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.     

Nutrient digestibility profile of premium (category III grade) animal protein by-products for temperate marine fish species (European sea bass, gilthead sea bream and turbot)

Three trials, with classical experimental designs for in vivo digestibility studies, were conducted to determine the apparent digestibility coefficient (ADC) of protein (ADCp), lipid (ADCl), energy (ADCe) and amino acids (AA) in selected animal by-products fed to European sea bass, Dicentrarchus labrax (Trial 1), gilthead sea bream, Sparus aurata (Trial 2), and turbot, Psetta maxima (Trial 3). In each trial, five experimental diets [including a reference diet (RD)] where fish meal (FM) was used as the sole protein source were fed ad libitum to the fish for a period of 4 weeks. Test diets were based on the FM RD and obtained by replacing 30% of the RD with a category III designated European animal by-products (fit for human consumption), namely: steam hydrolysed feather meal (HFM), enzyme-treated feather meal (EFM), poultry meat meal (PMM) and spray-dried haemoglobin meal (SDHM). Faecal material was collected using the 'Guelph system', and nutrient and energy digestibility coefficients were related to the measurement of chromic oxide (Cr₂O₃) incorporated into the diet at a rate of 0.5%. Without any exception, FM diets yielded the best digestibility values for all macro-nutrients and by all fish. Among the test ingredients, ADCp was consistently higher for PMM and SDHM in the three species (85.5%, 91.1% in sea bass; 79.2%, 82.8% in sea bream; and 78.4%, 74.8% in turbot). Conversely, ADCp of HFM and EFM were less efficiently digested (67.2%, 68.2% in sea bass; 21.5%, 21.7% in sea bream; and 46.6%, 36.0% in turbot). However, the novel processing method applied to feather meal did not considerably influence the digestibility of most of the nutrients in this feedstuff. The current investigation yielded valuable numerical ADC for EAA considered to be of prime importance in generating balanced diet formulations.     

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.     

Effect of a dietary phospholipid supplementation on growth and fatty acid composition of European sea bass (Dicentrarchus labrax L.) and turbot (Scophthalmus maximus L.) juveniles from weaning onwards

Two 40-day feeding trials using extruded diets were conducted to assess the effect of a dietary phospholipid (PL) supplementation on growth, survival and fatty acid composition of European sea bass (Dicentrarchus labrax) and turbot (Scophthalmus maximus) from weaning onwards. Two dietary treatments (FO and PL) were tested; both had an identical extruded basis (92.5% total diet weight) coated with a different lipid fraction (7.5% total diet weight). Diet PL contained 2% egg yolk PL (69% pure). In diet FO the PL was replaced by hydrogenated coconut oil. The isolipidic diets contained an equal amount of fish oil ethyl esters providing 1.6% (% diet dry weight) of n-3 highly unsaturated fatty acids (HUFA). A diet water stability test showed no effect of the PL supplementation on the leaching of the dietary fatty acids. In both fish species weight, but not survival, significantly increased as a result of PL supplementation. Weaning onto the experimental diets resulted in similar changes in the relative percent levels of fatty acids in both species. In general, the percentage of saturated fatty acids levelled off after a rapid increase, while monoenes increased after an initial decrease. Total n-3 polyunsaturated fatty acids (PUFA) decreased and total n-6 PUFA remained almost constant. The major effect of the dietary PL on fish fatty acid composition was a 50% increase in n-6 and n-3 HUFAs compared to the PL-free FO diet. The rise in n-6 HUFA may have reflected the higher moiety in the dietary PL. On the other hand this was not the case for the n-3 HUFA since they represented only low levels in the PL fraction (0.1%) compared to that provided by the ethyl esters (1.6%) suggesting a more efficient incorporation of the PL n-3 HUFA than of the ethyl ester n-3 HUFA. A second hypothesis is that the dietary PL may have favored the incorporation of the dietary ethyl ester n-3 HUFA.     

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.     

