Australia is home to many endemic genera of freshwater crayfish, including the cold-water adapted spiny crayfish, Euastacus. A recent study examining the species identities of Euastaus, including the mountain spiny crayfishes (Alpine crayfish E. crassus and Riek’s crayfish E. rieki), suggests that the species assigned to these crayfish by range maps are questionable and there may be three or four species of mountain spiny crayfish rather than two currently recognised (Austin et al. 2022). Suprisingly, there are little to no samples of Euastacus crayfish from Kosciusko National Park available, so again, it is unclear what species are present in the Park, nor what conservation actions are necessary to prevent their further decline. The first step in conservation action is identification of the units of conservation concern. Therefore, we collected 18 crayfish samples from the three river catchments of Kosciusko National Park (5 from creeks within the Snowy River catchment, 2 from creeks within the Murray River catchment and 11 from creeks wihin the Murrumbidgee River catchement) and conducted sequence analysis of the mitochondrial cytochrome c oxidase subunit (COI) gene to identify the species located within the Park using DNA barcoding based off of sequences deposited in GenBank and species identies from Austin et al. (2022). The species identifications and locations, implications for conservation and biogeography of these mountain spiny crayfishes are discussed.