Cytogenetic characterization of F1, F2 and backcross hybrids of the Neotropical catfish species Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum (Pimelodidae, Siluriformes)

The cytogenetic characteristics of Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum and their F1, F2 and backcross hybrids were assessed by using chromosome banding techniques. The diploid number of 56 chromosomes was constant in all species and lineages, with a karyotypic formula containing 20 metacentric, 12 submetacentric, 12 subtelocentric and 12 acrocentric chromosomes. Nucleolar organizer regions (NORs) were identified in two subtelocentric chromosomes in the parents and hybrids, with partial nucleolar dominance in F1 and F2 specimens. Heterochromatic blocks were detected in the terminal and centromeric regions of some chromosomes in all individuals. For parental and hybrid lineages, 18S ribosomal clusters corresponding to NORs and 5S ribosomal genes were identified in distinct pairs of chromosomes. The striking conservation in the chromosomal macrostructure of the parental species may account for the fertility of their F1 hybrids. Similarly, the lack of marked alterations in the chromosomal structure of the F1 hybrids could account for the maintenance of these features in post-F1 lineages

Tracing the genetic impact of farmed turbot Scophthalmus maximus on wild populations

The impact of escapees from aquaculture is of general concern for the sustainability of natural resources. Turbot Scophthalmus maximus is a marine flatfish of great commercial value whose land-based aquaculture started approx. 40 yr ago; hence, a low impact of escapees is expected on wild populations. However, enhancement of wild stocks using farmed turbot has been carried out along the Northeast Atlantic coasts in the last decades. Recently, a broad panel of single nucleotide polymorphism (SNP) markers (755 SNPs; 1 SNP Mb−1) has been used to evaluate the genetic structure of turbot throughout its distribution range, constituting the baseline to evaluate the impact of farmed fish in the wild. Two distinct origins were identified for farmed turbot (F_ORI1 and F_ORI2; FST = 0.049), which differentiated from wild populations after 5 generations of selection (average FST = 0.059), and consistent evidence of adaptation to domestication was de - tected. A notable proportion of fish of farmed ancestry was detected in the wild (15.5%), mainly in the North Sea, where restocking activities have taken place, determining genetic introgression in wild populations. Conversely, effects of land-based aquaculture appear negligible. A simulation exercise supported panels of 40 and 80 SNPs to identify fishes of F_ORI1 and F_ORI2 ancestry in the wild, respectively. Application to empirical data showed an assignment success (wild/farmed ancestry) of approx. 95% in comparison with the full SNP dataset. The SNP tools will be useful to monitor turbot of farmed ancestry in the wild, which might represent a risk, considering the lower fitness of farmed individualsThe project was funded by the 7th Framework Programme for research (FP7) under ‘Knowledge-Based Bio-Economy — KBBE’, Theme 2: ‘Food, Agriculture and fisheries, and Biotechnologies’ Project identifier: FP7-KBBE-2012-6-singlestage Grant agreement no.: 311920 ‘The development of tools for tracing and evaluating the genetic impact of fish from aquaculture: AquaTrace’ and the Spanish Regional Government Xunta de Galicia GRC2014/010. Ciência sem Fronteiras/CAPES − Brazil supported the fellowship for the stay of F.D.P. at USCS

IDENTIFICACIÓN MOLECULAR Y CARACTERIZACIÓN MORFOLÓGICA DE LARVAS L3 DE ANISAKIS SPP. (NEMATODA: ANISAKIDAE) EN SCOMBER COLIAS GMELIN, 1789 (PERCIFORMES: SCOMBRIDAE) DEL NORTE DE ARGENTINA

