A cross sectional study was conducted in the Serengeti ecosystem (wildlife-livestock interface) and central part of Tanzania (non-interface) area to determine the prevalence of foot and mouth disease (FMD) in Serengeti, Bunda, Kongwa and Iramba Districts. Seroprevalence investigation using 3ABC–ELISA technique indicated that the overall prevalence of antibodies against FMD virus was 66.3%. Significantly high prevalence was recorded in wildlife-livestock interface areas (71.5%) compared to non-interface areas (61.0%). District-wise, higher prevalence was recorded in Kongwa district (89.0%) followed by Serengeti (78.0%), Bunda (65.0%) and Iramba (33.0%). Species-wise, higher prevalence was found in bovines (69.8%), ovines (52.4%) and caprines (11.1%). From various risk factors, ecosystem distribution (X² = 4.9308, p = 0.0264) and species distribution (X² = 28.3236, P = 0.0001), the results indicated that FMD is highly prevalent in wildlife-livestock interface areas than in non-interface areas. However, uncontrolled livestock movement in Kongwa District resulted into much higher FMD prevalence than in districts where there is wildlife-livestock interface. The presence of antibodies against FMD virus in species other than cattle revealed that there is a need to consider other species in planning for FMD control.