Detection of four imperiled western North American freshwater mussel species from environmental DNA with multiplex qPCR assays

We developed multiplexed, species-specific, quantitative PCR assays for the detection of four freshwater mussel species native to western North America, , and , from environmental DNA (eDNA). These species have experienced dramatic declines over the last century and are currently threatened in many portions of their ranges. Therefore, improved tools for detecting and monitoring these species are needed. Species-specificity and sensitivity of assays were empirically tested in the lab, and multiplex assays were also validated with field collected eDNA samples. All assays were species-specific, sensitive, and effective for detection from eDNA samples collected from streams and rivers. These assays will aid in the detection, monitoring, management, and conservation of these vulnerable species.