Identification of the Eastern Massasauga Rattlesnake (Sistrurus catenatus) through genetic analysis of shed skin

Eastern massasauga rattlesnake ( Sistrurus catenatus ; hereafter EMR) populations have been severely affected by factors such as vegetative succession, habitat fragmentation, and water fluctuation. This has led to a great reduction in the number of populations and population sizes, with only half of the known historical populations remaining and many projected to be decimated in the next couple of decades. Current management efforts rely primarily on visual surveys to determine population status. However, EMR are very cryptic and the current resources to manage and survey populations are insufficient for proper species assessment. Here, we propose a non-invasive method to assess presence and absence of the EMR through the genetic analysis of shed skin. Using mitochondrial DNA amplification, EMR were conclusively identified and distinguished from other allopatric snake species. The viability of this method for species identification was supported by analysis of DNA from shed skins following one month of exposure at two EMR habitats. This methodology can be paired with citizen science projects to improve the current management of EMR populations throughout their range.