The African Jewelfish, Hemichromis letourneuxi, is an invasive, predatory cichlid fish introduced at least once to Florida. Its native range is in northern Africa. First encountered in Miami in the 1960s, it has since been found west and north within the State of Florida. It thrives in a wide range of aquatic habitats, including shallow, vegetated or rocky areas of canals, tidal creeks, rivers, and marshes. We generated mitochondrial DNA sequences from 6 Florida localities, and 1 Mexico population. The resulting tree describes recent historical relationships among populations. Nearly all Florida populations of African Jewelfish are intermixed, supporting a single invasion, with one possible exception: samples from the Tampa Bay locality are distinct and may represent a separate introduction. Samples obtained from the Mexico locality, previously identified as Hemichromis guttatus (Spotted Jewelfish), appear to be genetically identical to, and indistinguishable from the H. letourneuxi sampled in Florida localities.