Incidence and prevalence of Diplectanum aequans and its influence on the fitness of juvenile sea bass (Dicentrarchus labrax) in the Black Sea

High rates of unexplained mortalities (up to 70%) are anticipated in the cultured juvenile sea bass (Dicentrarchus labrax) in farms in the eastern Black Sea at least for the last 8 years. Diplectanum aequans, often blamed for the observed mortalities, is the only prevalent parasite impacting the health of sea bass in the brackish water of the Black Sea. To investigate the seasonal fluctuation of D. aequans prevalence, mean intensities and the potential effects of D. aequans on the fitness of sea bass, eight cages from one farm (Farm A) and four cages at another distinct site (Farm B) were surveyed monthly from May to October of 2008. Six hundred and sixty‐one juvenile sea bass from Farm A and 236 from Farm B were individually examined for the presence and intensity of the parasite. The prevalence of D. aequans in cultured sea bass (96.3 ± 11.11; Mean ± SD) ranged from 66.7% to 100% in June, the beginning of 6‐month growing season. The mean intensity was 6.08 ± 2.19, significantly higher than that (2.74 ± 0.87; P < 0.05) in December (P < 0.05), the end of the growing season. Fish with low fitness had significantly higher number of parasite than the fish with higher fitness (P < 0.005). Although D. aequans negatively influenced host's condition factor, the impact was not enough to lead mortalities in the cultured sea bass in the Black Sea.     

Peptide molecular weight distribution of soluble protein fraction affects growth performance and quality in European sea bass (Dicentrarchus labrax) larvae

Dicentrarchus labrax were fed from 10 to 40 days posthatching with six microdiets differing in the inclusion level (60 g kg^‐1 and 120 g kg^‐1) and type of protein hydrolysate (PH; yeast, YPH; pig blood, PBPH; pig red blood cells, PRBCPH). A microdiet containing 120 g kg^‐1 fish PH (FPH) was used as a control. PH differed in their amino acid (AA) profile and molecular weight distribution and therefore the tested microdiets too. The estimated content in FAA and di‐ and tripeptides in the FPH microdiet was 2 g kg^‐1 and 44 g kg^‐1, respectively. FAA estimated levels in YPH and PBPH microdiets were 26 g kg^‐1 and 53 g kg^‐1, whereas levels of di‐ and tripeptides were 30 g kg^‐1 and 60 g kg^‐1, respectively. The estimated levels of FAA in PRBCPH microdiets were 8 g kg^‐1 and 17 g kg^‐1, whereas estimated levels of di‐ and tripeptides were 11 g kg^‐1 and 22 g kg^‐1, respectively. Results revealed that FPH may be replaced by alternative PH from yeast and pig blood products, as fish fed those diets performed, in terms of growth, survival, digestive function and incidence of skeletal deformities, as well as those fed the FPH microdiet. Using YPH, PBPH and PRBCPH, the inclusion level of PH in microdiets might be reduced to a half with respect to current practices using FPH.     

Quantitative immunoenzymatic detection of viral encephalopathy and retinopathy virus (betanodavirus) in sea bass Dicentrarchus labrax

Viral encephalopathy and retinopathy disease caused by betanodavirus, genus of the family Nodaviridae, affects marine, wild and farmed species including sea bass, one of the most important farmed species in Europe. This work describes a reliable and sensitive indirect ELISA assay to detect betanodavirus in biological samples using a polyclonal antiserum (pAb 283) against the 283/I09 virus strain, the most common red‐spotted grouper nervous necrosis virus (RGNNV) genotype in the Mediterranean area, and a capture‐based ELISA using a monoclonal antibody (mAb 4C3) specific to a common epitope present on the capsid protein. Using adsorbed, purified VERv preparation, the detection limit of indirect ELISA was 2 μg mL^?1 (3 × 10⁵ TCID50 per mL), whereas for capture‐based ELISA, the sensitivity for the antigen in solution was 17 μg mL^?1 (35 × 10⁵ TCID50 per mL). The capture‐based ELISA was employed to detect VERv in brain homogenates of in vivo infected sea bass and resulted positive in 22 of 32 samples, some of these with a high viral load estimates (about 1.1 × 10⁸ TCID50 per mL). The ELISA system we propose may be helpful in investigations where coupling of viral content in fish tissues with the presence of circulating VERv‐specific IgM is required, or for use in samples where PCR is difficult to perform.     

