corning Corporation, Midland General Motors Research Laboratories, Warren International Research and Development Corporation, Mattawan Toxicity Research Laboratories, Ltd., Muskegon Minnesota: Medtronic, Inc., Coon Rapids Missouri: Environmental Health Laboratory, Monsanto Company, St. Louis New Jersey: Biodynamics, Inc., East Millstone Colgate Palmolive Research Center, Piscataway Cyanamid Foundation for Agricultural Development, Princeton Exxon Biomedical Sciences, Inc., East Millstone FMC Corporation Toxicology Laboratory, Somerville FMC Corporation, Princeton Lever Brothers Company, Edgewater Mobil Oil Corporation, Princeton Revlon Research Center, Inc., Edison New York: Eastman Kodak Company, Rochester Ohio: Ben Venue Laboratories, Inc., Bedford Hill Top Research, Inc., Miamiville North American Science Associates, Inc., Northwood Procter and Gamble Company, Cincinnati Toilet Goods Division, Procter and Gamble Company, Cincinnati Springborn Institute for Bioresearch, Inc., Spencerville WIL Research Laboratories, Inc., Ashland Pennsylvania: Biosearch, Inc. Philadelphia M.B. Research Laboratories, Inc., Spinnertown Pharmakon Research International, Inc., Waverly Rohm and Haas Company, Spring House Texas: Alcon Laboratories, Fort Worth Health and Environmental Sciences, Dow Chemical, U. S. A., Lake Jackson STILLMEADOW, Inc., Houston Virginia: Flow Laboratories, Inc., McLean Hazelton Laboratories America, Inc., Vienna Meloy Laboratories, Inc., Springfield Washington: Genetics Systems Corp., Seattle Hollister-Stier, Division of Miles Laboratories, Inc., Spokane Oncogen, Seattle Wisconsin: Hazelton Laboratories America, Inc., Madison Canada: Bio-Research Laboratories, Ltd., Senneville, Quebec 38-750 0 - 86 - 14 410 . Alternatives to Animal Use in Research, Testing, and Education Hospitals Arizona: Barrow Neurological Institute, St. Joseph’s Hospital and Medical Center, Phoenix California: Children’s Hospital of San Francisco, San Francisco Sutter Hospitals Medical Research Foundation, Sutter Community Hospitals, Sacramento Colorado: Denver General Hospital, Denver National Jewish Hospital, Denver District of Columbia: Research Foundation of the Washington Hospital Center Research Foundation of Children’s Hospital, Children’s Hospital Florida: Mount Sinai Medical Center, Miami Beach Illinois: Evanston Hospital Association, Evanston Michael Reese Hospital and Medical Center, Chicago Louisiana: Southern Baptist Hospital, New Orleans U.S. Public Health Service Hospital, Carville Maryland: Maryland Psychiatric Research Center, Catonsville Massachusetts: Beth Israel Hospital, Boston New England Deaconess Hospital, Boston St. Vincent Healthcare System, Inc., Worcester New England Medical Center Hospitals, Tufts- New England Medical Center, Boston Michigan: Henry Ford Hospital, Detroit Sinai Hospital of Detroit, Detroit Wayne County General Hospital, Westland Minnesota: Saint Paul-Ramsey Medical Center, St. Paul New Jersey: Newark Beth Israel Medical Center, Newark New York: Beth Israel Medical Center, New York Hospital for Special Surgery, New York Montefiore Hospital and Medical Center, Bronx Nassau Hospital, Mineola Nassau County Medical Center, East Meadow St. Luke’s-Roosevelt Institute for Health Sciences, New York St. Vincent’s Hospital and Medical Center of NY, New York Ohio: Akron City Hospital, Akron Children’s Hospital Research Foundation, Children’s Hospital, Columbus Children’s Hospital Research Foundation, Children’s Hospital Medical Center, Cincinnati Cleveland Research Institute, Cleveland Pennsylvania: Albert Einstein Medical Center, Northern Division, Philadelphia Graduate Hospital, Philadelphia Lehigh Valley Hospital Center, Allentown Skin and Cancer Hospital of Philadelphia, Philadelphia Joseph Stokes, Jr. Research Institute of the Children’s Hospital of Philadelphia Rhode Island: Miriam Hospital, Providence Tennessee: St. Jude Children’s Research Hospital, Memphis University of Tennessee Memorial Hospital and Research Center, Knoxville Texas: Scott and White Memorial Hospital, Temple Nonprofit Research Laboratories California: Cedar-Sinai Medical Research Institute, Los Angeles Huntington Institute of Applied Medical Research, Pasadena Lawrence Berkeley Laboratory, Berkeley Palo Alto Medical Research Foundation, Palo Alto Research and Education Institute, Inc., Harbor- UCLA Medical Center, Torrance SRI International, Menlo Park Whittier Institute for Diabetes and Endocrinology, La Jolla Connecticut: John B. Pierce Foundation Laboratory, New Haven Florida: Miami Heart Institute, Miami Beach Illinois: American Dental Association Research Institute, Chicago Life Sciences Division, IIT Research Institute, Chicago Kansas: Menninger Foundation, Topeka App. D—Laboratory-Animal Facilities Fully Accredited by AAALAC . 411 Louisiana: Division of Research, Alton Ochsner Medical Foundation, New Orleans USL New Iberia Research Center, New Iberia Maine: Jackson Laboratory, Bar Harbor Maryland: American Type Culture Collection, Rockville Massachusetts: Worcester Foundation for Experimental Biology, Inc., Shrewsbury Center for Blood Research, Inc., Boston Dana-Farber Cancer Institute, Boston Forsyth Dental Center, Boston Eunice Kennedy Shriver Center for Mental Retardation, Waltham Michigan: Michigan Cancer Foundation, Detroit Missouri: Midwest Research Institute, Kansas City Nebraska: Eppley Institute for Research in Cancer, Omaha New Mexico: Lovelace Biomedical and Environmental Research Institute, Inc., Albuquerque iNew Jersey: Institute for Medical Research, Camden New York: American Health Foundation, Naylor Dana Institute for Disease Prevention, Valhalla Cold Spring Harbor Laboratory, Cold Spring Harbor Memorial Sloan-Kettering Cancer Center, New York New York Blood Center, New York Trudeau Institute, Inc., Saranac Lake North Carolina: Chemical Industry Institute for Toxicology, Research Triangle Park Research Triangle Institute, Research Triangle Park Ohio: Battelle Memorial Institute, Columbus Cleveland Clinic Foundation, Cleveland Oklahoma: Oklahoma Medical Research Foundation, Oklahoma City Pennsylvania: Bushy Run Research Center, Export Federated Medical Resources, Honey Brook Institute for Cancer Research, Philadelphia Texas: Southwest Foundation for Research and Education, San Antonio Southwest Research Institute, San Antonio University of Texas Cancer Center, Houston Veterinary Resources Division, Science Park, The University of Texas Cancer Center, Bastrop Utah: University of Utah Research Institute, Salt Lake City Washington: Battelle, Pacific Northwest Laboratories, Richland Bob Hope International Heart Research Institute, Seattle Fred Hutchinson Cancer Research Center, Seattle Pacific Northwest Research Foundation, Seattle Virginia Mason Research Center, Seattle Laboratory-Animal Breeders Indiana: Engle Laboratory Animals, Inc., Farmersburg Harlan Sprague Dawley, Inc., Indianapolis Laboratory Supply Company, Indianapolis Maryland: M.A. Bioproducts, Inc., Walkersville Massachusetts: Charles River Breeding Laboratories, Inc., Wilmington Michigan: Charles River-Portage, Portage New Jersey: Carom Research Institute, Inc., Wayne Charles River Lakeview, Newfield H.A.R.E. Rabbits for Research, Marland Breeding Farms, Hewitt New York: Carworth Division, The Charles River Breeding Laboratories Inc., Kingston Charles River Research Primates Corporation, Port Washington Taconic Farms, Inc., Germantown Tennessee: Cumberland View Farms, Clinton Virginia: Hazelton Research Primates, Reston Hazelton Research Animals, Inc., Vienna Canada: Charles River Canada Incorporated, St. Constant, Quebec Appendix E International Agreements Governing Animal Use Convention on International Trade in Endangered Species In 1973, the Convention on International Trade in Endangered Species of Fauna and Flora (CITES) was signed by 61 nations. It has since been ratified by a total of 81 separate nations and has been enforced in the United States since 1977 (10,11). In addition to protecting animals from extinction, the Convention specifies in seven different places that the Management Authority must be “satisfied that any living specimen will be so transported and cared for as to minimize the risk of injury, damage to health or cruel treatment. ” CITES is administered on an inter- national basis by the International Union for the Con- servation of Nature and Natural Resources headquar- tered in Gland, Switzerland. Endangered plants and animals are listed in three Appendixes to the Conven- tion, according to level of endangerment. For purposes of monitoring, all primates have been included in Ap- pendix 11 (“Threatened”) except chimpanzees, which are classified as “Endangered. ” Under CITES provi- sions, the effect of the Appendix 11 classification has been to require export permits for all listed primates. The U.S. agency responsible for administration of CITES provisions is the Research Division of the Fish and Wildlife Service, Department of the Interior, which has additional responsibilities regarding international trade in endangered or threatened species under Sec- tion 7 of the Endangered Species Act of 1973. (For a brief discussion of how this act affects experimenta- tion in the United States, see ch. 13.) Current CITES Appendixes listings, by species of wildlife and family of plants, can be found in part 23 of title 50 of the Code of Federal Regulations; lists of endangered and threatened wildlife species and plant families affected by the Endangered Species Act are found in part 17. The Convention’s importance to research is twofold. First, it has limited trade in nonhuman primates and a few other species favored at one time or another in experiments (1). Second, continued review of the Convention by signatories has served as a forum for discussion of protection of laboratory animals. CITES signatories meet periodically in conferences, convened under CITES provisions, to discuss the required clas- sification of species according to the terms of the Con- vention. Under regulations promulgated by the Fish and Wildlife Service (50 CFR 23.31-,39), members of the public must be given notice of the U.S. negotiat- ing position at CITES conferences and an opportunity to provide information and comments on the proposed agenda, including at least one public meeting. Humane groups have used these meetings to raise the issue of humane treatment of laboratory-animal species in re- lation to the Convention’s articles (12). Recently, for example, CITES delegates were petitioned to ratify proposed interpretations of the Convention to reach that very question. The petition was ruled outside the terms of the Convention (9). Bans on Exporting Primates From time to time, nations with indigenous popula- tions of nonhuman primates that have been in demand for various types of traditional research have consid- ered or implemented prohibitions on their export, ei- ther to protect dwindling populations or because of high mortality rates suffered in transit. India ordered such a ban in 1955, for the latter reason. Because rhe- sus monkeys were in demand for testing polio vaccines at the time, India agreed to reopen trade with the United States on condition that the Surgeon General sign a certificate of need for each order of monkeys, with assurances that they be used humanely and only for medical research and vaccine production. The ban was reimposed by the Indian Government when it was revealed that military experiments, specifically pro- hibited under the agreement, were being done with some of the monkeys. Other countries have consid- ered similar bans or have imposed ceilings on exports. Bans were enacted in Malaysia and Bolivia in 1984, and a U.S. dealer was ousted from Bangladesh in 1979 for selling Rhesus monkeys for military research (5). Some commentators have been critical of U.S. estimates of need for nonhuman primates in research, finding them overstated, and have faulted the research community for attempts to circumvent export bans (13). Draft Convention of the Council of Europe The Council of Europe, headquartered in Stras- bourg, France, and with 21 member countries, was organized in 1949 to work for greater European unity, 412 App. E—international Agreements Governing Animal Use “ 413 to improve the conditions of life and develop humane values in Europe, and to uphold the principles of par- liamentary democracy (6). Historically, the Council has been concerned about the treatment of animals. It has drafted Conventions on the protection of animals in international transport (1968), on those kept for farming purposes (1976), on