Integrative taxonomy reveals the historically poorly defined armoured catfish Hypostomus variipictus (Ihering 1911), from the upper rio Paraná basin, Brazil (Siluriformes, Loricariidae).

In a recent expedition to the rio Grande basin, a tributary of the rio Paraná in southern Brazil, individuals of the armoured catfish genus Hypostomus with a peculiar and beautiful colour pattern composed of pale vermiculations on the head and four to five horizontal stripes on the flanks were collected. Initially, the specimens were identified as a colour morph of the pale-spotted H. margaritifer. However, when we compared their partial cytochrome C oxidase subunit I (COI) mitochondrial gene to sequences of some typically pale-spotted H. margaritifer, the striped specimens were genetically distinct. Further analysis of the striped individuals revealed that they are the poorly known but valid species Hypostomus variipictus, which was described by Ihering in 1911 from the rio Pardo, a tributary of the rio Grande, upper rio Paraná basin, in São Paulo State, Brazil. Since its descriptions, no robust taxonomic work has been published concerning this species. In this study, the newly sampled population was compared to the original description and to the holotype of H. variipictus, providing the foundation for a complete redescription, proper diagnosis, and first live colour illustration and description of the previously hidden H. variipictus.

Genetic evidence for a species complex within the piranha Serrasalmus maculatus (Characiformes, Serrasalmidae) from three Neotropical river basins based on mitochondrial DNA sequences.

Mitochondrial molecular markers (DNA sequences of D-loop, cytochrome b and cytochrome c oxidase I) were employed to characterize populations of the piranha Serrasalmus maculatus from Upper Paraná, Upper Paraguay and Tocantins River basins. D-loop sequences of S. maculatus population from Paraná-Paraguay River basin exhibited tandem repeats of short motifs (12 base pairs) and variable numbers depending on specimens, accounting for length variation. Concatenated mitochondrial sequences suggested that S. maculatus encompasses different mitochondrial DNA lineages. Although sampling was restricted to three river basins, phylogenetic analysis clearly indicated that the species currently recognized as S. maculatus presents high genetic variability. Maximum likelihood and Bayesian analysis clustered S. maculatus populations according to their locations. However, the highest genetic differentiation was identified between populations from Paraná-Paraguay system and Tocantins River basin. Three species delimitation analyses (PTP, GMYC, and ABGD) suggested that there are at least two species among the analyzed populations. The analysis of the mitochondrial sequences evidenced genetic differentiation among populations corresponding to related, but different species, suggesting that at least S. maculatus from the Tocantins River and Paraná-Paraguay River basins are most likely different species. Therefore, S. maculatus should be considered a species complex with morphologically cryptic diversity. An integrative revision is suggested.

Molecular characterization of the invasive aquatic macrophyte Hydrilla verticillata (Hydrocharitaceae) in Brazil.

Invasive populations of macrophytes are widely distributed and have been successfully introduced and established in freshwater habitats. Hydrilla verticillata was first recorded in 2005 in the Upper Paraná River floodplain and in 2007 at the Itaipu Reservoir (Brazil-Paraguay border, ca. 300 km downstream from its first record). However, its genetic variability within different sites in South America is unknown. We used nucleotide sequences corresponding to the trnL-trnF fragment cpDNA to genetically characterize populations of H. verticillata in different ecosystems of the Upper Paraná River basin. The results indicated an absence of genetic differentiation within and between populations of the basin, and even individuals collected 600 km apart belonged to the same haplotype. Moreover, H. verticillata populations of the Upper Paraná River basin also matched the dioecious biotype haplotype of the Southern United States and Asia. The identification of this single haplotype suggests that one founder genotype was introduced and established successfully in the Upper Paraná River basin, then, as a consequence of vegetative reproduction and the dispersal of propagules, spread to different habitats. However, firm conclusions about this inference can only be obtained with markers of biparental inheritance.

Performance of biomarkers metallothionein and ethoxyresorufin O-deethylase in aquatic environments: A meta-analytic approach.

