131 research outputs found

    Fuentes alternativas de proteínas en acuicultura. Disfunciones endocrinas, metabólicas e inmuno-histopatológicas.

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    RESUMEN Los alimentos formulados para el cultivo intensivo de peces carnívoros contienen, generalmente, un 40-50% de proteína que está normalmente proporcionada por las harinas de pescado. Sin embargo, mientras que la producción anual de harinas de pescado está estabilizada alrededor de 6-7 millones de toneladas, la acuicultura se expande a una tasa del 11% anual existiendo, de este modo, una seria preocupación acerca de la disponibilidad de esta fuente de alimento como materia prima en la formulación de dietas para su empleo en dicho sector. Dado que los ingredientes de origen vegetal están facilmente disponibles, en los últimos años han recibido la mayor parte de la atención como posibles sustitutos a las harinas de pescado. Para abordar esta problemática, se diseñaron dietas basadas en harinas de pescado (FM) o en ingredientes vegetales, de manera que sustituyeran las harinas de pescado en un 50% (PP50), 75% (PP75) y 100% (PP100). En todas las dietas el cociente entre aminoácidos esenciales y no esenciales estuvo próximo a 1,1, en base a los resultados obtenidos en una experiencia previa incluida en esta Tesis Doctoral. Juveniles de dorada (Sparus aurata) alimentados con la dieta PP100 exhibieron una pérdida severa de biomasa con respecto a los otros grupos experimentales, debido probablemente a una menor ingesta de alimento. Estos peces también exhibieron una menor retención lipídica así como un marcado efecto hipocolesterolémico. En cuanto al eje somatotrópico, se observó un aumento de los niveles circulantes de hormona de crecimiento (GH) con el mayor aporte de ingredientes vegetales, que estuvo acompañado por un descenso de las tasas de crecimiento, de los niveles circulantes de somatomedinas (IGF-I) y de la expresión hepática del receptor de la GH (GHR) y de IGF-I. Esta situación catabólica evidenció una insensibilización hepática a la acción anabólica de la GH en el grupo PP100. A medio plazo (6 meses) se observó una elevada deposición grasa en el hígado de los peces alimentados con la dieta PP100 así como un elevado número de vacuolas lipídicas en los enterocitos. Los niveles de lisozima en plasma no se vieron alterados por el tratamiento pero el estallido respiratorio en leucocitos aislados del riñón anterior y la actividad mieloperoxidasa en plasma aumentaron significativamente en los grupos PP75 y PP100. La actividad del complemento, por otra parte, aumentó significativamente en el grupo PP50 y disminuyó en los grupos PP75 y PP100. Como conclusión general, y junto con los ensayos de calidad sensorial realizados tras un ciclo completo de engorde (1 año), se determinó que la sustitución de harinas de pescado por ingredientes vegetales (hasta un 75%) es posible en una especie típicamente carnívora como la dorada. La actividad del eje somatotrópico también fue analizada en trucha arco iris (Oncorhynchus mykiss), en una experiencia similar a la anterior. Las tasas de crecimiento y la retención energética disminuyeron en los grupos PP75 y PP100, observándose un aumento progresivo en los niveles circulantes de GH. Sin embargo, no se observaron cambios ni en los niveles circulantes de IGF-I ni en la expresión hepática de GHR e IGF-I. Se encontraron dos bandas específicas de proteínas de unión a la IGF (IGFBPs) en el plasma de todos los peces analizados, aumentando progresivamente su concentración y su grado de glicosilación con el aumento de la cantidad de proteína vegetal de la dieta, lo que podría disminuir la disponibilidad de IGF libre. Así, la disminución del crecimiento pudo estar debido, al menos en parte, a una menor disponibilidad de la forma activa de la IGF (fracción libre) más que a una insensibilización hepática a la GH o a un defecto en la síntesis y liberación de IGF-I. __________________________________________________________________________________________________Aquafeeds for carnivore fish contain a 40-50% of crude protein, which is usually provided by fishmeal. However, while the annual production of fishmeal is stabilized around 6-7 million tonnes, the aquaculture industry expands at a rate of 11% per year. Thus, it exists a great concern about the availability of these raw materials to formulate aquafeeds for aquaculture. As plant ingredients are easily available, in the last years a great effort has been done to investigate the possible use of plant ingredients to replace fishmeal in aquaculture. To solve this problem, a diet with fish meal as the sole protein source was compared to diets with 50%, 75% and 100% of replacement in juveniles of gilthead seabream (Sparus aurata). The indispensable amino acid to dispensable amino acid ratio (IAA/DAA) was close to 1,1 in all diets as it was stablished in the first trial of this study. Growth performance, enzyme activities (amino acid catabolism and lipogenic pathways) and plasma metabolites (glucose, triglycerides and cholesterol) were measured. The hormone system that regulates fish growth was also study (plasma levels of growth hormone and IGF-I, hepatic expression of GH receptors and IGF-I). We also developed immuno-histochemical studies and parameters of the inespecific immunological system (plasma lysozyme, respiratory burst activity, plasma myeloperoxidase, complement) were also evaluated. As a conclusion, substitution of fishmeal by a mixture of plant protein sources compromised growth performance only at the 100% level, and decreased one of the immune defence mechanisms at above 75% level. The quality of the fillet in fish fed diet PP75 until they reached market size was not affected by dietary treatment which means that up to 5075% of fishmeal replacement seems to be feasible with IAA supplementation in a fish species having high dietary protein requirements. The activity of somatotropic axis was analysed in juveniles of rainbow trout (Oncorhynchus mykiss) in a similar growth trial. It is shown that a different regulation of this hormonal system is found in response to a partial or total replacement of fishmeal by plant ingredients compared to gilthead seabream