slaughter (1979), and on conservation of European wildlife and natural habitats (1979). In 1971, the Coun- cil adopted Recommendation 621, which contained three relevant proposals: G G G Establish a-documentation and information cen- ter on alternatives to animal use in testing and ex- perimentation. Establish tissue banks for research. Establishment of an Ad Hoc Committee of Experts to study the problems rising out of the abuse of live animals for experimental industrial purposes, The Committee was given the task of drafting in- ternational legislation setting out the conditions under which, and the scientific grounds on which, experiments on live animals may be authorized (15). The Ad Hoc Committee of Experts for the Protec- tion of Animals began its work on the Draft Conven- tion in 1978. In 1983, the committee presented a Draft Convention, guidelines for care and treatment, and a guidance note on data collection to the Council of Min- isters plenary sessions and seven working party meet- ings under three successive chairmen. The commit- tee was composed of experts from member countries. Observers from the United States and Europe, includ- ing representatives from several nongovernmental organizations (World Society for the Protection of Ani- mals, Federation of Veterinarians of the European Eco- nomic Community, European Federation of Pharma- ceutical Industries’ Associations, and the International Council for Laboratory Animal Science) were admitted to the committee’s meetings (2,14). The form of the Draft Convention follows an earlier one on the treatment of farm animals. Its preamble, restating the general objective of European unity in the context of protection of experimental animals, balances the need of “man in his quest for knowledge, health and safety . . . to use animals where there is a reasonable expectation that the result will be to ex- tend knowledge or be to the overall benefit of man or animal, just as he uses them for food, clothing and as beasts of burden” against the “moral obligation to respect all animals and to exercise due consideration for their capacity for suffering and memory.” As stated in the preamble, the general objective of the Conven- tion is ‘(to limit wherever practicable the use of ani- mals for experimental and other scientific purposes, in particular by seeking alternative methods to replace the use of animals” (2). Prospects for final ratification of the Draft Conven- tion remain unclear. Twice in 1983 the Council’s as- sembly failed to achieve the required two-thirds vote on the committee’s report to urge the Committee of Ministers to adopt it as soon as possible. Reported accounts stated that some delegates did not believe the Convention goes far enough in controlling animal experimentation. The assembly, however, rejected amendments that would have outlawed the use of ex- perimental animals (8). The Convention itself is are summarized below. General Principles divided into 10 parts, which Article 1 applies the Convention “to any animal be- ing used or intended for use in any experimental or other scientific procedure where that procedure may cause pain, suffering, distress, or lasting harm. It does not apply to any nonexperimental agricultural or clin- ical veterinary practice. ” “Animal” means, “unless otherwise qualified . . . any live non-human vertebrate, including free-living larval and/or reproducing larval forms, but excluding other foetal or embryonic forms. ” ‘(Procedure” is defined to include: . . . any experimental or other scientific use of an ani- mal which may cause it pain, suffering, distress or last- ing harm, including any course of action intended to, or liable to, result in the birth of an animal in any such condition, but excluding the least painful methods ac- cepted in modern practice (i.e., “humane” methods) of killing or marking an animal; a procedure starts when the animal is first prepared for use and ends when no further observations are to be made for that procedure; the elimination of pain, suffering, distress or lasting harm by the successful use of anesthesia or analgesia or other methods does not place the use of an animal outside the scope of this definition. Article 2 provides that a defined procedure can be performed on an animal for only one or more of the following purposes, subject to other restrictions con- tained in the Convention: the avoidance or prevention of disease, ill health or other abnormality, or their effects, in humans, vertebrate or invertebrate animals, or plants, including the production and the quality, efficacy, and safety testing of drugs, substances, or products; the diagnosis or treatment of disease, ill health or other abnormality, or their effects, in humans, vertebrate or invertebrate animals, or plants; the assessment, detection, regulation or modifi- cation of physiological conditions in humans, ver- tebrate and invertebrate animals, or plants; 38-750 0 - 86 - 15 414 . Alternatives to Animal Use in Research, Testing, and Education G the prolongation or saving of life of humans, ver- tebrate or invertebrate animals or plants; G the protection of the environment; Ž the production and quality control of foodstuffs; G the breeding of vertebrate or invertebrate animals; . scientific research; G education and training; or G forensic inquiries. Article 3 requires all member nations “to take all nec- essary steps to give effect to [its] provisions . . . and to ensure an effective system of control and supervi- sion” within 5 years of the Convention’s approval for ratification. Article 4 stipulates that ratification by a member country does not bar it from adopting stricter meas- ures to control experimental animal use. General Care and Accommodation Article 5 requires any animal to be used in a proce- dure to be provided with “accommodation, an envi- ronment, at least a minimum freedom of movement, food, water, and care all appropriate to its health and well-being. Any restriction on the extent to which an animal can satisfy its physiological and ethnological needs shall be limited as far as practicable.” Envi- ronmental conditions must be checked daily and as needed to prevent avoidable suffering. Conduct of Procedure Article 6 requires that procedures not be performed where “another scientifically satisfactory method, not entailing the use of an animal, is reasonably and prac- ticably available,” and asks member nations to “encour- age, if possible, scientific research into the development of methods which could provide the same informa- tion as that obtained in procedures.” Article 7 requires careful consideration of choice of species in procedures and that choices be explained, where required, to the responsible authority. Proce- dures should use the minimum number of animals, cause the least pain, suffering, distress, or lasting harm consistent with providing satisfactory results. Article 8 requires all procedures to be performed under general or local anesthetic or by other meth- ods designed to eliminate to the extent practicable pain, suffering, distress, or lasting harm unless the methods are judged to be more distressing than the procedure or are incompatible with the aim of the pro- cedure. Article 9 requires specific authorization of the au- thority where an animal may experience severe pain that is likely to endure. Authorization must be refused if the authority judges that the procedure is not of ex - ceptional importance for meeting the essential needs of humans or animals, including the solution of scien- tific problems. Article 10 declares that an animal under procedure remains subject to the provisions of article 5, except where those provisions are incompatible with the ob- ject of the procedure. Article 11 provides for a decision at the end of proce- dures whether the animal shall be kept alive or killed by a humane method, subject to the condition that it shall not be kept alive if, even though it has been re- stored to normal health in all other respects, it is likely to remain in lasting pain or distress. Such decisions must be made by a veterinarian or a person responsi- ble for the procedure. If an animal is not to be kept alive it should be killed by a humane method as soon as possible. Finally, the article provides that no ani- mal be used in more than one painful procedure un- less the second procedure is one in which the animal is subject throughout to general anesthesia, from which it is not allowed to recover, or the further pro- cedure will involve minor interventions only. Article 12 permits experimental animals to be set free as part of the procedure provided that the maxi- mum practicable care has been taken to safeguard the animal’s well-being. Procedures that involve setting the animal free are not permitted solely for educational or training purposes. Authorization Article 13 provides that procedures authorized by article 2 may be performed only by authorized per- sons or persons under their direct responsibility, or if the project is authorized by the legislation of a mem- ber country. Only persons deemed competent by the responsible authority may be so authorized. Breeding or Supplying Establishments The four articles contained in this part establish principles for breeders and suppliers of experimental animals, who would be required to: G G G G register and comply with article 5 (article 14); specify a competent person in charge with author- ity to administer or arrange for suitable care (ar- ticle 15); keep detailed records on breeding, shipment, and transfer, to be maintained at least 3 years from the date of last entry (article 16); and mark humanely for identification dogs and cats and maintain complete records to promote their identification (article 17). App. E—International Agreements Governing Animal Use G 415 User Establishments Under the provisions of the seven articles in this part, users (i.e., experimental facilities) would be re- quired to: register with national authorities and comply with article 5 (article 18); provide equipment and facilities appropriate for species used and to ensure that the procedures are performed as effectively as practicable with the minimum number of animals and the mini- mum degree of pain, suffering, distress, or last- ing harm (article 19); identify persons administratively responsible for care and equipment, provide sufficiently trained staff, and make adequate arrangements for veteri- nary advice and treatment (article 20); use only animals supplied by registered breeders or suppliers, subject to national exceptions (arti- cle 21); use only mice, rats, guinea pigs, golden hamsters, rabbits, dogs, cats, or quail originating in or ac- quired directly from registered breeding establish- ments, subject to national exemptions (member countries would add species to the list, particu- larly primates, as soon as there is a reasonable prospect of a sufficient supply of purpose-bred animals; straying domestic animals cannot be used and exemptions are not permitted) (article 22); conduct procedures outside their establishments only where authorized by the national authority (article 23); and keep records adequate to meet the requirements of article 27 and, in addition, to show the num- ber and species of all animals acquired, from whom acquired, and date of arrival, and to make such records available for inspections by the re- sponsible authority (article 24). Education and Training Article 25 specifies that professional and training procedures must be approved by responsible author- ities before being used and must be carried out by or under the supervision of a qualified person. Proce- dures are not permitted at or below the secondary level except when it is specifically directed to prepar- ing for a career involving treatment or care of ani- mals and the procedures entail no severe or enduring pain or suffering. Only the minimum measures abso- lutely necessary for the purpose are permitted, and only if their objective cannot be achieved by audio- visual or any other suitable methods. Article 26 re- quires that persons who carry out, take part in, or take care of animals used for procedures, including super- visors, must have adequate education and training. Statistical Information Article 27 requires each agreeing nation to collect and make public, where lawful, statistical information on animals in experimentation, including: G numbers and kinds of animals used; . numbers of animals, by categories, used in pro- cedures directly concerned with medicine and in teaching and learning; . numbers of animals, by