Abstract Marine turtle hybridization is usually sporadic and involves reports of only a few individuals; however, Brazilian populations have high hybridization rates. Here we investigated the presence of hybrids in morphologically identified immature hawksbills (Eretmochelys imbricata) along the South Western Atlantic (SWA). We sequenced one mitochondrial (D-Loop) and three nuclear DNA (RAG1, RAG2, and CMOS) markers to better understand the patterns and characteristics of hybrids. We identified 22 hybrids (n = 270), 11 of them at the extreme South of the SWA. Uruguay had the highest hybrid frequency in the SWA (~37.5%) followed by southern Brazil with 30%. These are common areas for loggerheads (Caretta caretta) but uncommon for hawksbills, and these hybrids may be adopting the behavior of loggerheads. By analyzing nuclear markers, we can infer that 50% of the sampled hybrids are first generation (F1) and 36% are the result of backcrosses between hybrids and pure E. imbricata (> F1). We also report for the first time immature E. imbricata x Lepidochelys olivacea hybrids at the Brazilian coast. Considering the high frequency of hybrids in the SWA, continuous monitoring should be performed to assess the fitness, genetic integrity, and extent of changes in the gene pools of involved populations.

ABSTRACT Microbiota, the microorganisms that colonize living organisms. Oysters rely exclusively on an innate immune system. This study characterized the cultivable gastrointestinal tract microbiota (GTM) of the oyster Crassostrea gasar and evaluated their influence on immune responses. Antibiotics were used to reduce or alter the microbiota during in vitro and in vivo assays. Haemocyte viability, concentration, and phagocytic capacity, as well as the production of reactive oxygen species (ROS), were evaluated in antibiotic-treated and control oysters. Microbiological analysis of the gastrointestinal tract was also performed; bacteria were selected using culture media and were identified by 16S ribosomal DNA. The results showed that the antibiotics eliminated bacteria in vitro but increased their concentrations in vivo. The GTM was composed of 26% amylolytic bacteria, 21% lipolytic bacteria, 18% proteolytic bacteria, 18% cellulolytic bacteria and 17% lactic acid bacteria; there were no differences in the amounts of these bacterial types between the control and treated oysters. Three major bacterial phyla, Proteobacteria, Firmicutes and Actinobacteria, and seven genera, Labrenzia, Pseudomonas, Halomonas, Shewanella, Vibrio, Bacillus, and Micrococcus, were detected. The concentration, viability, and phagocytic capacity of hemocytes and the production of ROS were not significantly altered by antibiotic treatment. In conclusion, the antibiotics altered the number of heterotrophic bacteria without changing hemocyte function, suggesting that this assay could be useful for verifying the influence of microbiota in host-parasite interactions.

O objetivo deste estudo foi detectar a presença potencial virulência de Vibrio cholerae isolado de estuários do Nordeste do Brasil. Amostras de água e sedimento foram coletadas e inoculadas sobre meio seletivo para víbrios (TCBS) e colônias com características morfológicas de V. cholerae foram isoladas. A identificação fenotípica seguiu chave dicotômica baseada em caraterísticas bioquímicas. Foram empregadas as técnicas de amplificação da polimerase em cadeia (PCR) utilizando o gene ompW e a de multiplex PCR para detecção de genes de virulência (ctx, tcp, zot e rfbO1). Os resultados da identificação das diferentes abordagens foram comparados. Nove cepas de V. cholerae foram identificadas fenotipicamente e cinco confirmadas através da detecção do gene ompW. A chave dicotômica utilizada foi eficiente para a confirmação da espécie. Os quatro estuários analisados apresentaram estirpes de V. cholerae, e nenhuma das cepas isoladas apresentaram genes de virulência.

The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

O objetivo deste trabalho foi estimar o nível e a distribuição da variabilidade genética em reprodutores brasileiros do camarão marinho (Litopenaeus vannamei). Nove dos maiores laboratórios de maturação do país foram analisados por meio de marcadores microssatélites codominantes e altamente polimórficos. Os resultados obtidos a partir da genotipagem de dez locos de microssatélites são indicativos de variabilidade genética compatível com aquela encontrada em populações naturais de L. vannamei no México e na América Central. Uma possível explicação é a origem altamente diversificada e relativamente recente dos plantéis disponíveis. A análise bayesiana detectou sinal correspondente a cinco populações originais. A distribuição das distâncias genéticas reflete parcialmente a localização geográfica, e essa informação será útil na formação de novos plantéis. Portanto, a variabilidade genética de L. vannamei entre nove dos maiores laboratórios de maturação nacionais pode ser considerada alta.

