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Tuesday, January 30 • 6:00pm - 9:00pm
Poster Display. Testing the Effectiveness of eDNA Procedures

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AUTHORS: Andrew Gregory, U.S Fish & Wildlife; Ashlee Nichter, Toledo Zoo; Hannah Olenik, Bowling Green State University Center for Undergraduate Research & Scholarship, Grase Lab

ABSTRACT: Fossorial species tend to be notoriously difficult to survey for, and so tend to be under sampled in species inventories. Using eDNA to screen for the presence of cryptic species at otherwise putatively suitable areas is a promising tool to increase survey effectiveness. eDNA is the use of genetics to survey an area for remnant DNA left behind after a species has visited a location. Prior to initiative an eDAN survey, it would be nice to verify the potential power of your proposed field and lab protocols to detect eDNA at a site. In this study, we report on the efficacy trials that were completed to optimize field and lab methods for surveying for eDNA of the Kirtland snake (Clonophis Kirtlandii) in northwest, Ohio. To test the threshold of how much tissue and water is needed to detect eDNA, I placed 0.1mg of snake tissue in 250, 500, and 1,000 mg of water. Environmental freshwater was obtained from a wetland located on the Bowling Green State University campus. Because Kirtland snake is a state threatened species, we used Black Rat Snake (Pantherophis obsoletus) tissue as a surrogate in our analysis. Both species are from the same tribe (Colubridae); thus use the same primers and have similar molecular chemistry. Water was extracted by filtration and extracted from filter paper using protocol of the FastDNATM SPIN Kit and then amplified at 6 microsatellite primers via PCR. We compared DNA amplification success rates among primers across water volumes and time of tissue exposure. We also compared the results of this analysis to water extracted form a tank that contained a live snake for 48 hours. We found that in all cases suitable genetic material was obtained for molecular genetic analysis. This was preliminary analysis prior to applying these methods in the field.

Tuesday January 30, 2018 6:00pm - 9:00pm CST
Ballroom C & Foyer