Crayfish are an aquatic fauna of conservation concern, yet regional studies are lacking on zoogeography and life history. We compared recent and historical species distribution data and assessed conservation standings of native and nonindigenous crayfish of the Potomac River Basin in West Virginia. From 2007–2011, a total of 1764 crayfish were collected from 159 sites. Data collection included species abundance, morphometrics, and life history parameters. Percentages of the number of individuals of each species of the total catch were 36.3% (Cambarus bartonii), 34.6% (Faxonius obscurus), 23.4% (Faxonius virilis), 3.6% (Procambarus cf. zonangulus) and 2.0% (Cambarus carinirostris). Cambarus bartonii was present throughout the drainage, F. obscurus was collected primarily from the North Branch, South Branch, and Cacapon river watersheds, and C. carinirostris was only collected in the South Branch watershed. Two nonnative species, F. virilis and P. cf. zonangulus, were only present in tributaries downstream of the Cacapon River watershed. Spinycheek crayfish (Faxonius limosus) were not collected during our survey, which suggests its possible extirpation from the West Virginia portion of its range. Our zoogeographic and life history data could serve as a baseline for future conservation-oriented monitoring efforts of the Potomac River watershed.