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      Temperature-Dependent Growth of Geomyces destructans, the Fungus That Causes Bat White-Nose Syndrome

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          Abstract

          White-nose syndrome (WNS) is an emergent disease estimated to have killed over five million North American bats. Caused by the psychrophilic fungus Geomyces destructans, WNS specifically affects bats during hibernation. We describe temperature-dependent growth performance and morphology for six independent isolates of G. destructans from North America and Europe. Thermal performance curves for all isolates displayed an intermediate peak with rapid decline in performance above the peak. Optimal temperatures for growth were between 12.5 and 15.8°C, and the upper critical temperature for growth was between 19.0 and 19.8°C. Growth rates varied across isolates, irrespective of geographic origin, and above 12°C all isolates displayed atypical morphology that may have implications for proliferation of the fungus. This study demonstrates that small variations in temperature, consistent with those inherent of bat hibernacula, affect growth performance and physiology of G. destructans, which may influence temperature-dependent progression and severity of WNS in wild bats.

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          Most cited references14

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          Overlapping confidence intervals or standard error intervals: What do they mean in terms of statistical significance?

          We investigate the procedure of checking for overlap between confidence intervals or standard error intervals to draw conclusions regarding hypotheses about differences between population parameters. Mathematical expressions and algebraic manipulations are given, and computer simulations are performed to assess the usefulness of confidence and standard error intervals in this manner. We make recommendations for their use in situations in which standard tests of hypotheses do not exist. An example is given that tests this methodology for comparing effective dose levels in independent probit regressions, an application that is also pertinent to derivations of LC50s for insect pathogens and of detectability half-lives for prey proteins or DNA sequences in predator gut analysis.
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            Physiology of Batrachochytrium dendrobatidis, a chytrid pathogen of amphibians.

            Batrachochytrium dendrobatidis is a pathogen of amphibians that has been implicated in severe population declines on several continents. We investigated the zoospore activity, physiology and protease production of B. dendrobatidis to help understand the epidemiology of this pathogen. More than 95% of zoospores stopped moving within 24 h and swam less than 2 cm before encysting. Isolates of B. dendrobatidis grew and reproduced at temperatures of 4-25 C and at pH 4-8. Growth was maximal at 17-25 C and at pH 6-7. Exposure of cultures to 30 C for 8 d killed 50% of the replicates. B. dendrobatidis cultures grew on autoclaved snakeskin and 1% keratin agar, but they grew best in tryptone or peptonized milk and did not require additional sugars when grown in tryptone. B. dendrobatidis produced extracellular proteases that degraded casein and gelatin but had no measurable activity against keratin azure. The proteases were active against azocasein at temperatures of 6-37 C and in a pH range of 6-8, with the highest activity at temperatures of 23-30 C and at pH 8. The implications of these observations on disease transmission and development are discussed.
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              Pan-European Distribution of White-Nose Syndrome Fungus (Geomyces destructans) Not Associated with Mass Mortality

              Background The dramatic mass mortalities amongst hibernating bats in Northeastern America caused by “white nose-syndrome” (WNS) continue to threaten populations of different bat species. The cold-loving fungus, Geomyces destructans, is the most likely causative agent leading to extensive destruction of the skin, particularly the wing membranes. Recent investigations in Europe confirmed the presence of the fungus G. destructans without associated mass mortality in hibernating bats in six countries but its distribution remains poorly known. Methodology/Principal Findings We collected data on the presence of bats with white fungal growth in 12 countries in Europe between 2003 and 2010 and conducted morphological and genetic analysis to confirm the identity of the fungus as Geomyces destructans. Our results demonstrate the presence of the fungus in eight countries spanning over 2000 km from West to East and provide compelling photographic evidence for its presence in another four countries including Romania, and Turkey. Furthermore, matching prevalence data of a hibernaculum monitored over two consecutive years with data from across Europe show that the temporal occurrence of the fungus, which first becomes visible around February, peaks in March but can still be seen in some torpid bats in May or June, is strikingly similar throughout Europe. Finally, we isolated and cultured G. destructans from a cave wall adjacent to a bat with fungal growth. Conclusions/Significance G. destructans is widely found over large areas of the European continent without associated mass mortalities in bats, suggesting that the fungus is native to Europe. The characterisation of the temporal variation in G. destructans growth on bats provides reference data for studying the spatio-temporal dynamic of the fungus. Finally, the presence of G. destructans spores on cave walls suggests that hibernacula could act as passive vectors and/or reservoirs for G. destructans and therefore, might play an important role in the transmission process.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                28 September 2012
                : 7
                : 9
                : e46280
                Affiliations
                [1 ]Department of Pathobiological Sciences, University of Wisconsin, Madison, Wisconsin, United States of America
                [2 ]Department of Ecology and Evolutionary Biology, University of Tennessee, Knoxville, Tennessee, United States of America
                [3 ]Department of Computer Science, University of Chicago, Chicago, Illinois, United States of America
                [4 ]Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany
                [5 ]US Geological Survey – National Wildlife Health Center, Madison, Wisconsin, United States of America
                Imperial College Faculty of Medicine, United Kingdom
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: MLV JGB DSB. Performed the experiments: MLV DSB. Analyzed the data: JGB WW MLV. Contributed reagents/materials/analysis tools: GW. Wrote the paper: MLV JGB DSB.

                [¤]

                Current address: Cooperative Wildlife Research Laboratory, Department of Zoology, Southern Illinois University, Carbondale, Illinois, United States of America

                Article
                PONE-D-12-16131
                10.1371/journal.pone.0046280
                3460873
                23029462
                9297f835-9b7e-4c0c-a22c-ab9a78a31114
                Copyright @ 2012

                This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

                History
                : 4 June 2012
                : 28 August 2012
                Page count
                Pages: 7
                Funding
                Financial support for this project was provided by the US Geological Survey, the US Fish and Wildlife Service, and Bat Conservation International. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology
                Microbiology
                Mycology
                Fungi
                Emerging Infectious Diseases
                Host-Pathogen Interaction
                Medical Microbiology
                Microbial Ecology
                Microbial Pathogens
                Microbial Physiology
                Veterinary Science
                Animal Types
                Wildlife
                Veterinary Diseases
                Veterinary Mycology
                Veterinary Microbiology

                Uncategorized
                Uncategorized

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