Historically, three obligate cave-dwelling species of Orconectes comprise an assemblage along the Cumberland Plateau of the southeastern United States, including Orconectes australis (with two subspecies, australis and packardi), O. incomptus, and O. sheltae. Using genetic data from three mitochondrial genes (cytochrome oxidase subunit I (COI), 12S, and 16S), we present evidence for four cryptic lineages excluding O. sheltae. The subspecies of Orconectes australis represent distinct taxonomic units with non-sister phylogenetic relationships and will be each recognized as separate species. Orconectes (Orconectes) barri is a new species of subterranean crayfish with a small distribution around the Kentucky Tennessee state border in Mississippian limestone. This species was uncovered during a previous thoroughly-sampled phylogeographic survey of the southern Appalachians. Additionally, we assign genetic barcodes (COI sequences) and conservation status to each species for management direction and identification of newly discovered populations.
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