The Guttural Toad, Sclerophrys gutturalis, has three established invaded regions: on Mauritius and Reunion islands, and in Constantia, a peri-urban area of Cape Town, South Africa. The native range of this toad covers much of central and southern Africa. Here we use mitochondrial DNA (mtDNA) to sample across the range of the natural distribution (from Ethiopia to South Africa) and compare ND2 and 16S sequences to those from animals sampled from each of the three invaded regions. We show that all individuals in invaded regions refer to the same mtDNA clade, which is naturally distributed in north-eastern South Africa, but not from adjoining Mozambique or southernmost Eastern Cape areas. Our findings corroborate previous reports of deliberate introductions from South Africa to Mauritius, and from Mauritius to Reunion. Similarly, our results suggest a single accidental translocation within South Africa from the northeast to Constantia. Our findings highlight the combination of anthropophilic behavior, and extreme long-distance dispersal occurring with accidental translocation for this species. We caution that accidental pathways are likely to continue into the future, withrnincreasing numbers of invasive populations of this species. Keywords: Amphibia, Anura, biogeography, phylogeography, invasive species, mitochondrial DNA, pathways.