Over 40 species of ectoparasites were identified from more than 100 road-killed animals in northwestern South Carolina. Fleas (Siphonaptera), flies (Diptera), lice (Phthiraptera), mites (Astigmata, Mesostigmata, and Prostigmata), and ticks (Ixodida) were routinely found on animal carcasses. Some of the ectoparasites found and the disease agents they could carry include Amblyomma americanum (Ehrlichia spp.), Ctenocephalides felis (Rickettsia typhi), Dermacentor variablilis (Rickettsia rickettsii), Haemaphysalis leporispalustris (Powassan encephalitic virus and Francisella tularenis), Neohaematopinus sciuropteri (Rickettsia prowazekii), and Pulex stimulans (Salmonella enteriditis). We emphasize the use of road-killed animals for the surveillance of disease agents.