Transplante interespecífico de espermatogônias-tronco de Brycon orbignyanus em Astyanax altiparanae (Teleostei: Characidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The endangered species Brycon orbignyanus: genetic analysis and definition of priority areas for conservation

Currently, biodiversity is threatened by several factors often associated with human population growth and the extension of areas occupied by human activity. In particular, freshwater fish fauna is affected by overfishing, deforestation, water pollution, introduction of exotic species and habitat fragmentation promoted by hydroelectric dams, among other environmental impact factors. Several action plans to preserve ichthyofauna biodiversity have been adopted; however, these plans frequently cover only a small number of species, and decisions are often made without strong scientific support. This study aimed to evaluate the genetic aspects of wild groups of Brycon orbignyanus, an endangered fish species, using microsatellites and D-loop regions to identify the genetic structure of the samples and to establish priority areas for conservation based on the genetic patterns of this species. The results indicated that the samples showed levels of genetic variability compatible with others studies with Neotropical fishes. However, the results obtained in the analysis of molecular variance (AMOVA) for microsatellites (F (ST) = 0.258) and D-loop (F (ST) = 0.234) and the interpopulation fixation index revealed that B. orbignyanus was structured in different subpopulations in the La Plata River basin; the areas with better environmental conditions also showed subgroups with higher rates of genetic variability. Future conservation actions addressing these sites should consider two different management units: the complex formed by the Ivinhema River, Upper Parana, Camargo Port and Ilha Grande groups; and the complex formed by the Verde River and Sucuriu River groups.

Growth, lipogenesis and body composition of piracanjuba (Brycon orbignyanus) fingerlings fed different dietary protein and lipid concentrations

Piracanjuba (Brycon orbignyanus) is a Brazilian migratory fast-growing omnivore, very appreciated as a sport fish, which is threatened to extinction in Southern Brazil due to stock over exploitation and dam building. Therefore, efforts have been made to raise this fish in captivity for reintroduction and aquaculture purposes. In the present study, the effects of different dietary protein and lipid concentrations on piracanjuba fingerlings growth performance, feed utilization, body composition, hepatosomatic index (HSI) and activity of the lipogenic enzymes fatty acid synthetase (FAS), glucose-6-phosphate dehydrogenase (G6PD) and malic enzyme (ME) were investigated using a 2 x 3 factorial experiment. Six casein-gelatin based diets were prepared combining two protein (30% and 32%) and three lipid concentrations (5.5%, 8.8% and 12.1%). Eleven fish, average weight 11.30 +/- 0.1 g, were held in each of 18 100-1 aquaria, supplied with recirculating freshwater. Each diet was randomly assigned to triplicate groups of fish and fed to apparent satiation, twice a day for 100 d. Piracanjuba fingerlings' daily weight gain (0.36-0.40 g), specific growth rate (1.43-1.51%), feed utilization and HSI were not influenced by dietary protein or lipid concentration. However, body composition was directly affected by dietary treatment. An increase in body fat and dry matter was observed as dietary lipid increased, for both dietary protein concentrations tested. The activity of FAS was depressed by increasing dietary fat levels but the G6PD activity did not differ among dietary treatments, although ME activity showed some regulation by dietary protein. These results indicate that an increase from 5.5% to 12.1% in the dietary lipid, at a dietary protein concentration of 30% or 32%, promotes body fat accumulation in piracanjuba fingerlings with no improvement in growth, suggesting that the lipid requirement for this species should be 5% or less, when raised for commercial purposes. However, the additional energy reserve from body fat accumulation could be desirable for piracanjuba fingerlings produced for stock enhancement. (C) 2003 Editions scientifiques et medicales Elsevier SAS and Ifremer/IRD/Inra/Cemagref. All rights reserved.

Water quality and zooplankton in tanks with larvae of Brycon Orbignyanus (Valenciennes, 1949)

