Gymnogeophagus setequedas is a rare and rheophilic species of tribe Geophagini, considered endangered in Brazilian red lists. Its previously known geographical distribution range was the Paraná River basin, in Paraguay, and a tributary of the Itaipu Reservoir in Brazil. Since its description no specimens have been collected in the original known distribution area. However, recent records of G. setequedas in the lower Iguaçu River, in a region considered highly endemic for the ichthyofauna, extended the known geographical distribution and may represent one of the last remnants of the species. The aim of this study was to estimate the genetic diversity and population structure of G. setequedas, using microsatellite markers and mitochondrial haplotypes, in order to test the hypothesis of low genetic diversity in this restricted population. Muscular tissue samples of 86 specimens were obtained from nine locations in the Lower Iguaçu River basin, between upstream of the Iguaçu Falls and downstream of the Salto Caxias Reservoir. Seven microsatellites loci were examined and a total of 120 different alleles were obtained. The number of alleles per locus (N_A) was 17.429, effective alleles (N_E) 6.644, expected heterozygosity (H_E) 0.675, observed (H_O) heterozygosity 0.592, and inbreeding coefficient (F_IS) 0.128. Twelve haplotypes in the D-Loop region were revealed, with values of h (0.7642) and π (0.00729), suggesting a large and stable population with a long evolutionary history. Thus, both molecular markers revealed high levels of genetic diversity and indicated the occurrence of a single G. setequedas population distributed along a stretch of approximately 200 km. The pattern of mismatch distribution was multimodal, which is usually ascribed to populations in demographic equilibrium. Nevertheless, the construction of a new hydroelectric power plant, already underway between the Salto Caxias Reservoir and Iguaçu Falls, could fragment this population, causing loss of genetic diversity and population decline, and for this reason it is necessary to maintain the Iguaçu River tributaries and downstream area from the Lower Iguaçu Reservoir free of additional dams, to guarantee the survival of this species.

Color patterns of the body are commonly used to distinguish and identify species of Trichomycterus. Therefore, variation in color pattern in a population can cause doubt concerning species identification. With the purpose to test the hypothesis of high variation in color pattern of Trichomycterus davisi (Haseman, 1911), 118 specimens were collected in a stream of a private Ecological Park in southern Brazil, of which 88 were used in the morphological analysis and 30 for DNA barcoding analysis. Three phenotypic classes were determined analyzing the distribution, size and shape of dark brown spots and blotches. The results of morphometric analysis indicate a tendency of association of those pigmentation patterns with the standard length, evidencing ontogenetic variation of color pattern in the species. The results of K2P intraspecific genetic distance (<0.72%), haplotypes network and Bayesian phylogenetic tree corroborate the existence of only one species with a high variable color pattern.

Freshwater crustaceans of the genus Aegla are endemic to southern South America. In Brazil, the greatest diversity of aeglid species occurs in river basins in the Rio Grande do Sul state. Nevertheless, recent studies have shown that the potential for discovering new species of Aegla in the Paraná state is huge. In recent years, for example, seven new species have been described for the state of Paraná. Furthermore, there is a need to solve problems related to cryptic species and refine taxonomic information about the group. Here we expand our current knowledge of the freshwater biodiversity of Aegla in the region by describing three new species. Aegla nanopedis sp. nov., Aegla abrupta sp. nov., and Aegla santosi sp. nov. were discriminated based on morphological and molecular evidence. Particular morphological characters of the carapace, chelipeds, epimeron and uropods distinguished the new taxa. Molecular results also support the separation of the new species from the closely related species A. jacutinga, A. castro and A. lata. The new species have limited distribution and are categorised as endangered (Aegla nanopedis sp. nov.) and vulnerable (Aegla abrupta sp. nov. and Aegla santosi sp. nov.) according to International Union for Conservation of Nature Red List Categories and Criteria. http://www.zoobank.org/urn:lsid:zoobank.org:pub:7A9A4BB-9B79-429B-B053-34A5D3667CD4

Freshwater crustaceans of the genus Aegla are endemic to southern South America, and present greater richness in the river basins of the state of Rio Grande do Sul, Brazil. However, recent studies have shown the enormous potential for discovering new species of Aegla in the state of Paraná. In the last six year the number of species recorded has almost doubled, increasing from 9 to 16. In the present work, we expand the current knowledge about the diversity of Aegla in a set of poorly explored drainage basins in the western region of the state and describe a new species, Aegla urussanga n. sp., based on morphological and molecular evidence. A combination of morphological characters supports the new species: orbital spine absent, non-elevated epigastric prominences, robust protogastric lobes with scales, anterior dorsal margin of epimeron 2 armed and with pronounced concavity, sub-rectangular palmar crest of chelipeds, internal margin of the ventral face of the ischium of the cheliped with a proximal stout spine, a distal stout spine, and up to three tubercles with spiniform scales. The molecular data, based on partial COI sequences, also support the distinction of the new species from others that occur in adjacent river basins. The species with the smallest genetic distance from A. urussanga n. sp. is Aegla castro Schmitt, 1942 which occurs in the Iguaçu River basin. The new species occurs in distinct streams, probably interconnected in the past and currently isolated by the Itaipu reservoir and is the second species of Aegla described from the complex of drainage basins called Paraná 3 basin.

The southern region of Brazil is characterized by high species diversity and endemism of freshwater fishes distributed across geographically isolated river basins. Microglanis cottoides has a widespread range across these river basins and occurs in sympatry with other endemic species of the genus (e.g. M. cibelae, M. eurystoma, and M. malabarbai). Herein we tested the monophyly of M. cottoides and presented for the first time information about the molecular phylogeny of species in the genus. The results suggest that M. cottoides currently forms a non-monophyletic group which includes populations endemic to the Uruguay River basin that are more closely related to M. malabarbai, and excludes M. cibelae, found to be nested within M. cottoides. Based on an integrative approach using morphological and molecular data, we propose M. cibelae as a junior synonym of M. cottoides, and the populations of the Uruguay River basin previously assigned to M. cottoides in fact belong to M. malabarbai. Our molecular phylogeny shows that M. cottoides is sister to M. parahybae, which is also a coastal species, and M. malabarbai is sister of M. garavelloi, both endemic to inland river basins. The time-calibrated phylogeny indicates that the separation between inland and the coastal clades occurred in the Tertiary period, and that the species within the coastal basins diverged in the Pliocene, which overlaps with the diversification times estimated for the two inland species as well. This pattern of diversification corroborates some previous studies with other fishes from the same region.