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.     

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.     

Dietary nucleotides enhance growth performance,feed efficiency and intestinal functional topography in European Seabass (Dicentrarchus labrax)

Nucleotides, nucleosides and nucleic acids (NU) have many critical functions in supporting life and increasing evidence suggests that exogenous supply can benefit the health of mammals and fish. For these reasons, a 6‐week feeding trial was conducted on juvenile European seabass (Dicentrarchus labrax) with diets containing 0%, 0.15% and 0.3% inclusion of a NU mixture (Laltide^®) derived from the yeast Saccharomyces cerevisiae. At the end of the study no significant differences were found in fish performance, although a tendency towards better performance was indicated in fish fed the Nu0.3 diet. In relation to histological assessment, a significantly greater perimeter ratio; internal to outer (IP/OP) was observed in the posterior intestine of fish fed supplemental NU. Microvilli heights in the posterior intestine were also shown to be significantly promoted in fish fed NU diets (p < 0.05). Goblet cell abundance was shown to be unaffected by the inclusion of NU in the diet (p > 0.05). Overall, this study indicates that orally administered NU may be effective promoters of gut functional topography with marginal associated improvements to fish performance. Nonetheless, longer exposure and/or commercial scale application, and in diets that were challenging in use of high inclusion levels of plant by‐products would potentially amplify improvements in production characteristics, in turn benefiting fish culturists.     

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.     

Methionine- and cystine-supplemented diets for juvenile sea bass (Dicentrarchus labrax)

Growth studies indicated that methionine-deficient diets can be supplemented with the free sulphur amino acids, methionine and cystine. The best growth rate was obtained with the diets for which the supplemented methionine:cystine ratio was 20:80. No significant effect on body composition of the methionine:cystine ratio could be detected. The serum free methionine increased with the supplemented methionine content in the diet. The serum free cystine did not exhibit such a relationship. The best growth rate and food conversion were obtained with the diet containing 0.91% methionine, 1.29% cystine and a methionine:cystine ratio of 36:64 on a sulphur molar basis.     

The effect of dietary ribonucleic acid incorporation in performance of European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the effect of dietary ribonucleic acid (RNA) on the performance of European sea bass juveniles. Two diets containing fish meal as the only nitrogen source were formulated with 39% or 50% of crude protein (LP and HP diet, respectively). Two other diets were formulated similarly to LP diet, but included yeast-RNA extract at 6.2% and 12.4% levels (LP6 and LP12 diets, respectively) by replacing dextrin. Total nitrogen content of the LP12 diet was equivalent to that of the HP diet. The four experimental diets were isoenergetic (22 kJ GE g⁻¹ DM). Triplicate groups of 25 fish (IBW=13 g) were fed the test diets for 10 weeks. The reduction of dietary protein level did not significantly affect growth rate but significantly increased feed intake and decreased feed efficiency. However, nitrogen retention (%N intake) was significantly improved with the decrease in dietary protein level. Dietary RNA incorporation did not affect feed intake or feed efficiency, although it reduced growth rate of fish fed the highest dietary RNA level. Nitrogen retention (%N intake) significantly decreased with the dietary supplementation of RNA. Dry matter, nitrogen and energy digestibilities were not significantly affected by dietary treatments. Final whole-body composition was also unaffected by dietary treatments, except for ash content which was significantly higher in the LP12 group. Hepatosomatic index was significantly increased by the dietary incorporation of RNA. The dietary supplementation of RNA had no protein sparing effect and significantly reduced energy retention. It may be concluded that dietary RNA supplementation had no nutritive value for European sea bass.     

Feed discrimination and selection in self-fed European sea bass Dicentrarchus labrax L.

Abstract This study was conducted to evaluate feed discrimination and preference of European sea bass Dicentrarchus labrax L., taking into account the effects of feed location, previous feeding experience and light conditions. Fish (63 g) were held in tanks and exposed to a 12L:12D photoperiod and ambient temperature. The following feeding practices were applied in triplicate: three‐choice feeding (self‐ feeding with feeds formulated for carnivorous, herbivorous and omnivorous fish rotated on a weekly basis) and monofeeding (self‐feeding with one of these feeds). After 9 weeks (phase I), fish previously held in monofeeding, were subjected to three‐choice feeding for 2 weeks (phase II). In phase I, fish discriminated between feeds after their rotational displacements and preferred the feed formulated for carnivorous species. This also supported the best growth. Fish used in phase II preferred the same feed. There was no preference for any location within tanks. Feeding was generally diurnal, although in some tests with the feeds for herbivores and omnivores, diurnal and nocturnal feed demands were similar and fewer in number than with the feed formulated for carnivores.     

