83 resultados para Dicentrarchus-labrax
Resumo:
En peces, al igual que en vertebrados superiores, la respuesta antiviral innata está mediada por el sistema interferón tipo I (IFN I), que actúa mediante la activación de genes denominados isg (interferon-stimulated genes), entre los que se incluye el gen isg15. La proteína codificada por este gen, denominada ISG15, presenta actividad antiviral actuando como citoquina, o mediante un mecanismo denominado ISGilación, que consiste en su unión covalente a proteínas víricas o celulares. Entre las enfermedades de etiología viral que afectan a la lubina (Dicentrarchus labrax), destaca la necrosis nerviosa viral, causada por el virus de la necrosis nerviosa (NNV, género Betanodavirus), que presenta dos segmentos de ARN monocatenario de polaridad positiva: ARN1 (polimerasa viral) y ARN2 (proteína de la cápside). Las diferencias en la secuencia de la región variable T4 del segmento ARN2 permiten su clasificación en 4 genotipos, siendo el genotipo RGNNV el único asociado a episodios de elevada mortalidad en lubina. El presente estudio contribuye a ampliar el conocimiento del papel del sistema IFN I en lubina frente a infecciones por betanodavirus, describiéndose la estructura del gen isg15, analizando su transcripción en respuesta a poli I:C y RGNNV; y evaluando su actividad in vitro. La lubina presenta un gen isg15, compuesto por dos exones y un intrón localizado en la región 5’-UTR. El ORF, de 474 pb, codifica una proteína de 157 aminoácidos constituida por dos dominios tipo ubiquitina y un motivo RLRGG en el extremo carboxilo terminal. La clonación del ORF en el plásmido pcDNA His/Max, y su posterior transfección en la línea celular E-11, ha permitido obtener una línea celular estable (DLISG15-E11), en la que se ha demostrado, mediante inmunofluorescencia indirecta, la localización citoplasmática de esta proteína. El análisis de transcripción muestra una respuesta más temprana e intensa tras la inducción por poli I:C que por RGNNV. Poli I:C estimula la expresión génica desde las 4 hasta las 24 h post-inyección (p.i.) en ambos órganos, y con una cinética similar. Sin embargo, ambos órganos difieren en el nivel de transcripción del gen tras la infección por RGNNV, produciéndose una estimulación temprana, desde las 12 h p.i., y de mayor nivel en el cerebro que en el riñón cefálico, órgano en el que la expresión comienza a las 72 h p.i. La actividad antiviral se ha evaluado mediante el análisis comparativo de la replicación de RGNNV, determinada mediante PCR cuantitativa, en células DLISG15-E11 y células control no transfectadas, no observándose diferencias significativas. Sin embargo, ISG15 podría actuar a otro nivel del ciclo de multiplicación viral, por lo que se están realizando ensayos de rendimiento vírico extracelular mediante titulación, y de ISGilación mediante Western-blot. El hecho de que el cerebro sea el principal órgano diana para la multiplicación de betanodavirus, y el papel antiviral atribuido a la ISG15, junto con los resultados obtenidos, contribuyen a dilucidar la importancia de este gen frente a las infecciones por RGNNV en lubina, incrementándose el conocimiento del papel del sistema del IFN I en la defensa frente a las infecciones vírcas y la relación patógeno-hospedador.