    The Dietary Lipid Content Affects the Tissue Gene Expression of Muscle Growth Biomarkers and the GH/IGF System of Pejerrey (<i>Odontesthes bonariensis</i>) Juveniles

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    Gene expression of growth hormone receptors (GHRs), insulin-like growth factors (IGFs), myostatin (MSTN) and myogenin (MyoG) was analyzed in juveniles pejerrey fed with graded levels of lipids (L): 6% (L6), 10% (L10), 25% (L25). After 14 weeks, no changes were found in liver GHR-I GHR-II and IGF-II mRNA levels whereas IGF-I decreased in L10 and L25. Muscle GHR-I gene expression increased in L25 whereas GHR-II, IGF-II and MyoG were higher in L6. IGF-I and MSTN expression was not affected by the different diets. Adipose IGF-I mRNA levels decreased in L10. Correlations between body weight and members of GH/IGF system in liver and skeletal muscle were found only in L10 group. Correlations found in L10 group between both liver and skeletal muscle GHR-I and IGF-I were lost in either L6 or L25 groups. Thus, fish fed with apparently unbalanced dietary lipid contents (6% and 25%) exhibit a compensatory regulation of systemic and local components of the GH/IGF axis. Furthermore, the marked inhibition of muscle MyoG gene expression in L25 might limit excessive lipid deposition and fish growth. Our data suggest that a dietary lipid contents of 10% would promote a particular adjustment of the endocrine and autocrine/paracrine GH/IGF system, stimulating body growth and perhaps muscle hyperplasia. On the other hand, a higher dietary lipid content would uncouple the GH/IGF system, reducing hepatic IGF-I, while slightly increasing hepatic GHR-I, probably to prompt lipolysis.Centro de Endocrinología Experimental y Aplicad

    Tissue-Specific Orchestration of Gilthead Sea Bream Resilience to Hypoxia and High Stocking Density

