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A B S T R A C TThis study evaluated the survival, post-larvae quality, and the population of bacteria in Litopenaeus vannamei after the addition of two strains of lactic-acid bacteria (2 and B6) experimentally infected by Vibrio harveyi. Fifteen hundred nauplii were distributed in 20 L capacity tanks with four replicates. The survival of control animals was lower (21%) than that of animals fed with the strains B6 (50%) and 2 (44%). Total bacterial population in the water and larvae, as well as of the Vibrio ssp. in water was not different among the treatments. No difference was observed in the population of Vibrio ssp. between the control larvae (5.5±0.5 log UFC/mL) and that fed with strain 2 (5.4±0.1 log UFC/mL). Shrimp from control and fed with strain 2 showed significantly higher bacterial population than those fed with strain B6 (1.2±0.2 log UFC/mL). It was detected the lower load of Vibrio ssp. bacteria with potential of pathogenicity after feeding with strain B6.Moreover, these larvae showed more active behavior and low number of necrosis in relation to the control group and to that fed with strain 2. R E S U M OEste trabalho avaliou a adição de duas cepas de bactérias lácticas (2 e B6) na sobrevivência, qualidade de pós-larva e na população de bactérias na larvicultura de Litopenaeus vannamei experimentalmente infectado por Vibrio harveyi. Mil e quinhentos náuplios foram distribuídos em tanques de 20 L com quatro repetições. A sobrevivência dos animais controle foi menor (21%) do que a dos alimentados com as cepas B6 (50%) e 2 (44%). Sobrevivência de misis após desafio com V. harveyi foi maior em B6 do que nos outros tratamentos. A população total de bactérias na água e nas larvas, bem como de Vibrio ssp. na água não foi diferente entre os tratamentos. Não houve diferença, também, entre a população de Vibrio ssp. em larvas do grupo controle (5,5±0,5 log UFC/mL) e larvas alimentadas com a cepa 2 (5,4±0,1 log UFC/mL). Camarões do grupo controle e alimentados com cepa 2 apresentaram maior população de bactérias do que os alimentados com cepa B6 (1,2±0,2 log UFC/mL). Foi comprovada a menor presença de bactérias entéricas com potencial de patogenicidade nos animais alimentados com a cepa B6, apresentando também comportamento mais ativo e menor número de necroses em relação ao controle e cepa 2.

Previous studies have shown that the gut microbiota of fish differs depending on host feeding habits and these populations may also be influenced by dietary ingredients. In this study, the bacterial populations of the intestinal tract of the opportunistic omnivore jundiá Rhamdia quelen and the typical omnivore tilapia Oreochromis niloticus were investigated in two experiments. In experiment I, the levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were compared in the proximal, mid and distal intestine using selective agar. Higher levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were observed in the tilapia than jundiá, in all intestinal regions. Jundiá contained higher levels of proteolytic bacteria and lipid digesting bacteria in the distal intestinal portion as compared with the mid intestine; both fish species also presented more amylolytic bacteria in the distal intestine. In experiment II, the amylolytic intestinal microbiota between the two species was compared after administration of diets containing different carbohydrate sources. Jundiá fed broken rice presented higher total culturable bacterial levels; however, dietary cassava bagasse and ground corn significantly elevated the population of amylolytic bacteria in tilapia (P > 0.05). PCR‐DGGE was also used to assess the bacterial communities in experiment II. A Cetobacterium spp. was detected in jundiá fed diets containing broken rice, and tilapia fed cassava bagasse, dextrin, broken rice and ground corn. Microbial differentiation was further demonstrated between jundiá and tilapia, because an uncultured bacterium was unique in tilapia and an uncultured spirochete was observed only in jundiá; the presence of these bacterial species was also influenced by dietary carbohydrate sources.