Devido à importância das condições das variáveis da água e do alimento disponível no desenvolvimento e sobrevivência das larvas de peixes, o objetivo deste estudo foi avaliar o efeito do uso de dois tratamentos alimentares (ração + zooplâncton e somente zooplâncton) e a qualidade da água, em tanques de larvicultura de Brycon orbignyanus. A transparência total da água (45 cm) esteve associada principalmente ao curto tempo de residência, fluxo contínuo e baixa profundidade. As concentrações de oxigênio dissolvido variaram de 1,32 a 7,00 mg.L-1 no tratamento contendo ração + zooplâncton, e de 1,82 a 7,60 mg.L-1 no tratamento contendo somente zooplâncton. Os nutrientes foram diretamente influenciados pela adição de ração no meio com exceção do nitrito. Rotifera apresentou a maior densidade, variando ao longo do período experimental de 8,7 x 10(5) a 1,3 x 10(6) org.m-3 entre os quatro tanques estudados, num total de dez espécies. A menor densidade foi observada para Cladocera, variando de 4,7 x 10(4) a 2,1 x 10(5) org.m-3 num total de seis espécies encontradas, sendo Diaphanosoma birgei a única espécie classificada como muito freqüente. O uso de ração junto com zooplâncton proporcionou melhor rendimento para larvas de B. orbignyanus.

The influence of triiodothyronine (T-3) on the early development of piracanjuba (Brycon orbignyanus)

This paper reports the triiodothyronine's (T-3) effects on the early growth and survival of piracanjuba (Brycon orbignyanus) produced from fertilized eggs hormone exposed The study was carried out in two phases In the first phase, eggs divided in 6 batches were Immersed in T-3 solutions 0 01, 0 05, 0 1, 0 5 ppm, 1 ppm and control (no T-3) After a 15-min immersion, eggs were transferred to incubators where larvae were kept up to 72 h after hatching Larval weight, length and yolk sac volume were determined every 12 h Sixty and 72 h after hatching, larvae exposed to 0 5 ppm T-3 were significantly heavier than the others, and those exposed to 1 ppm T-3 showed the lowest weight The yolk sac absorption was not affected In the second experimental phase, the resulting fry from the first phase were stocked into 3 boxes per treatment (5 larvae L-1) and fed with plankton, fish larvae and feed prepared in the hatchery (48% CP) in the first 3 days, plankton and feed from the 4th to the 10th day and only feed in the next (last) 5 days Fry weight, length and specific growth rate were determined at 1, 5, 10 and 15 days Survival was calculated in the last day In the 15th day, fry length did not differ among treatments but the weight of the control group was higher Higher survival in the T-3-treated groups suggested lower predation among fry The results allowed us to conclude that there was no expressive effect of T-3 on the growth, but it improved the survival of the piracanjuba progeny

PCR primed with minisatellite core sequences yields species-specific patterns and assessment of population variability in fishes of the genus Brycon

Minisatellite core sequences were used as single primers in polymerase chain reaction (PCR) to amplify genomic DNA in a way similar to the random amplified polymorphic DNA methodology. This technique, known as Directed Amplification of Minisatellite-region DNA, was applied in order to differentiate three neotropical fish species (Brycon orbignyanus, B. microlepis and B. lundii ) and to detect possible genetic variations among samples of the threatened species, B. lundii , collected in two regions with distinct environmental conditions in the area of influence of a hydroelectric dam. Most primers generated species-specific banding patterns and high levels of intraspecific polymorphism. The genetic variation observed between the two sampling regions of B. lundii was also high enough to suggest the presence of distinct stocks of this species along the same river basin. The results demonstrated that minisatellite core sequences are potentially useful as single primers in PCR to assist in species and population identification. The observed genetic stock differentiation in B. lundii associated with ecological and demographic data constitute a crucial task to develop efficient conservation strategies in order to preserve the genetic diversity of this endangered fish species.

Parâmetros seminais, ultraestruturais, criopreservação e androgênese de espécies do gênero Brycon ameaçadas de extinção

Pós-graduação em Aquicultura - FCAV

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare is the causative agent of columnaris disease in freshwater fish, implicated in skin and gill disease, often causing high mortality. The aim of this study was the isolation and characterization of Flavobacterium columnare in tropical fish in Brazil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) and cascudo (Hypostomus plecostomus) were examined for external lesions showing signs of colunmaris disease such as greyish white spots, especially on the head, dorsal part and caudal fin of the fish. The sampling comprised 50 samples representing four different fish species selected for study. Samples for culture were obtained by skin and kidney scrapes with a sterile cotton swabs of columnaris disease fish and streaked onto Carlson and Pacha (1968) artificial culture medium (broth and solid) which were used for isolation. The strains in the liquid medium were Gram negative, long, filamentous, exhibited flexing movements (gliding motility), contained a large number of long slender bacteria and gathered into ‘columns'. Strains on the agar produced yellow-pale colonies, rather small, flat that had rhizoid edges. A total of four Flavobacterium columnare were isolated: 01 Brycon orbignyanus strain, 01 Piaractus mesopotamicus strain, 01 Colossoma macropomum strain, and 01 Hypostomus plecostomus strain. Biochemical characterization, with its absorption of Congo red dye, production of flexirubin-type pigments, H2S production and reduction of nitrates proved that the isolate could be classified as Flavobacterium columnare.

