Effects on mortality and stress response in European sea bass, Dicentrarchus labrax (L.), fed mannan oligosaccharides (MOS) after Vibrio anguillarum exposure

The effects of dietary mannan oligosaccharides (MOS; 4 g kg^?1; Bio‐Mos, Alltech Inc, USA) in diets for European sea bass, Dicentrarchus labrax (L.), juveniles in relation to disease and stress resistance, combining intestinal infection with Vibrio anguillarum and stress challenge by confinement, were assessed in this study. After 8 weeks of MOS supplementation, fish were exposed to a pathogen challenge test against V. anguillarum by direct gut inoculation combined with a confinement stressor panel. Cumulative mortality of fish fed MOS caused by anally inoculated V. anguillarum decreased from 66% to 12.5% and from 54.1% to 25% in infected and infected + stressed fish, respectively, compared to fish fed control diet. Results for European sea bass revealed a positive effect of MOS dietary inclusion on disease resistance, in terms of cumulative mortality, against gut inoculated V. anguillarum, as well as reduced effects of stress on microbiota diversity. Both of these findings, together with the enhanced innate immune response and the higher gut mucus production and density of eosinophil granulocytes in gut mucosa obtained in previous studies after MOS supplementation ( Torrecillas et al. 2007, 2011a,b ) suggest that general reinforcement of the innate immune system, and particularly of the intestinal barrier efficiency, is the main defence mechanism of European sea bass fed MOS against pathogenic microorganisms.     

Evaluation of zinc supplementation in European sea bass (Dicentrarchus labrax) juvenile diets

This study aimed to investigate the effect of increased zinc dietary levels on the growth performance, feed utilization, immune status and induced wound healing in European sea bass (Dicentrarchus labrax). Fish weighing 10 g were fed five diets containing organic zinc at 30, 70, 110 and 150 mg kg^?1 diet and one inorganic zinc source (zinc oxide) at 150 mg kg^?1 diet for a period of 12 weeks. No significant (P>0.05) differences were found in the growth performance parameters (final weight, specific growth rate), immunological indices tested (respiratory burst activity in whole blood) and the wound‐healing process. Zinc accumulation in the skin, vertebrae and liver increased significantly (P<0.01) with increased dietary zinc levels but not in muscle. The highest zinc concentrations were obtained in skin tissue, followed by vertebrae, liver and muscle tissue. Using the concentration of zinc in skin as a response criterion, broken‐line analysis showed that the supplementation of 148 mg organic Zn kg^?1 diet seemed to be the optimum dietary zinc supplementation level for sea bass juveniles.     

Growth, digestibility and nutrient utilization of rainbow trout (Oncorhynchus mykiss) and European sea bass (Dicentrarchus labrax) juveniles fed different dietary soybean oil levels

A shortage of marine raw ingredients, such as fish oil, is predicted in the near future. The use of suitable alternative lipid sources, such as vegetable oils, is crucial for sustainable growth of the aquaculture sector. Three isonitrogenous (50% Crude Protein) and isoenergetic (19 kJ g⁻¹) diets, in which fish oil was gradually replaced by soybean oil (0, 25 and 50% of the oil fraction) were tested for 12 weeks, using European sea bass (Dicentrarchus labrax) and rainbow trout (Oncorhynchus mykiss) juveniles. No dietary effects (p>0.05) on growth performance, feed conversion, digestibility or body composition were observed with either species. The results obtained suggest the inclusion of soybean oil up to 50% of the dietary lipid is possible, in diets for sea bass and/or rainbow trout juveniles.     

