Aquatic <scp>eDNA</scp> can advance monitoring of a small‐bodied terrestrial salamander and amphibian pathogen

Successful conservation efforts for threatened species depend on accurate characterization of their distribution, habitat use, and threats. However, surveillance can be challenging with small size, cryptic coloring, or elusive behavior. Environmental DNA (eDNA) monitoring provide a sensitive noninvasive alternative to traditional techniques by detecting trace shed target into environment. We evaluated the efficacy eDNA four-toed salamander (Hemidactylium scutatum), terrestrial throughout much its range. Additionally, we integrated ranavirus, widespread pathogen ectothermic vertebrates, efficiently evaluate potential disease threat H. scutatum. designed novel species-specific quantitative PCR (qPCR) assay scutatum, multiplexed previously developed ranavirus qPCR assay, validated tests in silico, vitro, situ. collected aquatic weekly 8 weeks from pools historical reports scutatum (n = 6) contemporary associated uplands 4). identified at low quantities (< 12 copies/reaction) multiple sampling days two historic presence. Ranavirus was widespread; nine tested positive including where detected. increased, peaked, then decreased (i.e., were not sporadic time) several but never rose abundance levels ranavirosis other studies. Our results suggest that is distributed sporadically pools; recommend future prioritize increased frequency quantity sample collection per site visit include repeat surveys candidate sites. Further work characterize movement, temporal trends, relationship infectious virion, decay needed help advance this tool.