Date of Award

Spring 4-29-2025

Document Type

Thesis

Degree Name

Bachelor of Science (BS)

Department

Biology

College

College of Science

First Advisor

Scott L. Parker

Second Advisor

Michelle M. Barthet

Abstract/Description

The Pine Snake (Pituophis melanoleucus) is listed in the South Carolina Wildlife Action Plan as a “species of greatest conservation need.” However, little is known about their current abundance and distribution within South Carolina due to their low population density and secretive lifestyle. Commonly used presence-absence survey methods, such as drift-fence trap arrays, often fail to reliably detect Pine Snakes even when they are present in each habitat. To address this issue, we developed novel primers to detect Pine Snakes using environmental DNA (eDNA), aiming to increase the efficiency of detecting these snakes in critical habitats. eDNA is increasingly utilized to complement traditional ecological survey techniques for identifying rare and endangered species. We developed Pituophis-specific primer pairs targeting the NADH dehydrogenase subunit 4 (ND4) gene and assessed their specificity and sensitivity in both laboratory and environmental applications. Specificity tests confirmed selective amplification of Pituophis DNA versus that of conspecifics using skin and tissue samples. Sensitivity assays established a limit of detection threshold of 25 ng of tissue or skin DNA. eDNA samples were collected from three sectors, each approximately 200 hectares, within a study site on the South Carolina Coastal Plain known to host a Pine Snake population, to evaluate the effectiveness of the primers in detecting Pine Snake presence and distribution. Our findings may enhance Pine Snake conservation planning by providing an efficient, non-invasive tool for detecting Pine Snake populations statewide.

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