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      Heterogeneous Occupancy and Density Estimates of the Pathogenic Fungus Batrachochytrium dendrobatidis in Waters of North America

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          Abstract

          Biodiversity losses are occurring worldwide due to a combination of stressors. For example, by one estimate, 40% of amphibian species are vulnerable to extinction, and disease is one threat to amphibian populations. The emerging infectious disease chytridiomycosis, caused by the aquatic fungus Batrachochytrium dendrobatidis ( Bd), is a contributor to amphibian declines worldwide. Bd research has focused on the dynamics of the pathogen in its amphibian hosts, with little emphasis on investigating the dynamics of free-living Bd. Therefore, we investigated patterns of Bd occupancy and density in amphibian habitats using occupancy models, powerful tools for estimating site occupancy and detection probability. Occupancy models have been used to investigate diseases where the focus was on pathogen occurrence in the host. We applied occupancy models to investigate free-living Bd in North American surface waters to determine Bd seasonality, relationships between Bd site occupancy and habitat attributes, and probability of detection from water samples as a function of the number of samples, sample volume, and water quality. We also report on the temporal patterns of Bd density from a 4-year case study of a Bd-positive wetland. We provide evidence that Bd occurs in the environment year-round. Bd exhibited temporal and spatial heterogeneity in density, but did not exhibit seasonality in occupancy. Bd was detected in all months, typically at less than 100 zoospores L −1. The highest density observed was ∼3 million zoospores L −1. We detected Bd in 47% of sites sampled, but estimated that Bd occupied 61% of sites, highlighting the importance of accounting for imperfect detection. When Bd was present, there was a 95% chance of detecting it with four samples of 600 ml of water or five samples of 60 mL. Our findings provide important baseline information to advance the study of Bd disease ecology, and advance our understanding of amphibian exposure to free-living Bd in aquatic habitats over time.

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          Global emergence of Batrachochytrium dendrobatidis and amphibian chytridiomycosis in space, time, and host.

          Batrachochytrium dendrobatidis (Bd) is a chytrid fungus that causes chytridiomycosis in amphibians. Only named in 1999, Bd is a proximate driver of declines in global amphibian biodiversity. The pathogen infects over 350 species of amphibians and is found on all continents except Antarctica. However, the processes that have led to the global distribution of Bd and the occurrence of chytridiomycosis remain unclear. This review explores the molecular, epidemiological, and ecological evidence that Bd evolved from an endemic ancestral lineage to achieve global prominence via anthropogenically mediated spread. We then consider the major host and pathogen factors that have led to the occurrence of chytridiomycosis in amphibian species, populations, and communities.
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            Molecular Detection of Vertebrates in Stream Water: A Demonstration Using Rocky Mountain Tailed Frogs and Idaho Giant Salamanders

            Stream ecosystems harbor many secretive and imperiled species, and studies of vertebrates in these systems face the challenges of relatively low detection rates and high costs. Environmental DNA (eDNA) has recently been confirmed as a sensitive and efficient tool for documenting aquatic vertebrates in wetlands and in a large river and canal system. However, it was unclear whether this tool could be used to detect low-density vertebrates in fast-moving streams where shed cells may travel rapidly away from their source. To evaluate the potential utility of eDNA techniques in stream systems, we designed targeted primers to amplify a short, species-specific DNA fragment for two secretive stream amphibian species in the northwestern region of the United States (Rocky Mountain tailed frogs, Ascaphus montanus, and Idaho giant salamanders, Dicamptodon aterrimus). We tested three DNA extraction and five PCR protocols to determine whether we could detect eDNA of these species in filtered water samples from five streams with varying densities of these species in central Idaho, USA. We successfully amplified and sequenced the targeted DNA regions for both species from stream water filter samples. We detected Idaho giant salamanders in all samples and Rocky Mountain tailed frogs in four of five streams and found some indication that these species are more difficult to detect using eDNA in early spring than in early fall. While the sensitivity of this method across taxa remains to be determined, the use of eDNA could revolutionize surveys for rare and invasive stream species. With this study, the utility of eDNA techniques for detecting aquatic vertebrates has been demonstrated across the majority of freshwater systems, setting the stage for an innovative transformation in approaches for aquatic research.
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              Improving occupancy estimation when two types of observational error occur: non-detection and species misidentification.

              Efforts to draw inferences about species occurrence frequently account for false negatives, the common situation when individuals of a species are not detected even when a site is occupied. However, recent studies suggest the need to also deal with false positives, which occur when species are misidentified so that a species is recorded as detected when a site is unoccupied. Bias in estimators of occupancy, colonization, and extinction can be severe when false positives occur. Accordingly, we propose models that simultaneously account for both types of error. Our approach can be used to improve estimates of occupancy for study designs where a subset of detections is of a type or method for which false positives can be assumed to not occur. We illustrate properties of the estimators with simulations and data for three species of frogs. We show that models that account for possible misidentification have greater support (lower AIC for two species) and can yield substantially different occupancy estimates than those that do not. When the potential for misidentification exists, researchers should consider analytical techniques that can account for this source of error, such as those presented here.
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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
                2014
                15 September 2014
                : 9
                : 9
                : e106790
                Affiliations
                [1 ]Oregon State University, Environmental Science Graduate Program, Corvallis, Oregon, United States of America
                [2 ]US Geological Survey, Oregon Water Science Center, Portland, Oregon, United States of America
                [3 ]Biological Sciences, University of Southern California, Los Angeles, California, United States of America
                [4 ]Oregon State University, Department of Integrative Biology, Corvallis, Oregon, United States of America
                [5 ]Astrobiology Program, National Aeronautics and Space Administration Headquarters, Washington DC, United States of America
                [6 ]US Forest Service, Pacific Northwest Research Station, Corvallis, Oregon, United States of America
                [7 ]US Geological Survey, National Research Program, Reston, Virginia, United States of America
                Universität Zurich, Switzerland
                Author notes

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

                Conceived and designed the experiments: TC CWA MAV JK. Performed the experiments: TC CWA MAV JK. Analyzed the data: TC. Contributed reagents/materials/analysis tools: TC CWA RP DHO MAV ARB JK. Wrote the paper: TC CWA RP DHO MAV ARB JK.

                Article
                PONE-D-14-10225
                10.1371/journal.pone.0106790
                4164359
                25222122
                f1ac8aed-6796-4ffe-afb3-ff0fc4888c47
                Copyright @ 2014

                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
                : 17 March 2014
                : 6 August 2014
                Page count
                Pages: 11
                Funding
                Support was provided by the National Science Foundation GK-12 Graduate STEM Fellows in K-12 Education Program (Award #0948041) to TC, US Forest Service – Pacific Northwest Research Station and US Geological Survey Amphibian Research and Monitoring Initiative (ARMI). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Biogeography
                Ecology
                Microbial Ecology
                Mycology
                Organisms
                Earth Sciences
                Marine and Aquatic Sciences
                Aquatic Environments
                Freshwater Environments
                Ecology and Environmental Sciences
                Conservation Science
                Research and Analysis Methods
                Mathematical and Statistical Techniques
                Probability Estimation
                Custom metadata
                The authors confirm that all data underlying the findings are fully available without restriction. Data are available from: USGS Multitaxa Database, http://www.werc.usgs.gov/ProductDetails.aspx?ID=4323.

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                Uncategorized

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