The preservation of natural environments guarantees the conservation of biodiversity and ecosystem processes. Biomonitoring programs in preserved sites can be carried out using molecular biomarkers, which reflect possible stresses that exist in the monitored location. The metallothionein (MT) proteins and isoenzyme Cytochrome P4501A (CYP4501A) are among the most used biomarkers and reflect the detoxification of metal and organic xenobiotics, respectively. This study aimed to assess the performance of these biomarkers in natural aquatic environments using a meta-analytic approach. The data search was conducted in ISI Web of Science™, considering papers published until August 2016. Studies included in this research needed to compare reference or control sites and sites under stress and be conducted in situ. In general, both biomarkers were useful when comparing control sites with sites under stress. Moreover, when the data were categorized into groups of organisms, mainly bivalves and fishes, there were differences between the groups and between the monitored environments, marine or freshwater. The use of these biomarkers in fish is suitable for freshwater environments, and bivalves are suitable for marine environments. We concluded that the concomitant use of vertebrate and invertebrate bioindicators is useful to develop an effective biomonitoring program and to avoid biases due the physiology of the selected bioindicator.

Extraction of DNA from micro-tissue for bat species identification.

Bat populations are declining worldwide. Accurate identification is essential to promote species' conservation. However, minimal morphological differences and a high rate of cryptic species make identification difficult, unless voucher specimens are kept, a controversial issue today. The objective of this work was to standardize a method of extracting non-lethal DNA using bats' uropatagium micro-tissue, aiming the molecular identification of species that occur in the region of Maringá PR. The method standardized was efficient, and does not cause serious damage to bats. For future field studies, collection of micro-tissue and morphometry of the specimens will be sufficient for accurate identification.

Long-wavelength sensitive opsin (LWS) gene variability in Neotropical cichlids (Teleostei: Cichlidae).

Cichlid fishes are an important group in evolutionary biology due to their fast speciation. This group depends widely of vision for feeding and reproduction. During the evolutionary process it plays a significant role in interspecific and intraspecific recognition and in its ecology. The molecular basis of vision is formed by the interaction of the protein opsin and retinal chromophore. Long-wavelength sensitive opsin (LWS) gene is the most variable among the opsin genes and it has an ecological significance. Current assay identifies interspecific variation of Neotropical cichlids that would modify the spectral properties of the LWS opsin protein and codons selected. Neotropical species present more variable sites for LWS gene than those of the African lakes species. The LWS opsin gene in Crenicichla britskii has a higher amino acid similarity when compared to that in the African species, but the variable regions do not overlap. Neotropical cichlids accumulate larger amounts of variable sites for LWS opsin gene, probably because they are spread over a wider area and submitted to a wider range of selective pressures by inhabiting mainly lotic environments. Furthermore, the codons under selection are different when compared to those of the African cichlids.

Long-wavelength sensitive opsin (LWS) gene variability in Neotropical cichlids (Teleostei: Cichlidae)

ABSTRACT Cichlid fishes are an important group in evolutionary biology due to their fast speciation. This group depends widely of vision for feeding and reproduction. During the evolutionary process it plays a significant role in interspecific and intraspecific recognition and in its ecology. The molecular basis of vision is formed by the interaction of the protein opsin and retinal chromophore. Long-wavelength sensitive opsin (LWS) gene is the most variable among the opsin genes and it has an ecological significance. Current assay identifies interspecific variation of Neotropical cichlids that would modify the spectral properties of the LWS opsin protein and codons selected. Neotropical species present more variable sites for LWS gene than those of the African lakes species. The LWS opsin gene in Crenicichla britskii has a higher amino acid similarity when compared to that in the African species, but the variable regions do not overlap. Neotropical cichlids accumulate larger amounts of variable sites for LWS opsin gene, probably because they are spread over a wider area and submitted to a wider range of selective pressures by inhabiting mainly lotic environments. Furthermore, the codons under selection are different when compared to those of the African cichlids.