categories, used in pro- cedures for the protection of humans and their environment; and G numbers of animals, by categories, used in pro- cedures required by legislation. Article 28 specifies that, subject to its own secrecy laws, each nation must submit information annually in the form set out in Appendix B to the Secretary Gen- eral of the Council, who is required to publish it. Each nation is invited to send the name and address of the corresponding authority, to be included in the Secre- tary General’s compilation of statistics. Recognition of International Procedures Article 29 binds agreeing nations to share informa- tion on results of procedures and to provide mutual assistance in order to avoid unnecessary repetition of procedures for the purposes of satisfying national leg- islation on health and safety. Final Provisions Articles 30 through 36 specify the manner and con- ditions under which the Convention will become rati- fied and effective (i.e., 6 months after four member states express their consent to be bound and, for any ratifying or acceding state after that, 6 months after written ratification or accession), and reserve a mem- ber state’s right to reservation, partial application, or denunciation (2). Appendix A of the Draft Convention Appendix A, Guidelines on Accommodation and Care of Animals, contains detailed specifications for physi- cal facilities, holding-room environments and environ- mental control, and care. Though the specifications are comprehensive, article 5 does refer to them as “suggested” (3). Appendix B of the Draft Convention Appendix B consists of Statistical Tables and Guid- ance Notes for Their Completion in Fulfillment of the Requirements in Articles 27 and 28 of the Draft Euro- pean Convention for the Protection of Vertebrate Ani- 416 . Alternatives to Animal Use in Research, Testing, and Education reals Used for Experimental and Other Scientific Pur- poses. The appendix would require submission by agreeing nations of experimental-animal data, re- ported to the Secretary General for each calendar year under the general classifications established by the referenced articles. The method of data collection is left to each member nation (4). Guidelines of the Council for International Organizations of Medical Sciences Through the World Health Organization (WHO), headquartered in Geneva, Switzerland, more than 150 nations exchange information and share resources for laboratory-animal science training, technical informa- tion, consultative support, and other activities. In 1985, in the culmination of a 3-year effort initi- ated in 1982, the Council for International Organiza- tions of Medical Sciences (CIOMS), an international nongovernmental organization representative of many branches of medicine and cognate disciplines that was established under the auspices of WHO and UNESCO in 1949, issued International Guiding Principles for Bio- medical Research Involving Animals (7). Modeled after the Tokyo revision of the Declaration of Helsinki by the World Medical Association in 1975 and CIOMS’s Proposed International Guidelines for Biomedical Research Involving Human Subjects, issued in 1982, the CIOMS International Guiding Principles are intended to provide a conceptual and ethical frame- work for whatever regulatory measure each country chooses to adopt with respect to animal use (7). The International Guiding Principles enumerate 11 basic principles, as follows (7): 1. II. 111, IV, The advancement of biological knowledge and the development of improved means for the pro- tection of the health and well-being both of man and of animals require recourse to experimen- tation on intact live animals of a wide variety of species. Methods such as mathematical models, computer simulation and in vitro biological systems should be used wherever appropriate. Animal experiments should be undertaken only after due consideration of their relevance for hu- man or animal health and the advancement of biological knowledge. The animals selected for an experiment should be of an appropriate species and quality, and the minimum number required, to obtain scientifi- cally valid results. V. Investigators and other personnel should never fail to treat animals as sentient, and should re- VI. VII. VIII. IX. x. XI< gard their proper care and use and the avoidance or minimization of discomfort, distress, or pain as ethical imperatives. Investigators should assume that procedures that would cause pain in human beings cause pain in other vertebrate species although more needs to be known about the perception of pain in animals. Procedures with animals that may cause more than momentary or minimal pain or distress should be performed with appropriate sedation, analge- sia, or anesthesia in accordance with accepted veterinary practice. Surgical or other painful pro- cedures should not be performed on unanesthe - tized animals paralysed by chemical agents. Where waivers are required in relation to the provisions of article VII, the decisions should not rest solely with the investigators directly con- cerned but should be made, with due regard to the provisions of articles IV, V, and VI, by a suita- bly constituted review body. Such waivers should not be made solely for the purposes of teaching or demonstration. At the end of, or when appropriate during, an experiment, animals that would otherwise suf- fer severe or chronic pain, distress, discomfort, or disablement that cannot be relieved should be painlessly killed. The best possible living conditions should be maintained for animals kept for biomedical pur- poses. Normally the care of animals should be under the supervision of veterinarians having ex- perience in laboratory animal science. In any case, veterinary care should be available as re- quired. It is the responsibility of the director of an in- stitute or department using animals to ensure that investigators and personnel have appropri- ate qualifications or experience for conducting procedures on animals. Adequate opportunities shall be provided for in-service training, includ- ing the proper and humane concern for the ani- mals under their care. Additional special provisions accompany the basic principles. These deal with sources of supply of ani- mal subjects; transport conditions; housing, including space allocation, hygienic standards, and protection against vermin; environmental conditions, including temperature, humidity, lighting, and social interaction; nutrition appropriate to the species; provision of veterinary care; and the keeping of records (7). The CIOMS statement also urges that the develop- ment and use of alternatives be actively encouraged. Specifically mentioned are nonbiological methods— such as the study of structure-activity relationships or App. E—International Agreements Governing Animal Use G 417 computer modeling—and biological methods, includ- ing the use of micro-organisms, in vitro preparations, and sometimes animal embryos (7). Organization for Economic Cooperation and Development The Organization for Economic Cooperation and De- velopment (OECD) is a group of nations whose mem- bership accounts for two-thirds of the world’s chemi- cal production, including the United States, Canada, Japan, and most of the countries of Western Europe. It also embraces six organizations that have a major role in international efforts to regulate chemicals (6). In 1979-80, an international group of experts con- vened under the OECD’s Special Program on the Con- trol of Chemicals drafted and recommended for the Council’s approval OECD Principles of Good Labora- tory Practice. The Council approved the document in 1981 (OECD, Guidelines for Testing of Chemicals, C(81)30 (Final), Annex 2). Though the main purpose for adopting the Princi- @es was to promote international harmonization of chemical-testing practices and thereby help safeguard the integrity of test results required under health and environmental safety laws, the document is patterned very much after good laboratory practice regulations adopted in 1978 by the U.S. Food and Drug Adminis- tration (see ch. 13). Following the Principles’ general command would certainly have an impact on use of test animals, but they do not contain the same detailed language on animal care, management, and housing that domestic regulations do, nor are any sanctions to be levied for failure to observe them. Appendix E References 1. Caufield, C., “Animal Treaty is No Protection,” iVevv Scien- Iisl 1417:43, 1984. 2. Council of Europe, Ad Hoc Committee of Experts for the Protection of Animals, Final Activity Report, Addendum I: Draft European Convention for the Protection of Ver- tebrate Animals Used for Experimental and Other Scien- tific Purposes and Draft Explanatory Report (Strasbourg, France: May 18, 1983). 3. Council of Europe, Ad Hoc Committee of Experts for the Protection of Animals, Final Activity Report, Adden- dum 11: Draft European Convention for the Protection of Vertebrate AnimaJs Used for Experimental and Other Scientific Purposes, Appendix A-Guidelines on Accom- modation and Cam of Am”mals (Article 5 of the Draft Con- vention) (Strasbourg, France: May 18, 1983). 4. Councii of Europe, Ad Hoc Committee of Experts for the 5! 6, 7. 8. 9. 10. 11. 12. 13. Protection of Animals, Final Activity Report, Addendum III: Draft European Convention for the Protection of Ver- tebrate Animals Used for Experimental and Other Scien- tific Purposes, Appendix E—Statistical Tables and Guid- ance Notes for Their Completion in Fulfillment of the Requirements in Articles 27 and 28 of the Draft Conven- tion (Strasbourg, France: May 18, 1983). Gravitz, M., ‘(Primates Get Top Priority,” The Animals Agenda 5(5):4-5, 1984. Held, J., “Animals in Research: An International Over- view,” Cal. Vet. 1:93-95, 1983. Howard-Jones, N., “A CIOMS Ethical Code for Animal Ex- perimentation,” WHO Chronicle 39:51-56, 1985. Japan Science Council, On the Adoption of Guidelines for Anima] Experimentation, 80th General ,Meeting, Nov. 5, 1980. Lazarowitz, A., Management Authority, Division of Re- search, Fish and Wildlife Service, U.S. Department of the Interior, Washington, DC, personal communication, Au- gust 1984. Leavitt, E. (cd,), Animals and Their Legal Rights (Wash- ington, DC: Animal Welfare Institute, third cd., 1978), Appendix. Nay, A., et a]., Animal Welfare Laws in Foreign Coun- tries (Washington, DC: Library of Congress, 1976). Humane Society News, “Update: The Good With the Bad,” 28:24, 1983. Rowan, A, N., Of Mice, Models, & Men: A Critical Evalu- ation of Animal Research (Albany, NY: State University of New York Press, 1984). 14. Vallier, G., “European Concepts on the Use of Labora- tory Animals in Relationship With Animal Welfare Prob- lems,” Dev. Bio/. Stand. 45:189-195, 1980. 