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    Two different O-2 levels (normoxia: 75-85% O-2 saturation; moderate hypoxia: 42-43% O-2 saturation) and stocking densities (LD: 9.5, and HD: 19 kg/m(3)) were assessed on gilthead sea bream (Sparus aurata) in a 3-week feeding trial. Reduced O-2 availability had a negative impact on feed intake and growth rates, which was exacerbated by HD despite of the improvement in feed efficiency. Blood physiological hallmarks disclosed the enhancement in O-2-carrying capacity in fish maintained under moderate hypoxia. This feature was related to a hypo-metabolic state to cope with a chronic and widespread environmental O-2 reduction, which was accompanied by a differential regulation of circulating cortisol and growth hormone levels. Customized PCR-arrays were used for the simultaneous gene expression profiling of 34-44 selected stress and metabolic markers in liver, white skeletal muscle, heart, and blood cells. The number of differentially expressed genes ranged between 22 and 19 in liver, heart, and white skeletal muscle to 5 in total blood cells. Partial Least-Squares Discriminant Analysis (PLS-DA) explained [R2Y(cum)] and predicted [Q2Y(cum)] up to 95 and 65% of total variance, respectively. The first component (R2Y = 0.2889) gathered fish on the basis of O-2 availability, and liver and cardiac genes on the category of energy sensing and oxidative metabolism (cs, hif-1 alpha, pgc1 alpha, pgc1 beta, sirts 1-2-4-5-6-7), antioxidant defense and tissue repair (prdx5, sod2, mortalin, gpx4, gr, grp-170, and prdx3) and oxidative phosphorylation (nd2, nd5, and coxi) highly contributed to this separation. The second component (R2Y = 0.2927) differentiated normoxic fish at different stocking densities, and the white muscle clearly promoted this separation by a high over-representation of genes related to GH/IGF system (ghr-i, igfbp6b, igfbp5b, insr, igfbp3, and igf-i). The third component (R2Y = 0.2542) discriminated the effect of stocking density in fish exposed to moderate hypoxia by means of hepatic fatty acid desaturases (fads2, scd1a, and scd1b) and muscle markers of fatty acid oxidation (cpt1a). All these findings disclose the different contribution of analyzed tissues (liver >= heart > muscle > blood) and specific genes to the hypoxic- and crowding stress-mediated responses. This study will contribute to better explain and understand the different stress resilience of farmed fish across individuals and species

    Molecular cloning, tissue expression and regulation of Liver X Receptor (LXR) transcription factors of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss)

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    Fish are important sources of high quality protein, essential minerals such as iodine and selenium, vitamins including A, D and E, and omega-3 fatty acids in the human diet. With declining fisheries worldwide, farmed fish constitute an ever-increasing proportion of fish in the food basket. Sustainable development of aquaculture dictates that diets will have to contain increasing levels of plant products that are devoid of cholesterol, but contain phytosterols that are known to have physiological effects in mammals. Liver X receptors (LXR) are transcription factors whose activity is modulated by sterols, with activation inducing cholesterol catabolism and de novo fatty acid biosynthesis in liver. Transcriptomic analysis has shown that substitution of fish meal and oil with plant products induces genes of cholesterol and fatty acid metabolism in salmonids. Here we report the cloning of LXR cDNAs from two species of salmonid fish that are important in aquaculture. The full-length cDNA (mRNA) of LXR obtained from salmon was shown to be 3766 bp, which included a 5’-untranslated region (UTR) of 412 bp and a 3’-UTR of 1960 bp and an open reading frame (ORF) of 1394 bp, which specified a protein of 462 amino acids. The trout LXR full-length cDNA was 2056 bp, including 5’- and 3’-UTRs of 219 and 547 bp, respectively, and an ORF of 1290 bp, which specified a protein of 427 amino acids. The protein sequences included characteristic features of mammalian LXRs, including the DNA binding (DBD), containing P-box, ligand binding (LBD) and activation function-2 (AF-2) domains, D-box, D (hinge) region, and eight cysteines that belong to the two zinc fingers. Phylogenetic analysis clustered the salmonid LXRs together, more closely with zebrafish and more distantly from medaka and stickleback. A pair-wise comparison among vertebrate LXR sequences showed the amino acid sequence predicted by the salmon LXR ORF showed greatest identity to that of trout 97%, and 97%, 87% and 81% identity to LXRs of zebrafish, frog and human (LXRα). The trout LXR ORF showed 96%, 92% and 82% identity to LXRs of zebrafish, frog and human (LXRα). Surprisingly, the expression of LXR was lowest in liver of all tissues examined and in salmon the greatest expression was observed in pyloric caeca with liver showing intermediate expression. It is likely that tissue expression was affected by the physiological status of the sampled animals. Certainly, nutritional, environmental and/or developmental regulation was evident in salmon, where the expression of LXR in liver was higher in fish in seawater than in freshwater, and higher in fish fed fish oil compared to fish fed vegetable oil in adult salmon