Molecular markers in studies on fish parasites (Platyhelminthes): Review / Marcadores moleculares aplicados a estudos de Platyhelminthes parasitos de peixes: revisão

ABSTRACT. Studies with molecular markers are currently more common for all groups of living organisms. Molecular techniques used in Platyhelminthes parasites of fishes do not merely reveal complex life cycles, but are important for species distinction and the elucidation of the phylogenetic hypothesis. Current research verified which molecular markers were mainly used phylogenetic studies on Platyhelminthes parasites of fish so that subsidies for further phylogenetic studies in Icthyoparasitology could be provided. Data base of CAPES Journals platform was employed for bibliometric analysis comprising the keywords "fish" and "phylogeny" associated with "Cestoda", "Digenea" or "Monogenea". Information retrieved was quantified and tabulated. Most studies were on Monogenea (43%), followed by Digenea (37%) and Cestoda (18%). Ribosomal molecular markers were the most used in the phylogenetic studies for fish parasites. Due to the advance of molecular biology techniques and of bioinformatics, with more robust phylogenetic analysis, the use of these techniques in other areas such as Ichytioparasitology is on the increase. In fact, molecular phylogenetics and morphological structures analysis have efficiently contributed towards the understanding of phylogenetic relationships among the groups.

Estudos com marcadores moleculares são cada vez mais comuns em todos os grupos de seres vivos. Para os platelmintes parasitos de peixes, as técnicas moleculares possibilitam desvendar ciclos de vida complexos, sendo importantes também na distinção de espécies e na elucidação de hipóteses filogenéticas. Neste sentido, este trabalho teve como objetivo verificar quais são os principais marcadores moleculares utilizados nos estudos de platelmintos parasitos de peixes, visando fornecer subsídios para futuros estudos filogenéticos na Ictioparasitologia. Para a análise bibliométrica foi utilizado o banco de dados dos Periódicos da CAPES, tendo como palavras-chave "fish" e "phylogeny" associadas a "Cestoda", "Digenea" e "Monogenea". As informações obtidas nos trabalhos foram tabuladas e quantificadas. Dos 143 trabalhos obtidos 43% foram com monogenéticos, 37% com digenéticos e 18% com cestoides. Os marcadores moleculares ribossomais foram os mais utilizados nos estudos filogenéticos com estes parasitos de peixes. Com o avanço das técnicas de biologia molecular e da bioinformática, com análises filogenéticas mais robustas, é crescente a utilização destas técnicas na Ictioparasitologia. A filogenética molecular, juntamente com a análise de estruturas morfológicas tem contribuído de maneira mais eficiente para o entendimento das relações de parentesco, entre estes grupos de parasitos de peixes.

Opsin genes: research perspectives with Neotropical cichlids (Perciformes: Cichlidae) and their relevance in floodplain studies / Genes opsins: perspectivas para pesquisas com ciclídeos Neotropicais (Perciformes: Cichlidae) e sua relevância para estudos em planícies de inundação

Vision not only plays an important role in the behavior and exploration capacity of new ecology niches but also influences the evolution of species exposed to the heterogeneity of light. Floodplain environments have high habitat heterogeneity and, thus, different light gradients. Cichlids are a group of vertebrates that has stirred interest in evolutionary studies due to their morphological and behavioral diversity and their widely used vision. The molecular basis of vertebrates' vision occurs through the interaction of opsin proteins and retinal chromophores. Proteins are expressed by opsin genes where each is responsible for absorbing certain light wavelengths. Current review analyzes the main characteristics of opsin genes family and the possibility of using them in floodplain and Neotropical cichlids studies. Opsins may have different levels of expression and molecular polymorphisms according to the dispersion of the species. They are also related to such behavior as sexual selection, nourishment and exploration of new habitats. Floodplains are natural experiments and dynamic environments that provide a wide range of habitats. In fact, the integration of studies in floodplains and the opsin genes in Neotropical cichlids seems to be a promising and still unexplored area in Neotropical regions.

A visão desempenha um importante papel no comportamento e capacidade de explorar novos nichos, e também influencia o processo evolutivo de espécies que estão sujeitas à heterogeneidade de luz. Ambientes de planície de inundação têm alta heterogeneidade de habitats, exibindo diferentes gradientes de luminosidade. Um grupo de vertebrados que desperta interesse para estudos evolutivos são os ciclídeos, pela sua diversidade morfológica e comportamental, além de usarem amplamente a visão. A base molecular da visão dos vertebrados ocorre pela interação de proteínas opsinas e cromóforos retinais. Essas proteínas são expressas pelos genes opsins, cada uma responsável por absorver determinado comprimento de onda de luz. Esta revisão aborda as principais características destes genes e a possibilidade de sua utilização em estudos de planície de inundação e ciclídeos neotropicais. Os opsins podem apresentar diferentes níveis de expressão e polimorfismos de acordo com a dispersão das espécies estudadas. São relacionados com comportamentos de seleção sexual, forrageamento, e capacidade de explorar novos habitats. As planícies de inundação representam experimentos naturais, sendo ambientes dinâmicos e que apresentam vários tipos de habitats. Assim, a integração de estudos de planície de inundação e genes opsins em ciclídeos parece ser uma área promissora e ainda inexplorada em regiões neotropicais.

