Ophidiomyces ophidiicola is an emerging fungal pathogen associated with infections in snakes across North America. Although documented in Pennsylvania, O. ophidiicola has not been found at Powdermill Nature Reserve (PNR) in southwestern Pennsylvania, where the snake assemblage has been studied since 2002 and several species have recently declined. We surveyed for O. ophidiicola and putative ophidiomycosis at PNR. We screened five species of free-ranging, wild snakes (n=34) for suspected ophidiomycosis by visually checking for dermatitis and swabbing for the presence of O. ophidiicola DNA. We found a moderate prevalence of snakes with skin lesions (n=15) but a low prevalence of snakes with O. ophidiicola DNA in traditional PCR assays (n=2). Both positive snakes belonged to the same species and only one presented with lesions. When quantitative PCR screens were performed on duplicate swabs, 19 snakes were positive for O. ophidiicola DNA, with positive individuals in two species. Mark-recapture methods revealed seasonal variability in disease dynamics for sampled snakes. One individual presented with less than five skin lesions and tested negative in May 2020, had more than five lesions with a high fungal DNA load in June 2020, and no lesions with a low fungal DNA load in July 2020. We also found that snakes sampled from under the same cover object at the same time either all tested positive or all negative, including one instance involving two species. Our results underscore the value of using multiple screening techniques for O. ophidiicola surveillance and repeated sampling of individuals to understand the dynamics of ophidiomycosis in wild populations as compared to single method and single timepoint approaches.

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