Phosphorus requirement of European sea bass (Dicentrarchus labrax L.) juveniles

A growth trial was conducted to estimate the phosphorus requirement of European sea bass juveniles. Six experimental isonitrogenous and isoenergetic semi‐purified diets (casein based) were formulated to contain 0.48%, 0.65%, 0.77%, 0.86%, 1.05% and 1.25% phosphorus (diets D1, D2, D3, D4, D5 and D6 respectively). Dicalcium phosphate was used as dietary phosphorus source. Twelve groups of 25 fish of 10 g initial body weight were allocated to 55‐L tanks in a thermoregulated water recirculating system. Each experimental diet was assigned to duplicate groups of these fish. The trial lasted for 10 weeks and fish were fed two times a day, 6 days a week, to apparent visual satiation. At the end of the trial, final weight of fish fed diet D1 was significantly lower than that of the other groups, except of fish fed diet D4. Mortality of fish fed diet D1 was significantly higher than that of fish fed the other diets, except for diet D6. Feed efficiency and protein efficiency ratios were significantly lower with diet D1 than with the other diets. Nitrogen retention (% nitrogen intake) of fish fed diet D1 was significantly lower than in the other groups, except that of fish fed diet D4. Energy retention (% energy intake) was not significantly different among groups. At the end of the trial there were no differences in whole body composition among groups. Whole body phosphorus content averaged 0.72% (on a fresh weight basis) and was not significantly affected by dietary phosphorus content. Phosphorus retention averaged 6.1 g kg^?1 weight gain and was not significantly different among groups. Phosphorus retention (% phosphorus intake) was significantly higher in fish fed diets D2 and D3 than in fish fed higher dietary phosphorus levels. Expressed per unit body weight per day, phosphorus retention was not significantly different among groups fed diets D2 to D6, while phosphorus losses linearly increased with dietary phosphorus intake. Results of this trial indicate that the phosphorus requirement of sea bass juveniles was satisfied with a diet containing 0.65% of phosphorus.     

Improved feed utilization,intestinal mucus production and immune parameters in sea bass (Dicentrarchus labrax) fed mannan oligosaccharides (MOS)

In the present study, the effects of dose dietary administration of mannan oligosaccharides (MOS) (Bio‐Mos; Alltech Inc.) on growth, digestibility, liver morphology, lipid and carbohydrate metabolism enzymes, organoleptic properties, immune parameters and gastrointestinal tract mucus production were assessed. MOS dietary inclusion significantly reduced lipogenic enzymes activity in liver and resulted in a lower lipid vacuolization and more regular‐shaped morphology of hepatocytes around the sinusoidal spaces, which could be associated to a better energy and feed utilization because of the improvement of feed utilization and the reduction of feed intake found in the current experiment. Moreover, MOS inclusion enhanced phagocytic activity of head kidney leukocytes in fish fed 4 and 6 g kg^?1 MOS diets at days 30, 45 and 60. Gut quantitative morphological analyses showed a marked dose‐dependent enhancement in the number of cells secreting acid mucins by unit of area which could be related to the previous improvement in resistance to bacterial infection. No effects on sensorial parameters and biochemical composition of flesh were detected.     

Effects of dietary mannan oligosaccharides on growth,body composition and intestine of the sea bass (Dicentrarchus labrax L.)

The effect of Bio‐Mos incorporated into feed formulation on the growth, feed utilization, body composition and histological response of sea bass (Dicentrarchus labrax) was investigated. Final fish weight, feed conversion ratio (FCR), specific growth rate (SGR), protein efficiency ratio (PER), protein productive value (PPV) and energy utilization (EU) of sea bass feed Bio‐Mos supplemented diet and the control diet were determined at the end of experiment. Data present mean ± SD from triplicate determination (n = 3) for a 75‐day feeding trial. Different concentrations (0.5, 1.0, 2.0. 3.0 and 4.0 g kg diet^?1) of Bio‐Mos supplementation and a control without Bio‐Mos incorporation were fed to the fish, feed in the form of dry diets. Average final weight was significantly higher at 1.0 g kg diet^?1 of Bio‐Mos and lowest in control fish group; SGR significantly higher at the 1.0 g kg diet^?1 of Bio‐Mos. In addition,, PER, PPV and EU showed the same trend. The chemical body composition, crude protein, dry matter, crude fat and ash were significant (P ≤ 0.01) compared to control groups. We concluded that 1.0 g kg diet^?1 of Bio‐Mos improved the growth, feed efficiency, histology intestine and survival of sea bass.     

Influence of dietary levels of soybean meal on the performance and gut histology of gilthead sea bream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.)