Does exposure to glyphosate lead to an increase in the micronuclei frequency? A systematic and meta-analytic review.

Glyphosate-based herbicides are among the most used pesticides worldwide. Reviews on the safety of glyphosate have been conducted by several regulatory agencies and researches centers, many times with contradictory results. This study is a systematic meta-analytical review of experimental studies on the relationship between exposure to the glyphosate (GLY) and its formulations with the formation of micronuclei (MN) to establish a quantitative estimate of the environmental risks. The natural logarithm (ln) of the estimated response ratio was calculated from 81 experiments. A meta-analysis was performed on the complete data set, and individual meta-analyses were conducted after stratification by test system, class of vertebrate, exposure route, gender, endpoints, type of literature, formulation, GLY dose and exposure time. A forest plot showed an overall positive association between GLY exposure and its formulations and MN, corroborated by the cumulative effects size. Different responses were observed on mammalian and non-mammalian. Interesting results was noticed in exposure route where oral administration of GLY presented no significance. Exposure by intraperitoneal injection presented the highest MN formation. Pure GLY caused fewer effects than to commercial mixtures, but both presented mutagenic effects. The studies with males presented significant responses, while studies with females were not significant. The cumulative effects size was not clearly related to GLY dose, and was negatively related to exposure time. It can be attributed to different test systems, exposure routes and protocols analyzed. In conclusion, our results support the hypothesis that exposure to GLY and its formulations increases the frequency of MN formation.

Prospecting molecular markers to distinguish Cichla kelberi, C. monoculus and C. piquiti / Prospecção de marcadores moleculares para distinção de Cichla kelberi, Cichla monoculus e Cichla piquiti

Peacock bass, a fish of the genus Cichla, is an exotic species from the upper river Paraná floodplain in which the species Cichla kelberi and C. piquiti have been confirmed, coupled to the specie C. monoculus upstream in the Capivara and Taquaruçu dams. The introduction of this genus has caused negative impacts on the diversity of native species. Current research prospects DNA sequences capable of distinguishing the three species and provide molecular data for the taxonomic characterization of the species in the upper Paraná River basin. Sequencing of nuclear (tmo4c4, dlx2 and bmp4) and mitochondrial (cox1, cytb) loci were done from fish of the three species of the genus Cichla reported in the literature of the upper Paraná River basin. Sequence analysis provided molecular differentiation for the species through the usage of loci cytb, dlx2 and cox1. Since the latter only distinguished C. piquiti from the other Cichla species, the loci bmp4 and tmo4c4 were not adequate to accomplish our aim.

O tucunaré é um peixe pertencente ao gênero Cichla, sendo exótico na planície de inundação do alto rio Paraná, onde foi confirmada a presença das espécies Cichla kelberi e C. piquiti e, logo a montante, C. monoculus, nas represas de Capivara e Taquaruçu. A introdução deste gênero tem ocasionado impacto negativo à diversidade de espécies nativas. Visando providenciar dados moleculares para auxiliar na caracterização taxonômica das espécies deste gênero na bacia do alto rio Paraná, os objetivos deste trabalho foram prospectar sequências de DNA capazes de distinguir as três espécies. Procedeu-se o sequenciamento de loci nucleares (tmo4c4, dlx2 e bmp4) e mitocondriais (cox1, cytb) de peixes das três espécies do gênero Cichla relatadas na literatura da bacia do alto rio Paraná. As análises das sequências possibilitaram a diferenciação molecular para estas espécies pela utilização dos loci cytb, dlx2 e cox1; este último, somente para distinguir C. piquiti das demais espécies de Cichla. A utilização dos loci bmp4 e tmo4c4 não foi adequada para este propósito.