15. Weiderkehr, M., “The Council of Europe’s Conventions,” Council of Europe Forum, March 1982, p. 7. For this assessment, OTA commissioned 10 reports on various topics concerning alternatives to animal use in research, testing, and education. The manuscripts of these contract reports are available in three volumes from the National Technical Information Service, 5285 Port Royal Road, Springfield, VA, 22161. Volume I: Overview “Survey and Estimates of Laboratory Animal Use in the United States, ” Kurt Enslein, Health Designs, Inc., Rochester, NY. ‘(Ethical Considerations,” Arthur H. Flemming, Depart- ment of Philosophy, The University of Chicago, Chi- cago, IL. “Scope of ‘Alternatives’: Overview of the State of the Art, ” Roland M. Nardone and Lucille Ouellette, De- partment of Biology, Catholic University, Washing ton, DC. “Overview of Computer Use in Research, Testing, and Education,” Paul N. Craig, Shady side, MD. Volume II: Research “Alternatives to Animal Use in Biomedical Research,” Eileen M. Cline, Springfield, VA. “Alternatives to the Use of Animals in Behavioral Re- search)” Gordon G. Gallup, Jr., Department of Psy- chology, State University of New York at Albany, Al- bany, NY. “Alternatives to Animal Use in Veterinary Medicine,” Bennie I. Osburn and faculty, School of Veterinary Medicine, University of California, Davis, CA. Volume 111: Testing and Economics “Animal Testing for Safety and Effectiveness,” Thomas D. Sabourin, Betsy D. Carlton, Robin T. Faulk, and L. Barry Goss, Environmental and Health Sciences Division, Battelle Columbus Laboratories, Columbus, OH. “Animal Testing and Alternatives,” Meyer, Failer, and Weisman, P, C., Washington, DC. “Economic and Policy Considerations,” Henry R. Hertz- feld and Thomas D. Myers, Washington, DC. 418 Appendix G Acknowledgments OTA would like to thank the members of the advisory panel who commented on drafts of this report, the contractors who provided material for this assessment, and the many individuals and organizations that sup- plied information for the study. In addition, OTA acknowledges the following individuals for their review of drafts of this report: Robert F. Acker National Foundation for Infectious Diseases James Aftosmis E.I. du Pent de Nemours and Co., Inc. Donald G. Ahearn Georgia State University Gwynn C. Akin Syntex Corporation Robert L. Alkire Society of Toxicologic Pathologists Douglas L. Archer Food and Drug Administration John L. Bartholomew U.S. Army Michael Balls Fund for the Replacement of Animals in Medical Experiments Edward M. Barrows Georgetown University George W. Beran Iowa State University Richard N. Bergman University of Southern California Emmanuel M. Bernstein Psychologists for the Ethical Treatment of Animals Keith A. Booman Soap and Detergent Association Arnold P. Borsetti Food and Drug Administration Joseph F. Borzelleca Medical College of Virginia Richard C. Bostwick Merck and Company, Inc. Richard P. Bradbury Food and Drug Administration John E. Burris National Research Council Jack L. Carter Biological Sciences Curriculum Study Charles B. Cleveland Pharmaceutical Manufacturers Association Eileen M. Cline Springfield, VA Thomas G. Coleman University of Mississippi School of Medicine Frances K. Conley Stanford University Medical School Charles E. Cover E.I. du Pent de Nemours and Co., Inc. Geraldine V. Cox Chemical Manufacturers Association Paul N. Craig Shady Side, MD Arthur L. Craigmill University of California, Davis Lester M. Crawford Food and Drug Administration Lloyd E. Davis The University of Illinois Mary Dawson University of Strathclyde Charles DeLisi National Cancer Institute Kennerly H. Digges National Highway Traffic Safety Administration Rebecca Dresser Baylor College of Medicine Ronald Dubner National Institute for Dental Research Sarah Wells Duffy U.S. House of Representatives James L. Dwyer Millipore Corporation 419 420 G Alternatives to Animal Use in Research, Testing, and Education David M. Ferguson ICI Americas, Inc. Kenneth D. Fisher Federation of American Societies for Experimental Biology Michael Allen Fox Queen’s University Gordon G. Gallup, Jr. State University of New York at Albany Roger W. Galvin Animal Legal Defense Fund of Washington, DC William I. Gay National Institutes of Health Michael A. Giannelli The Fund for Animals, Inc. Robert P. Giovacchini The Gillette Company Dawn G. Goodman American College of Veterinary Pathology L. Barry Goss Battelle—Columbus Laboratories Sidney Green Food and Drug Administration Lowe]] M. Greenbaum Medical College of Georgia Earl W. Grogan National Research Council Francis J. Haddy Uniformed Services University for the Health Sciences Richard E. W. Halliwell University of Florida School of Medicine Harlyn O. Halvorson Brandeis University Thomas E. Harem Stanford University Medical School F. Gene Hampton National Science Teachers Association David G. Hattan Food and Drug Administration George A. Hedge West Virginia University Medical Center Lee A. Heilman American Association for the Accreditation of Laboratory Animal Care Joe R. Held Pan American Zoonoses Center John R. Herbold Department of Defense Henry R. Hertzfeld Washington, DC Karen M. Hiiemae The University of Illinois at Chicago Larry Horton Stanford University Peter J. Hyde International League for Animal Rights Robert Kainz Walkersville, MD Gerald S. Kanter Albany Medical College Donald Kennedy Stanford University Keith F. Killam, Jr. University of California, Davis Robert W. Krauss Federation of American Societies for Experimental Biology Sienna LaRene Michigan Humane Society C. Max Lang The Pennsylvania State University Thomas W. Langfitt The University of Pennsylvania Victor G. Laties University of Rochester School of Medicine and Dentistry Chung Lee Northwestern University Medical School Joel L. Mattsson Dow Chemical U.S.A. Charles R. McCarthy National Institutes of Health App. G—Acknowledgments G 421 Basil E. McKenzie Ortho Pharmaceutical Corporation Donald E. McMillan University of Arkansas for Medical Sciences Donald R. Meyer The Ohio State University Joel A. Michael Rush-Presbyterian-St, Luke's Medical Center Robert J. Moolenaar American Industrial Health Council John A. Moore Environmental Protection Agency Ray E. Moseley University of Arkansas for Medical Sciences Thomas H. Moss Case Western Reserve University Laila A. Moustafa World Health Organization Arnauld E. Nicogossian National Aeronautics and Space Administration Sharon J. Northup Travenol Laboratories, Inc. Mike G. Norton British Embassy Karl Johan Obrink Uppsala Biomedicinska Centrum F. Barbara Orlans Scientists Center for Animal Welfare Bennie I. Osburn University of California, Davis Robert E. Osterberg Food and Drug Administration Alex Pacheco People for the Ethical Treatment of Animals Douglas L. Park Food and Drug Administration Paul C. Rambaut National Aeronautics and Space Administration B. Randall, IV Congressional Research Service Walter C. Randall Loyola University Medical Center Tom Regan North Carolina State University Conrad B. Richter National Institute of Environmental Health Sciences Carol F. Rodgers U.S. House of Representatives Bernard E. Rollin Colorado State University Walter G. Rosen National Research Council Carl F. Rothe Indiana University School of Medicine Harry C. Rowsell University of Ottawa H. Rozemond Staatstoezicht op de Volksgezondheid Thomas D. Sabourin Battelle—Columbus Laboratories Jonathan D. Sackner Philadelphia, PA William M. Samuels American Physiological Society Robert A. Scala Exxon Corporation Trevor H. Scott World Society for the Protection of Animals Fred R. Shank Food and Drug Administration Kenneth J. Shapiro Psychologists for the Ethical Treatment of John F. Sherman Association of American Medical Colleges Charles E. Short Animals New York State College of Veterinary Medicine Lee R. Shun University of California, Davis Robert S. Shurtleff Springfield, MA Evan B. Siegel The Proprietary Association Sidney Siegel National Library of Medicine 422 Alternativws to Animal Use in Research, Testing, and Education Richard C. Simmonds Uniformed Services University Sciences Peter Singer Monash University Cheryl L. Sisk Michigan State University Kendric C. Smith of the Health Stanford University Medical School Judy A. Spitzer Louisiana State University Medical Center Dennis M. Stark The Rockefeller University Donald G. Stein Clark University Marshall Steinberg Society of Toxicology Christine Stevens Society for Animal Protective Legislation Irving 1. A. Tabachnick Schering Corporation Dennis J. Taylor Rhodes and Taylor J. W. Thiessen Department of Energy Robert Thomas U.S. Department of Energy Ethel Thurston American Fund for Alternatives to Animal Re- search Charles S. Tidball George Washington University Medical Center Richard J. Traystman The Johns Hopkins Hospital Bruce L. Umminger National Science Foundation James Vorosmarti, Jr. Department of Defense William J. Waddell University of Louisville School of Medicine James R. Walker The University of Texas Medical Branch at Galveston John S. Wassom Oak Ridge National Laboratory William L. West Howard University College of Medicine James A. Will University of Wisconsin Research Animal Resources Center James Willett National Institutes of Health Robert P. Williams Baylor College of Medicine Steven M. Wise Attorneys for Animal Rights (Boston), Inc. Earl H. Wood Mayo Foundation and Mayo Medical School Constantine J. Zervos Food and Drug Administration Appendix H Glossary of Acronyms and Terms Glossary of Acronyms AAALAC —American Association for Accreditation of Laboratory Animal Care AALAS —American Association for Laboratory Animal Science AAMC —Association of American Medical Colleges AAVMC –Association of American Veterinary Medical Colleges ACC –Animal Care Committee (Canada) ACP —American College of Physicians ACUC —Animal Care and Use Committee ADAMHA–Alcohol, Drug Abuse, and Mental Health Administration (PHS, DHHS) AFAAR —American Fund for Alternatives to Animal Research ALD —Approximate Lethal Dose ALDF –Animal Legal Defense Fund AMD —Aerospace Medical Division (U.S. Air Force) APA —American Psychological Association APHIS —Animal’ and Plant Health Inspection Service (USDA) APS –American Physiological Society ARC —Animal Research Committee ASPCA —American Society for the Prevention of Cruelty to Animals AUCC —Association of Universities and Colleges of Canada AVMA —American Veterinary Medical Association BID –bureau, institute, or division (NIH) BIOSIS —Biosciences Information Service CAAT -Center for Alternatives to Animal Testing (The Johns Hopkins University) CALAS -Canadian Association for Laboratory Animal Science CBO -Congressional Budget Office (U.S. Congress) CCAC —Canadian Council on Animal Care CDC --Centers for Disease Control (PHS, DHHS) CERCLA —Comprehensive Environment Response, Compensation, and Liability Act CFHS —Canadian Federation of Humane Societies CFR -Code of Federal Regulations CIIT --Chemical Industry Institute of Toxicology CIOMS CITES CPSC CTFA CT&T DHEW DHHS DOD DOE DOT EPA EWST FAA FASEB FDA FIFRA FOA FOIA FR FRAME FTC GAO GLP IACUC ILAR IRAC IRB ISEF LADB LC5O LD5O LSRO MRI NAL --Council of International Organizations of Medical Sciences --Convention on International Trade in Endangered Species —U.S. Consumer Product Safety Commission —Cosmetic, Toiletry, and Fragrance Association -Chemical Times and Trends —U.S. Department of Health, Education, and Welfare (see DHHS) —U.S. Department of Health and Human Services —U.S. Department of Defense —U.S. Department of Energy —U.S. Department of Transportation —U.S. Environmental Protection Agency —Ethics and Values in Science and Technology (NSF program) —U.S. Federal Aviation Administration —Federation of American Societies for Experimental Biology –Food and Drug Administration (PHS, DHHS) —Federal Insecticide, Fungicide, and Rodenticide Act —Friends of Animals, Inc. —Freedom of Information Act —Federal Register —Fund for Replacement of Animals in Medical Experiments —U.S. Federal Trade Commission -General Accounting Office (U.S. Congress) —Good Laboratory Practices —Institutional Animal Care and Use Committee —Institute for Laboratory Animal Resources (NRC) —Interagency Research Animal Committee —Institutional Review Board –International Science and Engineering Fair —Laboratory Animal Data Bank —median lethal concentration —median lethal dose –Life Sciences Research Office (FASEB) —magnetic resonance imaging —National Agricultural Library 423 424 G Alternatives to Animal Use in Research, Testing, and Education NAS NASA NBS NCI NCTR NIDA NIEHS NIH NIHARC NIMH NIOSH NLM NRC NSF NTIS NTP NWHL OECD OHER ONR OPRR OSHA OSTP OTA OTS PHS PMA PRI QSAR RCRA R&D RRF RTECS —National Academy of Sciences —National Aeronautics and Space Administration —National Bureau of Standards (Department of Commerce) –National Cancer Institute (NIH) —National Center for Toxicological Research (FDA) —National Institute on Drug Abuse (ADAMHA) –National Institute of Environmental Health Sciences (NIH) –National Institutes of Health (PHS, DHHS) —National Institutes of Health Animal Research Committee –National Institute of Mental Health (ADAMHA) —National Institute for Occupational Safety and Health (CDC) –National Library of Medicine (NIH) —National Research Council —National Science Foundation —National Technical Information Service (Department of Commerce) –National Toxicology Program (NIEHS) —National Wildlife Health Laboratory —Organization for Economic Cooperation and Development -Office of Health and Environmental Research (DOE) –Office of Naval Research (Navy) —Office for Protection from Research Risks (NIH) -Occupational Safety and Health Administration (U.S. Department of Labor) -Office of Science and Technology Policy (Executive Office of the President) -Office of Technology Assessment (U.S. Congress) –Office of Toxic Substances (EPA) –U.S. Public Health Service (DHHS) —Pharmaceutical Manufacturers’ Association —Primate Research Institute (University of New Mexico) -quantitative structure-activity relationships –Resource Conservation and Recovery Act —research and development –Registered Research Facility —Registry of Toxic Effects of Chemical Substances (NIOSH) SBIR –Small Business Innovation Research (program) SPCA –Society for the Prevention of Cruelty to Animals SSR —Society for the Study of Reproduction TDB –Toxicology Data Bank (NLM) TSCA —Toxic Substances