Development and assessment of an environmental DNA (eDNA) assay for a cryptic Siren (Amphibia: Sirenidae)

Environmental DNA (eDNA) assays have become a major aspect of surveys for aquatic organisms in the past decade. These methods are highly sensitive, making them well-suited for monitoring rare and cryptic species. Current efforts to study the Rio Grande Siren in southern Texas have been hampered due to the cryptic nature of these aquatic salamanders. Arid conditions further add to the difficulty in studying this species, as many water bodies they inhabit are ephemeral, sometimes constraining sampling efforts to a short window after heavy rain. Additionally, sirens are known to cease activity and reside underground when ponds begin to dry or as water temperatures increase. Conventional sampling efforts require extensive trap-hours to be effective, which is not always possible within the required sampling window. This study presents the development of a novel eDNA assay technique for this elusive species using conventional PCR and Sanger sequencing and compares eDNA sampling results with simultaneous trapping at multiple sites to assess the relative effectiveness of the procedure. Rio Grande Siren detection via eDNA sampling was significantly higher at all sites compared to trapping, confirming the utility of this assay for species detection. This methodology gives promise for future work assessing the distribution and status of the Rio Grande Siren and has potential for use on other southern Texas amphibians.