In situ assessment of a neotropical fish to evaluate pollution in a river receiving agricultural and urban wastewater.

We aimed to assess the quality of a midsize river that receives agricultural and urban wastewater. Nuclear abnormalities (NA), comet assays of blood and gills, and gill histopathology were evaluated in fish Astyanax aff. paranae during the summer and winter 2011 at three sites in Paraná State, Brazil: (1) a biological reserve (Rebio-reference area); (2) an agricultural site; (3) a downstream site that accumulates agricultural and urban effluents. We found the highest effects of pollutants in fish at the downstream site during the summer. The agricultural site showed an intermediate damage rate, and fish at Rebio generally had the least damage, with the exception of NA. Despite conflicting results from the biomarkers used, we observed an increase in damage associated with the accumulation of pollutants. Pesticides are probable xenobiotics in the agricultural area. Additionally, metals and substances such as pharmaceuticals and ammonia may be present at the downstream site.

A introdução do gênero Cichla [Block e Schneider, 1801] na planície de inundação do alto rio Paraná / The cichla [Block e Schneider, 1801] genus in floodplains of the upper Paraná river / La introducción del género Cichla [Block y Schneider, 1801] en la llanura de inundación del alto río Paraná

As espécies que compõem o gênero Cichla Block e Schneider, 1801 são piscívoras, possuem grande plasticidade fenotípica e oferecem cuidados para sua prole, motivos estes que as tornam invasoras de alto impacto ambiental. O conhecimento taxonômico e da diversidade genética são importantes para o monitoramento populacional das espécies introduzidas. O objetivo deste trabalho foi caracterizar as espécies e esboçar um histórico da presença do gênero Cichla na planície de inundação do alto rio Paraná evidenciando a problemática de sua introdução nesta região. Análises de diversidade genética em populações introduzidas na bacia do rio Paraná de Cichla kelberi [Kullander e Ferreira, 2006] e C. piquiti [Kullander e Ferreira, 2006] têm demonstrado a origem dos espécimes introduzidos assim como a sua hibridação...

The species constituting the Cichla [Block e Schneider, 1801] genus are piscivorous, present great phenotypic plasticity and provide care for their offspring, reasons that characterise these intruders as of high environmental impact. The taxonomic and genetic diversity knowledge is important for population monitoring of the introduced species. The aim of this study is to characterize the species and draw a history of the presence of the Cichla genus in the floodplain of the Upper Paraná River, showing the problems of its introduction in such region. The analyses of genetic diversity of C. kelberi [Kullander e Ferreira, 2006] and C. piquiti [Kullander e Ferreira, 2006] on populations introduced to the Paraná River basin have demonstrated the origin of specimens introduced as the hybridization of these two species...

Las especies que componen el género Cichla Block y Schneider, 1801 son piscívoras, tienen gran plasticidad fenotípica y ofrecen cuidados a su prole, motivos estos que las vuelven invasoras de alto impacto ambiental. El conocimiento taxonómico y la diversidad genética son importantes para el monitoreo poblacional de las especies introducidas. El objetivo de este trabajo ha sido caracterizar las especies y esbozar un histórico de la presencia del género Cichla en la llanura de inundación del alto Río Paraná evidenciando la problemática de su introducción en esta región. Análisis de la diversidad genética en poblaciones introducidas en la bacía del Río Paraná de Cichla kelberi [Kullander y Ferreira, 2006] y C. piquiti [Kullander y Ferreira, 2006], ha demostrado el origen de los especímenes introducidos, así como su hibridación...

Molecular identification of Cichla (Perciformes: Cichlidae) introduced in reservoirs in Southern Brazil / Identificação molecular de Cichla (Perciformes: Cichlidae) introduzidas nos reservatórios do Sul do Brasil