Diversidad genética de piracanjuba usada en programas de repoblación con marcadores microsatélites

El objetivo de este trabajo fue estimar la diversidad genética de un lote de Brycon orbignyanus usado en programas de repoblación, a través de marcadores microsatélites. Se analizaron muestras de 44 reproductores, de 70 larvas y de 69 alevinos, con la amplificación de cinco loci descritos para Brycon opalinus. El número de alelos, la heterozigosidad observada (Ho) y esperada (He), el índice de Shannon (IS), la diversidad genética de Nei (DGN), el coeficiente de endogamia (Fis), la distancia (DG) e identidad genética (IG), el número efectivo de alelos, el test del equilibrio de Hardy-Weinberg (EHW) y el desequilibrio de ligación fueron calculados. Reproductores y progenie tuvieron un número similar de alelos en los loci evaluados. La Ho media, IS, DGN, DG e IG mostraron que existe menor distancia genética entre parentales y larvas y una disminución de variabilidad genética en los alevinos. Fueron observados desvíos en EHW y desequilibrio de ligación en seis pares de loci. El Fis mostró exceso de heterocigotos en parentales y larvas y déficit de heterocigotos en los alevinos. El lote de reproductores está en proceso de pérdida de alelos y hubo disminución de la variabilidad genética entre la fase de larva y alevino.

Optimization of heterologous microsatellites in Piracanjuba

Abstract: The objective of this work was to evaluate 41 microsatellite markers for heterologous amplifications in piracanjuba (Brycon orbignyanus). Some markers were tested for the first time. Loci were optimized for PCR conditions and applied to a sample of 49 individuals. Thirty-one loci resulted in PCR product formation, whereas ten loci yielded intelligible polymorphic patterns in the evaluated sample and can be used for amplifications in this species. From the evaluated markers, four loci (BoM1, BoM13, Bh6, and Bh16) are valid to be applied in the study of piracanjuba.

Estudo dos restos de peixes dos sítios arqueológicos da área de influência da usina hidrelétrica Machadinho, RS, Brasil

Durante o resgate do material arqueológico dos sítios da região de Piratuba, SC, e de Aratiba, Machadinho e Maximiliano de Almeida, RS, área de influência da Usina Hidroelétrica de Machadinho, foram observadas grandes quantidades de ossos e escamas de peixes incorporando os restos alimentares encontrados nestes locais. Utilizando uma coleção osteológica de referência pudemos identificar restos de Salminus maxillosus Valenciennes, 1850, Brycon orbignyanus (Valenciennes, 1849), Pogonopoma obscurum Quevedo & Reis, 2002, Hemiancistrus fuliginosus Cardoso & Malabarba, 1999, Prochilodus lineatus (Valenciennes, 1836), Schizodon sp., Leporinus sp., Hoplias sp., Hypostomus sp. e Crenicichla sp. Com base em medições realizadas em exemplares de coleções, foram obtidas regressões a partir das dimensões do osso pré-maxilar em Crenicichla spp. e do esporão peitoral em Hemiancistrus fuliginosus, Pogonopoma obscurum e Hypostomus spp. correlacionadas com o comprimento padrão e peso dos espécimes. A partir das estimativas de comprimento padrão e peso das peças ósseas encontradas foi possível formular hipóteses sobre a tecnologia de pesca utilizada pelos habitantes destes sítios.

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare é o agente etiológico da columnariose em peixes de água doce, ocasionando enfermidade na pele e nas brânquias, provocando freqüentemente um grande número de mortalidade. O objetivo deste estudo foi o isolamento e a caracterização de Flavobacterium columnare em peixes tropicais no Brasil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) e cascudo (Hypostomus plecostomus) foram examinados externamente com relação a sinais característicos de columnariose, como manchas acinzentadas na cabeça, região dorsal e pedúnculo caudal dos peixes. A amostragem compreendeu a coleta de 50 exemplares de peixes, representando as quatro diferentes espécies escolhidas para este estudo. Amostras para o isolamento foram obtidas através de raspado com swab estéril das lesões e do rim dos peixes clinicamente diagnosticados como acometidos por columnarios e imediatamente semeados em meios de culturas artificiais (líquido e sólido) próprios para o estudo de Flavobacterium segundo Carlson e Pacha (1968). No meio líquido, houve o desenvolvimento de microrganismos que observados em gota pendente apresentaram a forma de bacilos finos, longos, móveis por deslizamento. Através da coloração de Gram, apresentaram morfologia de bacilos finos, Gram negativos, agrupados em colunas. em meio sólido, as colônias eram pequenas, cinza-amareladas, com borda em forma de raiz. No total, foram obtidos quatro isolamentos: 01 cepa de Brycon orbignyanus; 01 cepa de Piaractus mesopotamicus; 01 cepa de Colossoma macropomum; e 01 cepa de Hypostomus plecostomus. A caracterização bioquímica das amostras, como absorção do vermelho Congo, produção de flexirrubina, produção de H2S e redução do nitrato, sugere que os isolamentos poderiam ser classificados como Flavobacterium columnare.