Control Act UNEP —United Nations Environment Program UNESCO —United Nations Educational, Scientific, and Cultural Organization USDA —U.S. Department of Agriculture VA —U.S. Veterans’ Administration WHO —World Health Organization WRPRC —Wisconsin Regional Primate Center Glossary of Terms Research Acute Toxicity Test: Tests that are used to detect the toxic effects of single or multiple exposures to a sub- stance occurring within 24 hours. These are fre- quently the first tests performed to determine the toxic characteristics of a given substance. One of the most common acute toxicity tests is the LD5O test. Alternatives to Animal Use: For purposes of this assessment, OTA has chosen to define ‘(alternatives” as encompassing any subjects, protocols, or technol- ogies that replace the use of laboratory animals al- together; reduce the number of animals required; or refine existing procedures or techniques so as to minimize the level of stress endured by the animal. These technologies involve the continued, but modi- fied, use of animals; use of living systems; use of chem- ical and physical systems; and use of computers. American Association for Accreditation of Labo- ratory Animal Care (AAALAC): A voluntary private organization that, by April 1985, provided accredi- tation for 483 institutions. AAALAC accreditation is based on the provisions of the NIH Guide for the Care and Use of Laboratory Animals, and is recognized by the PHS. Ames Test: The most commonly used test for mutage- nicity, it tests “reverse mutation” by exposing an al- ready mutated strain of micro-organism to potential mutagens. If the mutation is reversed the micro- organisms regain their ability to produce the amino acid histidine and will proliferate in a histidine- deficient culture medium. However, when used alone the Ames test does not seem to be as predictive of human carcinogenicity as are animal tests. Analgesic: An agent that relieves pain without caus- ing loss of consciousness. Anesthetic: An agent that causes loss of the sensation of pain, usually without loss of consciousness. Anes- thetics may be classified as topical, local, or general. App. H—Glossary of Acronyms and Terms 425 Animal: For purposes of this assessment, animal is de- fined as any nonhuman member of five classes of vertebrates: mammals, birds, reptiles, amphibians, and fish. Within this group, two kinds of animals can be distinguished, warm-blooded animals (mammals and birds) and cold-blooded animals (reptiles, am- phibians, and fish). Under this definition, inver- tebrates are not considered to be animals. Animal Care and Use Committee (ACUC): An institu- tional committee that oversees housing and routine care of animals. The committee may also review research proposals. The committee’s membership generally includes the institution’s attending veter- inarian, a representative of the institution’s admin- istration, users of research animals, and one or more nonscientist and lay members. Animal and Plant Health Inspection Service (APHIS): A branch of USDA that, among other duties, is charged with the enforcement of the Animal Wel- fare Act. Enforcement of the act is directed through four regional offices and is carried out by 286 APHIS Veterinary Medical Officers (inspectors) who spend about 6 percent of their time inspecting over 1,200 research facilities (many of which have multiple sites). Animal Use The use of animals for research purposes. Three aspects of animal use are dealt with in this assessment: in behavioral and biomedical research; in testing products for toxicity; and in the education of students at all levels. This assessment does not cover animal use for food and fiber; animal use to obtain biological products; or animal use for sport, entertainment, or companionship. Animal Welfare Act: This act, passed in 1966 and amended in 1970, 1976, and 1985, was originally an endeavor to stop traffic in stolen animals that were being shipped across State lines and sold to research laboratories. Amendments to the act have expanded its scope to include housing, feeding, transportation, and other aspects of animal care. However, the act bars regulation of the conduct of research and test- ing by USDA. Animals covered by the act, as cur- rently enforced, are dogs, cats, hamsters, rabbits, guinea pigs, nonhuman primates, and marine mam- mals. The Animal Welfare Act is enforced by APHIS. Animal Welfare Enforcement Report: Annual re- port submitted to Congress by APHIS, based on data collected from the Annual Report of Research Facil- ity forms. Animal Welfare Groups: There are a number of groups concerned with animal rights and animal welfare-e. g., the ASPCA, FOA, and AFAAR. These groups cover a broad spectrum of ethical concerns about animal use, they may question the objectives as well as the means of research, but they generally find common ground in the principle of humane treatment of animals. Annual Report of Research Facility: This is required under the regulations stemming from the Animal Welfare Act. Research facilities must submit these annual reports, detailing animal use, to APHIS for evaluation. (Elementary and secondary schools are exempt, as are facilities using exempt species.) APHIS presents data collected from these reports to Con- gress in its annual Animal Welfare Enforcement Report. Anticruelty Statutes: Laws passed by States that pro- hibit active cruelty, and in some cases passive cru- elty (neglect), to animals. Some of these laws acknowl- edge the potential application of anticruelty statutes to research animals, but most of them exempt “scien- tific experiments” entirely. Twenty States and the District of Columbia regulate research to some ex- tent. Twenty-one States have some provisions in their codes requiring the teaching of “kindness” to or the “value” of animals, and a few place restrictions on animal experimentation in secondary schools. Behavioral Research: Research into the movements and sensations by which living things interact with their environment, with the purpose of better under- standing human behavior. A further goal of behav- ioral research is the better understanding of animal species of economic or intrinsic interest to humans. Behavioral research differs from biomedical research in that it is difficult to study behavioral phenomena in isolation; therefore continued, but modified, use of animals holds most promise for this area of research. Biological Model A surrogate or substitute for a proc- ess or organ of interest to an investigator. Animals or alternatives can serve as biological models. Biological Testing: The repetitive use of a standard biological test situation or protocol employing differ- ent chemicals or different test parameters. Such test protocols are more stereotyped than those used in research, and may be more amenable to the institu- tion of a computerized data retrieval system. Biomedical Research: A branch of research devoted to the understanding of life processes and the appli- cation of this knowledge to serve humans. A major user of animals, biomedical research affects human health and the health care industry. It is instrumen- tal in the development of medical products such as drugs and medical devices, and in the development of services such as surgical and diagnostic techniques. Biomedical research covers abroad spectrum of dis- ciplines, such as anatomy, biochemistry, biology, endocrinology, genetics, immunology, nutrition, on- cology, and toxicology. Carcinogen: An agent or process that significantly in- 426 Ž Alternatives to Animal Use in Research, Testing, and Education creases the incidence of abnormal, invasive, or un- controlled cell growth in a population. Carcinogens fall into three classes: chemicals, viruses, and ioniz- ing radiation. A variety of screening assays have been developed to detect chemical carcinogens, including the Salmonella-mediated mutagenesis assay (Ames test), the sister chromatid exchange assay, and tradi- tional laboratory animal toxicity tests. Cell Culture: Growth in the laboratory of cells isolated from multicellular organisms. Each culture is usually of one type. Cell culture may provide a promising alternative to animal experimentation, for example in the testing of mutagenicity, and may also become a useful adjunct in repeated dose toxicity testing. Center for Alternatives to Animal Testing (CAAT): Established by the Johns Hopkins University in 1981 to search for alternatives to animal use, CAAT puts out publications and supports intramural and extra- mural research. The Center is sponsored by the CTFA and corporate donors as well as consumer and in- dustrial groups. Chick Embryo Chorioallantoic Membrane Assay: A test used to determine the irritancy of a substance. A test sample is placed on the chorioallantoic mem- brane formed on top of a chick embryo. The mem- brane is then evaluated for response to the test sub- stance and the embryo is discarded. This test may be a promising alternative to the Draize Test. Chronic Toxicity Test: Repeated dose toxicity test with exposure to a test substance lasting at least 1 year, or the lifetime of the test species. Comprehensive Environment Response, Com- pensation, and Liability Act (CERCLA): Known as “Superfund,” CERCLA authorizes the Federal Gov- ernment to cleanup or otherwise respond to the re- lease of hazardous wastes or other pollutants that endanger public welfare. Crossover Test: A useful laboratory or clinical method whereby an animal serves as its own control by first receiving a drug or a placebo and then receiving the reverse. This kind of test has potential applications in anesthesiology, endocrinology, radiology, and vari- ous other fields. Computer Simulation: The use of specially devised computer programs to simulate cells, tissues, fluids, organs, and organ systems for research purposes; to develop mathematical models and algorithms for use in toxicity testing; and to simulate experiments tradi- tionally done with animals, for educational purposes. Data Sources: Can provide an alternative to animal testing by disseminating information generated from prior use of animals. The TDB and RTECS are two such sources, as was the LADB. Descriptive Toxicology A branch of toxicology deal- ing with phenomena above the molecular level. De- scriptive toxicology relies heavily on the techniques of pathology, statistics, and pharmacology to dem- onstrate the relationship between cause and effect— e.g., that certain substances cause liver cancer in cer- tain species within a certain time. It is most often used in regulatory schemes requiring testing. Distress: Usually the product of pain, anxiety, or fear. However, distress can also occur in the absence of pain. For example, an animal struggling in a restraint device may be free from pain, but maybe in distress. Distress can be eased with tranquilizers. Draize Eye Irritancy Test: A test that involves plac- ing a single dose of a test substance into one eye of four to six rabbits (the other eye remains untreated) and observing its irritating effects. A promising alter- native to this test is the chick embryo chorioallan- toic membrane assay. Education: The aspect of education dealt within this assessment is the use of animals and alternatives in the teaching of life sciences to secondary school stu- dents, university students, health professionals and preprofessionals, and research scientists. Federal Environmental Acts: A number of these have been passed to protect human health and the environment from the adverse effects of toxic sub- stances, and to regulate the release of such sub- stances into the environment. Among these acts are FIFRA, TSCA, the Clean Air Act, the Clean Water Act, RCRA, CERCLA, and the Consumer Product Safety Act, Animal testing provides much of the data needed for the enforcement of these acts. Federal Government Use of Animals for Research Six Cabinet departments and four Federal agencies conduct intramural research and testing involving animals. They are: USDA, Department of Commerce, DHHS, DOD, Department of the Interior, DOT, CPSC, EPA, NASA, and the VA. Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA): Designed to protect the human envi- ronment from the adverse effects of pesticides and their use, FIFRA regulates various aspects of pesti- cide use by means of registration, labeling, and the setting of maximum residue levels. It also established procedures for safe application, storage, and disposal of pesticides. Good Laboratory Practices (GLP): Rules adopted by FDA in 1978 requiring that all regulated parties con- ducting nonclinical laboratory studies keep records and permit audits of such studies. The GLP rules also contain specific provisions for animal housing, feeding, and care, In 1983, EPA issued similar GLP App. H—Glossary of Acronyms and Terms G 427 rules for its toxic substances and pesticides research programs. Guidelines for Animal Care and Use: Various or- ganizations outside the Federal Government have adopted their own guidelines-e.g., the APA’s Guide- lines for Ethical Conduct in the Care and Use of Ani- mals, which is the most comprehensive and has been endorsed by FASEB; the APS’s Guiding Principles in the Care and Use of Animals; and the AVMA’s Ani- mal Welfare Guiding Principles. For Federal guide- lines, see Interagency Research Animal Committee, NIH Guide for the Care and Use of Laboratory Ani- mals, and PHS Policy. Hepatotoxicity: The quality of exerting a destructive or poisonous effect upon the liver. Homology The correspondence among organisms of structures and functions derived from a common evolutionary origin (e g., a common gene structure). Immunoscintigraphy The use of external radioimag- ing techniques to locate tumors and to identify cer- tain noncancerous diseases. Institute for Laboratory Animal Resources (ILAR): A component of the National Research Council, ILAR performs periodic surveys on the use of laboratory animals. Interagency Research Animal Committee (IRAC): This committee was formed by 14 Federal entities in recognition of a need for an interagency body knowledgeable about the welfare of research ani- mals. IRAC meets regularly to discuss research needs and has written principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research and Training. These Principles, which incorporate nine injunctions on animal welfare, are intended to serve as a model for Federal agencies in developing pol- icies on animal use. Invertebrate Any nonplant organism without a spinal column-e. g., worms, insects, and crustaceans. In- vertebrates account for 90 percent of the Earth’s non- plant species. For the purposes of this assessment, invertebrates are not considered to be animals. In vitro: Literally, in glass; pertaining to a biological process or reaction taking place in an artificial envi- ronment, usually a laboratory. Human and animal cells, tissues, and organs can be cultured in vitro. In vitro testing may hold some promising alterna- tives to animal testing-e.g., in testing for eye irrita- tion and mutagenicity. In vivo: Literally, in the living; pertaining to a biologi- cal process or reaction taking place in a living cell or organism. Laboratory Animal Data Bank (LADB): Founded by NCI and NLM in the late 1970s, the LADB was sup- posed to provide a computer-based registry of re- search and testing data. However, the data were limited and consequently LADB had few users. It was terminated in 1981 because of lack of funding. It ex- ists today only as an archival reference, LC50:An acute toxicity test used to screen substances for their relative toxicity. LC50 is calculated to be the lethal concentration for half of the animals exposed to a test substance. Exposure may be by breathing vapor or immersion in liquid (e.g., fish in water). LD50: An acute toxicity test used to screen substances for their relative toxicity. LD50 is calculated to be the lethal dose for half of the animals exposed to a test substance. Exposure is often by ingestion. Mechanistic Toxicology An approach to testing that focuses on the chemical processes by which a toxic effect occurs. Mechanistic toxicology testing relies heavily on physiology, biochemistry, and analytical chemistry techniques to monitor these processes. Micro-organism: A minute microscopic or submicro- scopic living organism, such as bacteria, viruses, and protozoa. Mutagen: An agent that induces chemical changes in genetic material. Chemicals, viruses, and ionizing radiation can be mutagenic. Most carcinogens are mutagens, therefore many screening tests to detect carcinogens are designed to detect the mutagenic potential of the compound. Some mutagens are not direct-acting, requiring metabolic activation in the body before they exert their mutagenic potential. National Toxicology Program (NTP): NTP was char- tered in 1978 as a cooperative effort by DHHS. Par- ticipants in NTP are NIH (through its agencies NCI and NIEHS), FDA (through NCTR), and CDC (through NIOSH). The stated goals of NTP include the expan- sion of toxicological information; expansion of num- hers of chemicals to be tested; the validation, devel- opment, and coordination of tests to meet regulatory needs; and the communication of programs, plans, and results to the public. Neurotoxicity: The quality of exerting a destructive or poisonous effect on nerve tissue. NIH Guide for the Care and Use of Laboratory Ani- mals: Revised in 1985, the Guide lays out detailed standards for animal care, maintenance, and hous- ing. Its provisions apply to all research supported by NIH, and it is used by most animal research facil- ities, both within and outside the Federal Govern- ment. AAALAC and PHS also use it when assessing research facilities for accreditation. Nonliving Systems: Inanimate chemical or physical systems used in testing. Oncology: The study of tumors. 428 G Alternatives to Animal Use in Research, Testing, and Education Organ Culture: The attempt to isolate and maintain animal or human organs in in-vitro culture. Long- term culture of whole organs is not generally feasi- ble, but they can be sustained in cultures for short periods (hours or days). Pain: Discomfort resulting from injury or disease. Pain can also be psychosomatic, the product of emotional stress. Pain can be induced by mechanical, thermal, electrical, or chemical stimuli, and it can be relieved by analgesics or anesthetics. Pharmacokinetic Studies: A branch of toxicity test- ing that provides information about the mechanics of absorption. PHS Policy on Humane Care and Use of Labora- tory Animals by Awardee Institutions: Revised in 1985, the Policy applies to PHS-supported activi- ties involving animals (including those of NIH). It re- lies on the NIH Guide for the Care and Use of Lab- oratory Animals, and uses institutional committees for the assessment of programs and maintenance of records. Pound Release Laws: State laws that provide for the seizure, holding, and humane disposal of stray and unwanted animals, Most States permit the release of unowned animals to research institutions that have met specified conditions. These laws have been closely scrutinized. In the past 10 years and nine States have passed laws prohibiting the release of stray ani- mals to research institutions. The most far-reaching of these laws takes effect in Massachusetts in 1986. Also referred to as “pound seizure laws.” Protocol: The plan of a scientific experiment or treat- ment. Reduction Considered an alternative to animals when fewer animals are used in research and education through changed practices, sharing of animals, or better design of experimental protocols. Refinement: An alternative to animal use by better use and modification of existing procedures so that animals are subject to less pain and distress. Exam- ples of such refinements are the administration of anesthetics and tranquilizers, humane destruction, and the use of noninvasive imaging techniques. Registry of Toxic Effects of Chemical Substances (RTECS): An annually published compendium, ex- tracted from the literature, of known toxic and biological effect of chemical substances. RTECS is published by NIOSH under the provisions of the Oc- cupational Safety and Health Act of 1970. Repeated-Dose Toxicity Test: Repeated or prolonged exposure to measure the cumulative effects of expo- sure to a test substance. These tests involve chronic, subchronic, or short-term exposure to a test sub- stance. Replacement: An alternative to animal use, replac- ing methods using animals with those that do not. Examples include the use of a placenta instead of a whole animal for microsurgical training, the use of cell cultures instead of mice and rats, the use of non- living systems, and the use of computer programs. Research: The development of new knowledge and technologies, often with unpredictable but poten- tially significant results. Uncertainty, missteps, and serendipity are inherent in the research process. Re- search is distinguished from testing by the ways in which animals are used, and the identity of the in- vestigators. There are more research procedures than there are tests, and researchers are more likely to develop their own procedures. Research Facility Under the Animal Welfare Act, any individual, institution, organization, or postsecond - ary school that uses or intends to use live animals in research, tests, or experiments. Facilities that re- ceive no Federal support for experimental work and that either purchase animals only within their own State or that maintain their own breeding colonies are not considered research facilities under the act, however. Resource Conservation and Recovery Act (RCRA): This act was passed to protect public health and the environment through the regulation of the manage- ment and handling of hazardous waste and through the control of solid waste disposal. Resusci-Dog: A plastic mannequin linked to a com- puter. The Resusci-Dog can simulate an arterial pulse and pressure can be applied to its ribcage for cardiac massage or cardiopulmonary resuscitation. Sequential Design Test: The comparison of treatment groups at set stages of experimentation. Further ex- perimentation at higher doses is undertaken only if there is no significant difference between the two groups. This kind of test has potential application in anesthesiology, endocrinology, nutrition, and other fields. Serial Sacrifice: The sequential killing of animals to examine the occurrence and progress of induced effects. Short-Term Toxicity Test: Repeated dose toxicity test that involves exposure to a test substance over a pe- riod of 2 to 4 weeks. Speciesism: A term used by some animal rights activ- ists, referring to the denial of animal rights as a moral breach analogous to racism or sexism. State Environmental Acts: Legislation passed by States to regulate pesticides, air quality, water, and waste products. These laws are often the simple adoption or recodification of existing Federal laws. Subchronic Toxicity Test: Repeated dose toxicity test App. H—Glossary of Acronyms and Terms G 4 2 9 of intermediate duration, with exposure to a test sub- stance for 3 to 6 months. Testing: Standardized procedures that have been dem- onstrated to predict certain health effects in humans and animals. Testing involves the frequent repeti- tion of well-defined procedures with measurement of standardized biological endpoints. A given test may be used to test many different substances and may use many animals. Testing is used to establish the efficacy, safety, and toxicity of substances and pro- cedures. Tissue Culture: The maintenance in vitro of isolated pieces of a living organism. The various cell types are still arranged as they were in the original organ- ism and their differential functions are intact, Toxic Substances Control Act (TSCA): This act au- thorizes EPA to regulate substances that present an unreasonable risk of injury to health or to the envi- ronment. The act also requires the reporting or de- velopment of data to assess the risks posed by a given substance. Toxicity Testing: The testing of substances for toxic- ity in order to establish conditions for their safe use. There are now more than 50,000 chemicals on the market and 500 to 1,000 new ones are introduced each year. The Federal agencies with the largest role in toxicity testing are FDA, EPA, CPSC, and OSHA. Toxicology Data Bank (TDB): Made public by the NLM in 1978, the TDB provides toxicity information on more than 4,000 chemicals and substances. TDB information is based on conventional published sources. Tranquilizer: An agent that quiets, calms, and reduces anxiety and tension, with some alteration of the level of consciousness. T-test: An estimate of the difference between the mean values of one parameter of two treatments. This can be a powerful measure when the number of compari- sons is small, but the potential for error increases as the number of parameters grows. Veterinary Medicine: The maintenance and improve- ment of the health and well-being of animals, par- ticularly the 30 to 40 different species of animals of ‘economic, ecological, and environmental importance. Veterinary medicine is closely allied