ABSTRACT The genera Hemigrammus and Moenkhausia have been traditionally diagnosed mainly by the former having lateral line completely pored whereas the latter having a lateral line with a few pored scales. Those features have been used to diagnose species of both genera in the upper Paraná River floodplain. Specimens with the diagnostic features of Moenkhausia bonita, collected in the upper Paraná River floodplain, exhibited different developmental levels of the lateral line, making it difficult to distinguish them from specimens of Hemigrammus sp. We analyzed the gene encoding cytochrome C oxidase I (COI) and intron 1 of the nuclear gene S7 to investigate the genetic similarities between the called Hemigrammus marginatus and M. bonita and to confirm their identities. Molecular sequences of other Moenkhausia species were analyzed for genus delimitation tests. The results reveal genetic similarities of M. bonita specimens with different developmental levels of the lateral line, and also distinguish between M. bonita and Hemigrammus sp. Species delimitation tests revealed that specimens from the upper Paraná River floodplain were M. bonita and were distinct from other Moenkhausia species. The developmental level of the lateral line is not a consistent characteristic that distinguishes between Moenkhausia and Hemigrammus species.

Astyanax fishes are among the most important food-web components of South America rivers. In the Iguaçu River basin, the Astyanax genus is represented mainly by endemic species. For millions of years, that hydrographic basin has been geographically isolated from the Paraná River basin by the Iguaçu Falls. Recently, a species from the Upper Paraná River basin identified as Astyanax bimaculatus was revised and described as a new species named Astyanax altiparanae Garutti & Britski, 2000. Fauna endemism and geographic isolation triggered interest in investigations to evaluate the identification and genetic relatedness among two A. altiparanae populations from the Upper Paraná River basin and the population identified as A. bimaculatus in the Iguaçu River, upstream from the Iguaçu Falls. Mitochondrial DNA sequences and RAPD markers revealed high genetic diversity within each population, as well as low genetic distance, high gene flow, and high mitochondrial DNA similarity among all three populations. In conjunction with morphological similarities, these results demonstrated that the population presently known as Astyanax bimaculatus in the Iguaçu River should actually be stated as Astyanax altiparanae. Furthermore, it could be inferred that the A. altiparanae population is not endemic and most likely it was recently introduced in the Iguaçu River basin, maintaining the ancestral genetic identity.