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The use of low fishmeal/fish oil in marine fish diets affects dietary essential fatty acids (EFAs) composition and concentration and, subsequently, may produce a marginal deficiency of those fatty acids with a direct impact on the fish intestinal physiology. Supplementation of essential fatty acids is necessary to cover the requirements of the different EFAs, including the ones belonging to the n-6 series, such as arachidonic acid (ARA). ARA, besides its structural role in the configuration of the lipid classes of the intestine, plays an important role in the functionality of the gut-associated immune tissue (GALT). The present study aimed to test five levels of dietary ARA (ARA0.5 (0.5%), ARA1 (1%), ARA2 (2%), ARA4 (4%), and ARA6 (6%)) for European sea bass (Dicentrarchus labrax) juveniles in order to determine (a) its effect in selected distal intestine (DI) lipid classes composition and (b) how these changes affected gut bacterial translocation rates and selected GALT-related gene expression pre and post challenge. No differences were found between distal intestines of fish fed with the graded ARA levels in total neutral lipids and total polar lipids. However, DI of fish fed with the ARA6 diet presented a higher (P more...

The aim of this study was to evaluate the effects of increasing dietary arachidonic acid (ARA) levels (from 1 up to 6 % of total fatty acids) on European sea bass (Dicentrarchus labrax) juveniles growth performance, tissue fatty acid profile, liver morphology as well as long chain polyunsaturated fatty acids (LC-PUFA) biosynthesis, triglyceride and cholesterol synthesis and lipid transport. A diet with total fish oil (FO) replacement and defatted fish meal (FM) containing a 0.1 g ARA g-1 diet was added to the experimental design as a negative control diet. Dietary ARA inclusion levels below 0.2 g ARA g-1 diet significantly worsened growth even only 30 days after the start of the feeding trial, whereas dietary ARA had no effect on fish survival. Liver, muscle and whole body fatty acid profile mainly reflected dietary contents and ARA content increased accordingly with ARA dietary levels. Tissue eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) levels were positively correlated among them. Hepatic lipid vacuolization increased with reduced dietary ARA levels. Expressions of fatty acyl desaturase 2 and 3-hydroxy-3-methylglutaryl-coenzyme genes were up-regulated in fish fed the negative control diet compared to the rest of the dietary treatments denoting the influence of ARA on lipid metabolism. Results obtained highlight the need to include adequate n-6 levels and not only n-3 LC-PUFA levels in European sea bass diets. Keywords: Dicentrarchus labrax, arachidonic acid, growth performance, tissue fatty acid profile, lipid metabolism more...

The aim of the present study was to determine the combined effect of both stress and EFA deficiency on several biological and biochemical parameters. Fish were fed during 15 weeks two isocaloric and isoproteic diets: a control diet based on fish oil and formulated to meet the n-3 HUFA requirements for this species (1.5% of n-3 HUFA) and a deficient diet containing beef tallow and formulated to be deficient in n-3 HUFA. Each experimental diet was evaluated both at high and low stocking densities (10 and 3.2 kg m−3 of initial density, respectively). High stocking density produced a chronic stress situation with elevation of plasma cortisol levels. It also caused a reduction in hepatosomatic index and liver lipid contents, increasing the oleic acid/n-3 HUFA ratios in the polar lipids. Fish fed the EFA deficient diet at low stocking density showed common deficiency symptoms. High stocking density in fish fed the EFA deficient diet induced a higher degree of EFA deficiency symptoms leading to mortality, liver steatosis, liver lipid deposition, reduced muscle lipid and reduced n-3 HUFA contents, which particularly affected EPA, but not DHA, suggesting a preferential retention of the latter fatty acid, specially in the phosphoglycerides fraction. more...

Skeletal deformities constitute a major problem for aquaculture industry by decreasing the final value of the fish. An analysis of skeletal malformations in 11,640 fish was performed considering families and triplicates per family. Thirtynine different skeletal abnormalities were detected, such as lordosis, vertebral fusion, absence of one or both operculum, bent-jaw, etc. Moreover, a new unusual complex spinal column deformity consisting of a consecutive repetition of lordosis, scoliosis and kyphosis (LSK) from the head to the caudal fin was described. This syndrome was statistically associated with the family structure (Z2H−family,LSK=3.49; p more...

High stocking density has been shown to produce a wide variety of effects on cultured fish populations, such as alterations in behavior and poor feed utilization, resulting in mortality and poor growth. High stocking density has also been reported to produce chronic stress and mobilization of energy sources in fish. There are few studies focusing on the effect of high stocking density on one of the most important marine fish species for Mediterranean aquaculture, gilthead seabream (Sparus aurata). Consequently, this study investigates the effect of high stocking density on juveniles of this species, focusing on effects of stocking density on growth, biochemical composition, immune status and hematology. more...

Studies on the alternative complement pathway (ACP) activity in fish have suggested the particular relevance of this pathway as a non-specific immune mechanism being its activation directly related with the availability of certain nutrients. In the present study ACP activity in gilthead seabream fed different dietary levels of a-tocopherol and n-3 HUFA deficiencies was studied under different stress conditions (overcrowding and repetitive chasing). A reduction in ACP activity was found when diets with deficiencies in α-tocopherol and n-3 HUFA were assayed at any of the experimental conditions assayed. Levels of plasma cortisol were similar to those of fish after crowding stress subjected to dietary deficiencies at low stocking density, suggesting that dietary deficiencies produced a chronic elevation of cortisol. When fish were subjected to repetitive chasing, plasma cortisol from fish fed control diet showed a progressive increase whereas plasma cortisol levels from both dietary deficient groups showed a transitory peak 4 days after the beginning of the repetitive chasing. However, ACP activity levels of fish fed deficient diets remained lower than the control group and constant during chasing period. These results suggest that both a-tocopherol and n-3 HUFA nutritional deficiencies rather than stress are an important determinant of fish complement activity. more...

Gilthead sea bream, Sparus aurata, were subjected to different stressful situations and the haemolytic activity mediated by the alternative complement and the agglutinating serum activity were tested as indicators of immunocompetence. The results show that both parameters were significantly compromised after chronic or repeated acute stress as well as in groups fed with vitamin E or Ω3 highly unsaturated fatty acids (HUFA) deficient diets and in infected fish. These results suggest that both haemolytic and agglutinating activity are suitable indicators for immunocompetence as they are non-specific responses induced by non-specific stressors. In addition, a comparison is performed with other immune indicators subjected to the same stressors. more...