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Item Bias in Rating of Rodent Distress during Anesthesia Induction for Anesthesia Compared with Euthanasia(2018-03) Baker, Brittany A.; Hickman, Debra L.; Department of Medicine, School of MedicineSelection of an appropriate method of euthanasia involves balancing the wellbeing of the animal during the procedure with the intended use of the animal after death and the physical and psychologic safety of the observer or operator. The recommended practices for anesthesia as compared with euthanasia are very disparate, despite the fact that all chemical methods of euthanasia are anesthetic overdoses. To explain this disparity, this study sought to determine whether perception bias is inherent in the discussion of euthanasia compared with anesthesia. In this study, participants viewed videorecordings of the anesthesia of either 4 rats or 4 mice, from induction to loss of consciousness. Half of the participants were told that they were observing anesthesia; the other half understood that they were observing euthanasia. Participants were asked to rate the distress of the animals by scoring escape behaviors, fear behaviors, respiratory distress, and other distress markers. For mice, the participants generally rated the distress as high when they were told that the mouse was being euthanized, as compared with the participants who were told that the mouse was being anesthetized. For rats, the effect was not as strong, and the distress was generally rated higher when participants were told they were watching anesthesia. Because the interpretation of distress showed bias in both species—even though the bias differed regarding the procedure that interpreted as distressing—this study demonstrates that laboratory animal professionals must consider the influence of potential perception bias when developing policies for euthanasia and anesthesia.Item Evaluation of Low versus High Volume per Minute Displacement CO₂ Methods of Euthanasia in the Induction and Duration of Panic-Associated Behavior and Physiology(MDPI, 2016-08) Hickman, Debra L.; Fitz, Stephanie D.; Bernabe, Cristian S.; Caliman, Izabela F.; Haulcomb, Melissa M.; Federici, Lauren M.; Shekhar, Anatatha; Johnson, Philip L.; Department of Cellular & Integrative Physiology, IU School of MedicineCurrent recommendations for the use of CO ₂ as a euthanasia agent for rats require the use of gradual fill protocols (such as 10% to 30% volume displacement per minute) in order to render the animal insensible prior to exposure to levels of CO ₂ that are associated with pain. However, exposing rats to CO ₂ , concentrations as low as 7% CO ₂ are reported to cause distress and 10%-20% CO ₂ induces panic-associated behavior and physiology, but loss of consciousness does not occur until CO ₂ concentrations are at least 40%. This suggests that the use of the currently recommended low flow volume per minute displacement rates create a situation where rats are exposed to concentrations of CO ₂ that induce anxiety, panic, and distress for prolonged periods of time. This study first characterized the response of male rats exposed to normoxic 20% CO ₂ for a prolonged period of time as compared to room air controls. It demonstrated that rats exposed to this experimental condition displayed clinical signs consistent with significantly increased panic-associated behavior and physiology during CO ₂ exposure. When atmospheric air was then again delivered, there was a robust increase in respiration rate that coincided with rats moving to the air intake. The rats exposed to CO ₂ also displayed behaviors consistent with increased anxiety in the behavioral testing that followed the exposure. Next, this study assessed the behavioral and physiologic responses of rats that were euthanized with 100% CO ₂ infused at 10%, 30%, or 100% volume per minute displacement rates. Analysis of the concentrations of CO ₂ and oxygen in the euthanasia chamber and the behavioral responses of the rats suggest that the use of the very low flow volume per minute displacement rate (10%) may prolong the duration of panicogenic ranges of ambient CO ₂ , while the use of the higher flow volume per minute displacement rate (100%) increases agitation. Therefore, of the volume displacement per minute rates evaluated, this study suggests that 30% minimizes the potential pain and distress experienced by the animal.Item Indiana State Board of Health. Monthly Bulletin, 1907 Vol. 9 No. 8(8/1/1907) Farrand, Livingston, 1867-1939; Rucker Jr., J. B.; Knopf, S. Adolphus (Sigard Adolphus), 1857-1940; Barnard, H. E.; Fisher, IrvingItem Isoflurane and Carbon Dioxide Elicit Similar Behavioral Responses in Rats(MDPI, 2020-08-16) Kulkarni, Satyajit; Hickman, Debra; Laboratory Animal Resource Center, IU School of MedicineEuthanasia in rodents is an ongoing topic of debate due to concerns regarding the aversive nature of gases with anesthetic properties such as carbon dioxide (CO2) and isoflurane. The aim of this study was to expand upon previously published work evaluating the aversiveness of CO2 by introducing an isoflurane treatment group in parallel. Aversion was tested using a forced exposure setup and an aversion-avoidance setup. In the first part of the study, 12 naïve female Sprague–Dawley rats were exposed during four consecutive days, once to each of four treatments: isoflurane, fox urine, oxygen, and CO2. In the second part of the study, 24 naïve female Sprague–Dawley rats and 12 rats from the first experiment were exposed to CO2, isoflurane, or