Combined effects of dietary mannan oligosaccharides and total fish oil substitution by soybean oil on European sea bass (Dicentrarchus labrax) juvenile diets

This study evaluates the effects of dietary mannan oligosaccharides (MOS) on growth, tissue composition, fatty acid profiles and liver morphology of European sea bass (Dicentrarchus labrax) fed diets containing either soybean oil (SBO; SBOMOS) or fish oil (FO; FOMOS) as unique oil source for 8 weeks. Results showed that MOS supplementation enhanced specific growth rate, regardless of the oil source used, and that dietary oil source reduced fish length, regardless of dietary MOS supplementation. Dietary MOS favoured lipid accumulation in muscle and anterior intestine when supplemented in FO‐based diets compared to fish fed SBO diet and reduces it in liver in relation to lower hepatocyte area, particularly in fish fed SBOMOS diet. Dietary MOS favoured liver and not muscular ∑n‐3 PUFA, DHA, EPA and ARA deposition, when combined with FO but not when included in SBO‐based diets. Thus, MOS dietary supplementation favours fish performance and helps to minimize the side effects derived from high dietary SBO supplementation on liver lipid accumulation and hepatocyte vacuolization, which could be of especial interest on long‐term feeding trials; however, the effects on favoured deposition ∑n‐3 PUFA are limited to FO‐based diets.     

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.     

Requirement of n-3 long chain polyunsaturated fatty acids for European sea bass (Dicentrarchus labrax) juveniles: growth and fatty acid composition

European sea bass juveniles (14.4±0.1 g mean weight) were fed diets containing different levels of fish oil then of n-3 highly unsaturated fatty acids (n-3 HUFA) for 12 weeks. The fish performance as well as fatty acid (FA) composition of neutral and polar lipids from whole body after 7 and 12 weeks feeding were studied. The requirements of juvenile sea bass for n-3 highly unsaturated fatty acids (n-3 HUFA) were studied by feeding fish diets containing six different levels of n-3 HUFA ranging from 0.2% to 1.9% of the diet, with approximately the same DHA/EPA ratio (1.5:1).

The growth rate at the end of the trial showed significant differences. Fish fed low dietary n-3 HUFA (0.2% DM of the diet) showed significantly lower growth than the diet 3 (0.7%), then no further improvement (P>0.05) of growth performance was seen by elevating the n-3 HUFA level in the diet up to 1.9% (diet 6). No difference in feed efficiency, protein efficiency ratio or protein retention was observed among treatments, nor in protein and total lipid content. However, the n-3 HUFA levels in diets highly influenced fish fatty acid composition in neutral lipid, while polar lipid composition was less affected. Comparison of polar lipid content after 7 or 12 weeks indicated that DHA remained stable at the requirement level, while arachidonic acid decreased with time. Results of this experiment suggest that the requirement for growth of n-3 HUFA of juvenile sea bass of 14 g weight is at least 0.7% of the dry diet.     

Deposition of conjugated linoleic acid in market size sea bass (Dicentrarchus labrax) and its effects on performance,composition and fillet sensory and texture attributes

The effect conjugated linoleic acid (CLA) accumulation in fish fillet on sensory and texture attributes has been evaluated. Fish were fed diets with graded levels of CLA, which were administered for different durations prior to harvest. Fish were sampled for fillet proximate composition and fatty acid profile for sensory and texture parameters at the end of the trial. Growth performance, feed conversion ratio and muscle proximate composition were not affected by dietary treatments. Muscle fatty acid profile was not affected by dietary treatments despite the increase in saturated fatty acids in fish fed diet with 10 g kg⁻¹ of CLA for 16 weeks before harvest. Besides, the deposition levels of CLA isomers increased gradually depending on the CLA treatment, reaching its maximum level in fish fed 10 g kg⁻¹ CLA diet for 20 weeks before harvest. Fish fed 10 g kg⁻¹ were juicier (P < 0.05) compared with control group, whereas fish fed 5 g kg⁻¹ CLA diet were less (P < 0.05) chewy than fish fed control diet regarding sensory attributes. Texture parameters were not significantly affected by dietary CLA. These results suggest that market size European sea bass could successfully incorporate CLA isomers with some minor positive effects on sensory analyses.     

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.     

Effect of broodstock diets on lipid classes and their fatty acid composition in eggs of gilthead sea bream (Sparus aurata L.)

Total lipid content, total lipid fatty acids, lipid classes and their fatty acid compositions were studied in eggs of gilthead sea bream from two different broodstocks. The two broodstock diets used had the same total lipid content, but differed with respect to their fatty acid levels. The lipid class composition of eggs from different origins was not significantly different. The predominant polar lipids were phosphatidylcholine, phosphatidylethanolamine and sphingomyelin, and the main neutral lipid classes were cholesterol esters and triacylglycerols. The fatty acid composition of total lipids and main lipid classes in the two groups of eggs showed marked differences, reflecting the influence of fatty acid levels in the broodstock diets.     