with veterinary. research. Veterinary Research: A branch of biomedical re- search devoted to the understanding of the life proc- esses of animals and the application of this knowl- edge to serve animals as well as humans. Index Index AAALAC, 16, 49 accreditation, process of, 344-345, 401-411 animal use and, 335-337 AAVMC and, 351 Federal agencies and, 386, 387, 388, 389, 392, 393 GLPs and, 293 AALAS, 208, 344, 345-346 AAMC, 204-206 AAVMC, 207, 350-351 ACC, 361-362 ACP, 349 ACUC, 14, 15-16, 292, 386, 391-392 ADAMHA, 45, 92, 295, 337 AFAAR, 210, 268, 269 Agriculture, Department of (U.S.). See USDA Alabama, 288, 307, 319 Alaska, 288, 307, 308, 319 Alberta, University of, 360 Alcohol, Drug Abuse, and Mental Health Adminis- tration. See ADAMHA American Association for Accreditation of Labora- tory Animal Care. See AAALAC American Association for Laboratory Animal Science. See AALAS American College of Physicians. See ACP American Fund for Alternatives to Animal Research. See AFAAR American Physiological Society. See APS American Psychological Association. See APA American Society for the Prevention of Cruelty to Animals. See ASPCA American Veterinary Medical Association. See AVMA Ames test, 186, 187-188, 250 Animal Care Committee. See ACC Animal Care and Use Committee. See ACUC Animal and Plant Health Inspection Service. See APHIS Animal Research Committee. See ARC Animal rights “consistency argument” and, 77-78 “interest theory” and, 76-77 legal action and, 314-316 “speciesism” and, 5, 79 “will theory” and, 76 Animal use and data accumulations, 5, 43-49, 58-65 economics of, 12, 29, 99, 115, 116, 123, 128, 151, 155, 206, 209-210, 213, 243-253 in education, 3, 199-214, 321-322 ethics of, 6, 71-82, 200, 202 in Federal Government, 14-15, 43-49 guidelines for, 13, 14, 15-16, 31, 33, 152, 157-167, 176, 200-202, 291, 293-296, 335-352, 361, 383-393, 395-400 humane treatment and, 6, 78-79 IACUCs and, 340-344 international agreements governing, 412-417 limitations of OTA study on, 50-52 modified, 7-8, 113, 114-118, 126-132, 175-176, 208-209 and pain, 103-105 patterns of, 43-66 in pharmacokinetics, 153 philosophical traditions of, 74-75 as a policy issue, 26-29, 31-32 product liability and, 157 public concern about, 3, 149, 157, 175, 181, 266, 293-294, 323-324, 339 regulation of, 13-18, 46, 157-167, 201-202, 203, 275-298, 305-322, 335-352, 359-375, 386-393 regulatory practices and, 12, 157-167, 175, 181-182, 188-189, 248, 278, 280, 283-289, 291-292, 297-298 religious traditions of, 71-74 in research, 3, 89-108 restricted necessity and, 80-81 in science fairs, 200-202 in testing, 3, 149-168 trends in, 5, 16-17, 57-65, 157 in the United Kingdom, 203-204 utilitarian principle of, 6, 79-81, 82 Animal use, alternatives to computer systems as, 7-8, 11-12, 124-126, 136-138, 228-238 economics of, 12, 189, 249-250, 265-266 in education, 10-11, 208-214 funding for, 13, 213-214, 259-270 Health Research Extension Act of 1985 and, 291-292 ‘ IACUCs and, 341 living systems as, 4, 7-8, 118-123, 133-136, 175, 177-179, 183, 184-186, 189-190, 209-210 nonliving systems as, 7-8, 124, 136, 180-181, 210-214 organ culture as, 119-120 OTA’s definition of, 39 as a policy issue, 18-23 reduction as, 4, 10-11, 39, 114-116, 126-128 refinement or replacement as, 4, 10-11, 39 in research, 6-8, 113-138 trends in, 188-190 See also Reduction; Refinement; Replacement Animal Welfare Act of 1966. See Laboratory Animal Welfare Act 433 434 . Alternatives to Animal Use in Research, Testing, and Education Animals benefits to, of research, 102 classifications of, 37-38 definitions of, 4, 37-38, 306 estimates of numbers and, 5, 43, 206, 207 importation of, 56 moral status of, 71-82 sharing of, in research, 114-115, 128 students’ attitudes toward, 200 Anticruelty laws, 305-314, 318 APA, 346-347 APHIS, 14, 32-33 administration of the Animal Welfare Act by, 283-289, 291, 293 animal use data and, 5, 29, 30, 31, 50, 53, 54, 57, 58-65, 295-297 criticisms of, 297 Federal Government and, 43-44, 46-49, 293, 388, 393 GLPs and, 293 NSF and, 392 U.S. Surgical Corporation inspections by, 324, 325 Approximate lethal dose (ALD), 182 APS, 347-348 ARC, 390 Aristotle, 74 Arizona, 288, 307, 309, 319, 320 Arkansas, 158, 288, 307, 319 ASPCA, 269, 309 Association of American Medical Colleges. See AAMC Association of American Veterinary Medical Col- leges. See AAVMC Association of Universities and Colleges of Canada, 361 Augustine, Saint, 73 Australia, 17, 324, 359-360 AVMA, 207, 344-345, 350 Barth, Karl, 74 Biosciences Information Service (BIOSIS), 224, 231, 238 Bristol Myers Company, 13, 266 British Columbia, University of, 360 Brown, Alex & Sons, 56 Bureau of Standards, U.S. Department of Com- merce. See National Bureau of Standards (NBS) CAAT, 13 animal use alternatives and, 189 funding for alternatives by, 266-267 private funding of, 264 California, 288, 289, 307 pound release laws in, 319, 320 regulations in, 166, 167, 316-317, 321 RRFs in, 287 California, University of Southern, 346, 351 Calvin, John, 74 Canada, 17, 176, 201-202, 268, 359, 360-362 Canadian Council on Animal Care. See CCAC Canadian Federation of Humane Societies. See CFHS Catholic University, 210 CCAC, 360-362 CDC, 295 animal use by, 9,45 economics of testing and, 251 funding for NTP by, 264-265 guidelines of, 337 regulatory activities by, 158, 165 Cell culture, 118 as alternative, 4, 121-122, 133, 175, 177-179, 183, 184-186, 189-190 and polio vaccine, 91 training in use of, 210 See also In vitro techniques; Living systems Center for Alternatives to Animal Testing. See CAAT Centers for Disease Control, See CDC CERCLA, 163-164 CFHS, 361 Chemical Industry Institute of Toxicology. See CIIT Chemical Times and Trends (CT&T), 287 CIIT computer-based registries and, 238 and data sharing, 176, 252 literature prepared for, 219 Clarke Institute of Psychiatry, 360 Colgate-Palmolive Company, 268 Colorado, 166, 286, 288, 307, 312-313, 316, 319, 320 Colorado State University, 320, 342 Commerce, Department of (U.S.), 44, 277, 297, 386 Comprehensive Environmental Response, Compen- sation, and Liability Act. See CERCLA Computer systems as animal alternative, 7-8, 11-12, 124-126, 136-138, 180-181, 182-183, 185, 211-214, 228-238 FRAME and, 189 NIH funding for, 261-262 policy issues and, 22 in research, 13 Congressional Budget Office (CBO), 280 Connecticut, 288, 307, 315, 316-317, 318, 319, 323-328 Consistency argument, 77-78 Consumer Product Safety Commission. See CPSC Index G 4 3 5 Cornell University, 213, 264, 269 Cosmetic, Toiletry, and Fragrance Association (CTFA), 13, 266, 268 Council of International Organizations of Medical Science (CIOMS), 295, 416-417 CPSC animal use by, 9, 46 regulations in, 157-158, 164, 390-391 and research funding, 23 Culpability, 306, 308, 313 Dalhousie University, 360 Data accumulation, 151, 223-228 animal use and, 5, 8, 49-65, 180-181, 188-189 epidemiologic, 155, 187 by Federal agencies, 43-49, 161-163, 165, 166 patent claims and, 247-248 as a policy issue, 29-31, 43 species choice and, 98 by States, 165-166 Data analysis, 152, 222 of Ames tests, 187 animal use and, 52-65 of computer simulations, 138 economy of animal use and, 245-246 GLPs and, 293 reduction in animal use by, 126-128, 175, 177, 181 Databases, 11-12, 219-228 computer simulation and, 125-126 EPA use of, 163-164 epidemiologic, 124 and information sharing, 224-238 and micro-organism tests, 186 reduction of animal use and, 176, 181 Data sharing, 10, 11-12, 23-26, 220, 221-228 by APHIS, 295-297 computer systems and, 228-238 by FDA, 295-297 LADB and, 233-238 by NIH, 295-297 on-line, 229, 231-238 proprietary interests and, 176, 252-253 Delaware, 288, 307, 319 Denmark, 18, 359, 363-366 Department of Health, Education, and Welfare, U.S. (DHEW), 277, 279 Department of Health and Human Services, U.S. (DHHS), 14, 33, 269, 295 animal use by, 45, 49 animal use guidelines of, 339 animal use regulation by, 388-390 databases and, 163-164 funding by, 157, 264-265 LADB and, 235 Descartes, Rene, 74-75 District of Columbia, 15, 276, 288, 290, 307, 316, 319, 336 DNA technology, 91, 121, 123, 125, 185-186, 188, 261 DOD and animal use, 15, 44-45, 49 animal use regulations in, 16, 26, 158, 386-388 Dodge Foundation, Geraldine R., 13, 264, 269 DOE, 45,” 162, 388 DOT animal use by, 9, 45 animal use regulation and, 158, 164, 297, 386 Dow-Corning, 131 Draize eye irritancy test, 8 alternatives to, 183-184, 259, 266 CPSC modification of, 391 funding for alternatives to, 13, 268 methodology of, 154 and policy options, 21 restrictions on, 27-28 trends in use of, 157 Drug Enforcement Agency, U. S., 324 Duke University, 125 Economics alternatives to animal use and, 13, 189 of animal use, 12, 13, 89, 115, 116, 123, 128, 151, 181, 206, 209, 243-253, 265-266 of the Animal Welfare Act, 278, 279, 281, 287 data sources and, 220, 221, 223 of GLPs, 294 of IACUCS) 342 of LADB, 235 pound release laws and, 319 proprietary interests and, 252-253 regulations and, 251 of research, 245-248, 250-251 and species choice, 99 of testing, 155, 157, 184, 248-251, 252-253 Education alternatives to animal use in, 10-11, 208-214 animal use in, 30, 199-214 animal use regulation in, 321-322 computer simulations in, 211-214 funding of, 268-269 overlap with research, 202 Environmental Protection Agency. See EPA EPA animal use by, 9, 46 and data accumulation, 163-164, 221, 229 funding for toxicological research by, 23, 265 GLPs and, 292-294 and the LD5O test, 19 literature prepared for, 219 — — — —. . 436 c Alternatives to Animal Use in Research, Testing, and Education protocol restriction and, 28 and regulation of animal use, 13, 15, 157, 161-164 and testing economics, 251 testing guidelines and, 152, 248, 384-385 Epidemiology database use in, 124 protocol as alternative to animal use, 163, 181 Ethics of animal use, 6, 71-82, 200, 202 and economics, 247 of embryo use in research, 133 of invertebrate use, 134 Ethics and Values in Science and Technology (EVIST), 262 FAA, 277 FDA, 292-295 animal use by, 9, 45 animal use regulation in, 386, 388-389 baldness prevention and, 92 CPSC contracts with, 390 and data collection, 31, 47-49 and data sharing, 176, 295-297 economics of, 251 funding by, 13, 23, 264-265 GLPs and, 292-293 guidelines for) < 337, 383 hepatitis B vaccine and, 91 and the LD5O test, 19 literature prepared for, 219 product liability testing requirements by, 167 protocol restriction and, 28 regulation of animal use by, 13, 14 regulatory activities by, 157, 158-160, 165 testing guidelines and, 152, 383 and toxicological testing, 150-151 Federation of American Societies for Experimental Biology (FASEB), 235-236, 238, 346, 348 Florida, 286, 288 anticruelty laws in, 307, 308, 314 pesticide program in, 166 pound release laws in, 319, 320 regulation of animal use in, 321-322 RRFs in, 287 FOIA and data sharing, 24-25, 223 GLPs and, 293 regulation of animal use and, 297 unpublished information and, 221-222 USDA and, 290 Food and Drug Administration, U.S. See FDA FRAME, 153-154, 189-190, 267 France, 220 Francis, Saint, 74 Friends of Animals, Inc. v. U.S. Surgical Corpora- tion, 315, 316, 323-328 Freedom of Information Act. See FOIA FTC, 9, 158, 165 Fund for Replacement of Animals in Medical Ex- periments. See FRAME GAO APHIS and, 286 military research and, 293 USDA and, 289-290 Genetics. See DNA technology; RNA replication Georgia, 288, 307, 308, 319 Germany, Federal Republic of, 359, 366-367 Good Laboratory Practices (GLPs), 292-294 Guide for the Care and Use of Laboratory Animals (NIH). See NIH, guidelines by Guidelines, for animal use, 13, 14, 16, 31, 33, 152, 157-167, 176, 200-202, 291, 293-296, 335-352, 361, 383-393, 395-400, 412-417 Harrison, R. G., 120 Harvard University, 115 Hatch, Orrin G., 3 Hawaii, 288, 307, 309, 318, 319 Health Professions Education Assistance Amend- ments of 1985, 13, 269, 291 Health Research Extension Act of 1985, 14, 281, 291-292 Humane treatment, principle of, 6, 78-79 Hume, David, 74-75 IACUC, 15, 337-344 Idaho, 288, 307, 309, 319 ILAR, 5 and data collection, 31, 50, 53-55, 56, 57, 58-59, 202-203 and LADB, 233-234, 236 NSF funding of, 262 Illinois, 288, 307, 319 regulations in, 314, 316-317, 321 RRFs in, 287 Illinois, University of, 267 India, 56 Indiana, 288, 307, 308, 319 Industry data sharing and, 219, 221-222 economics and, 252-253 research funding by, 13, 22 Information centers, 220 Information, unpublished, 219-220, 221-222, 224-228 Insects, 186-187 Index G 4 3 7 Institute for ILAR Institutional IACUC Institutional Interagency Laboratory Animal Resources. See Animal Care and Use Committee. See Review Board. See IRB Research Animal Committee. See IRAC