The success of evaluating areawide control of the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), in urban landscapes hinges on detailed understanding of colony movement and plasticity of the breeding system. Most previous studies of colony affiliation and breeding systems of C. formosanus have been conducted in relatively undisturbed park-like areas. However, disturbance in the form of landscaping, construction, and nearby treatment may impact termite colony dynamics in urban habitats. Therefore, we used microsatellite genotyping to identify the number of colonies, assess colony movement, and investigate the breeding structure of colonies surrounding the Supreme Court Building in New Orleans, LA. During 4 yr, 18 distinct colonies were identified in the study area. In contrast to earlier studies in park-like areas, which indicated stable foraging areas, colonies in this study moved frequently into and out of inground stations. Five colonies were simple families; two of these colonies were headed by inbred nestmate pairs, and three of these colonies were headed by outbred unrelated pairs. Thirteen colonies were extended families headed by fewer than five neotenic reproductives. During the study, the predominant breeding system shifted; simple family colonies either moved or were eliminated, and they were replaced with new extended family colonies. In one case, a generation turnover within a colony from simple to extended family was observed.