Approximately 390 native North American crayfish species are known, representing nearly two-thirds of the world&s crayfish fauna. The majority of these species occur in the southeastern United States. North Carolina supports a substantial proportion of that diversity with 41 described indigenous crayfish species, 12 of which are endemic, and 3 introduced species, many of which are of significant conservation interest. In the late 1990s, the North Carolina Wildlife Resources Commission (NCWRC) began a focused effort to inventory and establish baselines for monitoring populations of both native stream-dwelling and burrowing crayfishes and invasive non-native species. During 2004–2005, that effort was completed for the Hiwassee, Little Tennessee, Savannah, French Broad, Watauga, New, Catawba, and Broad river basins in western North Carolina. Twenty-four stream-dwelling species and 5 burrowing species were collected from 199 stream sites and 58 burrowing sites. New records for many species, including a new river basin record for Cambarus reduncus (Sickle Crayfish), and new county records for C. howardi (Chattahoochee Crayfish), C. dubius (Upland Burrowing Crayfish), and C. nodosus (Knotty Burrowing Crayfish), were determined during these surveys. Small range expansions were documented for Orconectes virilis (Virile Crayfish; not native to North Carolina) and for Procambarus acutus (White River Crawfish; introduced outside its native range in North Carolina). We failed to detect the non-native O. rusticus (Rusty Crayfish), and P. clarkii (Red Swamp Crawfish) at or near previously reported localities. Observations of life-history traits, such as reproductive condition, fecundity, and habitat use were recorded. Specimens were also provided to crayfish taxonomists to help resolve certain taxonomic problems and to assist in the completion of new species descriptions. Data collected during this and previous NCWRC inventories, as well as data obtained from the North Carolina State Museum of Natural Sciences, North Carolina Division of Water Quality, Ohio State University, and other cooperators, were incorporated into a detailed GIS database. This database was used to identify data gaps to guide sampling efforts and to assess species and population status. In the future, this GIS database should provide a useful tool in monitoring the status of native crayfish populations and the spread of invasive species, and informing conservation and management decision making.