Distributions and phylogeographic data of rheophilic freshwater fishes provide evidences on the geographic extension of a central-brazilian amazonian palaeoplateau in the area of the present day Pantanal Wetland

The analysis of the distribution patterns presented by examples of freshwater fishes restricted to headwater habitat: the anostomid Leporinus octomaculatus, the characins Jubiaba acanthogaster, Oligosarcus perdido, Moenkhausia cosmops, Knodus chapadae, Planaltina sp., the loricariid Hypostomus cochliodon, and the auchenipterid Centromochlus sp. provided evidences of a relatively recent shared history between the highlands of the upper rio Paraguay and adjoining upland drainage basins. Restricted to headwater of the uplands in the upper rio Paraguay and adjoining basins, these species provide biological evidence of the former extension of the central Brazilian plateau before the origin of the Pantanal Wetland. Disjunction took place due to an ecological barrier to these rheophilic taxa represented tectonic subsidence related to the origin of the Pantanal Wetland. Molecular analysis of Jubiaba acanthogaster revealed that the sample from the upper rio Xingu basin are the sister-group of a clade that includes samples from the upper rio Arinos (upper rio Tapajós) plus the upper rio Paraguay basin, supporting the assumption that the origin of the upper rio Paraguay basin causing vicariance between this basin and the upper rio Tapajós is the least vicariant event in the evolutionary history of the group.

A análise do padrão de distribuição apresentado por exemplos de peixes de água doce neotropicais restritos à habitats reofílicos de cabeceira: o anostomídeo Leporinus octomaculatus, os caracídeos Jubiaba acanthogaster, Oligosarcus perdido, Moenkhausia cosmops, Knodus chapadae, Planaltina sp., o loricariídeo Hypostomus cochliodon e o auquenipterídeo Centromochlus sp. fornecem evidências de uma história compartilhada relativamente recente entre as terras altas do alto rio Paraguai e bacias de terras altas vizinhas. Restritas às cabeceiras das áreas altas nas bacias do alto rio Paraguai e bacias vizinhas, estas espécies fornecem evidências biológicas da pretérita extensão do Planalto Central Brasileiro anteriormente a origem tectônica da planície alagável do Pantanal. Disjunções ocorreram devido a barreira ecológica para estes taxons reofílicos representada pela subsidência tectônica associada à origem do Pantanal. A análise molecular de Jupiaba acanthogaster revelou que amostras da bacia do alto rio Xingu consistem no grupo-irmão de um clado mais inclusivo que inclui amostras do alto rio Arinos (alto rio Tapajós) mais o alto rio Paraguai, suportando a suposição de que a origem do alto rio Paraguai causou vicariância entre esta bacia e o alto rio Tapajós no último evento vicariante na história evolutiva do grupo.

Can DNA barcoding accurately discriminate megadiverse Neotropical freshwater fish fauna?

BACKGROUND: The megadiverse Neotropical freshwater ichthyofauna is the richest in the world with approximately 6,000 recognized species. Interestingly, they are distributed among only 17 orders, and almost 80% of them belong to only three orders: Characiformes, Siluriformes and Perciformes. Moreover, evidence based on molecular data has shown that most of the diversification of the Neotropical ichthyofauna occurred recently. These characteristics make the taxonomy and identification of this fauna a great challenge, even when using molecular approaches. In this context, the present study aimed to test the effectiveness of the barcoding methodology (COI gene) to identify the mega diverse freshwater fish fauna from the Neotropical region. For this purpose, 254 species of fishes were analyzed from the Upper Parana River basin, an area representative of the larger Neotropical region. RESULTS: Of the 254 species analyzed, 252 were correctly identified by their barcode sequences (99.2%). The main K2P intra- and inter-specific genetic divergence values (0.3% and 6.8%, respectively) were relatively low compared with similar values reported in the literature, reflecting the higher number of closely related species belonging to a few higher taxa and their recent radiation. Moreover, for 84 pairs of species that showed low levels of genetic divergence (<2%), application of a complementary character-based nucleotide diagnostic approach proved useful in discriminating them. Additionally, 14 species displayed high intra-specific genetic divergence (>2%), pointing to at least 23 strong candidates for new species. CONCLUSIONS: Our study is the first to examine a large number of freshwater fish species from the Neotropical area, including a large number of closely related species. The results confirmed the efficacy of the barcoding methodology to identify a recently radiated, megadiverse fauna, discriminating 99.2% of the analyzed species. The power of the barcode sequences to identify species, even with low interspecific divergence, gives us an idea of the distribution of inter-specific genetic divergence in these megadiverse fauna. The results also revealed hidden genetic divergences suggestive of reproductive isolation and putative cryptic speciation in some species (23 candidates for new species). Finally, our study constituted an important contribution to the international Barcoding of Life (iBOL.org) project, providing barcode sequences for use in identification of these species by experts and non-experts, and allowing them to be available for use in other applications.