Species of peacock bass were introduced in several watersheds in South America and worldwide, mainly due to its importance to sport fishing, by being a fighting fish. A recent revision of the genus Cichla showed that the species introduced in reservoirs of the South, Southeast and Northeast regions of Brazil are two new species, described as Cichla kelberi (yellow peacock bass) and Cichla piquiti (blue peacock bass), erroneously identified as C. monoculus and C. ocellaris. With the purpose to identify the populations of Cichla in Paranapanema and Paraná rivers, a total of 323 base pairs (bp) of the mtDNA control region were sequenced, obtained from 84 specimens of Cichla in six different localities (Tapajós river, Solimões river, Capivara, Taquaruçu and Rosana reservoirs in the Paranapanema river, and in the upper Paraná river floodplain). The analyses revealed the genetic diversity of Cichla monoculus, introduced into the Capivara reservoir, originally from the region of Manaus (Amazonas State), and spread in the reservoirs downstream (Taquaruçu and Rosana). The occurrence of the same haplotypes in the three reservoirs suggests one single introduction. This study confirmed the introduction of Cichla in the Capivara reservoir and showed the genetic diversity of Cichla in the Paranapanema river.

Espécies de tucunaré foram introduzidas em inúmeras bacias hidrográficas da América do Sul e em outras regiões do planeta, principalmente pelas suas características esportivas, de peixe lutador. Revisão recente das espécies do gênero Cichla mostraram que as espécies que foram introduzidas nos reservatórios das regiões Sul, Sudeste e Nordeste, são duas espécies novas, descritas como Cichla kelberi (tucunaré amarelo) e Cichla piquiti (tucunaré azul) identificadas erroneamente como C. monoculus e C. ocellaris. Com o objetivo de identificar as populações de Cichla presentes no rio Paranapanema e Paraná, foram sequenciadas um total de 323 pares de bases (pb) da região controle (mtDNA) obtidas de 84 espécime de Cichla em seis localidades diferentes (rio Tapajós, rio Solimões, Reservatórios de Capivara, Taquaruçu, Rosana localizados no rio Paranapanema e na bacia do alto rio Paraná. Os dendrogramas e as análises das populações revelaram fortes evidências de que Cichla monoculus foi introduzida no reservatório de Capivara, proveniente da região de Manaus e se dispersou para os reservatórios localizados a jusante (Taquaruçu e Rosana). A ocorrência dos mesmos haplótipos nos três reservatórios sugerem uma única introdução. Este trabalho confirma a introdução de Cichla no reservatório de Capivara e revela a diversidade genética das espécies presentes no rio Paranapanema.

Molecular variants in populations of Bryconamericus aff. iheringii (Characiformes, Characidae) in the upper Paraná river basin / Variantes moleculares em populações de Bryconamericus aff. iheringii (Characiformes, Characidae) da bacia do alto rio Paraná

There are evidences that Bryconamericus aff. iheringii represents a species complex. DNA molecular markers have been effective in studies on phylogeny, taxonomy, and identification of cryptic species. In this study, partial sequences of genes of ATPase 6 and 8 were used to assess genetic diversity within and among populations of B. aff. iheringii of sub-basins of Tibagi, Pirapó and Ivaí rivers, belonging to the Upper Paraná river basin. The analysis of the sequences of genes pointed out high genetic diversity in B. aff. iheringii from the sub-basins studied with genetic distance values comparable to those found among different species. There was a division of the individuals into five groups. The comparison with other species of Bryconamericus that have sequences available in GenBank confirmed that the individuals studied have relevant values of genetic distance, found among different species. Nevertheless, with the available data it is not possible to refute the hypothesis that the populations correspond to a group resulting from hybridization or that there might have been introgression of mitochondrial DNA among different species.

Há indícios de que Bryconamericus aff. iheringii represente um complexo de espécies. Os marcadores moleculares de DNA têm sido eficazes em estudos de filogenia, taxonomia e identificação de espécies crípticas. Neste estudo, as seqüências parciais de genes da ATPase 6 e 8 foram utilizados para avaliar a diversidade genética dentro e entre populações de B. aff. iheringii das sub-bacias dos rios Tibagi, Pirapó e Ivaí, pertencentes a bacia do Alto Rio Paraná. As análises das seqüências dos genes apresentaram alta diversidade genética em B. aff. iheringii das sub-bacias estudadas, com valores de distâncias genéticas semelhantes às encontradas entre espécies diferentes. Houve uma divisão dos indivíduos em cinco grupos. A comparação com outras espécies de Bryconamericus que têm seqüências disponíveis no GenBank confirmou que os indivíduos estudados possuem valores relevantes de distância genética encontrados entre espécies diferentes. No entanto, com os dados disponíveis não é possível descartar a hipótese de que as populações correspondem a um grupo resultante de hibridação, nem que houve introgressão de DNA mitocondrial entre espécies diferentes.

