Overharvesting and habitat loss resulted in the extirpation of river otters (Lontra canadensis) across much of Kentucky and efforts in the mid-1990s by the Kentucky Department of Fish and Wildlife Resources have led to a resurgence of otter populations across the state. Since recovery, limited data on the biology of these animals have been collected, including levels of infection by endoparasites. We performed necropsies on 170 river otters obtained during the 2006–2009 harvest seasons. We found nematode parasites present in the fascial layers of the hind leg of 5 juvenile river otters for a minimum infection level for the Kentucky population of 0.0294. Infected animals were recorded in the Green, Kentucky, and Mississippi River watersheds. All nematodes were females of the genus Dracunculus, which cannot be assigned to the species level based on morphological characteristics. These observations are the first documented case of Dracunculus infection in river otters in Kentucky.