Se realizó la identificación molecular de larvas de nemátodos en el tercer estadío del Scomber colias Gmelin, 1789 recogidos en la costa norte de Argentina, y la caracterización morfológica y morfométrica de estos parásitos. Los órganos internos y los músculos de 31 muestras de S. colias fueron analizadas. La identificación genética de larvas se realizó por PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) mediante la amplificación de un fragmento de ADN nuclear (ITS1, ITS2 y 5,8S) y el escote con enzimas de restricción específicas (HinfI, HhaI) para la identificación. Para el análisis morfológico y morfométrico, los especímenes se observaron en el microscopio trinocular y en microscopía electrónica de barrido. Alrededor de 369 nematodos fueron recolectados en el 68% de los peces analizados. Todos los parásitos se encontraron en los órganos internos del huésped, y no se detectó formas larvarias en los músculos. La identificación molecular produjo cuatro diferentes patrones Anisakis pegreffii Campana-Rouget & Biocca, 1955; Anisakis typica (Diesing, 1860); un híbrido de Anisakis spp; y otras larvas que no muestran patrones moleculares correspondientes a Anisakis. Se observaron diferencias morfológicas y morfométricas entre las larvas de A. pegreffii y A. typica. Excepto el diente larvario, todas las estructuras de A. pegreffii tenían dimensiones mucho mayores que los encontrados en A. typica. Además, el extremo posterior de A. pegreffii se estrecha más lentamente y el mucrón acompaña esta conicidad. En A. typica el extremo posterior es más robusto y el mucrón es muy afilado. Esta es la primera contribución a la identificación molecular con la caracterización morfológica de Anisakis spp. en S. colias de Argentina

Molecular identification of Pseudoplatystoma sp. fish fillets by Multiplex PCR

Nuclear and mitochondrial genes were used as molecular markers for verifying the iden-tity of fish fillets marketed as pintado (Pseudoplatystoma corruscans). Based on THE polymorphisms of nuclear DNA (RAG2, globin, and EF1 genes) and mitochondrial regions (16S), we examined whether the fillets originated from inbred species of pintado or from hybrids derived from crosses between cachara (P. reticulatum) and pintado. Nuclear genes from both species were detected in the analyzed fillets (n = 29). This clearly iden-tified these fish as interspecific hybrids (or F1/first filial generation) of the type “cacha-pinta,” resulting from a cross between female cachara and male pintado. These results demonstrate that monitoring fish fillet trading is crucial for detecting discrepancies between the marketed species and related information declared on the label. Species that are frequently hybridized, such as pintado and cachara, require special attention.Título PT: Identificação molecular de filés de peixe Pseudoplatystoma sp. por PCR-MultiplexMarcadores moleculares (PCR-Multiplex de genes nucleares e mitocondriais) foram utilizados para verificar a identidade molecular de filés de peixe comercializados como pintado (Pseudoplatystoma corruscans) com base em polimorfismos de regiões do DNA nuclear (genes RAG2, globina e EF1) e mitocondriais (16S) para verificar se os filés pertenciam a espécie pura de pintado ou se eram híbridos derivados do cruzamento entre cachara (Pseudoplatystoma reticulatum) e pintado (Pseudoplatystoma corruscans). Os filés analisados (n = 29) apresentaram genes nucleares de ambas espécies P. corruscans e P. reticulatum, e desta forma, foram identificados como híbridos interespecíficos ou F1 (primeira geração filial) do tipo “cachapinta” resultante do cruzamento entre uma fêmea de cachara e um macho de pintado. Estes resultados mostram que o monitoramento da comercialização de filés de peixe é fundamental para identificar situações onde existem diferenças entre as espécies comercializadas e as informações declaradas no rótulo. Espécies em que a hibridação tem sido frequentemente realizada como é o caso do pintado e da cachara, merecem atenção especial

Caracterização citogenética e molecular das espécies pintado (Pseudoplatystoma corruscans), cachara (Pseudoplatystoma reticulatum) e seus híbridos utilizados na piscicultura brasileira

