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DEVELOPMENT OF EDNA DETECTION PROTOCOLS FOR THE EASTERN HELLBENDER IN A TRIBUTARY OF THE WEST BRANCH SUSQUEHANNA RIVER

    Wands, Tara, Clean Water Institute, Lycoming College, 1 College Place, Williamsport, PA, 17701, wantara@lycoming.edu; Kaunert, Matt, Clean Water Institute, Lycoming College, 1 College Place, Williamsport, PA, 17701, kaunert@lycoming.edu; Andrew, David, Biology Department Lycoming Clean Water Institute 1 College Place Williamsport PA, 17701, andrew@lycoming.edu.

    The distribution of the Eastern hellbender (Cryptobranchus alleganiensis) in the Susquehanna Watershed has been previously studied through traditional rock-flipping assays. This method is beneficial in that hellbenders can be physically observed and potentially tagged, but rock-flipping assays are time consuming, cost ineffective, and pose increased physical risk to animals and researchers. A relatively novel and less invasive method for hellbender detection is through quantitative PCR (qPCR) of environmental DNA (eDNA). The goal of this this study is to develop eDNA collection, isolation, and detection protocols tailored to the resources available at Lycoming College and implement them in a tributary of the West Branch Susquehanna River to detect hellbender populations. Preliminary findings suggest that the protocols used are effective in hellbender eDNA detection studies. Positive controls, negative controls, and designed standards are consistent with duplication. In early testing, hellbender DNA was detected in spiked water samples and samples from known hellbender habitat. Future work includes implementing these protocols on previous rock-flipping survey sites and additional sites to update the knowledge of hellbender distribution within this tributary.

    eastern hellbender, eDNA