Phylogenetic relationships of rabies viruses recovered from black-backed jackals (Canis mesomelas) and domestic dogs (Canis familiaris) in northern South Africa were investigated to determine whether the black-backed jackal is an emerging maintenance host species for rabies in this region. A panel of 123 rabies viruses obtained from the two host species between 1980 and 2006 were characterised by nucleotide sequencing of the cytoplasmic domain of the glycoprotein gene and the non-coding G-L intergenic region. Through phylogenetic analysis a viral cluster specific to black-backed jackals and spanning a 5-year period was delineated in western Limpopo. Virus strains associated with domestic dogs prevail in densely populated communal areas in north-eastern Limpopo and in south and eastern Mpumalanga. The data presented in this study indicated the likelihood that black-backed jackals are capable of sustaining rabies cycles independent of domestic dogs. It is proposed that wildlife rabies control strategies, in synergy with domestic animal vaccination should be considered for effective control of rabies in South Africa.