Interest theory, 76-77 Interior, Department of the (U.S.), 45, 49, 285, 386, 390 International Science and Engineering Fair OSEF), 201 Invertebrates as animal alternative, 122-123, 133-135, 177, 179, 209 ethical use of, 134 in toxicity tests, 185 In vitro techniques, 157 as animal alternative, 10, 118-122, 124, 126, 150, 177-179, 182-183, 185, 186, 188, 210 delays in implementing, 189 economics of, 250 funding of, 261, 265 mathematical model of, 267 for patents, 248 and policy options, 22 See also Cell culture; Living systems; Organ cul- ture; Tissue culture In vivo techniques, 120, 157, 184, 186 in education, 210 in research, 117, 124, 188 Iowa, 288, 289, 307, 316-317, 319 IRAC, 16, 295 DOD and, 387 DOE and, 388 guidelines of, 337-339 NASA and, 391 IRBs, 340, 342 Japan, 150 animal use regulation in, 359, 362-363 data sharing by, 223 Johns Hopkins University, The, 13, 23, 57, 264 Kansas, 288, 289, 307, 309, 316-317, 319, 321 Kant, Emmanuel, 76, 81 Kentucky, 288, 307, 319 Laboratory Animal Data Bank (LADB), 11-12, 176, 229, 233-238 Laboratory Animal Welfare Act, 13-14, 16, 46, 49, 275, 276-291, 295 amending of, 16, 26, 32-34 1970 amendments to, 278-279, 281-283 1976 amendments to, 279-283 1985 amendments to, 12, 14, 16, 280-281, 386 animal rights and, 315, 316 and APHIS, 29-30, 31, 283-290, 291-294 CPSC and, 391 criticisms of, 297-298 and data collection, 5 economics of, 278, 279, 281, 287 and Federal agencies, 31, 44, 269, 289-290, 387-392 litigation and, 290-291 research facilities and, 276-277, 278-279, 281-287 State duplication of, 317 Taub case and, 310-312 Labor, Department of (U.S.), 164 LC50 test, 166-167 LD50 test, 8, 80-81 alternatives to, 268, 351 in chemical testing, 151 cost of, 250 CPSC and, 164, 391 data uses, 166-167 in Denmark, 365 FDA and, 388-389 FRAME and, 153 methodology of, 153 modification of, 19, 21, 175, 181-183, 189-190 restrictions on, 28 in science fairs, 201 in Switzerland, 359 trends in use of, 157 Life Sciences Research Office, 235 Limit test as alternative to LD5O test, 182 CPSC and, 391 methodology of, 153 Litigation, 167-168, 175, 247, 253, 290-291, 323-328 Living systems, 7-8, 118-123, 126, 133-136, 177-179, 209-210 See also Cell culture; In vitro techniques; Organ culture; Tissue culture Louisiana, 288, 307, 319 Louisiana State University, 56 Maine, 288, 307, 308, 318, 319 Maryland, 288, 293, 307, 308, 310-312, 319 Maryland, University of, 57 Maryland v. Taub, 310-312, 313, 335 Massachusetts, 15, 287, 288, 307, 313, 316-318, 319, 321-322 Massachusetts Institute of Technology, 260 Mean lethal dose test. See LD50 test Michigan, 287, 288, 307, 314, 316-317, 319, 320 Microorganisms, 123, 177, 179, 186 Microsurgery, 204, 205, 210 Midgeley, Mary, 78 Minnesota, 288, 307, 316, 319 . 438 . Alternatives to Animal Use in Research, Testing, and Education Mississippi, 288, 307, 319 Missouri, 288, 289, 307, 319 Montana, 288, 307, 319 Moore, Marie A., 263 Moral theory, 71-82 Nace, George, 56 NASA animal use by, 46, 115 animal use regulations of, 16, 386, 391-392 species substitution by, 116 use of micro-organisms by, 123 National Agricultural Library (NAL), 12, 21, 24, 238, 281 National Bureau of Standards (NBS), 222, 238 National Cancer Institute. See NCI National Center for Toxicological Research. See NCTR National Institute of Environmental Health Sciences. See NIEHS National Institutes of Health. See NIH National Institute for Occupational Safety and Health. See NIOSH National Library of Medicine. See NLM National Science Foundation. See NSF National Technical Information Service (NTIS), 24, 235 National Toxicology Program. See NTP National Wildlife Health Laboratory, 390 NCI data accumulation and, 11, so epidemiologic studies and, 181 estimations of animal use by, 57-58 funding by, 23, 233, 235, 264-265, 270 guidelines and, 152 historical data and, 176 NCTR AAALAC and, 389 animal use and, 158 data accumulation by, 47-49 FDA contracts with, 390 funding of NTP by, 264-265 and policy options, 21 Nebraska, 288, 307, 319 Netherlands, 359, 367-368 Nevada, 288, 307, 319 New Hampshire, 288, 307, 318, 319 New Jersey, 287, 288, 307, 316, 318, 319, 322, 325, 327, 328 New York, 286, 288, 289 enforcement of anticruelty laws in, 307, 309, 314 Friends of Animals in, 324 hazardous waste regulations in, 166 on-line databases in, 231 pound release laws in, 319, 320 regulation of animal use in, 316-317, 321-322 RRFs in, 287 New York State College of Veterinary Medicine, 4 NIEHS, 157, 264-265 NIH, 13, 15, 295 animal use by, 45, 57-58 animal use regulation in, 15-16, 386, 389-390 animal use survey and, 202-203 and Animal Welfare Act’s enforcement, 33 data accumulation and, 31, 50, 295-297 economics of testing and, 251 funding by, 13, 22-23, 46, 259-265, 269-270 guidelines by, 15-16, 335-337, 345-346, 351, 391, 392 Health Research Extension Act of 1985 and, 291-292 IRAC and, 295 literature prepared for, 219 and policy options, 19, 21 regulation of research by, 317 review of IACUCs by, 341 and Taub case, 310 NIH Guide for the Care and Use of Laboratory Animals. See NIH, guidelines by NIOSH animal use by, 45, 164-166 funding by, 23, 264-265 RTECS and, 229-231, 237 NLM databases of, 11-12, 229 data sharing by, 24, 26, 223, 281 and LADB, 233-235, 237-238 RTECS and, 231 Nonliving systems, 7-8, 124, 136, 180-181, 210-214 North Carolina, 288, 307, 316-317, 319 North Carolina State University, 57 North Dakota, 288, 307, 319 Norway, 359, 368-369 Nozick, Robert, 71 NSF animal use by, 46 animal use regulation in, 392 funding by, 13, 22, 259, 262-263 NTP animal use and, 158 computer-based registries and, 238 and data sharing, 24 funding by, 13, 264-265 funding of, 157 and policy options, 21 and test batteries, 250 unpublished data and, 224 Occupational Safety and Health Administration. See OSHA Index . 4 3 9 OECD data sharing and, 223 FDA and, 388 and policy options, 22 testing guidelines use by, 152, 156, 384, 417 unpublished information and, 228 Ohio, 268, 288 on-line databases in, 231, 233, 235 regulations in, 307, 316-317, 319 RRFs in, 287 Oklahoma, 288, 307, 316-317, 319 Oregon, 166, 288, 307, 319 Organ culture, 119-120, 133, 177, 179 See also In vitro techniques; Living systems Organization for Economic Cooperation and De- velopment. See OECD OSHA, 9, 157, 162, 166 Pain Animal Welfare Act and, 279 definition of, 4-5 funding for relief of, 270 in research animals, 103-105, 117-118, 130-132, 176, 209 Patents, 247-248 Paul, Saint, 73 Pennsylvania regulations in, 307, 308, 310, 316-317, 318, 319, 321 RRFs in, 287, 288 . Pennsylvania, University of, 263 Pharmaceutical Manufacturers’ Association. See PMA Pharmacokinetics, 153, 157, 185, 190 PHS, 13, 15, 26 and data collection, 31 economics of animal use and, 244 guidelines by, 295, 335-339, 340-341, 395-400 Health Research Extension Act of 1985 and, 281, 291 recognition of AAALAC by, 345 VA and, 393 PHS Policy on Humane Care and Use of Laborato- ry Animals. See PHS, guidelines by Plants, as animal alternatives, 123, 133, 135-136 PMA AAALAC and, 344 computer-based registries and, 238 and data sharing, 176 Policy issues and animal use alternatives, 18-34 data accumulation and, 43 options for, 18-34 Policy on Humane Care and Use of Laboratory Animals by Awardee Institutions (PHS). See PHS, guidelines by Pound release laws, 318-320 Primates, 5, 49, 52, 89-93, 132, 151, 243, 291, 294, 351, 412 Primate Research Institute, 114 Product liability and animal use, 157, 167-168, 189 and economics, 249 testing and, 175 Protocols for animal use, 105-108 computer systems and, 138, 228-230 and data quality, 221, 222, 228-230 EPA and, 161, 163 experimental, 149, 156 FDA and, 158-159 IACUCs and, 342 for pain relief, 117-118, 130-132, 176 and product liability, 168 for replacing animals, 8-10, 107, 114-116, 126-128 Public Health Service. See PHS Purdue University, 352 Quantitative structure-activity relationships (QSAR), 180 Quintana, Pierre, 327, 328 Reduction of animal use, 4, 10, 11, 39, 114-116, 126-128, 175-179, 186-187, 188-190, 209 computer systems and, 228 data analysis and, 126-128 definition of, 4 funding for, 263-264, 268, 269-270 LD5O test and, 182 protocols and, 8-10 see also Refinement; Replacement Refinement of animal use, 4, 10, 11, 39, 182, 183, 188-190, 209 definition of, 4 funding for, 263-264, 270 limit test as, 182 Regan, Tom, 77, 81-82 Registered Research Facilities (RRFs), 287, 288, 291 Registry of Toxic Effects of Chemical Substances. See RTECS Regulations animal rights and, 315-316 of animal use, 13-18, 46, 157-167, 201-202, 203, 275-298, 305-322, 335-352, 359-375, 386-393 criticisms of, 297-298 economics and, 251, 252-253 Federal preemption of State, 33-34, 311-313 funding for alternatives and, 264 of nonanimal organisms, 179 440 . Alternatives to Animal Use in Research, Testing, and Education product liability and, 167-168 unpublished data and, 221-222 Regulatory practices Animal Welfare Act and, 278, 279, 280, 283-290 APHIS and, 286-289 effect on animal use of, 12, 157-167 Health Research Extension Act of 1985 and, 291-292 modifications of animal use laws and, 297-298 patents and, 248 testing methods and, 150, 157-167, 175, 181-182, 188-189 Replacement of animals in testing, 4, 10-11, 39, 128-130, 179-181, 182-183, 185-190, 209-214 definition of, 4 funding of, 259-269 plants as, 135-136 in research protocols, 107 Resusci-Dog as a, 4, 213, 269 vertebrates as, 133-135 Research alternatives to animal use in, 6-8, 13, 113-138 animal use in, 3, 89-108 Animal Welfare Act and, 276-291 anticruelty laws’ applicability to, 310-314 benefits to animals from, 102 economics of, 245-248, 250-251 and education, 202 funding of, 22-23, 259-264, 269 Health Research Extension Act of 1985 and, 291-292 IACUCs and, 340-344 pain reduction in animals used in, 117-118, 130-132 pound release laws and, 318-320 species choice in, 94-99, 103 and testing, 149 Restricted necessity, 80-81 Resusci-Dog, 213, 269 Revlon Inc., 265-266 Rhode Island, 288, 307, 316-317, 318, 319 Rickaby, Joseph, 73 RNA replication, 122, 125 Rockefeller University, 13, 23, 189, 265-266, 267 Rowan, Andrew N., 56, 57 RTECS, 229-231, 237 Saunders & Co., W.B., 55 Schweitzer, Albert, 74 Science, 117 Science fairs, 200-202 Singer, Peter, 76-77, 79 Small Business Innovation Research Program, 263 Smith, Kline & French Laboratories, 346, 352 Snell, Inc., Foster D., 55-56 Society for the Study of Reproduction (SSR), 349 South Carolina, 288, 307, 319 South Dakota, 288, 307, 316, 319 “Speciesism,” 6, 79 Superfund, See CERCLA Sweden, 359, 369-371 Switzerland economics of test ban in, 251 information center in, 220 regulations in, 28, 369, 371-372 Taub, Edward, 310, 312 Tennessee, 220, 288, 307, 316, 319, 321 Testing animal use in, 3, 8-10, 149-168, 175-190 data accumulation and, 8 economics of, 248-251, 252-253 funding for alternatives to animals in, 264-268 Government’s role in, 157-167 methods of, 150-157 overlap with research, 149 standardized methods in, 152, 156 Texas, 286, 288, 289, 307, 319 constitutionality of anticruelty laws in, 309 hazardous waste regulations in, 166 RRFs in, 287 Thomas Aquinas, Saint, 73, 81 Tissue culture, 120-122, 133, 210 See also In vitro techniques; Living systems Toxicology Data Bank, 229 Treasury, Department of (U.S.), 277 T-test, 115, 135 Tufts University, 57 United Kingdom, 268 animal use regulation in, 17, 18, 203-204, 359, 373-375 and data collection, 29 licensing animal users in, 28 United Nations Environment Program, 224 Unrestricted necessity, 81 USDA, 5, 33, 46, 53 animal use by, 9, 44 animal use regulation and, 386, 387, 388, 392 and the Animal Welfare Act, 14, 44, 276-281, 283-289, 291, 293-295, 298 and Federal agency use, 43, 289-290 FOIA and, 290 GLPs and, 293 pound release laws and, 318 /ndex G 4 4 7 regulatory activities by, 158, 159, 165 Taub case and, 310, 312 U.S. Surgical Corporation and, 324, 325 See also APHIS U.S. Surgical Corporation, 315, 316, 323-328 Utah, 288, 307, 316, 319, 321 Utilitarian principle, 6, 79-81, 82 Varana, Rudolph, 327 Vermont, 288, 307, 318, 319 Veterans’ Administration animal use by, 46, 49 policy of, 16 regulation in, 386, 392-393 Virginia, 288, 307 constitutionality of anticruelty laws in, 309 regulations in, 306, 314, 316, 319 Virginia, University of, 114 Washington (State), 288, 307, 319 West Virginia, 288, 307, 319 Will theory, 76 Winkler v. Colorado, 312-313 Wisconsin, 288, 307 hazardous waste regulations in, 166 pound release laws in, 319, 320 regulations in, 321 Wisconsin Regional Primate Research Center (WRPRC), 351-532 Wisconsin, University of, 57, 351 World Health Organization (WHO), 176, 219 Wyoming, 288, 307, 319, 321 Zoonoses, 94