both gases. In the forced exposure study, there were no significant differences between CO2 and isoflurane treatments except in line crosses. Overall, rats were more active in the isoflurane and CO2 treatments compared to the control groups, suggesting that isoflurane and CO2 are similarly aversive. In the aversion-avoidance study, rats previously exposed to isoflurane left the dark chamber significantly earlier compared to naïve rats during exposure to isoflurane. We also show that learned aversion to isoflurane is sustained for at least 15 days after initial exposure. Given this result, we suggest that CO2 is superior to isoflurane when euthanizing rodents with prior exposure to isoflurane. Overall, these results confirm previous studies which suggest that care should be taken when considering the serial use of isoflurane as an anesthetic.Item Minimal Exposure Times for Irreversible Euthanasia with Carbon Dioxide in Mice and Rats(American Association for Laboratory Animal Science, 2022) Hickman, Debra L.; Laboratory Animal Resource Center, School of MedicineWhen using an anesthetic overdose to euthanize laboratory rodents, a secondary method of euthanasia is recommended to ensure that the apparent death is irreversible. This secondary method usually is accomplished through the collection of tissues that are required to complete the research project. However, frequently laboratory rodents must be euthanized because they cannot be used for studies; in these cases, caretakers must perform a secondary method of euthanasia. Performing physical methods of euthanasia, even on unconscious rodents, can contribute to compassion fatigue in these persons. The current study was designed based on existing literature regarding minimal exposure times for preweanling rats and mice euthanized with carbon dioxide. The study evaluated the minimal time that adult rats and mice must remain in 100% carbon dioxide for death to be irreversible on removal. Adult rats (14 stocks and strains) and mice (more than 40 stocks and strains) were euthanized using a 50% volume per minute displacement rate of carbon dioxide for 2 min. The cages were then left undisturbed for predetermined times, ranging from 0 to almost 12 min. Upon removal from the cage, the animals were stimulated to determine whether they could be resuscitated. If an animal recovered, it was euthanized by using a physical method of euthanasia, and a duration that was 30 s longer than the previous predetermined time was assessed using other animals. The study demonstrated that exposure times of at least 3 min in carbon dioxide reliably result in irreversible euthanasia of mice but that exposure times of at least 10.5 min in carbon dioxide were required to ensure irreversible euthanasia of rats. Although an irreversible death can be attained with carbon dioxide, the use of appropriate species-specific exposure times is critical.Item Self-determination and Care at the End of Life Opinion(2006-10-13T17:02:02Z) German National Ethics Council; BioethicsDying is an individual process which, as such, cannot be removed from the sphere of the individual's self-determination. At the same time, however, it involves a range of different ethical obligations, legal demands and religious expectations, which call for thorough discussion and evaluation in all their aspects. The National Ethics Council has intensively discussed the issues involved in dealing responsibly with dying. It has perused a large volume of material, obtained expert opinions, consulted with doctors and other medical specialists, and held meetings in Augsburg and Munster at which it exposed itself to public debate. The outcome is enshrined in the Opinion now presented. Self-determination and care at the end of life continues the examination of the themes addressed in the Opinion The advance directive published in June 2005. The present analysis, in conjunction with the clarification of terminology here proposed, may facilitate interpretation of the recommendations set out in that Opinion.Item Wellbeing of Alcohol-preferring Rats Euthanized with Carbon Dioxide at Very Low and Low Volume Displacement Rates(American Association for Laboratory Animal Science, 2019-01-01) Hickman, Debra L.; Laboratory Animal Resource Center, IU School of MedicineThe 2013 AVMA Guidelines on Euthanasia recommend the use of very-low or low flow rates of 100% carbon dioxide to euthanize small rodents. Although inhalation of high concentrations of carbon dioxide are generally recognized as painful in humans, whether the use of these low-flow methods of euthanasia increase potential distress for rats is unclear. This study compared physiologic and behavioral markers of animal wellbeing for rats euthanized by using 10% volume displacement per minute (VD/min), 30% VD/min, and 70% VD/min of 100% carbon dioxide. Rats were recorded during euthanasia for subsequent behavioral scoring, and blood samples were taken after euthanasia for assessment of blood glucose and serum corticosterone levels. In this study, rats euthanized with 10% or 30% VD/min of 100% carbon dioxide demonstrated increases in various behaviors, such as rearing and standing, concurrent with increases in serum corticosterone. Rats euthanized with 70% VD/min of 100% carbon dioxide did not exhibit these changes. The results suggest that a euthanasia method of 70% VD/min of 100% carbon dioxide may minimize potential pain and distress and thus be more humane for rats, as compared with very-low- and low-flow methods of carbon dioxide euthanasia.