Browsing by Author "Werne, Josef P."

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    Characterization of diverse bacteriohopanepolyols in a permanently stratified, hyper-euxinic lake
    (Elsevier, 2022-06) O'Beirne, Molly D.; Sparkes, Robert; Hamilton, Trinity L.; van Dongen, Bart E.; Gilhooly, William P., III; Werne, Josef P.; Earth Science, School of Science
    Bacteriohopanepolyols (BHPs) are a diverse class of bacterial lipids that hold promise as biomarkers of specific microbes, microbial processes, and environmental conditions. BHPs have been characterized in a variety of terrestrial and aquatic environments, but less is known about their distribution and abundance in extreme environmental systems. In the present study, samples taken from the water column and upper sediments of the hyper-euxinic, meromictic Mahoney Lake (Canada) were analyzed for BHPs. Analyses show distinct BHP distributions within the oxic mixolimnion, the chemocline, and the euxinic monimolimnion. Bacteriohopanetetrol (BHT) and unsaturated BHT are the dominant BHPs found in the oxic mixolimnion and at the chemocline, whereas a novel BHP (tentatively identified as diunsaturated aminotriol) dominates the euxinic monimolimnion. Along with the novel BHP structure, composite BHPs (i.e., BHT-cyclitol ether and BHT-glucosamine) were observed in the euxinic monimolimnion and sediments, indicating their production by anaerobic bacteria. Complementary metagenomic analysis of genes involved in BHP biosynthesis (i.e., shc, hpnH, hpnO, hpnP, and hpnR) further revealed that BHPs in Mahoney Lake are most likely produced by bacteria belonging to Deltaproteobacteria, Chloroflexi, Planctomycetia, and Verrucomicrobia. The combined observations of BHP distribution and metagenomic analyses additionally indicate that 2- and 3-methyl BHTs are produced within the euxinic sediments in response to low oxygen and high osmotic concentrations, as opposed to being diagnostic biomarkers of cyanobacteria and aerobic metabolisms.
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    The use of dithiothreitol for the quantitative analysis of elemental sulfur concentrations and isotopes in environmental samples
    (Elsevier, 2018) Kurek, Martin R.; Gilhooly, William P., III; Druschel, Gregory K.; O'Beirne, Molly D.; Werne, Josef P.; Earth Science, School of Science
    Determining the concentration and isotopic composition of elemental sulfur in modern and ancient environments is essential to improved interpretation of the mechanisms and pathways of sulfur utilization in biogeochemical cycles. Elemental sulfur can be extracted from sediment or water samples and quantified by converting to hydrogen sulfide. Alternatively, elemental sulfur concentrations can themselves be analyzed using HPLC and other methodologies; however, the preparation and analysis times can be long and these methods are not amenable to stable isotopic analysis. Current reduction methods involve the use of costly and specialized glassware in addition to toxins such as chromium chloride or cyanide to reduce the sulfur to hydrogen sulfide. The novel reduction method presented here uses dithiothreitol (DTT) as a less toxic reducing agent to obtain both elemental sulfur concentrations and isotopic composition from the same sample. The sample is dissolved in an aqueous or organic liquid medium and upon reaction with DTT, the elemental sulfur is volatilized as hydrogen sulfide and collected in a sulfide trap using an inexpensive gas extraction apparatus. The evolved sulfide concentrations can easily be measured for concentration, by absorbance spectrophotometery or voltammetry techniques, and then analyzed for sulfur isotopic composition. The procedure is quantitative at >93% recovery to dissolved elemental sulfur with no observed sulfur isotope fractionation during reduction and recovery. Controlled experiments also demonstrate that DTT is not reactive to sulfate, sulfite, pyrite, or organic sulfur.