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Freshwater species are declining rapidly but more complete data are needed for determining the extent and cause(s) of population declines and extirpations. Integrating newer survey techniques, freely available data, and traditional field work may allow for more effective assessment of population decline. We used detailed historical species records and environmental DNA (eDNA) survey methods to identify changes in population distribution of a long‐lived, imperiled stream salamander, the eastern hellbender (Cryptobranchus alleganiensis alleganiensis: Cryptobranchidae). We used logistic regression with Bayesian inference to test whether selected environmental variables may be good predictors of hellbender population persistence and extirpation. Hellbenders persisted in only 42% of the 24 historical record sites. The best fit model indicated electrical conductivity (EC) was the strongest predictor of hellbender population persistence (EC < 278 μS/cm) and extirpation. Conductivity was strongly negatively correlated with canopy cover within the total watershed (r = −0.83, n = 21, p < 0.001) and riparian buffer of the watershed (r = −0.77, n = 21, p < 0.001). Electrical conductivity tends to increase following deforestation, and may inhibit sperm motility and thus limit recruitment of hellbenders and other aquatic vertebrate species with external fertilisation. By integrating historical data, eDNA, field data, and freely available high resolution remote sensing data, our study design allowed for rapid assessment of predictors of and changes in hellbender distribution over a relatively broad geographic area. This cost‐ and time‐effective approach may be used for evaluating other rare aquatic species.

Investigation into the causes underlying the rapid, global amphibian decline provides critical insight into the effects of changing ecosystems. Hypothesized and confirmed links between amphibian declines, disease, and environmental changes are increasingly represented in published literature. However, there are few long-term amphibian studies that include data on population size, abnormality/injury rates, disease, and habitat variables to adequately assess changes through time. We cultured and identified microorganisms isolated from abnormal/injured and repressed tissue regeneration sites of the endangered Ozark Hellbender, Cryptobranchus alleganiensis bishopi, to discover potential causative agents responsible for their significant decline in health and population. This organism and our study site were chosen because the population and habitat of C. a. bishopi have been intensively studied from 1969–2009, and the abnormality/injury rate and apparent lack of regeneration were established. Although many bacterial and fungal isolates recovered were common environmental organisms, several opportunistic pathogens were identified in association with only the injured tissues of C.a. bishopi. Bacterial isolates included Aeromonas hydrophila, a known amphibian pathogen, Granulicetella adiacens, Gordonai terrae, Stenotrophomonas maltophilia, Aerococcus viridans, Streptococcus pneumoniae and a variety of Pseudomonads, including Pseudomonas aeruginosa, P. stutzeri, and P. alcaligenes. Fungal isolates included species in the genera Penicillium, Acremonium, Cladosporium, Curvularia, Fusarium, Streptomycetes, and the Class Hyphomycetes. Many of the opportunistic pathogens identified are known to form biofilms. Lack of isolation of the same organism from all wounds suggests that the etiological agent responsible for the damage to C. a. bishopi may not be a single organism. To our knowledge, this is the first study to profile the external microbial consortia cultured from a Cryptobranchid salamander. The incidence of abnormalities/injury and retarded regeneration in C. a. bishopi may have many contributing factors including disease and habitat degradation. Results from this study may provide insight into other amphibian population declines.

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