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Welcome to the technical sessions schedule for the 2015 SEAFWA Annual Meeting.

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Tuesday, November 3 • 4:00pm - 4:20pm
Evaluation of Cytochrome b PCR-RFLP to Distinguish DNA of Southeastern U.S. Carnivore Species

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Dusty J. Harrell, Jean Fantle-Lepczyk, Todd D. Steury –Auburn University

Non-invasively collected fecal samples are an important method of monitoring wildlife species. However, similarity in feces appearance amongst sympatric species can complicate identification and thus negatively impact accuracy of estimates generated. In order to efficiently and unambiguously identify carnivore species from fecal DNA samples, Bidlack et al (2007) developed a PCR-RFLP protocol to distinguish among a suite of western North American carnivores. While the technique was effective for the seven species included in the study, we investigated how the technique would apply to carnivore species in the southeastern US. We collected tissue and scat samples from Alabama populations of Bidlack et al’s (2007) seven carnivore species, as well as samples from three additional carnivore species (black bear, domestic dog, and domestic cat) that are locally common and that could potentially complicate the identification process. We extracted DNA using QiaAMP stool and tissue DNA extraction kits by Qiagen. We used polymerase chain reaction (PCR) to amplify a 196 base pair polymorphic region of cytochrome b from the extractions. We then evaluated how three restriction enzymes (HpaII, DdeI, HpyCH4V) would cut the amplified region at species-specific locations. Using this technique, we were able to distinguished among DNA from coyote, grey fox, striped skunk, domestic dog, opossum, and river otter. However, bobcat and domestic cat cut in the same location, as did black bear and raccoon. Thus, this method can potentially be an accurate and inexpensive method of identifying carnivore scat in the southeastern U.S. However care must be taken to consider limitations.

Tuesday November 3, 2015 4:00pm - 4:20pm EST
Ballroom Salon A

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