Resumo:
In the last few years two factors have helped to significantly advance our understanding of the Myxozoa. First, the phenomenal increase in fin fish aquaculture in the 1990s has lead to the increased importance of these parasites; in rum this has lead to intensified research efforts, which have increased knowledge of the development, diagnosis, and pathogenesis of myxozoans. The hallmark discovery in the 1980s that the life cycle of Myxobolus cerebralis requires development of an actinosporean stage in the Oligochaete. Tubifex tubifex, led to the elucidation of the life cycles of several other myxozoans. Also, the life cycle and taxonomy of the enigmatic PKX myxozoan has been resolved: it is the alternate stage of the unusual myxozoan. Tetracapsula bryosalmonae, from bryozoans. The 18S rDNA gene of many species has been sequenced, and here we add 22 new sequences to the data set. Phylogenetic analyses using all these sequences indicate that: 1) the Myxozoa are closely related to Cnidaria (also supported by morphological data), 2) marine taxa at the genus level branch separately from genera that usually infect freshwater fishes; 3) taxa cluster more by development and tissue location than by spore morphology; 4) the tetracapsulids branched off early in myxozoan evolution, perhaps reflected by their having bryozoan. rather than annelid hosts; 5) the morphology of actinosporeans offers little information for determining their myxosporean counterparts (assuming that they exist), and 6) the marine actinosporeans from Australia appear to form a clade within the platysporinid myxosporeans. Ribosomal DNA sequences have also enabled development of diagnostic tests for myxozoans. PCR and in situ hybridisation tests based on rDNA sequences have been developed for Myxobolus cerebralis. Ceratomyxa shasta. Kudoa spp,, and Tetracapsula bryosalmonae (PKX). Lectin-based and antibody tests have also been developed for certain myxozoans, such as PKX and C. shasta. We also review important diseases caused by myxozoans. which are emerging or re-emerging. Epizootics of whirling disease in wild rainbow trout (Oncorhynchus mykiss) have recently been reported throughout the Rocky Mountain states of the USA. With a dramatic increase in aquaculture of fishes using marine netpens, several marine myxozoans have been recognized or elevated in status as pathological agents. Kudoa thyrsites infections have caused severe post-harvest myoliquefaction in pen-reared Atlantic salmon (Salmo salar), and Ceratomyxa spp., Sphaerospora spp., and Myxidium leei cause disease in pen-reared sea bass (Dicentrarchus labrax) and sea bream species (family Sparidae) in Mediterranean countries.
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A aquacultura desempenha, actualmente, um papel fundamental no abastecimento de proteína animal de elevada qualidade. Nesta dissertação foi elaborado um projecto de uma unidade de piscicultura semi-intensiva de enguia, robalo, dourada e linguado e em regime extensivo a ostra. A unidade está localizada na Ria de Aveiro, na propriedade Ilha do Poço, com uma área total de 54,3 hectares. A execução de um projecto de aquacultura é um trabalho multidisciplinar que necessita a mobilização de diferentes recursos e conhecimentos. Os projectos de aquicultura estão directamente relacionados com o local de instalação da unidade e podem ser negativamente afectados pela poluição, por factores económicos locais ou por factores sociais que coloquem em risco o sucesso desta actividade económica. O principal objectivo desta dissertação foi o estudo de crescimento das espécies, taxa de alimentação, qualidade da água à entrada e saída da aquicultura, o consumo de oxigénio pelas espécies e o oxigénio necessário para oxidar a amónia em nitrato, o estudo de arejamento pelo vento, a necessidade de arejamento mecânico nos tanques de engorda e por fim a análise de investimento do projecto. Os valores obtidos para o tempo de crescimento das espécies até atingir o peso comercial foram, para a enguia 2,25anos, para a dourada 1,5 anos, para o robalo 2 anos, para o linguado 2,25 anos e para a ostra 1ano. Os índices de conversão alimentar (ICA) obtidos para as espécies foram os seguintes: 1 para a enguia, 1,24 para o robalo, 1,2 para a dourada e 1,58 para o linguado. Relativamente à concentração de amónia nos tanques de engorda, a espécie robalo apresenta o maior valor (0,3mg/L) por tanque. O arejamento natural dos tanques apenas é conseguido se o vento tiver uma velocidade de 20m/s. Por essa razão, e para garantir o arejamento requerido, serão necessários pelo menos 46 arejadores mecânicos com potência de 6kW a funcionar 24/24 horas. Para a taxa ponderada obteve-se um valor de 11,80%. O valor obtido para o VAL foi de 2.036.862 € e para a TIR, de 27,95%. O tempo necessário para que o investidor efectue o reembolso do capital investido no projecto é de 5,5 anos. O índice de rentabilidade tem um valor de 1,83 o que significa que o projecto é rentável.
