A study was conducted to investigate the seroprevalence and associated risk factors of Chlamydia abortus infection in cattle and some selected wildlife species at selected interface areas at the periphery of the Great Limpopo Transfrontier Conservation Area in Zimbabwe. Three study sites were selected based on the type of livestock–wildlife interface, porous livestock–wildlife interface (unrestricted), non-porous livestock–wildlife interface (restricted by fencing), and livestock–wildlife non-interface (totally absent or control). Sera were collected from cattle aged =¿2 years representing both female and intact male. Sera were also collected from selected wild ungulates from Mabalauta (porous interface) and Chipinda Pools (non-interface) areas of the Gonarezhou National Park. Sera were tested for antibodies to Chlamydia abortus using a CFT. A X 2 test was used to assess differences between categories and p¿<¿0.05 was considered as significant. In cattle, the overall seroprevalence was 32.7% (327/1011; 95% CI 29.5–35.3). A significantly lower seroprevalence was recorded for the porous interface (24.2%) compared to the non-porous (42.5%) (p¿<¿0.0001) and the non-interface (36.2%) (p¿=¿0.001). Overall, the wet season recorded a significantly (p¿=¿0.015) higher seroprevalence than the dry season. In wildlife, antibodies were detected in buffaloes (47.7%) and impalas (43.8%) but not in kudus. Buffaloes from Chipinda Pools (53.4%) had a significantly (p¿=¿0.036) higher seroprevalence than those from Mabalauta (26.1%). The results established the presence of chlamydiosis in cattle and selected wildlife and that independent infections may be maintained in buffalo populations. Further studies are required to clarify chlamydiae circulating between cattle and wildlife.