Lagos bat virus (LBV) is a phylogroup 2 lyssavirus which causes rabies when it infects the brain of terrestrial mammals1. Vaccination against classical rabies virus (phylogroup 1) does not protect against LBV as there is little or no serological cross protection across these lyssavirus phylogroups2. To understand risk factors for public and domestic animal health, an understanding of viral infection dynamics in reservoir hosts is required3. The straw-coloured fruit bat, Eidolon helvum, is a natural reservoir host of LBV1. Three LBV isolates obtained from wild E. helvum were available, including the type virus isolated from a bat from Lagos Island, Nigeria in 1956. Prior to investigating viral infection dynamics in this host, we conducted a pilot study to determine if each of these isolates would cause disease should it reach the brain of E. helvum. Each of three groups of captive-bred E. helvum known to have never been exposed to phylogroup 1 or 2 lyssaviruses (or any viruses that serologically cross react with them) were experimentally inoculated intracerebrally with one of the LBV isolates. Equal titres of virus were inoculated across all isolates using a standard site and rate of inoculation. A fourth group was mock-infected using virus-free tissue culture fluid. While mock-infected animals remained healthy throughout, each LBV isolate produced a distinct disease phenotype with a clinical duration of less than 24 hours. The incubation period and the pattern of viral spread to the salivary glands also appeared to be isolate-specific.