Ophidiomycosis (snake fungal disease; SFD) is a disease of conservation concern caused by the fungus Ophidiomyces ophidiicola that threatens the health of snake populations worldwide. Gaps exist in our knowledge about the prevalence of this disease across landscapes. In our study, we compared the prevalence of ophidiomycosis between a low-impacted forest site (n = 93) and a highly disturbed remediated landfill (n = 53) in Anderson County, Tennessee, USA. Free-ranging snakes were examined for the presence of skin lesions that are consistent with ophidiomycosis and were swabbed to detect O. ophidiicola DNA using quantitative PCR (qPCR). Apparent ophidiomycosis (qPCR-positive and skin lesions present) was diagnosed at both sites, but there was no significant difference in prevalence between the two sites (24.7% at the forest site; 22.6% at the landfill site). Apparent ophidiomycosis was most prevalent in Racers (38%; Coluber constrictor) and Ring-Necked Snakes (26%; Diadophis punctatus). There was no difference in ophidiomycosis status between sites for the most-sampled species: Racers, Black Rat Snakes (Pantherophis obsoletus), and Ring-necked Snakes (Diadophis punctatus). Our study represents the first report of ophidiomycosis at focal sites in Tennessee. The findings suggest that O. ophidiicola may be ubiquitous across the landscape and point to the need for further study of diverse habitat types for the prevalence of O. ophidiicola.