Genetic characterization of native and invasive Plagioscion squamosissimus (Perciformes, Sciaenidae) populations in Brazilian hydrographic basins.

The genetic diversity of invasive and native populations of Plagioscion squamosissimus (Heckel, 1840) from the Paraná, Parnaiba and Araguaia-Tocantins river basins was assessed by using Random Amplified Polymorphic DNA (RAPD) markers. Genetic data confirmed the hypothesis of low genetic variability within and among P. squamosissimus populations introduced in the upper Paraná River basin, and indicated that they likely originated from a common ancestor. Moreover, the data demonstrated that, in agreement with available historical records, the P. squamosissimus populations established in the Paraná River basin were derived from a population native to the Parnaíba River basin. The genetic data presented here are of potential future application for the management of the invasive P. squamosissimus populations and for the preservation of the genetic legacy of native fish.

Genetic divergence among invasive and native populations of Plagioscion squamosissimus (Perciformes, Sciaenidae) in Neotropical regions.

The genetic divergence among invasive and native populations of Plagioscion squamosissimus from four Neotropical hydrographic basins was assessed using the hypervariable domain of the mitochondrial DNA (mtDNA) control region. Plagioscion squamosissimus is native to the neighbouring hydrographic basins of the Parnaíba and Amazon Rivers, and the latter includes the Araguaia-Tocantins drainage, but it is invasive in other basins due to introductions. The mtDNA nucleotide polymorphism supported the hypothesis that the Amazon and Parnaíba populations constitute the same species and are separated into two independent evolutionary lineages. Absence of nucleotide polymorphism was observed within and among P. squamosissimus populations invasive to the uppper and middle Paraná River basins. Nucleotide divergence was null or low comparing the Paraná invasive populations with the populations native to the Parnaíba River basin, whereas it was significantly high compared to Tocantins populations. These results ascertain that P. squamosissimus populations invasive to the upper Paraná River basin and to the middle Paraná River basin downstream of the Itaipu dam are derived from the Parnaíba River basin. The genetic data presented are potentially useful to assist further studies on P. squamosissimus taxonomic and geographic distribution, development of ecological guidelines for managing populations invasive to the upper Paraná River basin and for preservation of native fish diversity.

Mitochondrial D-loop nucleotide diversity in Astyanax (Osteichthyes, Characidae) from the upper Paraná and upper Paraguay River basins.

The morphological discrimination between the species Astyanax altiparanae and A. asuncionensis of the upper Paraná River and Paraguay River basins, respectively, has always been difficult. Two D-loop haplogroups of A. altiparanae are known, one with the presence (AltoPR) or the absence (AltoPR-D) of a 32-bp block similar to that in A. asuncionensis. We examined these samples to characterize A. altiparanae and verify whether A. asuncionensis occurred in the upper Paraná River prior to the submergence of the Sete Quedas Falls when Itaipu reservoir was impounded. D-loop sequences were analyzed in A. altiparanae of the upper Paraná and Iguaçu Rivers and those of A. asuncionensis of the upper Paraguay River. The haplogroup AltoPR was found at all sites of the upper Paraná and Iguaçu Rivers, whereas AltoPR-D occurred in the Itaipu reservoir, floodplain and in the Tietê and Grande Rivers. Two haplogroups of A. asuncionensis were identified and both did not have the 32-bp block. However, AltoPR and AltoPR-D differed from one another in 5.1% of their bases and between 8.9 and 12.5% with regard to the haplogroups of the upper Paraguay basin. Further, AltoPR-D occurred in the Grande River upstream the Marimbondo Falls and other older reservoirs than Itaipu. The results reject the hypothesis of the establishment of A. asuncionensis and suggest that the haplogroup AltoPR-D existed in the upper Paraná River before the impounding of the Itaipu reservoir. Moreover, morphological similarity and high genetic variation within the altiparanae/asuncionensis group suggest the existence of a cryptic species complex.

Molecular characterization and phylogenetic relationships among species of the genus Brycon (Characiformes: Characidae) from four hydrographic basins in Brazil.

