Parentage Analysis and Conservation Genetics Educational Material for the Eastern Hellbender Salamander, Cryptobranchus alleganiensis alleganiensis

Populations of the Eastern hellbender salamander, Cryptobranchus alleganiensis alleganiensis, are declining, making this a species of special concern in New York State and under consideration for Federal Endangered Species listing. As a result of this decline, the New York State Department of Environmental Conservation and the Buffalo Zoo initiated a headstarting program with an egg mass found in the Allegheny River drainage. The juveniles being raised by the Zoo will be released back into the watershed and so understanding the genetic diversity and parentage of these hellbenders will inform the reintroduction efforts. Furthermore, in order to determine how to conserve hellbenders, the structure of their populations must be studied to determine the genetic diversity present. Microsatellite markers are a powerful tool used to study the genetic makeup of a population. Primers developed for the Eastern hellbender salamander were used to amplify four separate microsatellite regions of hellbender DNA. The optimal annealing temperatures of these primers were determined and 49 juvenile hellbenders at the Buffalo Zoo were genotyped. Genotypes were then used to conduct a parentage analysis with the COLONY software. The parentage analysis indicated approximately 16 parents (nine fathers and seven mothers). However, this result had very little statistical support. It is unlikely, based on hellbender reproductive biology, for this extreme number of parents (although allelic diversity indicates that there are at least four parents). Genotyping a larger group of juveniles may provide a more accurate parent estimate. Finally, educational material, in the form of a lesson plan and activity, was developed and tested for use in high school biology classes. This activity will be a resource for teaching genetics. It may also serve as a way to spread the importance of conservation genetics and introduce students to a unique and rare species of salamander.