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QuEChERS method was evaluated for extraction of 16 PAHs from fish samples. For a selective measurement of the compounds, extracts were analysed by LC with fluorescence detection. The overall analytical procedure was validated by systematic recovery experiments at three levels and by using the standard reference material SRM 2977 (mussel tissue). The targeted contaminants, except naphthalene and acenaphthene, were successfully extracted from SRM 2977 with recoveries ranging from 63.5–110.0% with variation coefficients not exceeding 8%. The optimum QuEChERS conditions were the following: 5 g of homogenised fish sample, 10 mL of ACN, agitation performed by vortex during 3 min. Quantification limits ranging from 0.12– 1.90 ng/g wet weight (0.30–4.70 µg/L) were obtained. The optimized methodology was applied to assess the safety concerning PAHs contents of horse mackerel (Trachurus trachurus), chub mackerel (Scomber japonicus), sardine (Sardina pilchardus) and farmed seabass (Dicentrarchus labrax). Although benzo(a)pyrene, the marker used for evaluating the carcinogenic risk of PAHs in food, was not detected in the analysed samples (89 individuals corresponding to 27 homogenized samples), the overall mean concentration ranged from 2.52 l 1.20 ng/g in horse mackerel to 14.6 ± 2.8 ng/ g in farmed seabass. Significant differences were found between the mean PAHs concentrations of the four groups.
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Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Engenharia do Ambiente,perfil Gestão e Sistemas Ambientais
Resumo:
Due to their toxicity, especially their carcinogenic potential, polycyclic aromatic hydrocarbons (PAHs) became priority pollutants in biomonitoring programmes and environmental policy, such as the European Water Framework Directive. The model substances tested in this study, namely benzo[b]fluoranthene (B[b]F), considered potentially carcinogenic to humans and an effector carcinogenic PAH to wildlife, and phenanthrene (Phe), deemed a non-carcinogenic PAH, are common PAHs in coastal waters, owning distinct properties reflected in different, albeit overlapping, mechanisms of toxicity. Still, as for similar PAHs, their interaction effects remain largely unknown. In order to study the genotoxic effects of caused by the interaction of carcinogenic and non-carcinogenic PAHs, and their relation to histopathological alterations, juvenile sea basses, Dicentrarchus labrax, a highly ecologically- and economically-relevant marine fish, were injected with different doses (5 and 10 μg.g-1 fish ww) of the two PAHs, isolated or in mixture, and incubated for 48 h. Individuals injected with B[b]F and the PAH mixture exhibited higher clastogenic/aneugenic effects and DNA strand breakage in blood cells, determined through the erythrocytic nuclear abnormalities (ENA) and Comet assays, respectively. Also, hepatic histopathological alterations were found in all animals, especially those injected with B[b]F and the PAH mixture, relating especially to inflammation. Still, Phe also exhibited genotoxic effects in sea bass, especially in higher doses, revealing a very significant acute effect that was accordant with the Microtox test performed undergone in parallel. Overall, sea bass was sensitive to B[b]F (a higher molecular weight PAH), likely due to efficient bioactivation of the pollutant (yielding genotoxic metabolites and reactive oxygen species), when compared to Phe, the latter revealing a more significant acute effect. The results indicate no significant additive effect between the substances, under the current experimental conditions. The present study highlights the importance of understanding PAH interactions in aquatic organisms, since they are usually present in the aquatic environment in complex mixtures.