Genetic structure of the Atlantic Rainforest tree species Luehea divaricata (Malvaceae).

The Atlantic Rain Forest is one of the most important Brazilian biomes and a hotspot for biodiversity that is characterized by its high level of endemism, where new species are still being described. Luehea divaricata (Malvaceae) is commonly found in riparian forests areas of the Atlantic forest. Because of the importance of this species in reforestation programs, we used nine pairs of microsatellite loci to study the genetic variability of this species along its distribution area and verify if fragmentation is compromising the survival of these populations. A total of 50 alleles were obtained with an average observed and expected heterozygosity of 0.53 and 0.67, respectively. Seven of the nine populations studied showed a heterozygosity deficit. Most of the genetic diversity was found within populations; while the level of genetic differentiation was moderated (6.84) between populations. Different levels of gene flow between the populations were detected. Positive and significant values of Fis were found for seven populations. The signal test for excess of heterozygosity indicated that a recent genetic bottleneck occurred in the fragmented populations. The dendrogram constructed by the UPGMA method revealed the formation of seven clusters, which was confirmed by the Bayesian analysis for number of K clusters. The presence of several pairs of loci in linkage disequilibrium confirms that these populations experienced a loss of genetic diversity caused by genetic drift. The results showed that it is necessary to develop management strategies for the conservation of these populations of L. divaricata as the viability of the next generations are severely compromised.

A recent transposition of river involving Paraná and São Francisco basins: effects on the genetic variability and structure of the neotropical fish Parauchenipterus galeatus (Siluriformes, Auchenipteridae).

This study analyzed sequences of the control region of mitochondrial DNA (D-loop) in three populations of Parauchenipterus galeatus from the basins of the Paraná, São Francisco, and Piumhi rivers, of which the last river being a region that suffered transposition river. A fragment of 850 base pairs was obtained with a total of 65 polymorphic sites. The data discuss aspects related to the genetic distance between the populations through the phylogenetic reconstruction methods (neighbor-joining, maximum-likelihood, and Bayesian analysis). Moreover, the data suggest that the Piumhi River population (transposition region) has recently gone through a significant bottleneck effect, which must be directly related to the anthropic action that occurred in this region, since the drainage the old existing swamp was necessary for the construction of the transposition channel potentially leading this population in to the current lack of genetic diversity.

Nucleotide diversity of Hemigrammus cf. marginatus (Characiformes, Characidae) in the upper Paraná river floodplain / Diversidade nucleotídica de Hemigrammus cf. marginatus (Characiformes, Characidae) na planície de inundação do alto rio Paraná

Characidae is the largest and more diversified family from Characiformes and presents several classification problems, with several genera currently allocated as incertae sedis, such as the genus Hemigrammus. The upper Paraná river floodplain is an environment with high fish diversity. There is at least one species of Hemigrammus, however there are divergences among some authors about the number and the identification of the species from this genus. Therefore the goal of this study was to characterize, using a molecular approach, individuals of Hemigrammus from the upper Paraná river floodplain and to compare them with individuals from the type locality of Hemigrammus marginatus, since this is the only species distributed in this floodplain. For this, the DNA was extracted and a partial region from the mitochondrial genes ATPase 6 and ATPase 8 were amplified and sequenced. The results evidenced the existence of two species of Hemigrammus in the floodplain, although impossible to be distinguished only through morphological traits. High nucleotide diversity among individuals from the upper Paraná river in relation to those from the type locality was also observed, indicating that both species of Hemigrammus present in the upper Paraná river floodplain are not Hemigrammus marginatus.

