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.     

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.     

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.     

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.     

Effects of successive size grading on the growth of juvenile European sea bass (Dicentrarchus labrax L.) at three temperatures

Juvenile European sea bass from the same fish stock were selected by successive size grading processes using 2, 3 and 4 mm bar graders at 79, 96 and 99 days post hatching, thus forming three groups (n=300) consisting of similar‐sized fish that differed by time of each group formation. The growth patterns of fish groups were studied at three temperatures during 5 weeks of rearing. Three‐way anova followed by the Tukey multiple comparison test (P<0.05) showed a high dependence of growth on the temperature applied. The smallest size and weight of fish were detected in all groups reared at 19 °C compared with fish held at 21 and 23 °C respectively. Differences in coefficients of variation of lengths were small and insignificant between groups and temperatures. Growth in the length of sea bass juveniles during the test period was a linear function of time and no differences were observed in growth rate among groups at a particular temperature. Growth rates of fish were 0.71 ± 0.02, 0.62 ± 0.01 and 0.52 ± 0.02 mm day^?1 at 23, 21 and 19 °C respectively. These results indicated that the variations in body size of juveniles in the test period were not the result of differences in the growth potential of individuals.     

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.     

Cryopreservation of turbot Scophthalmus maximus (L.) sperm: fertilization and hatching rates

The present paper assesses the fertilization and hatching rates of an artificial fertilization series (n=1153) using fresh and cryopreserved sperm from 49 specimens of turbot Scophthalmus maximus (L.), carried out to confirm the results of a previous study, with the ultimate aim of transferring these cryo‐preservation techniques to commercial hatcheries. No significant differences were found between the fertilization rates of the two groups (fresh and cryopreserved sperm) when their respective fertility rates were >69.2%, which was the case in 75% of all fertilizations. Likewise, no significant differences in hatching rates were found. In order to use the cryopreserved sperm more efficiently, a key concern for commercial use of this technique, we also experimented with a lower sperm:diluent ratio (1:1) than used previously at our centre. We also compared the traditional 0.5‐mL straws with 2‐mL cryotubes able to contain a higher volume of sperm, finding no significant differences in the resulting fertilization and hatching rates. In conclusion, the use of cryopreservation for turbot sperm presents major advantages for broodstock management in commercial hatcheries.     

Antipredator responses to overhead fright stimuli in hatchery-reared and wild European sea bass (Dicentrarchus labrax L.) juveniles

In this study, shoals of hatchery‐reared and wild sea bass juveniles (Dicentrarchus labrax L.) were tested for differences in their antipredator responses towards a visual (shadow) and a mechanical (dummy bill) overhead stimulus. Two behavioural variables – distance from the bottom and freezing duration – were measured during post‐stimulus phases of each test and compared between wild and hatchery‐reared shoals, composed of 10 juveniles each. The results showed that in both hatchery‐reared and wild juveniles, stimulus exposure elicited a significant decrease in the mean shoal distance from the bottom. Similarly, individuals from both groups engaged a freezing reaction, but the mean freezing duration was significantly higher in wild‐ than in hatchery‐reared juveniles. Results are discussed in the light of their relevance for the enhancement of restocking programmes.     

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.     

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.     

Effects of 2-phenoxyethanol on survival of normal juveniles and malformed juveniles having lordosis or nonfunctional swimbladders of European sea bass (Dicentrarchus labrax L., 1758)

