A retrospective serosurvey was carried out between 2009 and 2012 to detect antibodies to Brucella spp. in free-ranging African wildlife ungulates from five selected game parks in Zimbabwe. Samples were drawn from wildlife-livestock interface and non-interface areas in Zimbabwe. A total of 270 serum samples from four different species, namely African buffalo (Syncerus caffer) (n=106), impala (Aepyceros melampus) (n = 72), black rhinoceros (Diceros bicornis) (n= 45) and white rhinoceros (Ceratotherium simum) (n = 47), were tested. The percentage of positive samples was 17.0% in buffalo (18/106; 95% CI: 9.72% – 24.1%) and 1.4% in impala (1/72; 95% CI: 0% – 4.2%). No antibodies to Brucella spp. were detected in the two rhinoceros species. The difference in the percentage of seropositive cases between buffalo and impala was significant (p< 0.05). Seropositivity to Brucella spp. was higher (19.1%) in adult buffalo compared with juveniles and sub-adults younger than six years (5.9%). Further, seropositivity was marginally higher (20.4%) in animals from wildlife-livestock interface areas than in those from non-interface areas (13.45%; OR = 1.45) although the difference was not statistically significant. The study showed that brucellosis could be more widespread in buffalo and may circulate in this species independently in the absence of contact with cattle, whilst rhinoceros may be considered less susceptible to brucellosis. The role of the wildlife-livestock interface in the epidemiology of brucellosis in wildlife and livestock is probably overstated but needs to be explored further.