We examined comparative skin permeability by measuring cutaneous water loss and epidermal lipid content of two sympatric snake congeners, the Southern Copperhead (Agkistrodon contortrix) and the Western Cottonmouth (A. piscivorus). Samples of shed epidermis from each snake species were taken to first investigate species-specific differences. Shed samples from multiple regions along the dorsal and ventral integument were then used to examine potential variation in skin permeability and lipid content among different regions along the body for each species. Rates of cutaneous water loss were significantly higher for A. piscivorus. In addition, cutaneous water loss rates of A. piscivorus differed among region (i.e., anterior, mid, and posterior) and surface (i.e., dorsal and ventral) with the highest rates of cutaneous water loss from the mid-dorsum. Lipid content did not differ between species. However, lipid content varied between the dorsal and ventral surfaces of both species, with greater lipid content on the dorsal surface. Our results support a species-specific difference in cutaneous water loss between A. contortrix and A. piscivorus and may reflect species-specific adaptations to their differences in microhabitat preference.