Estrogen binding sites in the ovary of the European sea bass (Dicentrarchus labrax L.)§

The binding characteristics of the European sea bass estrogen receptor (ER) were determined in ovarian cytosolic and nuclear extracts. Results were consistent with the presence of a single class of high affinity and low capacity ER, the first to be characterized in teleost ovary. This will contribute to our understanding of changes in ER affinity and abundance during sex differentiation in this economically important species.     

Food sources,digestive efficiency and resource allocation in the sea cucumber Holothuria forskali (Echinodermata: Holothuroidea): Insights from pigments and fatty acids

Various research projects in Europe and North Africa have recently intended to breed temperate holothurians to alleviate fishing pressure on natural populations. However, to date little is known about the nutritional requirements of East Atlantic and Mediterranean species. In this study, we propose a “natural population”‐oriented approach to characterize food sources, digestive efficiency and resources allocation based on the composition of pigments and fatty acids (FA) in gut contents and tissues (muscles, gonads and digestive tract walls) of wild individuals of the species Holothuria (Panningothuria) forskali (Delle Chiaje, 1823) sampled in Brittany (France). Our study reveals that neither green nor red algae enter the diet of H. forskali in spring and that the only fresh vegetal material found in gut contents is brown algae (very likely diatoms). The high nutritional quality of gut contents however contrasts with the detrital nature of the ingested food sources, suggesting that a trophic upgrading of organic matter occurs before digestion. In addition, unusual FA (i.e. only present in a few groups of living species) such as long‐chain monounsaturated FA (especially the FA 23:1ω9) were found in large proportions in muscles and gonad and their effect on sea cucumber fitness needs further investigation.     

Assessment of homozygosity and fertility in meiotic gynogens of the European sea bass (Dicentrarchus labrax L.)

Analysis of 5–6 microsatellite loci was used to measure the increment of homozygosity in two meiogynogenetic progenies (A and B) of sea bass with respect to their mother. In progeny A and B, 20% and 12% of the meiogynogens retained heterozygosity for all investigated maternal loci, respectively, while complete homozygosity was observed only in 6% of B and in none of A, indicating the occurrence of significant allelic recombination during meiosis. The overall increment of homozygosity for the investigated loci was 27% for A and 47% for B. Although survival at hatching of meiogynogens was about half that of controls, they subsequently grew as controls and displayed the same onset of puberty and reproductive potential at adulthood during three consecutive years when crossed between themselves (11 crosses) or with control fish (13 crosses). In particular, meiogynogenetic females (n=24) underwent vitellogenesis and yielded eggs of good quality upon stimulation with LH–RH analogue similarly to normal fish. Sperm released by meiogynogenetic males (n=23) was equivalent to that of controls in terms of volume, quality and fertilization capability.Second generation meiogynogens were obtained by chromosome set manipulation from meiogynogenetic females and found to be morphologically normal at 3 years of age. Interestingly, under our culture conditions, the percentage of meiogynogenetic males in the second generation was 7% as opposed to 39% in the first generation.     

Effects of different dietary oils in sea bass (Dicentrarchus labrax) nutrition

Five diets having the same proximate composition but containingdifferent types of supplemental oils, singly or in combination, were used forgrowing sea bass from 95 g to about 200 g in smallseacages. The oils tested were olive oil, soybean oil and fish oil. The dietsformulated contained EPA and DHA levels ranging from 0.88 to 1.35% of the diet.Growth parameters and fish body composition were not significantly affected bythe type of oil used. The same was generally apparent for liver andhematological characteristics. The content of phospholipids in EPA and DHA washighest in the livers of fish fed diets supplemented with fish oil. A positivecorrelation was found between dietary and liver n-6 PUFA. Histological sectionsindicated extended pathological symptoms (intensive liver degeneration andhemorrhages, changes in the gill structure) in the fish receiving dietssupplemented only with plant oils. These symptoms existed but to a smallerfrequency and degree in livers of fish fed diets supplemented with plant andfish oil, while were not apparent in those fed the fish oil diet.     