Brycon is one of the main genera of Neotropical freshwater fish. In Brazil, Brycon species have been found in many hydrographic basins, such as the Amazon, Paraná, Paraguay, and Araguaia-Tocantins basins. We examined the phylogenetic relationships among the species Brycon orbignyanus, B. hilarii, B. cf. pesu, B. cephalus, B. falcatus, and B. gouldingi, using mitochondrial and nuclear molecular markers. Specimens of B. orbignyanus were collected in the Paraná River. Specimens of B. hilarii were collected in the Manso River. Specimens of B. cephalus were obtained from a fish farm, and specimens of B. cf. pesu, B. falcatus and B. gouldingi were sampled in the Araguaia-Tocantins basin. DNA extraction was carried out using the phenol/chloroform method. Molecular polymorphism studies of Brycon species were carried out with the inter-simple sequence repeat (ISSR) technique, using the total DNA of six specimens of each species. In DNA amplification of B. cf. pesu, eight specimens were used. The partial sequence of mitochondrial cytochrome b was amplified by PCR. The PCR products were used directly in sequencing reactions. Each ISSR primer produced from 7 to 14 scorable and reproducible bands. The (GGAC)(3)A and (GGAC)(3)C primers produced the greatest number of species-specific bands. A 264-bp fragment, corresponding to the partial region of mitochondrial DNA cytochrome b, was sequenced and used for analysis. According to the phylogenetic tree obtained from the data, these Brycon species can be divided into two clades: one comprised only B. cf. pesu, and the second comprised the remaining Brycon species. We conclude that ISSR primers can be used for the identification of species-specific bands in fish, such as Brycon spp.

Effects of flooding regime on the feeding activity and body condition of piscivorous fish in the Upper Paraná River floodplain.

Flood pulses affect floodplain enrichment via the incorporation of nutrients and terrestrial biomass. As a result, they positively affect the body condition of aquatic organisms. This paper evaluates whether the absence of floods (resulting from dam control) affects the feeding activity and body condition of piscivorous fish. In addition, whether piscivores respond similarly to alterations in the flooding regime was assessed. Five piscivorous species were selected (Acestrorhynchus lacustris, Hoplias aff. malabaricus, Plagioscion squamosissimus, Rhaphiodon vulpinus, and Salminus brasiliensis). The fish were captured in four distinct years and in three river subsystems with differentiated water level fluctuations (Ivinheira = not regulated; Baía = regulated by the Paraná River level; Paraná = regulated by dams). Feeding activity and body condition were evaluated using the mean values of the standard residuals generated by regression models between body and stomach weights and standard length and body weight (all log-transformed). Differences among years and subsystems were evaluated via two-way analysis of variance. Pearson and Spearman correlations were performed between flooding attributes (duration, amplitude, timing, and daily variability) and feeding activity and body condition. Feeding activity differed across subsystems, whereas body condition varied across years, depending on the subsystem. Hoplias aff malabaricus (an ambusher adapted to starvation) presented feeding activity independent of the flooding regime and also presented better body condition in times of high water levels. Rhaphidon vulpinus exhibited variations in feeding activity but did not present alterations in body condition. The other species presented poorer body condition in years or subsystems with regular floods. Correlations identified that the duration and timing of floods had negative effects on body condition, whereas amplitude and mean annual water level improved feeding activity. Therefore, regular floods affect the feeding activity and body condition of piscivorous fish, and the response of each species depends on the existence or absence of pre-adaptation to long periods of starvation. Consequently, considering the role of the flood pulse in determining the availability of feeding resources for early life stages of piscivores, control of the flood pulse by dams is also disadvantageous for piscivorous fish.

Inter-annual variations in the abundance of young-of-the-year of migratory fishes in the Upper Paraná River floodplain: relations with hydrographic attributes.

In this study, we identified and characterized the hydrographic attributes related to the success of recruitment of migratory fishes in the Upper Paraná River floodplain. To achieve our objectives, we analyzed inter-annual variations in the abundance of young-of-the-year (YOY; index of recruitment) of six migratory species and their relations with hydrographic attributes. Recruitment was related to the intensity, duration (in different fluviometrical levels), elasticity, number of pulses, greater uninterrupted overflow and delay of the floods (all obtained using the PULSO software). Collections of fish were conducted in the period between January 1987 and November 2007 in distinct environments (river channels, secondary channels and connected and disconnected floodplain lakes) distributed along three subsystems (Paraná, Baía and Ivinheima). Relations between recruitment and the attributes of interest were determined through analysis of covariance. In the studied period, the highest abundances of YOY were registered in 2007, followed by 1992, 1993, 2005 and 1988. The abundance of YOY was positively correlated with an intensity of high water levels (potamophase) and the duration of potamophase 1 and negatively with the duration of low water levels (limnophase) and a delay of flood. Higher hydrometric levels (540 and 610 cm for Paraná and 325 and 450 cm for Ivinheima) and greatest uninterrupted overflow presented different relations (significant interactions) among subsystems, but all with positive effects on recruitment. Results evidenced that recruitment responded better when floods started in January with potamophase intensities above 610 cm and water levels above 450 cm over a period of 50 days and repeated every two years (or > 610 cm for 38 days and repeated every two or three years). Therefore, artificial control of the floods at intervals of two or three years by manipulating the discharge of dams located upstream from the floodplain in a way that promotes potamophases with the potential to ensure recruitment of migratory species may become an important tool for conservation of migratory fish species in the floodplain.

