Examination of the morphological and genetic features of spined loach (Cobitis sp.) collected from the Oyodo River system, southern Kyushu Island, Japan, indicated that they represented a new taxon. The specimens were clearly distinguished from other known Japanese Cobitis species by their mitochondrial DNA sequences. In addition, the shape of the adult male lamina circularis differed to those of the C. sp. ‘yamato’ complex and C. biwae.