The Aphyosemion calliurum species group is poorly diagnosed by chromatic and meristic characteristics leading various authors to propose different species as members. We used partial sequence of the mitochondrial cytochrome b gene to characterize all species that have been at one time or another included in the A. calliurum species group. Results obtained allowed a clear definition of the group which is composed of 10 species: A. ahli, A. australe, A. calliurum, A. celiae, A. edeanum, A. franzwerneri, A. heinemanni, A. lividum, A. pascheni, and a herein newly described species A. campomaanense.