Genetic divergence between Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum (Siluriformes: Pimelodidae) in the Paraná River Basin.

Pseudoplatystoma corruscans (Spix and Agassiz, 1829) and Pseudoplatystoma reticulatum (Eingenmann and Eigenmann, 1889) are large migratory catfishes of high biological importance and great commercial value in South America. Because fertile crossbreeds can be artificially produced in hatcheries, a high genetic proximity between these two Pimelodidae species is conceivable. Possible escape of crossbred specimens from pisciculture stations is a serious environmental concern. Despite their importance, knowledge of P. corruscans and P. reticulatum biology, ecology, population diversity and genetics is limited. In the present work, the genetic divergence between P. corruscans and P. reticulatum populations from the Paraná River Basin was analyzed on the basis of polymorphisms in ISSR fragments and in the hypervariable sequence of the mitochondrial DNA (mtDNA) control region. Estimates of intraspecific haplotype (h > 0.5) and nucleotide diversities (pi < 0.01) indicate that P. corruscans and P. reticulatum have survived a historical population decline, followed by a demographic expansion. The interspecific polymorphisms within the mtDNA control region and ISSR fragments were suitable as diagnostic molecular markers and could be used to discriminate the two species. A unique Pseudoplatystoma specimen, captured in the Upper Paraná River Floodplain, was identified by these DNA diagnostic markers as a hybrid P. reticulatum x P. corruscans, which possibly escaped from pisciculture. The integrity of the natural population of P. corruscans in the Upper Paraná River is at risk of genetic introgression or homogenization due to the presence of hybrids and the transposition of P. reticulatum upstream through the Canal da Piracema at Itaipu Dam. Data presented herein improve the understanding of the genetic relatedness between P. corruscans and P. reticulatum and represent potential tools for future programs of conservation and surveillance of genetic introgression events and the genetic integrity of these populations.

Molecular variability in Brycon cf. pesu Müller and Troschel, 1845 (Characiformes: Characidae) from the Araguaia-Tocantins Basin.

Brycon pesu is a small-sized fish distributed throughout the Amazon and Orinoco Basins and other coastal basins of northeastern South America. Brycon cf. pesu specimens from the Araguaia-Tocantins Basin are currently separated into two morphotypes, Brycon sp1 and Brycon sp2, owing to different coloration of their anal fin. Brycon sp2 has a reddish margin stripe on the anal fin which morphologically distinguishes it from Brycon sp1. In the present research, nuclear and mitochondrial markers were used to test the hypothesis that the Brycon sp1 and Brycon sp2 morphotypes are distinct species. Specimens from the two morphotypes were collected from the Lajeado Hydroelectric Plant and the Palmas River in the Araguaia-Tocantins Basin. Thirty-five loci obtained by the amplification of five inter-simple sequence repeat primers were analyzed but no species-specific bands were detected. Electrophoretic profiles obtained from 5S rDNA non-transcribed spacer amplification failed to show any differentiation in morphotypes. These results were corroborated by nucleotide sequence analysis of the mtDNA control region, in which 24 polymorphic nucleotide sites, representing a polymorphism rate of only 5%, were detected. The low rates of polymorphism detected by inter-simple sequence repeat, non-transcribed spacer and mtDNA D-loop markers strongly reject the hypothesis that the two morphotypes Brycon sp1 and Brycon sp2 represent distinct species within Brycon cf. pesu. Further studies are needed to obtain conclusive data on the notion that the coloration of the anal fin is an intraspecific polymorphism, possibly related to environmental factors.

Diversity and genetic distance in populations of Steindachnerina in the upper Paraná river floodplain of Brazil.

Whereas four species of the genus Steindachnerina occur in the Paraná river basin, S. insculpta was the only endemic species of the region under analysis, which is the third lower section of the upper Paraná river. Among other factors, this species has been characterised by the absence of spots in the basal region of the dorsal fin. However, various specimens with this characteristic appeared in the region after the construction of the Itaipu Hydroelectric Plant in 1982. An analysis of the genetic variability of Steindachnerina populations with or without spots is provided. Specimens were collected in different sites of the floodplain of the upper Paraná river and samples were compared by random amplified polymorphic DNA (RAPD) technique and morphological analyses. Ninety-eight amplified loci with nine random primers were analysed in 19 specimens of each phenotype. Data for genetic distance showed great divergences between the two phenotypes and indicate two different species. Spotted specimens may be identified as S. brevipinna, found in the region downstream Sete Quedas Falls. The species must have overcome the geographical barrier during the building of the Itaipu hydroelectric dam that submerged the waterfalls and which became an obstacle between the upper and middle Paraná river some 150 km downstream. Since phenotypes do not share dominant alleles, absence of gene flow has been suggested.