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.     

A pharmacokinetic study of flumequine in sea bass, Dicentrarchus labrax (L.), after a single intravascular injection

The pharmacokinetic properties of flumequine following a single intravascular injection (10 mg kg^–1 fish) were studied in sea bass, Dicentrarchus labrax (L.), 120 g held at 18 °C. The absorption half life ( t _1/2α) and the elimination half life ( t _1/2β) of the drug were calculated to be 1.05 and 10.71 h, respectively. Tissue penetration of flumequine seemed to be moderate because both the apparent volume of distribution of the drug at steady-state ( V _d(ss)) and the extensive apparent volume of the central compartment ( V _c) were found to be small (1.51 and 0.626 L kg^–1). The mean residence time ( MRT ) was short (09.73 h) and the total clearance (CL_T) of the drug was rapid (0.156 L kg^–1 h^–1).     

Behaviour of European sea bass,Dicentrarchus labrax L., in cages – impact of early life rearing conditions and management

European sea bass, Dicentrarchus labrax L., groups reared under different conditions during larval stages (mesocosm and intensive rearing) were monitored during on‐growing in sea cages until marketable size (350–400 g). Four caged groups were followed for a period of 17 months each at a pilot scale farm, where vertical distribution behaviour was monitored. This was performed while fish were under calm conditions, during feeding and when stress events occurred. Also during two specific periods: (i) spawning and (ii) high water temperature. Clear differences in the behavioural pattern of swimming depth, displacement and used space between the groups were observed. The individuals from the mesocosm rearing were more sensitive to human presence, showing stronger reactions (speed of displacement and vertical distribution). Most pronounced differences were observed during the ‘extreme’ warm period and the reproductive season. Within the seasons, European sea bass responded during feeding and stress showing a tendency to move deeper compared to calm conditions. The increased displacements were longer during feeding and stress.     

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.     

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.     

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.     

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.     

Treatment of vibriosis in European sea bass larvae, Dicentrarchus labrax L., with oxolinic acid administered by bath or through medicated nauplii of Artemia franciscana (Kellogg): efficacy and residual kinetics

European sea bass larvae were challenged by bath with Listonella anguillarum strain 332A, 2.5×10(7) CFUmL(-1) for 1h. Fish either received no treatment or oral treatment with Artemia franciscana (Kellog) nauplii enriched with oxolinic acid, or bath treatments with oxolinic acid. Medication commenced 1day following challenge and was performed on days 1, 3 and 5 post-challenge at a dosage of 20mgL(-1) for 2h for bath treatments, while two doses each of 750 nauplii per fish were administered daily for five consecutive days in oral treatments. Cumulative mortality reached 96% for the unmedicated challenged group, 32% in the group receiving bath treatments and 17% in the group receiving medicated nauplii. Pharmacokinetic parameters of oxolinic acid were calculated in sea bass larvae, for both treatments. Steady-state concentrations of oxolinic acid of 48.0 and 75.2μgg(-1) were achieved for bath treatment and oral treatment, respectively, while the elimination half-life was calculated to be 25.1h for bath treatment and 21.7h for oral treatment.     

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'.     

Acid–base balance in sea bass (Dicentrarchus labrax L.) in relation to water oxygen concentration

The influence of water oxygen concentration on the acid–base balance of sea bass was evaluated. Fish weighing 200–250 g were cultured under different dissolved oxygen concentrations of 64%, 97%, 150% and 250% saturation (92.7, 140.5, 217.5 and 362.7 mmHg respectively) under mild hypoxia, normoxia, mild hyperoxia and high hyperoxia conditions. The results showed that high hyperoxia and mild hypoxia conditions modified some blood parameters significantly when compared with fish held under the normoxia condition, while no differences were shown with respect to the acid–base balance of fish cultured under normoxia and mild hyperoxia conditions. This testifies that the mild hyperoxia condition does not produce physiological disturbances in the acid–base status of sea bass and it could be considered a favourable condition in sea bass land‐based farming, mainly in comparison with the mild hypoxia condition, responsible for other physiological problems.     

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.     

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.     

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.     

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.     

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.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Annual cycle of plasma insulin and glucose of sea bass.Dicentrarchus labrax,L.

Annual plasma insulin and glucose cycles were studied inDicentrarchus labrax maintained in either seawater (37.8) or brackish water (3.5). In both media, the highest insulin levels were found during the prespawning period (August–November) coincident to increases in weight and a decrease in plasma glucose. During spawning (December–April) and postspawning (May–July) periods, the decrease in insulin occurred at the same time as a reduction in growth and an increase of plasma glucose. Temperature and salinity conditions impeded spawning in the brackish water group, in which a minor weight loss was regained more quickly than in the sea water group; insulin levels were also higher.