The captive North American cheetah (Acinonyx jubatus) population is not self-sustaining because of high prevalences of unusual diseases and poor reproductive success. Cheetahs are commonly moved between zoos for breeding purposes to maintain genetic diversity within the captive population, and movement may exacerbate infertility and disease. Fecal corticoids were analyzed by radioimmunoassay to measure the stress response of cheetahs to movement between facilities. Fecal samples were collected from 15 cheetahs for 14 days before movement and for at least 30 days after movement. For each cheetah, premovement fecal corticoid concentrations were used to determine baseline and then compared with trends in postmovement concentrations. In general, postmovement corticoid concentrations either increased (n = 8), did not change (n = 2), or decreased (n = 5). Although individual animal differences occurred, corticoid concentrations increased for most animals moved on-exhibit and decreased in animals moved off-exhibit. Animals moving on-exhibit had an 18-times greater risk of having corticoids elevated more than two standard deviations above baseline for 30 days after movement compared with animals that moved off-exhibit. In addition, greater day-to-day variation in corticoids occurred in animals moved on-exhibit. In general, animals with initially low baseline corticoid concentrations had a greater postmovement corticoid response than cheetahs with initially high baseline levels. These results indicate that some cheetahs have a prolonged stress response when moved between facilities, and the magnitude and character of this response is influenced by the exhibit environment.