Lagos bat virus (LBV) belongs to genotype (gt) 2 of the lyssavirus genus in the family Rhabdoviridae, order Mononegavirales. This virus causes fatal rabies encephalitis in vertebrate animals and has only been reported from the African continent except for an imported case from African origin identified in France. The prototype lyssavirus is in fact rabies virus (gt 1) for which a variety of different vaccines are commercially available. These vaccines, however, do not provide protection against the gt 2 viruses. Genotype 2 viruses have not been well studied to date and the true risk for humans and animals is uncertain. The aim of this study was to investigate the epidemiology and pathogenicity of this uniquely African virus. In this project, our surveillance in South Africa reported six new LBV cases after this virus was not reported for the previous 12 years prior to this study. These results indicated that the incidence of this virus is greatly underestimated due to lack or absence of surveillance or ineffective diagnostic abilities of laboratories in Africa. Molecular epidemiological analysis of previously identified and new gt 2 isolates from this study indicated a high intragenotypic nucleotide and amino acid sequence diversity with respect to the Nucleo-, Phospho-, Matrix- and Glycoprotein genes. Based on these analyses, it has been proposed that two virus isolates that were previously reported as gt 2 LBV, may in fact constitute a new lyssavirus genotype. These findings emphasize the need to investigate different criteria for lyssavirus classification. As more lyssaviruses are discovered and with rapid progress in full genome sequencing, diversity becomes accentuated and challenges the criteria upon which lyssavirus taxonomy is based. As a compliment to these genetic findings, our study of viral pathogenicity in a murine model, identified that the pathogenicity of phylogroup II viruses has previously been underestimated. LBV poses a potential risk to humans and animals and future vaccine strategies should ideally include protection against phylogroup II viruses.