    Long -term feeding with high plant protein based diets in gilthead seabream (Sparus aurata, L.) leads to changes in the inflammatory and immune related gene expression at intestinal level

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    [EN] Background: In order to ensure sustainability of aquaculture production of carnivourous fish species such as the gilthead seabream (Sparus aurata, L.), the impact of the inclusion of alternative protein sources to fishmeal, including plants, has been assessed. With the aim of evaluating long-term effects of vegetable diets on growth and intestinal status of the on-growing gilthead seabream (initial weight = 129 g), three experimental diets were tested: a strict plant protein-based diet (VM), a fishmeal based diet (FM) and a plant protein-based diet with 15% of marine ingredients (squid and krill meal) alternative to fishmeal (VM+). Intestines were sampled after 154 days. Besides studying growth parameters and survival, the gene expression related to inflammatory response, immune system, epithelia integrity and digestive process was analysed in the foregut and hindgut sections, as well as different histological parameters in the foregut. Results: There were no differences in growth performance (p = 0.2703) and feed utilization (p = 0.1536), although a greater fish mortality was recorded in the VM group (p = 0.0141). In addition, this group reported a lower expression in genes related to pro-inflammatory response, as Interleukine-1 beta (il1 beta, p = 0.0415), Interleukine-6 (il6, p = 0.0347) and cyclooxigenase-2 (cox2, p = 0.0014), immune-related genes as immunoglobulin M (igm, p = 0.0002) or bacterial defence genes as alkaline phosphatase (alp, p = 0.0069). In contrast, the VM+ group yielded similar survival rate to FM (p = 0.0141) and the gene expression patterns indicated a greater induction of the inflammatory and immune markers (il1 beta, cox2 and igm). However, major histological changes in gut were not detected. Conclusions: Using plants as the unique source of protein on a long term basis, replacing fishmeal in aqua feeds for gilthead seabream, may have been the reason of a decrease in the level of different pro-inflammatory mediators (il1 beta, il6 and cox2) and immune-related molecules (igm and alp), which reflects a possible lack of local immune response at the intestinal mucosa, explaining the higher mortality observed. Krill and squid meal inclusion in vegetable diets, even at low concentrations, provided an improvement in nutrition and survival parameters compared to strictly plant protein based diets as VM, maybe explained by the maintenance of an effective immune response throughout the assay.The research has been partially funded by Vicerrectorat d'Investigacio, Innovacio i Transferencia of the Universitat Politecnica de Valencia, which belongs to the project Aquaculture feed without fishmeal (SP20120603). 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    Insulin-Like Growth Factor-Binding Proteins of Teleost Fishes

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    The insulin-like growth factor (Igf) binding protein (Igfbp) family has a broad range of physiological functions and a fascinating evolutionary history. This review focuses on the Igfbps of teleost fishes, where genome duplication events have diversified gene repertoire, function, and physiological regulation—with six core Igfbps expanded into a family of over twenty genes in some lineages. In addition to briefly summarizing the current state of knowledge on teleost Igfbp evolution, function, and expression-level regulation, we highlight gaps in our understanding and promising areas for future work
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