The present study was performed to determine the effect of soybean meal (SBM) on the performance and gut histology of gilthead sea bream and European sea bass. Three isonitrogenous and isolipidic extruded diets (crude protein, 470 g kg⁻¹ diet; crude fat, 200 g kg⁻¹ diet) were formulated containing 0 (0 SBM), 180 (180 SBM) and 300 (300 SBM) g kg⁻¹ diet SBM and tested on both species in two separate experiments. Fish at an initial average weight of around 18 g were randomly allocated to 800 L square tanks connected to a closed recirculating system. The trials lasted 80 days for sea bream and 89 days for sea bass. Fish were hand-fed to apparent satiation. Increasing the level of SBM had no significant effects on the specific growth rate, feed intake and feed conversion rate in both the species. In the sea bream distal intestine, lamina propria was moderately and diffusely expanded in some fish due to an increase in cell infiltration represented by mononuclear cells, this finding being more frequent in animals fed diet 300 SBM. No other morphological alterations in intestinal folds, enterocytes or other inflammation signs were noticed in the sea bream distal intestine of any group. No histological differences were found in sea bass in any experimental group.     

Partial replacement of fish oil by soybean oil on lipid distribution and liver histology in European sea bass (Dicentrarchus labrax) and rainbow trout (Oncorhynchus mykiss) juveniles

A 12‐week feeding trial was conducted to evaluate the effects of fish oil replacement by soybean oil, on lipid distribution and liver histology of two commercially important finfish species: rainbow trout (Oncorhynchus mykiss) and European sea bass (Dicentrarchus labrax). Sea bass (16.2 ± 0.5 g; mean ± SD) and rainbow trout (52.1 ± 0.5 g) juveniles were fed one of three isonitrogenous (500 g kg^?1 CP) and isoenergetic (19 kJ g^?1) diets, containing 0% (control, diet A), 25% (diet B) and 50% (diet C) soybean oil. At the end of the experiment, lipid deposition was evaluated in muscle, liver and viscera. Cholesterol and triglycerides levels were also determined in plasma. Tissue total, neutral and polar lipid composition (g kg^?1 total lipids) showed no significant differences within species, regardless the dietary treatment. The same trend was observed for plasma parameters (P > 0.05). Viscera were the preferential tissue of lipid deposition, with 252–276 and 469–513 g kg^?1 total lipid content in trout and sea bass, respectively. Dietary fish oil replacement had no effect on either hepatic lipid droplets accumulation or degree and pattern of vacuolization in the observed liver sections. These data suggest that both sea bass and trout can be fed diets containing up to 50% soybean oil without adverse effects on tissue lipid composition or liver histology.     

Influence of partial substitution of dietary fish oil by vegetable oils on the metabolism of [1-14C] 18:3n-3 in isolated hepatocytes of European sea bass (Dicentrarchus labrax L.)

The static or declining supply of fish oil from industrial fisheries demands the search of alternatives, such as plant (vegetable) oils, for diets in expanding marine aquaculture. Vegetable oils are rich in C₁₈ polyunsaturated fatty acids but devoid of the n-3 highly unsaturated fatty acids in fish oils. Previous studies, primarily with salmonids, have shown that including vegetable oils in their diets increased hepatocyte fatty acid desaturation. In the present study, we have investigated the effects of dietary partial substitution of fish oil (FO) with rapeseed oil (RO), linseed oil (LO) and olive oil (OO) on the desaturation /elongation and, -oxidation capacities of [1-¹⁴C]18:3n-3 in isolated hepatocytes from European sea bass (Dicentrarchus labrax L.), in a simultaneous combined assay. Fish were fed during 34 weeks with diets containing 100% FO, or RO, LO and OO, each included at 60% with the balance being met by FO, with no detrimental effect upon growth or survival. The highest total desaturation rates were found in hepatocytes of fish fed FO diet (0.52±0.08 pmol/h/mg protein) and OO diet (0.43±0.09 pmol/h/mg protein), which represented 3.2% and 2.7% of total [1-¹⁴C]18:3n-3 incorporated, respectively. In contrast, lowest desaturation rates were presented by hepatocytes of fish fed LO and RO diets (0.23±0.06 and 0.14±0.05 pmol/h/mg protein, respectively) represented 1.4% and 0.9% of total [1-¹⁴C]18:3n-3 incorporated, respectively. The rates of [1-¹⁴C]18:3n-3 β-oxidized were between 11-fold and 35-fold higher than desaturation. However, no significant differences were observed among β-oxidation activities in hepatocytes of fish fed any of the diets. The present study demonstrated that the European sea bass, as a carnivorous marine fish, presented a ‘marine’ fish pattern in the metabolism of 18:3n-3 to 20:5n-3 and 22:6n-3. This species appeared to have all the enzymic activities necessary to produce 22:6n-3 but presented only extremely low rates of fatty acid bioconversion. Furthermore, nutritional regulation of hepatocyte fatty acid desaturation was minimal, and dietary vegetable oils did not increase desaturase activities, and in RO and LO treatments the activity was significantly lower. This revised version was published online in July 2006 with corrections to the Cover Date.     

