A study was conducted to determine the serotypes of foot-and-mouth disease viruses (FMDV) circulating in African buffaloes (Syncerus caffer) from selected areas in Zambia. Sera and probang samples were collected between 2011 and 2012 and analysed for presence of antibodies against FMDV while probang samples were used to isolate the FMDV by observing cytopathic effect (CPE). Samples with CPE were further analysed using antigen ELISA. High FMD seroprevalence was observed and antibodies to all the three Southern African Territories (SAT) serotypes were detected in four study areas represented as follows: SAT2 was 72.7 percent; SAT1 was 62.6 percent; and SAT3 was 26.2 percent. Mixed infections accounted for 68.6 percent of those that were tested positive. For probang samples, CPE were observed in three of the samples, while the antigen ELISA results showed positivity and for SAT1 (n = 1) and SAT2 (n = 2). It is concluded that FMDV is highly prevalent in Zambian buffaloes which could play an important role in the epidemiology of the disease. Therefore livestock reared at interface with the game parks should be included in all routine FMDV vaccination programmes.