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Tuesday, January 30 • 10:20am - 10:40am
SYMPOSIA-06: Using eDNA Surveys to Detect a Fossorial Species of Conservation Concern, the Kirtland Snake (Clonophis kirtlandii) in Northwest Ohio

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AUTHORS. Hannah Olenik, Bowling Green State University; Matt Cross, Toledo Zoo; Andrew Gregory, Bowling Green State University

ABSTRACT. Kirtland’s Snake (Clonophis kirtlandii) is a fossorial species known to occupy moist grassland systems throughout the north-central Midwest. Since the early 1980’s populations of Clonophis kirtlandii may have declined by nearly 90%. Consequently, it is currently being considered by the USFWS for federal listing under provisions of the Endangered Species Act. However, because of the elusive nature of C. kirtlandii, ecological data—including basic population surveys for the species are lacking. This elusive nature is due to C. kirtlandii spending most of their lives in underground crayfish burrows. Consequently, a determination of the population status or actual rate of loss is nebulous. We used non-invasive eDNA surveys to sample for C. kirtlandii presence at fivesites in northwest Ohio that putatively had C. kirtlandii, three of which no detections had been made in the last few years. We extracted water from crayfish burrows during late July-October to correspond to C. kirtlandii natality periods, and concentrated crayfish burrow water filtrate on filter paper. We then extracted DNA from the filter paper using an MP Bio FastDNA Spin Kit. We screened each DNA sample using a set of molecular markers that have been shown to be polymorphic in C. kirtlandii. We then compared the amplified bands with control bands that were extracted using stock tissue and blood samples of C. Kirtlandii and seven other common Thamnophiine snakes. We successfully amplified Clonophis kirtlandii bands from six different sites. We also successfully amplified 24 bands of three other common Ohio snakes, as well as 32 bands of unknown snake species from our sample sites. This suggests that using water from crayfish burrows to detect Clonophis kirtlandii is a plausible field sampling protocol, and could help further progress methodologies for detecting multiple snakes and other fossorial species.

Tuesday January 30, 2018 10:20am - 10:40am CST