Effects of different dietary oils in sea bass (Dicentrarchus labrax) nutrition

Five diets having the same proximate composition but containingdifferent types of supplemental oils, singly or in combination, were used forgrowing sea bass from 95 g to about 200 g in smallseacages. The oils tested were olive oil, soybean oil and fish oil. The dietsformulated contained EPA and DHA levels ranging from 0.88 to 1.35% of the diet.Growth parameters and fish body composition were not significantly affected bythe type of oil used. The same was generally apparent for liver andhematological characteristics. The content of phospholipids in EPA and DHA washighest in the livers of fish fed diets supplemented with fish oil. A positivecorrelation was found between dietary and liver n-6 PUFA. Histological sectionsindicated extended pathological symptoms (intensive liver degeneration andhemorrhages, changes in the gill structure) in the fish receiving dietssupplemented only with plant oils. These symptoms existed but to a smallerfrequency and degree in livers of fish fed diets supplemented with plant andfish oil, while were not apparent in those fed the fish oil diet.     

Characterization of mevalonate metabolism in the sea bass (Dicentrarchus labrax L) liver

The activities of mevalonate kinase, mevalonate 5-phosphate kinase and mevalonate 5-pyrophosphate decarboxylase, were examined in sea bass (Dicentrarchus labrax L) liver. The activities of the three enzymes were studiedin vitro in relation to the influence of protein content, time of incubation, pH, temperature, mevalonate, ATP and Mg⁺⁺ concentration. Protein content in the assay medium affected the three enzymes differently. Mevalonate kinase, mevalonate 5-phosphate kinase, and mevalonate 5-pyrophosphate decarboxylase activities were linear up to 0.2, 0.4 and 0.8 mg protein, respectively. With respect to the time course studies, the enzymes also behaved differently. Mevalonate kinase activity increased over forty minutes, reaching a plateau thereafter, while mevalonate 5-phosphate kinase and decarboxylase increased over the entire assay period. All the three enzymes showed a maximum in activity at pH 7.5. The effect of reaction temperature showed that phosphorylation increased to maximum around 35°C for mevalonate kinase and 30°C for mevalonate 5-phosphate kinase while decarboxylation rates remained constant well until 30°C temperature decreasing afterwards. The enzymes behaved differently as a function of mevalonate concentration. Mevalonate 5-phosphate formed was maximal when the initial mevalonate concentration was 272 M, whereas mevalonate 5-pyrophosphate and CO₂ were formed maximally at mevalonate concentrations of 136 M and 68M, respectively. Optimal ATP concentration in the medium was 3 mM for decarboxylase and 6 mM for kinases, and Mg⁺⁺ requirements varied from 4 mM for decarboxylase to 6 mM for kinases.     

Enhancement of growth and feed utilization of the European sea bass (Dicentrarchus labrax) fed supplementary dietary salt in freshwater

The effects of increased dietary sodium chloride supplement on growth, feed efficiency (FE), feed dispensed rate and fillet composition and morphological indices of European sea bass (initial weight of 4.9 g) were investigated in freshwater using five different levels of supplemental salt (0 (control diet, with no salt supplemented), 1%, 3%, 5% and 9% of diet). Over the experimental period of 50 days, the final weight of dietary salt (DS) groups was found to be approximately 19% higher than the control group. The final weight, specific growth rate and FE were greatest at the moderate level (1–5%) of salt supplementation. Optimum level of DS inclusion was calculated as 3%. Feed efficiency was significantly enhanced with increasing salt level up to 5%. A negative relationship between daily feed intake and FE (r²=0.66) was observed. The average muscle ratio (MR) in groups fed 1–5% supplemented salt was around 51.4% whereas fish receiving control and 9% salty diets had 5.3% lower MR (48.7%) (P<0.05). However, no significant effects of salt inclusion were found on viscera or hepatosomatic indices. Muscle ash and dry matter content slightly rose with increasing DS in the diet, but with no statistical difference (P>0.05). Similarly, no significant changes were observed in either muscle protein or lipid content among the groups (P>0.05).     