Hematology and agglutination titer after polyvalent immunization and subsequent challenge with Aeromonas hydrophila in Nile tilapia (Oreochromis niloticus)Hematología y título de aglutinación después de inmunización polivalente y desafío con Aeromonas hydrophila a tilapias del Nilo (oreochromis niloticus)

This study evaluated the total and differential leukocyte counting and the phagocytic activity in Nile tilapia Oreochromis niloticus experimentally injected with Enterococcus sp. in the swim bladder. Fish were distributed in four treatments in triplicates of non-injected fish, fish injected with 1 ml of sterile saline solution 0.65%, and fish injected with 1 x 10(3) and 1 x 10(6) colony-forming units (CFU) of Enterococcus diluted in 1 ml sterile saline. Twenty-four hours after injection, the fish were anesthetized and the blood collected for white blood cell (WBC) counts, differential counting of WBC, and phagocytic activity of blood leukocytes. The increased numbers of WBC and lymphocytes were followed by decreased number of monocyte after infection. The percentages of phagocytic activities in the blood were 55.3 and 55.9%, respectively, in tilapia injected with 1 x 10(3) and 1 x 10(6) CFU/ml.

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A preliminary study to characterize filamentous bacteria, whose presence is related to high mortality of Litopenaeus vannamei larvae cultured in Santa Catarina State, Brazil, is reported. The extract of infected larvae was diluted in different concentrations, cultured in marine agar (Difco TM , Marine Agar 2216) and incubated at 30 °C for 48 hours. The biochemical characterization included hydrolytic reactions of starch, gelatin and tyrosine, growth in TCBS agar, growth in 0 and 37‰ salinity, pigment production in tyrosine agar, production of H 2 S, nitrate reduction, congo red reaction, oxidase and catalase. The isolated bacteria belong to the species Flexibacter maritimus, Gram-negative bacilli of 0.4-0.5 µm width and 15 µm length. Experiments were carried out on pathogenicity of F. maritimus in post-larvae of L. vannamei. Survival and symptoms in L. vannamei post-larvae 24 hours after inoculation with F. maritimus and its growth in marine agar were evaluated. Mortality was detected around 92,5% as well as symptoms like melanized lesions in several parts of body, discolouration of gills, bad formation of appendages and of the last abdominal segment, low motility and feeding reduction. The experimental infection results suggested that isolated bacteria of the genus Flexibacter are pathogenic to the shrimp Litopenaeus vannamei post-larvae. Keywords: Litopenaeus vannamei, hatchery, Flexibacter maritimus, experimental infection. Caracterização e infecção experimental de Flexibacter maritimus em pós-larvas de Litopenaeus vannamei ResumoNeste trabalho, relata-se um estudo preliminar para caracterizar bactérias filamentosas associadas a elevadas mortalidades de larvas e pós-larvas cultivadas no Estado de Santa Catarina, Brasil. Um extrato de larvas infectadas foi diluído em diferentes concentrações, cultivado em Agar Marine (Difco TM , Marine Agar 2216) e incubado a 30° C por 48 horas. A caracterização bioquímica inclui reações de amido, gelatina e tirosesine, crescimento em Agar TCBS,crescimento a 0 e 37‰ de salinidade, produção de pigmentos em Agar Tirosine, produção de H 2 S, redução de nitrato, reação vermelho congo, oxidase e catalase. A bactéria isolada apresentou características de Flexibacter maritimus, um bacilo Gram negativo de 0,4-0,5 µm de largura e 15 µm de comprimento. Os experimentos foram realizados sobre a patogenicidade de F. Maritimus, em pós-larvas de L. vannamei. Foram avaliados a sobrevivência e os sintomas das pós-larvas 24 horas após a inoculação com F. Maritimus, além do seu crescimento em Agar Marine. Foram encontradas mortalidades de 92,5% e sintomas próprios da enfermidade, tais como lesões melanizadas em várias partes do corpo, descoloração das brânquias e diminuição da motilidade e da alimentação. Com base nos resultados da infecção experimental, pode se concluir que a bactéria isolada do gênero Flexibacter é patogênica para pós-larvas de L. vannamei.Palavras-chave: Litopenaeus vannamei, pós-larvas, Flexibacter maritimus, infecção experimental. Mouriño, JLP. et al.

In one year, 76 first generation (F1) Brazilian sardine Sardinella brasiliensis breeders spawned 84 times spontaneously in captivity. This is the first report for the genus Sardinella. Following the pattern found in the natural environment, the number of spawns, as well as fertilization and hatching rates were higher in spring, summer and fall than in winter. Until now, winter spawning was only presumed to occur in a natural environment. Water temperature, and not photoperiod, was positively correlated with reproductive performance.