Lebiasina yepezi, a new Lebiasininae with a conspicuous color pattern, is described. The new species is endemic of the headwaters of the rio Negro, rio Branco, and rio Orinoco in the Serra Parima-Tapirapecó Mountains, at the border of Brazil and Venezuela. The new species is readily distinguished from all other Lebiasininae by the presence of four black longitudinal stripes on the trunk, and the triangular shaped dorsal surface of the mesethmoid, lacking lateral projections. The unusual color pattern is contrasted with those of other lebiasinin, as well as members of the pyrrhulinin genus Nannostomus. A close relationship between Lebiasina yepezi and the Gran Sabana (Venezuela) species is suggested based in color pattern features. The present contribution corresponds to a further refutation of the type locality of L. intermedia, as suggested on its description, since Lebiasinins, except L. bimaculata, L. boruca, and L. festae, do not occur in low land waters.