DNA barcoding reveals hidden diversity in the Neotropical freshwater fish Piabina argentea (Characiformes: Characidae) from the Upper Paraná Basin of Brazil.

BACKGROUND AND AIMS: We analyzed a small and wide geographically distributed Neotropical freshwater fish, the Piabina argentea from the Upper Paraná Basin, to check the hypothesis that this species is composed of more than one biological unit, since it has a limited dispersion, through the DNA barcode technique. MATERIALS AND METHODS: Partial mitochondrial COI and CytB gene sequences were obtained for 58 specimens drawn from 13 localities. RESULTS: Phylogenetic analysis revealed six major clusters of P. argentea. Kimura-two-parameter (K2P) genetic divergences among these six P. argentea clusters ranged from 2 to 5.6% and from 2.3 to 5.4% for COI and CytB genes, respectively, and these values were on average approximately nine times greater than intra-cluster K2P divergences. The fixation index (F(ST)) among clusters showed very high values and the haplotype network analysis displayed seven unconnected units. CONCLUSION: These results reinforce the hypothesis that the widely distributed P. argentea species concept as currently conceived actually represents more than one species (possibly six). These results demonstrate the efficacy of DNA barcoding for the discovery of hidden diversity in Neotropical freshwater fishes, and we conclude that barcoding is a useful tool for alpha taxonomy.

DNA barcodes discriminate freshwater fishes from the Paraíba do Sul River Basin, São Paulo, Brazil.

BACKGROUND AND AIMS: Considering the promising use of DNA barcoding for species identification, the importance of the freshwater fish fauna of the Paraíba do Sul River Basin, and its advanced stage of degradation, the present study evaluated the effectiveness of DNA barcoding to identify the fish species in this basin. MATERIALS AND METHODS: A total of 295 specimens representing 58 species belonging to 40 genera, 17 families, and 5 orders were sequenced. RESULTS: The DNA barcodes discriminated all species analyzed without ambiguity. The results showed a pronounced difference between conspecific and congeneric pair-wise sequence comparisons, demonstrating the existence of a "barcode gap" for the species analyzed. The nearest-neighbor distance analysis showed only three cases with Kimura two-parameter values lower than a 2% divergence threshold. However, the patterns of divergence observed in each case remained sufficient to discriminate each species, revealing the accuracy of DNA barcoding even cases with relatively low genetic divergence. At the other extreme, three species displayed high genetic sequence divergence among conspecifics. For two cases, Characidium alipioi and Geophagus proximus, barcoding proved effective at flagging possible new species. For another case, Astyanax bimaculatus, the use of DNA barcoding of the comparison of shared freshwater fish fauna between different basins revealed itself as highly useful in disclosing that the previously identified A. bimaculatus "cluster A" probably represents the species Astyanax altiparanae. CONCLUSION: The present study is among the first to assess the efficiency of barcoding for the Brazilian freshwater fishes. The results demonstrate the utility of barcoding to identify the fauna from this basin, contribute to an enhanced understanding of the differentiation among species, and to help flag the presence of overlooked species.