Replacement of fishmeal with rice protein concentrate in practical diets for European sea bass Dicentrarchus labrax reared at winter temperatures

As no information is available regarding the efficacy of using rice protein concentrate (RPC) to replace fishmeal (FM) in diets for European sea bass (Dicentrarchus labrax), a trial was conducted to determine the optimum inclusion for European sea bass juveniles. Diets were formulated to replace 25%, 50% and 75% of fishmeal with RPC. Additional high level inclusion diets (50% and 75%) were supplemented with lysine and methionine to determine if these were limiting factors. Fish (18.0 ± 0.06 g) were fed the experimental diets for 12 weeks. The final mean weight and specific growth rate (SGR) were significantly higher in fish fed the control diet (diet FM) and low level RPC (diet RPC25) than in fish fed the other diets. The supplementation of the amino acids had a positive effect on growth, elevating the final weight of the high level RPC (RPC75 + AA) group above that of the non‐supplemented group (RPC75). However, this was still not comparable to fish fed FM and RPC25 diets. No significant differences in body composition were observed. A digestibility study demonstrated an inverse relationship between elevating inclusion levels of RPC and apparent digestibility coefficients (ADCs) for dry matter and lipid, with significantly lower values than the control being observed with RPC inclusions above 25%. The mesenteric fat index followed this trend. Circulating leucocyte levels, leucocyte ratios and serum lysozyme activity remained unaffected by dietary treatment. However, compared with the control group, fish fed RPC75, RPC50 + AA RPC75 + AA displayed significantly lower haematocrit values. The present study demonstrates that the inclusion of RPC at 140 g kg^?1 (effectively replacing 25% of FM content) does not compromise European sea bass growth performance, body quality or basic haematological parameters. It is suggested that future studies including higher levels test the efficacy of additional amino acid supplementation (e.g. tryptophan).     

Interactive effects of dietary fishmeal level and plant essential oils supplementation on European sea bass,Dicentrarchus labrax: Growth performance,nutrient utilization,and immunological response

The inclusion of phytogenics in fish feed is a promising strategy to compensate for the negative performance effects of replacing fishmeal (FM) with vegetable sources. The present work assessed the interactive effects of different dietary FM levels (22.5 and 10% of formulation) and the supplementation of a commercial blend of anise, citrus, and oregano essential oils (Digestarom PEP M.G.E 150) on European sea bass, Dicentrarchus labrax growth performance, nutrient utilization, gut morphology, antioxidant status, and immunological response over a 60‐day growth trial. Results showed decreased growth and protein efficiency ratio and increased feed conversion ratio with a low dietary FM level. In contrast, supplementation of the phytogenic product demonstrated improved performance and nutrient utilization together with increased protein and energy retention. Supplementation with the plant essential oils fully compensated for the negative intestinal changes observed in sea bass fed a low‐FM diet but showed little improvement in fish immunological response, except for the 30% increase in lysozyme activity observed in fish fed the low FM‐supplemented diet compared to those fed the standard high‐FM diet. Overall, this study supports the use of this phytogenic product in low‐FM diets as a possible tool to decrease feed costs associated with FM without compromising fish performance, nutrient utilization, and health.     