Resumo:
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous priority pollutants that tend to be trapped in aquatic sediments due to their high hydrophobicity. Nonetheless, the differential toxicological effects and mechanisms between the various classes of PAHs and their mixtures, as they invariably occur in the environment, are scarcely known, especially under ecologically-relevant scenarios. This thesis aimed at establishing a bridge between the study of mechanistic pathways and environmental monitoring of carcinogenic and non-carcinogenic PAHs, by introducing ecological-relevance in the research with model PAHs. A first bioassay conducted in situ with the mussel Mytilus edulis demonstrated that, dredging operations in harbours increase PAH bioavailability, eliciting genotoxicity, and showed that established environmental guidelines underestimate risk. Subsequent ex situ bioassays were performed with the carcinogenic benzo[b]fluoranthene (B[b]F) and non-carcinogenic phenantrene (Phe), selected following preceding results, and revealed that low-moderate concentrations of these PAHs in spiked sediments induce genotoxic effects to the clam Ruditapes decussatus, therefore contradicting the general notion that bivalves are less sensitive to PAHs than vertebrates due to inefficient bioactivation. Also, it was demonstrated that passive samplers permit inferring on PAH bioavailability but not on bioaccumulation or toxic effects. On the other hand, sea basses (Dicentrarchus labrax), yielded a complex pattern of effects and responses, relatively to genotoxicity, oxidative stress and production of specific metabolites, especially when exposed to mixtures of the PAHs which led to additive, if not synergistic, effects. It was shown that Phe may elicit significant genotoxicity especially in presence of B[b]F, even though the low, albeit realistic, exposure concentrations diluted dose- and time-independent relationships. The present work demonstrated that environmental quality guidelines underestimate the effects of PAHs in realistic scenarios and showed that the significant genotoxic and histopathological effects caused by mixed PAHs may not be reflected by oxidative stress- or CYP-related biomarkers. Besides important findings on the metabolism of PAH mixtures, the work calls for the need to re-evaluate the criteria for assessing risk and for the disclosure of more efficient indicators of toxicological hazard.
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Visual scuba diving censuses were used to assess the effects of fishing prohibition on abundance and size structure of littoral fish populations by comparing the same benthic communities inside and outside the protected area of Medes Islands (NE Catalonia, Spain). The total number of species found was 43 in the reserve and 44 outside, but the mean value of species richness per sampling station was significantly higher in the protected area. However, diversity, heavily affected by the presence or absence of large schools of pelagic species, showed no significant differences between sites. The prohibition of fishing for 6 years is the first factor affecting the qualitative and quantitative structure of fish populations ('reserve effect'), and depth is the second factor. Thus, except in the cases of Serranus cabrilla and Mullus surmuletus, all other vulnerable species are highly sensitive to the protection measures. The size structure of all vulnerable species was found to be absolutely different at the reserve sites than in the unprotected zones, and the modal size classes of size frequency distributions were always higher in the reserve than outside. The reserve effect was significantly responsible of the differences observed in this change on size structure. Some highly vulnerable species, such as Epinephelus guaza and Sciaena umbra, have only been found in the protected area. Others, such as Sparus aurata, Diplodus cervinus and Dicentrarchus labrax, were much more frequent inside the reserve.
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This study investigated the effects of stocking density on the growth and fatty acid (FA) of Brycon insignis metabolism. Fingerlings (360) were distributed into eight ponds at two stocking densities (105 and 210 g/m(3)). The analysis of growth showed that the condition factor (K) and the coefficient of variation (CV) for body mass were not affected by stocking density. However, final body mass and length, specific growth rate (SGR), and weight gain (WG) were higher in the low stocking density group, which also presented a higher feed efficiency (FE) and survival (S). By contrast, muscle protein levels were higher in the high stocking density group. The plasma and muscle lipid content were not affected by stocking density, but fish reared at lower stocking density presented higher lipid concentration in the liver, with no differences in hepatosomatic index values. Even with the differences observed in metabolic and growth parameters, plasma cortisol was not affected by stocking density. The FA profile in the muscle and liver neutral fraction were not affected by stocking density, but the FA in the polar fractions differed between the two stocking densities. In the liver, total polyunsaturated fatty acids (PUFA) and PUFA n - 3 increased in higher stocking density, mainly due to an increase in docosahexaenoic acid (DHA). In addition, PUFA n - 6 were also increased in the higher stocking density group, mainly due to an increase in arachidonic acid (AA) and docosadienoic acid (22:2n - 6). In the muscle polar fraction, the saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) decreased in the animals from the higher stocking density group, and this reduction was compensated by an increase in PUFA n - 3 and PUFA n - 6, mainly the FA with 20-22 carbons (20:4n - 6: 22:4:n - 6; 22:5n - 6, 22:5n - 3, and 22:6n - 3). A different profile was observed for the C18 PUFAs, mainly 18:2n - 6 and 18:4n - 6, which were higher in the lower density stocking group. The data suggest that when living in high stocking density, B. insignis differentially utilizes the hepatic lipids as energy source and remodels the membrane fatty acids, with higher amounts of DHA in the polar muscle fraction compensated for by a decrease in MUFA. The zootechnical and physiological indices reveal that the lower stocking density group achieve overall better performance. (C) 2010 Elsevier B.V. All rights reserved.