The objectives of this study were to evaluate the effects of 2‐phenoxyethanol (2‐PE), which is an anaesthetic, on survival rates of normal juveniles and malformed juveniles having lordosis or nonfunctional swim bladders of European sea bass (Dicentrarchus labrax L., 1758) and to establish the LC₅₀ (the concentration lethal to 50% of test animals at concentrations of 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4 and 4.5 mL L⁻¹) and LT₅₀ (the time lethal to 50% of test animals after 10‐, 20‐, 30‐, 40‐, 50‐ and 60‐min time periods) of 2‐PE at 19±0.5°C, salinity 38 g L⁻¹, pH 7.4–7.8 and dissolved oxygen >8 mg L⁻¹. Between concentrations of 0.05 and 0.25 mL L⁻¹, 2‐PE did not cause any mortality or toxicity on normal, lordosis and nonfunctional swimbladder juveniles of sea bass during the 60‐min exposure period. On the other hand, significance in each group fish in their mortality rates between concentrations of 0.30 and 0.45 mL L⁻¹ was observed (P<0.05). The nonfunctional swimbladder juveniles showed lower LC₅₀ than normal and lordosis juveniles respectively. Also, nonfunctional swimbladders juveniles showed lower LT₅₀ than normal and lordosis juveniles respectively. At concentrations of 0.30, 0.35, 0.40 and 0.45 mL L⁻¹, induction times were found to be significantly different among the three groups (P<0.05). Recovery times were not found to be significantly different in two groups at concentrations of 0.30 and 0.40 mL L⁻¹ (P>0.05). The toxic effect of 2‐PE on sea bass juveniles increased depending on the exposure times (P<0.05). The most suitable concentrations of 2‐PE were 0.30–0.35 mL L⁻¹ between minutes 10 and 30, although the normal juveniles can resist to 0.45 mL L⁻¹ of 2‐PE concentration for 20 min. The 2‐PE showed toxicity in relation to the concentrations and exposure time combinations among the three groups in the order; nonfunctional swimbladder fish >lordosis fish >normal fish.     

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.     

Characterization of nodavirus and viral encephalopathy and retinopathy in farmed turbot, Scophthalmus maximus (L.)

An outbreak of nodavirus infection in turbot larvae is described with respect to histopathology, immunohistochemistry, cell culture cultivation, RT-PCR amplification and sequence analysis of the capsid protein gene RNA2. Affected turbot developed classical signs of viral encephalopathy and retinopathy (VER) with abnormal swimming behaviour and high mortality levels. In the acute stage of infection, light microscopy revealed vacuolation of the central nervous system (CNS), with positive immunohistochemical staining for nodavirus. Later in the infection, CNS lesions appeared more chronic and contained clusters of cells immunopositive for nodavirus. Bacterial overgrowth in the intestines of the fish may have provoked or influenced the course of the nodavirus infection. We were unable to propagate the virus in cell culture. While RT-PCR using primers designed to detect Atlantic halibut nodavirus gave negative results, further testing with primers complementary to a more conserved region of RNA2 resulted in amplification of a product of the expected size. The entire RNA2 segment was cloned and sequenced. Sequence alignment showed that the turbot nodavirus (TNV) was different from previously described fish nodaviruses. In addition, phylogenetic analysis based on an 823 nt region of the sequence indicated that TNV clustered outside the four established fish nodavirus genotypes, suggesting a fifth genotype within the betanodaviruses.     

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.     

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).     

Statistical properties and performance of pairwise relatedness estimators using turbot (Scophthalmus maximus L.) family data

The statistical properties and performance of four estimators of pairwise relatedness were evaluated in several scenarios using the microsatellite genotype data from a set of large known full‐sibships of turbot. All estimators showed a significant negative bias for the four kinships commonly used in these studies (unrelated: UR, half‐sibs, full‐sibs and parent–offspring), when allele frequencies of the reference population were estimated from the individuals analysed. When these frequencies were obtained from the base population from which all families proceeded, the bias was mostly corrected. The Wang (W) and Li (L) estimators were the least sensitive to this factor, while the Lynch and Ritland (L&R estimator) was the highest one. The error (mean around 0.130) was very similar in all scenarios for W, L and Queller and Goodnight (QG) estimators, while L&R was the highest error‐prone estimator. Parent–offspring kinship resulted in the lowest error, when using W, L and QG estimators, while UR resulted in the lowest error with the L&R estimator. Globally, W was the best‐performing estimator, although L&R could perform better in specific sampling scenarios. In summary, pairwise estimators represent useful tools for kinship classification in aquaculture broodstock management by applying appropriate thresholds depending on the goals of the analysis.