Caracterização molecular de isolados de Flavobacterium columnare de tilapia do nilo e piracanjuba por meio de RAPD-PCR

Flavobacterium columnare is a cosmopolite bacteria and it is one of the main problem in Brazilian aquaculture, causing high mortalities index and economic damage. The main factors that contribute to columnaris disease are inadequate water quality, excess handling, high density of fish and temperature variations. For a successful epidemiological study and disease control, it is essential to differentiate the F. columnare from other yellow pigmentation bacteria. The present study used molecular techniques to characterize, by RAPD-PCR, two strains of F. columnare isolated from Oreochromis niloticus and Brycon orbignyanus. Data were analyzed as binary (0 and 1) and a genetic similarity matrix was generated by Jaccard's coefficient. Cluster analysis was performed by the neighbor joining method. The RAPD-PCR technique confirmed to be a usefull tool to obtain genetic profiles from F. columnare isolates based on the oligonucleotides used and to verify genetic similarity.

Chemical transfer of warning information in non-injured fish

In this study we describe pacus, Piaractus mesopotamicus, detecting the presence of a predator by conspecific alerting pheromone. Moreover, we investigate whether this chemical information indicates the presence of a specific predator, or whether it indicates general disturbance. We exposed groups of pacus to the view of a predator fish (trahira, Hoplias malabaricus), a non-predator fish (piracanjuba, Brycon orbignyanus) or an aquarium without any fish (control), and then we transferred their water to isolated conspecifics. We set up six trials of each condition in which we analysed the dispersion and the distance from the visual stimulus in water-donor fish and the distance from the chemical stimulus in water-receiver pacus. This study showed that pacus visually identified the presence of another fish and recognised it as predator or non-predator. This is interpreted as an innate response. Such heterospecific detection affects the behaviour of pacus, which release chemicals that induce conspecifics to adopt a similar behavioural response. At least two chemicals might be involved, one of them possibly an alerting pheromone.

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare é o agente etiológico da columnariose em peixes de água doce, ocasionando enfermidade na pele e nas brânquias, provocando freqüentemente um grande número de mortalidade. O objetivo deste estudo foi o isolamento e a caracterização de Flavobacterium columnare em peixes tropicais no Brasil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) e cascudo (Hypostomus plecostomus) foram examinados externamente com relação a sinais característicos de columnariose, como manchas acinzentadas na cabeça, região dorsal e pedúnculo caudal dos peixes. A amostragem compreendeu a coleta de 50 exemplares de peixes, representando as quatro diferentes espécies escolhidas para este estudo. Amostras para o isolamento foram obtidas através de raspado com swab estéril das lesões e do rim dos peixes clinicamente diagnosticados como acometidos por columnarios e imediatamente semeados em meios de culturas artificiais (líquido e sólido) próprios para o estudo de Flavobacterium segundo Carlson e Pacha (1968). No meio líquido, houve o desenvolvimento de microrganismos que observados em gota pendente apresentaram a forma de bacilos finos, longos, móveis por deslizamento. Através da coloração de Gram, apresentaram morfologia de bacilos finos, Gram negativos, agrupados em colunas. em meio sólido, as colônias eram pequenas, cinza-amareladas, com borda em forma de raiz. No total, foram obtidos quatro isolamentos: 01 cepa de Brycon orbignyanus; 01 cepa de Piaractus mesopotamicus; 01 cepa de Colossoma macropomum; e 01 cepa de Hypostomus plecostomus. A caracterização bioquímica das amostras, como absorção do vermelho Congo, produção de flexirrubina, produção de H2S e redução do nitrato, sugere que os isolamentos poderiam ser classificados como Flavobacterium columnare.