Comparative pathogenicity study of ten different betanodavirus strains in experimentally infected European sea bass,Dicentrarchus labrax (L.)

Viral encephalopathy and retinopathy (VER), otherwise known as viral nervous necrosis (VNN), is a severe pathological condition caused by RNA viruses belonging to the Nodaviridae family, genus Betanodavirus. The disease, described in more than 50 fish species worldwide, is considered as the most serious viral threat affecting marine farmed species in the Mediterranean region, thus representing one of the bottlenecks for further development of the aquaculture industry. To date, four different genotypes have been identified, namely red‐spotted grouper nervous necrosis virus (RGNNV), striped jack nervous necrosis virus (SJNNV), tiger puffer nervous necrosis virus and barfin flounder nervous necrosis virus, with the RGNNV genotype appearing as the most widespread in the Mediterranean region, although SJNNV‐type strains and reassortant viruses have also been reported. The existence of these genetically different strains could be the reason for the differences in mortality observed in the field. However, very little experimental data are available on the pathogenicity of these viruses in farmed fish. Therefore, in this study, the pathogenicity of 10 isolates has been assessed with an in vivo trial. The investigation was conducted using the European sea bass, the first target fish species for the disease in the Mediterranean basin. Naive fish were challenged by immersion and clinical signs and mortality were recorded for 68 days; furthermore, samples collected at selected time points were analysed to evaluate the development of the infection. Finally, survivors were weighed to estimate the growth reduction. The statistically supported results obtained in this study demonstrated different pathogenicity patterns, underlined the potential risk represented by different strains in the transmission of the infection to highly susceptible species and highlighted the indirect damage caused by a clinical outbreak of VER/VNN.     

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.     

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).     

Dummy regression analysis for modelling the nutritionally tailored fillet fatty acid composition of turbot and sole using gilthead sea bream as a reference subgroup category

Farmed turbot and sole were sampled at different stages of the production cycle for analysis of fillet lipid content and fatty acid (FA) composition. The entire data set along with our own published data on gilthead sea bream were fitted to dummy regression equations with turbot and sole as dummy variables, gilthead sea bream as a reference subgroup category, and diet FA composition and fillet lipid content as independent variables. The relative contribution of each independent variable to the total variance was found to vary within and among FAs and fish species, but strong correlation coefficients (0.76 <  r² > 0.99) were found for almost all of the FA equations, including saturated FAs, monoenes and long‐chain polyunsaturated fatty acids (PUFA) of n‐3 and n‐6 series. Given the differences in lipogenic activities of the fish species, major interaction effects between fillet lipid content and dummy variables were found for monoenes and saturated FAs. The proposed equations (hosted at www.nutrigroup-iats.org/aquafat ) were able to fit different proportions of EPA, DPA and DHA underlying the fish species differences in FA desaturation/elongation pathways. The robustness of the model was proven with extra data from the three fish species, allowing a close linear association near to equality for the scatter plot of observed and predicted values.     

Effects of six alternative lipid sources on growth and tissue fatty acid composition in Japanese sea bass (Lateolabrax japonicus)

Ten-week experiment was carried out on Japanese sea bass (5.87 ± 0.02 g) to study the effects of replacement of fish oil with six alternative lipid sources: pork lard, PL; beef tallow, BT; poultry fat, PF; soybean oil, SO; corn oil, CO; and a mixed-fat (MF: tallow, 60%; soy oil, 20%; fish oil, 20%) on growth performance and fatty acid (FA) composition in fillet and liver. Seven isoenergetic and isonitrogenous experimental diets were formulated, containing 10% of added lipid. Fish oil was used in control diet, which was substituted by 50% with the alternative lipid sources in the other six diets.

Weight gain (WG), specific growth rate (SGR), Feed conversion ratio (FCR) feed intake and hepatosomatic index (HSI) of fish fed the experimental diets were not significantly different (P > 0.05). Protein efficiency ratio (PER) in fish fed the PF diet were significantly lower than those of fish fed SO and CO diets. Significant differences in carcass moisture and lipid contents of carcass and liver were observed among fish fed the dietary treatments. Generally, the fatty acid composition of fish fillets and livers reflected the dietary FA composition.     

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.