A hibridação artificial interespecífica de peixes é utilizada em diversos estabelecimentos voltados à piscicultura no país, com a finalidade de produzir indivíduos mais vantajosos e favoráveis para o cultivo. Porém, devido principalmente às dificuldades encontradas na identificação dos híbridos, esta prática pode determinar o surgimento de sérios problemas como a contaminação genética dos estoques de cultivo, a comercialização de produtos híbridos como espécies puras, a introdução de espécies exóticas e escapes de produtos de piscicultura para o ambiente natural e até eventos de introgressão genética e extinção das espécies nativas. Neste contexto, o presente trabalho teve por objetivo analisar geneticamente exemplares das linhagens parentais de Pseudoplatystoma corruscans (pintado) e Pseudoplatystoma reticulatum (cachara) e seus híbridos interespecíficos pintachara (macho de pintado x fêmea de cachara) e cachapinta (fêmea de cachara x macho de pintado), provenientes dos estoques de cultivo do CEPTAlICMBio, Pirassununga, SP, a fim de identificar e estabelecer marcadores genéticos para possibilitar sua identificação e diferenciação. Também foram analisadas geneticamente amostras de P. corruscans e P. reticulatum coletadas no Rio Paraguai, MS (bacia do Paraguai) e de P. corruscans capturados do rio Mogi- Guaçu, SP (bacia do Alto Paraná), a fim de identificar a possível ocorrência de híbridos entre estas espécies na natureza. As análises citogenéticas revelaram um número diploide de 2n=56 cromossomos para ambas as espécies parentais, com cariótipos caracterizados por cromossomos dos tipos 20m+12am+12st+12a e número fundamental (NF) igual a 100, indicando uma fórmula cariotípica conservada entre estas espécies. A análise dos padrões de heterocromatina pelo bandamento C revelou blocos heterocromáticos localizados nas porções...Artificial interspecific hybridization of fish is used in various establishments linked to fish farming in the country, in order to produce individuais more advantageous and favorable for cultivation. However, due mainly to difficulties in the hybrid products identification, this practice may determine the onset of serious problems such as genetic contamination of the breeder stocks, marketing of hybrids as pure species, introduction of exotic species and escapes of farmed products to the natural environment and sometimes leading to events of genetic introgression and extinction of native species. In such context, the present study aimed to analyze genetically copies of the parental lines of Pseudoplatystoma corruscans (pintado) and Pseudoplatystoma reticulatum (cachara) and their interspecific hybrids pintachara (female of pintado x male of cachara) and cachapinta (female of cachara x male of pintado), from the stocks manteined in CEPTAlICMBio, Pirassununga, SP, in order to characterize and establish genetic markers to allow their identification and differentiation. Samples of P. corruscans and P. reticulatum collected in the Paraguay river, MS (Paraguay basin) and P. corruscans captured in Mogi-Guaçu river, SP (Alto Paraná basin), also were genetically analyzed in order to identify the possible occurrence of hybrids between these species in nature. The cytogenetic analysis revealed a diploid number of 2n = 56 chromosomes for both parental species with karyotypes characterized by the formula 20m+12am+12st+12a and fundamental number (NF) of 100, indicating a karyotypic formula conserved among these species. The analysis of the heterochromatin C-banding patterns revealed heterochromatic blocks located in the pericentromeric and terminal portions of some chromosomes, the NOR was sim pie and located in one chromosome pai r and 5S ribosomal genes located on two different subtelocentric... (Complete abstract click electronic access below)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Marcadores moleculares na identificação de híbridos e introgressão genética em populações de Pseudoplatystoma corruscans e Pseudoplatystoma reticulatim

