Antibody response in sea bass (Dicentrarchus labrax L.) in relation to water temperature and oxygenation

The influence of water temperature and water oxygenation on the specific antibody response was evaluated by indirect ELISA in sea bass (Dicentrarchus labrax L.) immunized against human‐γ‐ globulins emulsified in Freund's complete adjuvant. A higher antibody response was observed in fish reared at 24 and 30 °C, than at 12 and 18 °C. For the fish group reared at 24 °C, the immunological observation was carried out until 300 days after immunization. At the end of this period, fish kept at 24 °C still showed immunological competence, although the antibody response began to decrease significantly from 120 days after the immunization. Fish reared under mild hyperoxia and normoxia conditions had higher antibody responses than fish reared under mild hypoxia conditions. Moreover, at the end of the experimental period of 56 days the antibody response of fish reared under hyperoxia conditions was significantly higher than the immune response detected in fish reared under normoxia and hypoxia conditions.     

Influence of environmental temperature and water oxygen concentration on gas diffusion distance in sea bass (Dicentrarchus labrax, L.)

Gas diffusion distance (GDD) of sea bass was measured in fish bred under farm conditions, at different dissolved oxygen concentrations (DO): normoxia condition (80–100% of the saturation value) and `mild' hyperoxia condition (120–130% of the saturation value). Measures were carried out two times in a year (beginning of summer and autumn) in order to evaluate the effect of water temperature on GDD at the two different dissolved oxygen concentrations. There was a significant influence of both dissolved oxygen concentration (p 0.001) and environmental temperature (p 0.001) on GDD. In summertime it was 1.75 m and 2.31 m for fish reared under normoxia and hyperoxia, respectively, and in autumn 2.51 m and 2.96 m for fish reared under normoxia and hyperoxia, respectively. When DO was reduced at the higher temperatures, GDD decreased as well. Results lead to the conclusion that GDD increased with the increasing of DO, both due to reduced water temperature and to the mild oxygen hypersaturation following application of pure oxygen. The advantage for fish may be found in the compromise between maximising O<sub>2</sub> diffusion at the gills and ions/water intake/loss, known as `osmoregulatory compromise'.     

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 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.     

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.     

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.     

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.     

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.     

Influence of tank volume on vitellogenesis and spawning performances in sea bass Dicentrarchus labrax L.

Sea bass, Dicentrarchus labrax (mean weight: 748±13 g), were maintained before and during vitellogenesis in 1, 3, 8, 16 and 32 m³ tanks, and then they were transferred to 2 m³ tanks, for the spawning season. During the first 2 months of the experiment, the growth rates were significantly lower in smaller tanks (1 m³). In August, the oocyte diameters were significantly lower in smaller tanks (1, 3 and 8 m³) than in larger (16–32 m³) tanks. At the end of the experiment, the fish mean weight in the 1 m³ tanks was significantly lower than in the 3 m³ tanks, but oocyte diameters and plasma oestradiol concentrations were not significantly different between the volumes. This shows a longer acclimation requirement in smaller volume rearing. Although all the females had not spawned, one spawn at least was collected in each volume. The variation in conditioning volume has not blocked the spawning process. The qualitative and quantitative characteristics of spawns were not significantly different between volumes. The conditioning volume of 3 m³ seems to be a minimal volume required to obtain good reproduction of sea bass.     

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.     

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.     

New perspective to control of tenacibaculosis in sea bass Dicentrarchus labrax L

This work was carried out to investigate the effects of injection of Tenacibaculum maritimum formalin‐killed cells (FKC), extracellular products (ECPs) and crude lipopolysaccharide (LPS) as well as 1% feed supplements of oil extracts of Echinacea purpurea and Origanum vulgare on sea bass immunity improvement to the favour of T. maritimum experimental infection control after 4 weeks of the experiment. Tenacibaculum maritimum isolated from naturally infected sea bass showed brown to yellowish‐brown lesions (sores) on gills, skin and/or fins and identified by different biochemical methods and polymerase chain reaction technique. Immune parameters namely, total protein, globulin and lysozyme activity, as well as the relative level of protection were improved by T. maritimum (FKC), (LPS), (ECPs), O. vulgare and E. purpurea, respectively compared with control. Histopathological examination of T. maritimum naturally infected sea bass indicated many pathological changes in gill, skin and musculatures. Present study could be concluded that application of T. maritimum (FKC), (LPS), (ECPs), O. vulgare or E. purpurea improved sea bass immunity to the favour of disease resistance against T. maritimum. Further investigations on the combination between the previous control methods and the vaccine application methods will be needed.     

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.     

Delayed gametogenesis and spawning of sea bass (Dicentrarchus labrax L.) kept under different photoperiod and temperature regimes

The present work investigates the importance of day length and temperature in the control of reproduction of sea bass, as well as the effectiveness of LHRHa and HCG in inducing spawning out of season in this species. A controlled regime was produced and seasonal cycles with high components of temperature and photoperiod were extended from the summer solstice for at least 6 months, followed by a short photoperiod regime for 3 months before a new increase in these components. Natural spawning in the control fish occurred more frequently in mid February, although it was also observed in January and early March. Temperature manipulation delayed the spawning one month with respect to the controls, although some of the animals entered into gonadal regression. Photoperiodic manipulation delayed maturation for three months with respect to controls but it was necessary to perform hormonal induction of spawning. Although LHRHa and HCG were both applied, only intraperitoneal injections of LHRHa were effective in inducing spawning of sea bass out of season when the temperatures were 17°C. Dephasing between the annual changes in photoperiod in relation to the coordination of the different events of the sexual cycle of sea bass is considered.