Partial substitution of fish oil with rapeseed, linseed and olive oils in diets for European sea bass (Dicentrarchus labrax L.): effects on flesh fatty acid composition, plasma prostaglandins E2 and F2α, immune function and effectiveness of a fish oil finishing diet

Triplicate groups of European sea bass (Dicentrarchus labrax L.), of initial weight 90 g, were fed four practical‐type diets in which the added oil was 1000 g kg^?1 fish oil (FO) (control diet), 600 g kg^?1 rapeseed oil (RO) and 400 g kg^?1 FO, 600 g kg^?1 linseed oil (LO) and 400 g kg^?1 FO, and 600 g kg^?1 olive oil (OO) and 400 g kg^?1 FO for 34 weeks. After sampling, the remaining fish were switched to the 1000 g kg^?1 FO diet for a further 14 weeks. Fatty acid composition of flesh total lipid was influenced by dietary fatty acid input but specific fatty acids were selectively retained or utilized. There was selective deposition and retention of docosahexaenoic acid (DHA; 22:6n‐3). Eicosapentaenoic acid (EPA; 20:5n‐3) and DHA were significantly reduced and linolenic (LNA; 18:3n‐3), linoleic (LA; 18:2n‐6) and oleic (OA; 18:1n‐9) acids significantly increased in flesh lipids following the inclusion of 600 g kg^?1 RO, LO and OO in the diets. No significant differences were found among different treatments on plasma concentrations of prostaglandin E₂ and prostaglandin F_2α. Evaluation of non‐specific immune function, showed that the number of circulating leucocytes was significantly affected (P < 0.001), as was macrophage respiratory burst activity (P < 0.006) in fish fed vegetable oil diets. Accumulation of large amounts of lipid droplets were observed within the hepatocytes in relation to decreased levels of dietary n‐3 HUFA, although no signs of cellular necrosis was evident. After feeding a FO finishing diet for 14 weeks, DHA and total n‐3 HUFA levels were restored to values in control fish although EPA remained 18% higher in control than in the other treatments. This study suggests that vegetable oils such as RO, LO and OO can potentially be used as partial substitutes for dietary FO in European sea bass culture, during the grow out phase, without compromising growth rates but may alter some immune parameters.     

Effects of conventional and organic feed on the mineral composition of cultured European sea bass (Dicentrarchus labrax)

European sea bass (Dicentrarchus labrax) is a widely consumed marine fish in Mediterranean areas, and different farming techniques are applied for fish culturing to satisfy the growing demand for seafood. The aim of this study was to investigate the effects of conventional and organic feed on the quality of cultured European sea bass (Dicentrarchus labrax) collected during the growth period. The concentrations of ash, moisture, essential macro‐elements (Ca, K, Mg, Na and P), micro‐elements (Co, Cr, Cu, Fe, Mn, Ni and Zn) and toxic elements (As, Cd, and Pb) were determined in feeds and in fillets of cultured fish. The results were compared to those obtained from wild sea bass. Results showed that the differences between organic diet‐fed and conventional diet‐fed sea basses varied in relation to the specific element measured and the growth period. The former showed higher concentration of Fe, Mg and Cr, and lower Na content. The amount of P, Na, Fe, Cu and Cr in wild sea bass was significantly (P ≤ 0.05) higher than that found in farmed fish. The levels of toxic elements in cultured sea bass were always within the allowed limit for fishery products. Wild samples had significantly (P ≤ 0.05) higher content of arsenic and lead than farmed sea bass. Cultured fish represent a valuable dietary source of essential macro‐ and micro‐elements. Controlled rearing systems and feedings were related to a decrease in the presence of some toxic metals in cultured fish compared with wild fish.     

Growth,development and survival of European sea bass (Dicentrarchus labrax) larvae cultured under different light spectra and intensities

We assessed the effects of light intensity and spectrum on the growth, development and survival of Dicentrarchus labrax larvae from 30 to 96 days after hatching. Twelve lighting regimes were applied using 0.3, 1 and 2 W/m² full spectrum white, blue, red and green light. By day 66 of exposure, growth performance was significantly better in larvae reared under green or red light compared with white or blue light (p < 0.05), while it was significantly better at 2.0 W/m² compared with 1.0 or 0.3 W/m² (p < 0.05). Larvae reared under white or blue light had significantly more deformities (p < 0.05) than did larvae reared under light of other spectra, while larvae reared under 0.3 W/m² light intensity had a significantly higher proportion of jaw malformations than reared under 1.0 and 2.0 W/m² (p ? 0.05). Dicentrarchus labrax larvae subjected to green light showed significantly lower survival compared with the other light spectra (p < 0.05). Moreover, light significantly affected brain aanat1a, aanatt1b, mellc, mt1, mt2, gh, tsh and crf mRNA expression. Therefore, the red and green light groups (or under 2.0 W/m²) could improve the growth performance and enhance secretion of melatonin, gh and tsh, and reduce the proportion of jaw malformations.     

