Assessment of Environmental DNA Surveys for the Cryptic Salamander Mussel ( Simpsonaias ambigua )

Simpsonaias ambigua (Salamander Mussel), is a small and thin shelled freshwater mussel often found in difficult to survey habitats, such as beneath slab stones, in the crevices of rock walls, or buried within roots of emergent vegetation and in undercuts of banks. The use of environmental DNA (eDNA - genetic material released from urine, waste, mucus, or sloughed cells) sampling may improve detection and assessment of presence / absence for this rare mussel in comparison to visual tactile techniques. This study completed side by side comparisons of traditional mussel searches and eDNA for a direct assessment of mussel detection efficiencies. Surveying was conducted in several waterbodies of different habitat characteristics with varying abundances of S. ambigua . Additionally, a broad assessment of S. ambigua presence was conducted throughout the proposed critical habitat reach of the Blanchard River in northwest Ohio, to assess if the species remained extant. All eDNA samples were also assessed for the presence of Necturus maculosus (Mudpuppy), the obligate host species for S. ambigua . The eDNA sampling successfully detected S. ambigua from multiple sites and watersheds where it was found with visual surveys. In some cases, eDNA detections occurred in locations where over 16 hours of search yielded only a single individual or fresh dead shells, supporting the sensitivity of eDNA for detection of rare species. Furthermore, probability of detection analysis suggests eDNA sampling can provide high detection efficiency with relatively low effort in comparison to visual searches. The development and validation of an eDNA protocol for the simultaneous detection of S. ambigua and its host salamander increases survey efficiency, reduces field costs, and can support future conservation efforts for listing drainages of extant populations and monitoring conservation goals.