KSEF RDE: Leveraging novel genetic and genomic technologies to understand and conserve a Kentucky state endangered giant salamander

Hellbenders (genus Cryptobranchus) are large, obligately aquatic salamanders that are critically endangered across their range in the eastern and central US, particularly in Kentucky. Recent research suggests that Cryptobranchus may comprise several unrecognized cryptic species, each more imperiled than currently realized. Effective and timely hellbender conservation hinges critically upon understanding: 1) where hellbender populations remain in the wild, 2) the boundaries between evolutionarily distinct lineages, 3) population-level parameters, such as effective population sizes, levels of genetic diversity, and rates of gene flow between populations, and 4) the effects of anthropogenic modification of watersheds and waterways on population viability. However, the secretive lifestyles of this salamander have confounded detailed elucidation of its geographic distribution. Additionally, existing genetic methods and analytical approaches are unable to provide much needed genome-wide perspectives on lineage boundaries and population dynamics in this group, impeding protection and repatriation efforts. Accordingly, our proposed research leverages several innovative approaches to confront these pressing challenges. We will employ emerging methods for aquatic environmental DNA detection to efficiently locate hellbender populations in waterways across Kentucky. We will develop and apply next-generation sequencing methods for genome analysis and multilocus species delimitation to reveal species boundaries and population genetic structure in this genus. Importantly, we will integrate these findings with rigorous Geographic Information System-based modeling of land use practices and watershed architecture in Kentucky to provide novel insights into the conservation biology of this enigmatic and rapidly declining salamander. Because of its central position in the hellbender's geographic range and the interface of several distinct watersheds, Kentucky stands as a potential keystone for understanding and conserving hellbenders on both local and regional scales. Our comprehensive and integrative approach to hellbender conservation, combining novel field, laboratory, and analytical approaches, aims to become a model for conservation of aquatic species in Kentucky and beyond.