Karyotyped specimens from three populations of spined loaches, genus Cobitis, that occurred in the Veleka (Bulgaria), Chernaya (Crimean Peninsula) and Southern Bug (Ukraine) Rivers in north-western Pontic region were subjected to enzyme electrophoresis (for 5 loci), comparative morphological studies, and phylogenetic analysis (based on PCR of a 1230 bp fragment of mtDNA and the cytochrome b gene). These studies resulted in the description of loaches from the Crimean Peninsula as a new species Cobitis taurica, while the taxonomic status of populations from the Veleka and S. Bug rivers appeared to be controversial and in need of further investigation.