Resumo:
Fatty acids have been used in marine biogeochemistry as food chain biomarkers, but in freshwater these studies are rare. In order to evaluate the fatty acid potential as biomarkers in freshwater, their profile was analyzed during vitellogenesis in two fish species, in both waterfall and reservoir environments of the Paraiba do Sul River Basin. Detrivorous Hypostomus affinis and omnivorous Geophagus brasiliensis seem to elongate and desaturate polyunsaturated fatty acids (PUFA) and transfer them to the ovaries` phospholipids. Waterfall Geophagus brasiliensis have more highly unsaturated fatty acids in the liver, but in the reservoir, accumulation mainly occurs in muscle and ovary triglycerides, suggesting trophic opportunism and a plasticity during vitellogenesis. In Hypostomus affinis, PUFA alteration occurs only in the reservoir, suggesting a high phytoplankton occurrence. Eutrophication and water speed is reflected in Hypostomus affinis ovaries by higher PUFAn3 and bacterial fatty acids. As in marine environments, analysis of mono- and polyunsaturated fatty acids during vitellogenesis can be used as a tool in food chain studies in freshwater.
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This article documents the addition of 473 microsatellite marker loci and 71 pairs of single-nucleotide polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Barteria fistulosa, Bombus morio, Galaxias platei, Hematodinium perezi, Macrocentrus cingulum Brischke (a.k.a. M.abdominalis Fab., M.grandii Goidanich or M.gifuensis Ashmead), Micropogonias furnieri, Nerita melanotragus, Nilaparvata lugens Stal, Sciaenops ocellatus, Scomber scombrus, Spodoptera frugiperda and Turdus lherminieri. These loci were cross-tested on the following species: Barteria dewevrei, Barteria nigritana, Barteria solida, Cynoscion acoupa, Cynoscion jamaicensis, Cynoscion leiarchus, Cynoscion nebulosus, Cynoscion striatus, Cynoscion virescens, Macrodon ancylodon, Menticirrhus americanus, Nilaparvata muiri and Umbrina canosai. This article also documents the addition of 116 sequencing primer pairs for Dicentrarchus labrax.
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The objective of this work was to determine the hematological parameters and the phagocytic capacity of peritoneal macrophages of fat snook related to sex, stage of gonadal maturation and seasonal cycle. Blood was collected from 135 animals (78 females and 57 males) and used for determinations of: erythrocyte number, hematocrit, hemoglobin, erythrocyte indices mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), total and differential leukocyte counts, and thrombocyte count. The phagocytic capacity and phagocytic index were determined after Saccharomyces cerevisiae inoculation in the peritoneal cavity of the animals. The hematological results according to sex showed that the erythrocyte, total leukocyte and thrombocyte counts were statistically higher in males than females, with the latter showing a higher MCV. Concerning to erythrocyte count, hematocrit and hemoglobin concentration analyzed separately by sex and stage of gonadal maturation, males were found to have significantly elevated values in the mature stage and decreased levels in the resting stage. The results of the erythrocyte and leukocyte series, thrombocytes and phagocytic activity related to seasonal cycle showed significant differences in both sexes, where hematocrit and hemoglobin concentration were lower in winter and higher in the other seasons, mean corpuscular volume was higher in the summer and lower in the winter and fall, total leukocytes and thrombocytes lower in the spring and higher in the fall, lymphocytes low in the winter and summer and high in the spring and phagocytic capacity and phagocytic index high in the summer and low in the winter and fall. The results showed that the hematological values in males are statistically higher than those in females, the erythrocyte values in males increase with the progression of gonadal maturation and that winter is the season of the year least favorable for hematological and phagocytic responses for survival of fat snook kept in captivity. The parameters studied could be utilized in the evaluation of the health status of this species in captivity. (C) 2012 Elsevier Ltd. All rights reserved.
