Specimens historically identified as Odontostilbe microcephala from the upper rio Paraná and Andean piedmont tributaries of the río Paraguay are reviewed and split in three species. We found that the distribution of O. microcephala is restricted to the Andean slope of the río Paraguay basin. The species is distinguished from congeners with subterminal mouth by the elongate body, usually 10-12 gill rakers on upper branch and smaller horizontal orbital diameter (24.6-32.8 % HL, mean 28.7%). Specimens from upper rio Paraná constitute two new species, diagnosed from other Cheirodontinae by the presence of mesopterygoid teeth, grouped on median portion and forming a continuous row. The new species are distinguished from each other by having premaxillary teeth with five cusps vs. nine cusps and by the number of lamellae in left and right sides of central median raphe of olfactory rosette with 20-21 vs. 11-12.