Effect of vitamin A on the skeletal morphogenesis of European sea bass,Dicentrarchus labrax (Linnaeus, 1758)

The purpose of this study was to examine the effects of vitamin A levels in the larval diet of Dicentrarchus labrax, (a) on the shape of juvenile skeleton and (b) on the development of pelvic fins. To exclude the side‐effects of skeletal abnormalities on skeleton shape, morphometric analysis was performed on normal individuals only. Geometric morphometric analysis revealed a significant effect of larval nutrition on the shape of juvenile skeleton (Wilk's λ=0.0188, P<0.001), 55 days after the application of the different nutritional regimes. Between‐groups shape differences mainly concerned the position of anterior dorsal pterygiophore, which was associated with a significant (P<0.05) increase of the number of dorsal lepidotrichia and hard spines at low levels of vitamin A (3–9 × 10³ IU kg^?1 dry food). At the lowest vitamin A levels (3 × 10³ IU kg^?1 dry food), the development of pelvic fins was abnormal, with severe bilateral (44±10%, mean±SD) or unilateral (22±9%) lack of the fin lepidotrichia, hard spines and their supporting elements. In the 82–94% of the unilaterally deformed individuals, deformation concerned the left pelvic fin. Concerning the shape of pelvic fins in the normally developed individuals, results demonstrated a significant effect of nutritional conditions on the size and position of all the elements of the pelvic fin area (basipterygium, lower metacleithrum, cleithrum, scapula) (Wilk's λ=0.3087, P<0.001). Our results demonstrate a significant nutritionally driven phenotypic plasticity in D. labrax, with dietary levels of vitamin A determining the skeleton shape and the left–right symmetry of pelvic fins.     

Response to domestication and selection for growth in the European sea bass (Dicentrarchus labrax) in separate and mixed tanks

Selective breeding of European sea bass (Dicentrarchus labrax) receives a growing interest, as the estimated heritability of growth is medium to high. In this study, we compared the offspring of four groups of sea bass sires, mated with the same wild dams: wild (W), first generation of domestication (D), first generation of mass selection for length (M), first generation of PROSPER-like selection for length (P). The comparison was done both in replicated tanks (separate rearing) and in mixed tanks (mixed rearing) where sire origins were recovered by genotyping of eight microsatellite markers. Weight, length and growth rate were measured from day 238 post-fertilization (69 g mean weight) to day 611 post-fertilization (390 g mean weight). Both in mixed and separate tanks, both selected groups (P, M) were larger than unselected groups (W, D). No difference was seen at any time between W and D, nor between M and P. The selection response estimate on weight was larger in mixed tanks when compared to separate tanks (+ 42% in mixed tanks, + 23% in separate tanks at day 611), yielding realized heritability estimates of 0.60 and 0.34, respectively, and confirming the excellent potential of the species for growth improvement through selective breeding. Both selection response and the amplification effect between mixed and separate tanks decreased as rearing density increased. Our hypothesis is that selection response is magnified by competition in mixed tanks, while sub-optimal rearing conditions lower the observed selection response, more in separate tanks (where selected thus larger fish are at a higher density than unselected ones) than in mixed tanks (where all fish experience the same density effects).     

Muscle cellularity in relation to somatic growth in the European sea bass Dicentrarchus labrax (L.)

Sea bass, Dicentrarchus labrax (L.), juveniles (1.32 g) grew at a rate of 4.2% per day, when fish were fed ad libitum and reared at optimal thermal conditions (ambient sea water temperature, 20-26°C). At 13°C, feeding and spontaneous activity were severely restricted and somatic growth was reduced to 0.6% per day. Over a period of 6 weeks, both muscle fibre hyperplasia and nuclear division were higher in the ambient-temperature group compared with the fish reared at 13°C. Despite the differences in growth rate and spontaneous activity, muscle fibre hyperplasia was paralleled by nuclear division in the lateral axial muscle in both temperature groups and the number of nuclei per myofibre did not differ significantly between the two temperature groups. It is concluded that at optimal thermal and feeding conditions, somatic growth of sea bass juveniles is mediated through an increase in the number of nuclei and muscle fibres.