Characidae é a maior e mais diversificada família de Characiformes e apresenta vários problemas de classificação, com inúmeros gêneros alocados atualmente como incertae sedis, dentre estes Hemigrammus. A planície de inundação do alto rio Paraná é um ambiente com alta diversidade de peixes. Existe neste ambiente pelo menos uma espécie de Hemigrammus, entretanto, existem divergências entre alguns autores quanto ao número e a identificação das espécies deste gênero. Portanto, o objetivo deste trabalho foi realizar a caracterização molecular de indivíduos de Hemigrammus da planície de inundação do alto rio Paraná e compará-los com exemplares da localidade-tipo de Hemigrammus marginatus, tendo em vista ser esta a única espécie identificada de Hemigrammus com distribuição na referida planície. Para isso, foi extraído o DNA, amplificado e sequenciado uma região parcial dos genes mitocondriais ATPase 6 e ATPase 8. Os resultados demonstraram a existência de duas espécies de Hemigrammus na planície de inundação do alto rio Paraná, embora impossíveis de serem diferenciadas apenas pelos caracteres morfológicos utilizados atualmente. Alta diversidade nucleotídica entre os exemplares do alto rio Paraná em relação aos da localidade-tipo também foi observada, indicando que ambas as espécies de Hemigrammus presente na planície de inundação do alto rio Paraná não são da espécie Hemigrammus marginatus.

Genetic variability of brycon orbignyanus (valenciennes, 1850) (characiformes: characidae) in cultivated and natural populations of the upper paraná river, and implications for the conservation of the species

This study aimed to evaluate the genetic variability of B. orbignyanus in cultivated and natural populations of the Upper Paraná River, using molecular RAPD markers and mtDNA control region. Specimens were collected in the Paraná River and in the Piracema fish farm in Maringá, State of Paraná, Brazil. RAPD primers produced 82 loci with consistent expression. The population from the Paraná River showed 28 polymorphic loci, whereas the population from the fish farm presented only 12. Data revealed genetic differentiation between the two populations, although not very pronounced. These results were corroborated by the principal coordinate analysis and by neighbor-joining clustering. The alignment of the D-loop sequences of B. orbignyanus indicated the existence of polymorphism only in the natural population. These data could be helpful for the formulation of management strategies and conservation of the genetic diversity of the species.

Identificação do gene mcyA em florações naturais de Radiocystis fernandoi, em um tributário do reservatório de Rosana, Brasil / Identification of the mcyA gene in natural blooms of Radiocystis fernandoi from a tributary of the Rosana reservoir, Brazil

As cianobactérias são conhecidamente produtoras de toxinas. Dentro de uma mesma espécie, podemos encontrar variedades tóxicas e não-tóxicas, impossíveis de diferenciação apenas pela morfologia. A principal toxina produzida pelas cianobactérias é a microcistina. Esta proteína é biossintetizada por um grupo de genes conhecidos como mcy. A detecção destes genes a partir de PCR permite a distinção das variedades tóxicas e não-tóxicas. Desse modo, o objetivo desse trabalho foi investigar a ocorrência de florações produtoras de toxinas em um rio tributário do reservatório de Rosana, via amplificação do gene mcyA por PCR. Foram coletadas duas amostras de água da subsuperfície. As duas amostras coletadas no rio do Corvo foram dominadas pela espécie Radiocystis fernandoi e apresentaram resultados positivos para a presença do gene mcyA, confirmando o potencial tóxico dessa espécie. Os resultados representam alerta sobre a qualidade da água do rio do Corvo. A técnica PCR foi eficiente para a rápida detecção de cianobactérias produtoras de toxinas, inclusive podendo ser utilizada antes mesmo do agravamento das condições ambientais pela produção de toxinas, além de apresentar baixo custo.

Cyanobacterias are known as toxin producers. Within the same species, toxic and non-toxic varieties can be found and it is impossible to differentiate them only by morphology. The most important toxin produced by cyanobacteria is microcystin. This protein is synthesized by a cluster of genes known as mcy. The detection of these genes by PCR allows the differentiation of the producing toxin strain from the non-producing toxin strain. Thus, the goal of this work was to investigate the occurrence of toxigenic blooms of cyanobacteria in the Corvo River through PCR amplification of mcyA gene. For this, two samples of blooms of cyanobacteria were collected in Corvo River. Both samples were dominated by Radiocystis fernandoi and presented positive results for the presence of the mcyA gene, which may confirm the potential toxigenicity for that species. These results are an alert about water quality in the Corvo River. Here we demonstrate that amplification of the mcyA gene by PCR is a fast, cheap and efficient method for the detection of toxin- producing cyanobacteria.