The objective of this work was to estimate the genetic variability level and distribution in Brazilian broodstocks of marine shrimp (Litopenaeus vannamei). Nine of the country's largest hatcheries were evaluated using codominant and highly polymorphic microsatellite markers. The results obtained from genotyping of ten microsatellite loci are indicative of genetic variability that is compatible with that found in wild populations of L. vannamei in Mexico and Central America. A possible explanation is the highly diversified and relatively recent origin of the available broodstocks. Bayesian analysis detected a signal for five founding populations. The distribution of genetic distances partially reflects geographical location, and this information will be useful for the creation of new broodstocks. Therefore, L. vannamei genetic variability among nine of the largest national hatcheries can be considered high.

A carcinicultura cresceu significativamente no Brasil ao longo dos últimos 10 anos, especialmente a produção do camarão branco do Pacífico, o exótico Litopenaeus vannamei. Em 2004, a atividade foi marcada por uma crise na produção, que despertou interesse na implantação de programas de melhoramento dos estoques de camarão. A importação de crustáceos foi banida do Brasil por uma Instrução Normativa de 1997, como uma medida de precaução sanitária. Desde então, a reposição de matrizes nas larviculturas passou a ser conduzida com estoques domesticados, gerando preocupações sobre o possível declínio da diversidade genética e sobre a possibilidade de que a diversidade genética existente pudesse garantir ganhos efetivos em programas de melhoramento. No presente trabalho, parâmetros genéticos, tais como número de alelos, número de alelos efetivos, heterozigosidade esperada e observada, coeficiente de consanguinidade, coeficiente de diferenciação genética e desvio do equilíbrio de Hardy-Weinberg, foram estimados para duas importantes larviculturas comerciais do Nordeste do Brasil, por meio da genotipagem de cinco marcadores microssatélites. O número de alelos efetivos (3 a 10,5) e as heterozigosidades médias observada e esperada (0,480 e 0,680) foram consistentes com aqueles relatados para populações de peneídeos de cativeiro e selvagens. Entretanto, valores positivos de F IS (0,381 para a larvicultura A e de 0,249 para a larvicultura B) mostraram uma deficiência significativa de heterozigotos (P<0,01). Apesar disso, é possível concluir que, mesmo após 10 anos de proibição na importação de crustáceos, tem sido possível manter um alto nível de variabilidade genética, possivelmente devido a origens múltiplas do estoque de fundadores dessa espécie no Brasil e da constante troca de reprodutores entre larviculturas.

The shrimp industry has grown significantly over the past 10 years in Brazil, especially the farmed production of the exotic Pacific white shrimp, Litopenaeus vannamei. In 2004, this industry was marked by a productivity crisis, which stirred interest towards genetic improvement of shrimp stocks. Shrimp breeders importation was banned in Brazil by a govern Normative Instruction in 1997, as a sanitary precaution. Since then, broodstock replacement in hatcheries has been based on domestic stocks, raising concerns on the decline of genetic diversity and if the existing diversity would allow effective genetic improvement programs. In the present research, genetic parameters such as number of alleles, effective allele number, expected and observed heterozygosities, inbreeding coefficient, genetic differentiation index and deviation from Hardy-Weinberg equilibrium have estimated of two important commercial hatcheries in Northeast Brazil, genotyping 5 microsatellite loci. Effective allele number (3 to 10.5) and average observed and expected heterozygosities (0.480 and 0.680) were consistent with those reported for cultured and wild Penaeid populations. However, F IS positive values (0.381 for hatchery A and 0.249 for hatchery B) reflected a significant heterozygous deficiency within hatcheries (P<0.01). Nevertheless, we concluded that even after ten years of limited genetic input, it has been possible to maintain a high level of genetic variability, possibly due to the wide diverse origin of the founder broodstocks and the constant breeders exchange among hatcheries.