African buffalo (Syncerus caffer) act as maintenance hosts for foot-and-mouth disease (FMD) in southern Africa. A single buffalo can become infected with all three of the endemic serotypes of FMD virus (SAT-1, SAT-2, and SAT-3) and pose a threat of infection to other susceptible cloven-hoofed animals. The floods of 2000 in southern Africa damaged the Kruger National Park (KNP) game fence extensively, and there were several accounts of buffalo that had escaped from the park. The VP1 gene, which codes for the major antigenic determinant of the FMD virus, was used to determine phylogenetic relationships between virus isolates obtained from the outbreaks and those previously obtained from buffalo in the KNP. These results demonstrate that buffalo were most probably the source of the outbreaks, indicating that disease control using fencing as well as vaccination is extremely important to ensure that FMD does not become established in domestic livestock.