Resumo:
O pacu, Piaractus mesopotamicus, é um teleósteo da Família Characidae, intensivamente cultivado no Brasil devido sua rusticidade, crescimento rápido e fácil adaptação. O conhecimento morfológico dos sistemas corpóreos, incluído órgãos linfóide, se faz necessário, para uma melhor produção no cultivo de peixes, fornecendo subsídios na manutenção dos estoques. O objetivo deste estudo foi descrever morfologicamente o rim e rim cefálico de Piaractus mesopotamicus, analisando os perfis celulares de cada órgão com o uso de microscopia de luz e microscopia eletrônica de transmissão. O resultado da análise macroscópica mostrou que a localização do rim e rim cefálico são as mesmas encontradas na maioria dos teleósteos. O rim apresentou uma forma em "H", onde a região média se expandia sobre a bexiga natatória. O rim cefálico se apresentou como uma dilatação na região cranial do rim, mostrando-se bem visível. Na microscopia eletrônica de transmissão também foram observadas similaridades ultraestruturais com outros teleósteos. Observando nossos resultados concluímos que histologicamente e ultraestruturalmente, os órgãos linfóides rim e rim cefálico de Piaractus mesopotamicus são similares aos de outros teleósteos.
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The objective of this study was to obtain information about the possible mechanisms related to poor reproductive performance in tropical rheophilic fish. To that effect, cages (Cs) and earthen ponds (EPs) were used as experimental systems to provide unsuitable and suitable conditions, respectively, for curimbata (Prochilodus lineatus) breeders. Fish were maintained under experimental conditions for 18 months, and during this period females were randomly sampled every two months for biometric analysis (n=30), blood (n=5/sampling) and ovary (n=5/sampling). After this period EPs females (EPFs) and Cs females (CFs) were submitted to the induced breeding experiments. The results showed that rearing curimbata for such long time in a cage at this stocking density, reduces its growth, plasma E2 levels and vitellogenesis. During vitellogenesis, the mean plasma estradiol levels of CFs were three times lower than those of EPFs (P<0.01). CFs presented poorer results than EPFs for all the examined parameters of reproductive performance. Taken together these data showed that the reduced estradiol levels during vitellogenesis (and the consequently less intense transition from the previtellogenic to vitellogenic phase) and reduced amounts of yolk are mechanisms associated with the formation of low quality oocytes and shortened and delayed breeding season in this species. Moreover, our data showed that the onset of vitellogenesis (six months before the spawning season) must be considered as a key period related to the formation of oocytes of good quality, and adequate management should be provided throughout the year.
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Rainbow trout Oncorhynchus mykiss triploids are regularly produced in fish farms to improve growth because the triploid females do not develop ovaries during the reproductive cycle. In this study, the tissue fatty acid allocations in triploid and diploid females were compared during the reproductive cycle to determine whether the ploidy influences the fatty acid profile of fish produced in aquaculture. The ovaries, liver, and white muscle fatty acid contents of diploid and triploid females were analyzed during the reproductive cycle. Diploid females tend to accumulate more polyunsaturated fatty acids than triploids during some phases of the reproductive cycle, and this profile was compensated by an increase in saturated and monounsaturated fatty acids in triploid females. Arachidonic acid (C20:4n6) was the main n6 polyunsaturated fatty acid in the ovaries of diploid females during the most advanced phases of the reproductive cycle, and docosahexaenoic acid (C22:6n3) was the main n3 polyunsaturated fatty acid. In triploid females, the percentage of both of these polyunsaturated fatty acids was lower than in diploid females during the most advanced phases of the reproductive cycle. In general, the lack of ovary development altered the hepatic synthesis of some fatty acids, mainly monounsaturated and polyunsaturated fatty acids, decreasing the content of the main fatty acids in the white muscle and, consequently, the mobilization of these specific fatty acids to the ovaries.