Compensatory growth response of European sea bass (Dicentrarchus labrax L.) under cycled starvation and restricted feeding rate

The compensatory growth response of the European sea bass (Dicentrarchus labrax) that faced cycling starvation and restricted ration was assessed. Juveniles (10.5 g) were stocked into 15 tanks at a density of 25 fish per tank. Five different feeding regimes were tested on triplicate groups of fish: C_Satiation: control fed for 60 days without deprivation, C_Restricted: 25% restricted feeding, S‐R: 1 day starvation then 4 days C_Restricted feeding, R‐F: 1 day C_Restricted feeding then 4 days C_Satiation feeding, and, finally, S‐F: 1 day starvation then 4 days C_Satiation feeding. The specific growth rate of fish in the C_Satiation (2.5 ± 0.06% day^?1), S‐F (2.5 ± 0.11% day^?1) and R‐F (2.4 ± 0.18% day^?1) were significantly higher than that of C_Restricted (2.2 ± 0.05% day^?1) or S‐R (2.0 ± 0.01% day^?1). Fish in S‐F group were able to achieve catching up with the C_Satiation. There was no significant difference in feed conversion rates but R‐F and S‐F consumed approximately 34% more feed than C_Satiation following the first re‐feeding day. Although, the highest lipid content was observed in C_Satiation (14.4%), S‐R (33.3% dry matter) had the highest water content compared with the C_Satiation and C_Restricted (37.8% and 36.9% dry matter respectively). In conclusion, it may be concluded that sea bass has rapid response to cycling starvation/re‐feeding and that a 25% restricted feeding ratio is insufficient to invoke a compensatory growth response in sea bass.     

Effect of lordosis on the swimming performance of juvenile sea bass (Dicentrarchus labrax L.)

The critical swimming speed (U_crit, cm s⁻¹) value of normal juvenile sea bass (Dicentrarchus labrax) (64.7±6.33) was significantly higher than that of fish with lordosis (60.3±6.66) (P<0.05). Both normal fish and those with lordosis showed increased endurance with increasing length during exercise at a fixed water velocity of 50 cm s⁻¹. This speed was used to investigate the separation possibility of deformed specimen from the normal developed fish based on their swimming performance, and 28% separation was achieved without losing any normal fish by the end of the 10th minute. To achieve a better separation of the deformities from the normal, first grading of the juveniles and then a swimming endurance test is advised.     

Saponins and phytosterols in diets for European sea bass (Dicentrarchus labrax) juveniles: effects on growth,intestinal morphology and physiology

This study aimed to elucidate the effects of two soy antinutrients, saponins and phytosterols, on growth, intestinal health and function of European sea bass juveniles. Seven fishmeal‐based diets were formulated: a control diet without antinutrients and six experimental diets containing low or high levels of soyasaponins (SapL and SapH, respectively), low or high levels of phytosterols (PhytL and PhytH, respectively) and low or high levels of soyasaponins + phytosterols (SapPhytL and SapPhytH, respectively). A feeding trial was conducted for 59 days after which blood was collected for plasmatic cholesterol quantification and intestinal samples were collected for histology, digestive enzymes activity and gene expression analysis. Histology was also performed at day 15. PhytL resulted in high distal intestine (DI) relative weight and decreased plasma cholesterol, while PhytH caused inflammatory changes in the DI. SapH depressed maltase and alkaline phosphatase activity in DI and the histological evaluation indicated some inflammatory changes. The SapPhytH resulted in decreased maltase activity in the DI and indications of inflammatory changes that were supported by the results on gene expression profiles. The antinutrients tested did not compromise growth, however, caused some gastrointestinal disturbance that may affect fish in the long term.