Population genetics of white-tailed deer (Odocoileus virginianus) have been influenced through human actions including the translocation of deer from across the United States in the 1900s and, recently, the creation of the captive-cervid industry, which uses animal husbandry to manipulate genetic variation. To assess the effects of these actions, I studied the genetic variation of free-range and captive populations of deer across the southcentral U.S. using a 14 microsatellite panel. In free-range populations I found genetic structure that divided deer west to east along the Mississippi River. Additionally, I found that captive populations were genetically distinct from geographically proximate free-range populations. However, after 2 generations of hybridization, this distinction disappeared. Finally, using both Bayesian clustering and multivariate approaches, I was able to identify a non-native individual from local free-range populations in southern Mississippi. Using these methods, wildlife managers can further investigate cases of hybridization between non-native deer and free-range populations.