• crustacean;
  • invasive species;
  • cave;
  • subterranean;
  • population genetic


This study examined the phylogeography and population demographics of Cambarus tenebrosus, which has an unusually large distribution for a freshwater crayfish species, encompassing the Interior Lowlands and Cumberland Plateau of the eastern United States. This facultative cave-dweller provides a unique perspective on the biologic connections between surface and subsurface freshwater ecosystems, which are considered to be highly imperiled due to pollution and habitat degradation. The 16S mitochondrial gene was sequenced for 233 individuals from 84 cave and 20 surface locations throughout the range, with most sampling concentrated around the Cumberland Plateau of the southern Appalachians, to assess conservation status of this species and examine the extent of gene flow between the two habitat types. Cave and surface populations formed a single monophyletic group relative to Cambarus striatus, and clades showed strong geographical associations, but lacked habitat structuring. Occupation of subterranean environments does not appear to be a recent event in the evolutionary history of the species. The large amount of genetic diversity within the species, coupled with its ability to inhabit surface and subsurface environments, suggests that this species may pose a threat as a possible invasive species in other karst-dominated landscapes.