Muscle activity as a key indicator of welfare in farmed European sea bass (Dicentrarchus labrax L. 1758)

Groups of adult sea bass were reared at either low (10 kg m^?3) or high (50 kg m^?3) stocking densities respectively for 84 and 116 days. To monitor the red muscle activity, about 20 fish from both densities were surgically implanted with EMG (Electromyograms) radio transmitters, after EMG calibration during exhaustive swimming exercise (U_crit test). Blood samples and morphometric measurements were also taken. EMG showed that the muscle activity of fish reared at 50 kg m^?3 was on average twofold higher than fish kept at lower density. Cortisol was significantly more elevated at higher density and haemoglobin, haematocrit and RBCC (red blood cells count) showed the same trend, while lysozyme decreased. Patterns for glucose and lactate were less clear. The results showed that the contemporary use of functional (EMG) and physiological (haematological and biochemical) profiles could give a more comprehensive view of the fish status validating the diagnosis of fish stress induced by culture practices.     

Effects of dietary Bergamot (Citrus bergamia) peel oil on growth,haematology and immune response of European sea bass (Dicentrarchus labrax) juveniles

The aim of the present study was to evaluate the effects of essential oil extracted from bergamot (Citrus bergamia) peel on growth performance and health status of European sea bass juveniles (5.10 + 0.05 g). Test diets supplemented with 0.0%, 0.5%, 1.0% and 2.0% bergamot peel oil (BPO₀, BPO_0.5, BPO₁ and BPO₂) were fed to experimental fish for 60 days. At the end of the experiment, the best growth performance was obtained in the BPO_0.5 and BPO₁ groups (p < .05). No significances were found for haematological parameters including red blood cell count, haematocrit and haemoglobin concentration at the end of the feeding experiment. There were significant differences between the total protein, cholesterol, alkaline phosphatase and glutamic oxaloacetic transaminase (p < .05) between the BPO0.5 and BPO1 supplemented groups and the control group with no BPO addition. The addition of BPO showed significant differences in immune parameters such as lysozyme activity, total myeloperoxidase content, superoxide dismutase and glutathione peroxidase activity between BPO₀ and BPO supplemented groups (p < .05). It might be concluded that dietary addition of 0.5%‐1% BPO could increase the growth performance and feed utilization with improved health status in sea bass juveniles.     

Fertilization strategies for Sea Bass Dicentrarchus labrax (Linnaeus, 1758): effects of pre‐incubation and duration of egg receptivity in seawater

Studying gamete biology can provide important information about a species fertilization strategy as well as their reproductive ecology. Currently, there is a lack of knowledge about how long sea bass Dicentrarchus labrax eggs can remain viable after being activated in seawater. The objectives of this study were to understand the effects of pre‐incubation of fresh and overripe sea bass eggs in seawater and to determine the duration of egg receptivity. Pooled eggs (fresh and overripe) from four females were pre‐incubated in seawater for 0 min (control), 0.5 min, 1 min, 3 min, 10 min and 30 min and then fertilized by pooled sperm from four males. The fresh eggs had a higher fertilization success than overripe eggs. Our results revealed a significant effect of pre‐incubation time for both the fresh (P < 0.01) and overripe eggs (P < 0.01). Fertilization success of eggs significantly declined for both these treatments after 3 min of pre‐incubation, which clearly indicates that sea bass eggs are able to be fertilized by sperm for up to 3 min after release into seawater. This study has particular importance for understanding fertilization strategies, reproductive potential, as well as reproductive ecology of sea bass.     

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.     

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.     

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.     

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 propolis enriched diet on growth performance and plasma biochemical parameters of juvenile European sea bass (Dicentrarchus labrax L.) under acute low‐temperature stress

To study the effect of propolis in crude form (CPP), prepared without any chemical refinement (CPP) on Dicentrarchus labrax under low‐temperature stress, sea bass juveniles were randomly divided into three groups: a control group fed with basal diet and two treatment groups fed with basal diet supplemented with 1.25 and 2.5 g kg⁻¹ of propolis. At the end of a 10‐week feeding trial, sea bass were exposed to low‐temperature stress at 12 °C for 24 h. The growth performance, RNA/DNA ratio and changes in serum biochemical parameters were investigated. Dietary intake of propolis stimulated the specific growth rate (SGR), feed conversion efficiency (FCE), RNA/DNA ratio and alkaline phosphatase (ALP) enzyme activity, while decreasing plasma triglycerides and aspartate aminotransferase (AST) activity. Supplement of 2.5 g kg⁻¹ CPP in diet significantly increased the mean SGR and FCE up to 9% and 13.4%, respectively, in comparison with the control group. Low‐temperature stress elevated serum triglycerides, glucose and cortisol levels in all groups; however, glucose and cortisol reached significantly lower end values in group receiving highest amount of propolis in diet. This study suggests that ingestion of basal diet supplemented with 2.5 g kg⁻¹ of propolis could prevent adverse effects of low‐temperature stress and promote the growth of sea bass.     