Levels of genetic diversity and structure, as well as the investigation of interspecific and introgressive hybridization events in wild populations, are amongst the major analyzes to be considered in conservation projects. Hybridization between native fish species, especially when caused by human actions, has led to increasing rates of non natural crosses between different taxa, which may cause ecological and genetic impacts. The Neotropical catfish species, Pseudoplatystoma reticulatum and Pseudoplatystoma corruscans, have been affected by the induced production of hybrids in aquaculture and by the contact of these hybrids with their populations. In this study, different genetic markers were developed (genes and microsatellites), in order to identify the occurrence of hybridization and introgression in South American rivers and to access the intra and inter populational genetic diversity of these species. EF1 and 18S nuclear gene markers were completely diagnostics in the identification of the species and their hybrids. Through a Roche 454 pyrosequencing, were developed and characterized 16 microsatellite loci for P. reticulatum, which presented 100% of transferability in P. corruscans and high polymorphism for the majority of the loci. Population analysis using these microsatellites revealed high variability for the both species in all rivers from Paraguay and Paraná hydrographic basins. The interpopulational genetic differentiation of these species within the Paraguay basin was low or absent, revealing a high gene flow and the possible existence of a panmitic population, resulting from the migration and reproduction of individuals from closely rivers. However, significant genetic differences were observed between geographically distant rivers and distinct hydrographic basins. In P. reticulatum, genetic differentiation was found between the Cuiabá and Miranda Rivers (Paraguai basin), and between Paraguay and Lower Paraná ...Níveis de diversidade e estruturação genética, assim como a investigação de eventos de hibridação interespecífica ou introgressiva em populações selvagens, estão entre as principais análises a serem consideradas em projetos de conservação. A hibridação entre espécies nativas de peixes, principalmente quando decorrente de ações humanas, tem levado a taxas crescentes de cruzamentos não naturais entre diferentes taxa, o que pode ocasionar impactos ecológicos e genéticos. As espécies de bagres Neotropicais, Pseudoplatystoma reticulatum e Pseudoplatystoma corruscans, têm sido afetadas pela produção induzida de híbridos na aquicultura e pelo contato destes híbridos com suas populações. Neste estudo foram desenvolvidos diferentes marcadores genéticos (genes e microssatélites) com o intuito de identificar a ocorrência de hibridação e introgressão em rios da América do Sul e para acessar a diversidade genética intra e interpopulacional destas espécies. Marcadores dos genes nucleares EF1 e 18S foram completamente diagnósticos na identificação das espécies e seus híbridos. Através de um pirosequenciamento Roche 454, foram desenvolvidos e caracterizados 16 microssatélites para P. reticulatum, os quais apresentaram 100% de transferabilidade em P. corruscans e alto polimorfismo para a maioria dos loci estudados. As análises populacionais utilizando estes microssatélites revelaram alta variabilidade para ambas espécies em todos os rios das bacias hidrográficas do Paraguai e Paraná. A diferenciação genética interpopulacional destas espécies dentro da bacia do Paraguai foi baixa ou inexistente, revelando um alto fluxo gênico e a possível existência de uma população panmítica resultante da migração e reprodução entre indivíduos de rios próximos. Porém, diferenças genéticas significativas foram observadas entre rios distantes geograficamente e entre bacias hidrográficas ...Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Genetic identification of interspecific hybrid of Neotropical catfish species (Pseudoplatystoma corruscans vs. Pseudoplatystoma reticulatum) in rivers of Mato Grosso do Sul State, Brazil

Hybrids of the species Pseudoplatystoma corruscansvs.Pseudoplatystoma reticulatum are extensively traded because they are more docile and precocious compared to their parents. This study aimed to identify the occurrence of hybrid 'Surubim' in rivers of the State of Mato Grosso do Sul, applying molecular techniques. Pseudoplatystoma corruscans, Pseudoplatystoma reticulatum and hybrid 'Surubim' were detected using multiplex PCR and PCR-RFLP in RAG2, GLOBIN, EF1α, 18S rRNA nuclear genes, and mitochondrial 16S rRNA gene. The results revealed the presence of F1 hybrids in rivers and the occurrence of genetic introgression, evidenced by the Post-F1 hybrids. Furthermore, discrepancies between the morphological and genetic identification were observed, demonstrating the effectiveness of using different molecular markers for the correct identification of these hybrids. The necessity of implementation of management and conservation projects to maintain the genetic integrity of native species populations threatened by hybridization in the rivers of Mato Grosso do Sul State is evident

Detection of post-F1 fish hybrids in broodstock using molecular markers: Approaches for genetic management in aquaculture

Herein, we have developed molecular markers for nuclear genes to use in multiplex-PCR and PCR-RFLP, with the goal of characterising hybrid lines derived from crosses between pintado Pseudoplatystoma corruscans and cachara P. reticulatum. These markers, together with others described previously, were used to perform molecular identification analyses as genetic subsidies for Brazilian aquaculture. These analyses were performed due to the problems of high mortality in the offspring reported by the aquaculturist. From a total of 16 broodstock samples, 13 were genetically identified as hybrids; surprisingly, nine of these hybrids were found to be post-F1 lineages. These data show that the fertility of these animals can seriously affect the cultivated stocks, thus causing financial damage in this aquaculture system. The establishment of PCR-RFLP and multiplex-PCR as molecular techniques allows for both the correct management of these animals and the routine monitoring of production and trade of fish hybrids in aquaculture. Consequently, such tools will enable a sustainable development in the aquaculture industry. © 2012 Blackwell Publishing Ltd