Sea bass Dicentrarchus labrax (L.) bacterial infection and confinement stress acts on F‐type lectin (DlFBL) serum modulation

The F‐lectin, a fucose‐binding protein found from invertebrates to ectothermic vertebrates, is the last lectin family to be discovered. Here, we describe effects of two different types of stressors, bacterial infection and confinement stress, on the modulation of European sea bass Dicentrarchus labrax (L.) F‐lectin (DlFBL), a well‐characterized serum opsonin, using a specific antibody. The infection of the Vibrio alginolyticus bacterial strain increased the total haemagglutinating activity during the 16‐day testing period. The DlFBL value showed an upward regulation on the first, second and last days and underwent a slight downward regulation 4 days post‐challenge. In contrast, the effect of confinement and density stress showed a decrease in the plasma concentration of lectin, ranging from 50% to 60% compared with the control. The modulation of DlFBL is in line with the hypothesis that humoral lectins could be involved and recruited in the initial recognition step of the inflammation, which leads to agglutination, and the activation of mechanisms responsible for killing of the pathogens.     

Growth parameters,innate immune response and resistance to Listonella (Vibrio) anguillarum of Dicentrarchus labrax fed carvacrol supplemented diets

The research was aimed to assess the effect of dietary carvacrol (0.025% and 0.05%) on sea bass (Dicentrarchus labrax) growth, immune response and resistance to Listonella anguillarum. Fish (69.2 ± 0.22 g) were fed the experimental diets for 9 weeks. Dietary carvacrol did not negatively affect fish survival, growth performance, feed intake and feed conversion ratio (P > 0.05) nor carcass yield and viscerosomatic, hepatosomatic and mesenteric fat index (P > 0.05). Serum and head kidney leucocytes were collected after 1, 4 and 8 weeks of feeding. Carvacrol significantly reduced serum proteins, immunoglobulins and lysozyme activity (P < 0.01) and moderately increased phagocytosis and pinocytosis of head kidney macrophages. The release of reactive oxygen species by leucocytes was reduced in carvacrol‐fed fish, even if significantly (P < 0.05) only in those fed 0.05% carvacrol for 1 week. Dietary carvacrol did not significantly affect the aspecific immune response, although a potential antioxidant activity might be speculated. Moreover, feeding carvacrol provided an appreciable resistance to a challenge with L. anguillarum, when a bacterial dose lower than the Lethal Dose₅₀ was used. Cumulative mortality in fish fed 0.025% carvacrol was significantly lower than that of untreated controls (75% Relative Per cent Survival).     

Appraisal of a high protein distiller’s dried grain (DDG) in diets for European sea bass,Dicentrarchus labrax fingerlings on growth performance,haematological status and related gut histology

High protein distiller's dried grains (HP‐DDG) are co‐products of ethanol production that uses prefractionation technology. An 8‐week growth trial was conducted to investigate the effect of partial replacement of soybean meal (SBM) by three levels of HP‐DDG (30%, 40% and 50%) on growth performance, physiological parameters and histological changes of the intestine of European sea bass, Dicentrarchus labrax. The results indicated that an increased dietary level of HP‐DDG of more than 30% significantly increases growth performance and improved the FCR of sea bass. In addition, replacement of SBM by HP‐DDG enhanced feed intake efficiency and the health status of fish. Haematology and serum biochemistry (haemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs), packed cell volume (PCV %) and humeral immune parameters including total protein, albumin, globulin, cholesterol, lysozyme activity and total antioxidant capacity significantly increased with increase HP‐DDG inclusion levels. The findings of this study indicated that HP‐DDG is a good complementary protein source for inclusion aquaculture diets and levels above 30% as a replacement of SBM did not compromise growth performance and physiological parameters of sea bass while enhancing some important indices of health status in this species.