α‐Tocopherol in weaning diets for European sea bass (Dicentrarchus labrax) improves survival and reduces tissue damage caused by excess dietary DHA contents

The objective of the present study was to investigate the combined effect of several dietary contents of vitamin E and polyunsaturated fatty acids (PUFA), mainly docosahexaenoic acid (DHA), on growth, survival, biochemical composition and tissue morphology of sea bass along early development. A feeding experiment was conducted in sea bass larvae using five different diets with the same proximate composition and different ratios of DHA concentrated fish oil [10, 30 and 50 g kg^?1 dry weight (DW)] and vitamin E (α‐tocopherol acetate) (1500 and 3000 mg kg^?1 DW). DHA was readily deposited in fish tissues and associated with higher sea bass mortalities probably because of increased peroxidation risks. Besides, the elevation of dietary DHA contents up to 5% severely increased the incidence of muscular lesions and the presence of ceroid pigment within hepatocytes. However, elevation of dietary vitamin E levels markedly reduced the incidence of these symptoms in sea bass, increasing the tissue content in several PUFA and improving growth and stress resistance. Moreover, when sea bass was fed diets containing high vitamin E levels, fish showed a significant improvement in growth when dietary DHA was raised from 1% to 3%. Therefore, in sea bass larvae, a ratio of 30 g kg^?1 DHA and 3000 mg kg^?1 vitamin E seems to be adequate to achieve a good larval performance and to avoid muscular lesions.     

Effect of different dietary levels of docosahexaenoic acid (DHA, 22:6w-3) on the DHA composition of lipid classes in sea bass larvae eyes

The effect of different dietary levels of docosahexaenoic acid (DHA, 22:6w-3) on the corresponding composition of lipid classes of the eyes of sea bass, Dicentraxrchus labrax, larvae was studied using Artemia nauplii enriched with different products: oil emulsions, liposomes, a dry microalga and baker's yeast. DHA was found to be a major constituent of phosphatidylserine and phosphatidylethanolamine of visual tissues. The different DHA dietary levels were markedly reflected in the fatty acid composition of the lipid classes of eyes, suggesting a dose-dependent relationship between DHA in the food and in visual tissue lipids of sea bass larvae. The possible implications of this dietary effect are discussed.     

Growth,development and survival of European sea bass (Dicentrarchus labrax) larvae cultured under different light spectra and intensities

We assessed the effects of light intensity and spectrum on the growth, development and survival of Dicentrarchus labrax larvae from 30 to 96 days after hatching. Twelve lighting regimes were applied using 0.3, 1 and 2 W/m² full spectrum white, blue, red and green light. By day 66 of exposure, growth performance was significantly better in larvae reared under green or red light compared with white or blue light (p < 0.05), while it was significantly better at 2.0 W/m² compared with 1.0 or 0.3 W/m² (p < 0.05). Larvae reared under white or blue light had significantly more deformities (p < 0.05) than did larvae reared under light of other spectra, while larvae reared under 0.3 W/m² light intensity had a significantly higher proportion of jaw malformations than reared under 1.0 and 2.0 W/m² (p ? 0.05). Dicentrarchus labrax larvae subjected to green light showed significantly lower survival compared with the other light spectra (p < 0.05). Moreover, light significantly affected brain aanat1a, aanatt1b, mellc, mt1, mt2, gh, tsh and crf mRNA expression. Therefore, the red and green light groups (or under 2.0 W/m²) could improve the growth performance and enhance secretion of melatonin, gh and tsh, and reduce the proportion of jaw malformations.     

Fatty acid composition,osmolality, Na+, K+‐ATPase activity,cortisol content and antioxidant status of rainbow trout (Oncorhynchus mykiss) in response to various dietary levels of eicosapentaenoic acid and docosahexaenoic acid

This study was conducted to evaluate the effect of dietary eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels on the fatty acid composition, salinity tolerance and antioxidant status of rainbow trout (Oncorhynchus mykiss). Four diets were formulated with total EPA and DHA contents of 5.41, 9.55, 13.97 and 17.88 g/kg (abbreviated as ED‐5.41, ED‐9.55, ED‐13.97 and ED‐17.88 respectively). Rainbow trout (initial weight of 90.61 ± 9.25 g) were fed the experimental diets for 8 weeks to accumulate significant differences in fatty acid composition and subsequently underwent salinity acclimation. Our results indicated that high dietary EPA and DHA significantly improved the EPA and DHA content in fish tissues. The serum osmolality of fish returned to their freshwater values in the ED‐9.55, ED‐13.97 and ED‐17.88 groups. The Na⁺, K⁺‐ATPase (NKA) activity of fish in the ED‐13.97 group changed dramatically to adapt the fish to the hypertonic environment. Moreover, there was no significant difference in the serum cortisol concentration and liver catalase (CAT) activity of fish in the ED‐13.97 group during salinity acclimation. The liver superoxide dismutase (SOD) activity in the ED‐13.97 group was significantly higher than that in ED‐5.41 and ED‐9.55 groups at the end of salinity acclimation. The muscle malondialdehyde (MDA) content in the ED‐13.97 group was significantly lower than that in the ED‐17.88 group before salinity acclimation and significantly lower than the ED‐5.41 and ED‐17.88 groups on day 7 of acclimation. The results of this study indicate that the rainbow trout in the ED‐13.97 group exhibited optimal salinity acclimation performance.     

硫辛酸对大口黑鲈氧化应激相关酶活性的影响

为研究硫辛酸对大口黑鲈抗氧化性能的影响,通过腹腔注射不同浓度硫辛酸评估其对大口黑鲈抗氧化酶活性的影响.该试验挑选60尾健康大口黑鲈随机分成3组,根据鱼体质量分别注射含有不同剂量的硫辛酸,设定对照组(0 mg/kg)、T20组(20 mg/kg)和T40组(40 mg/kg),注射6h、12h、24 h后分离其血清和肝脏...     

Importance of dietary arachidonic acid for the growth,survival and stress resistance of larval European sea bass (Dicentrarchus labrax) fed high dietary docosahexaenoic and eicosapentaenoic acids

Together with docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), arachidonic acid (ARA) is being considered to be an essential fatty acid in marine fish larval diets. The objective of the present study was to determine the importance of dietary ARA levels for larval European sea bass performance, when EPA and DHA are also present in the diet. Eighteen‐day‐old larvae were fed, for 14 days, gelatine‐based microdiets containing the following ARA levels: 0.3%, 0.6% or 1.2%. Elevation of dietary ARA up to 1.2% showed a positive correlation with larval survival and a significant improvement in the specific growth rates, body weight and total length. Arachidonic acid was efficiently incorporated into larval lipids, even at a higher proportion than that in the diets. Increased accumulation of ARA did not affect the incorporation of DHA or EPA from the diet into larval total lipids. A significant positive correlation was found between dietary ARA levels and survival after handling stress, indicating the importance of this fatty acid in sea bass larvae response to acute stressors. The results show the importance of ARA for sea bass larvae, but higher dietary levels should be tested to determine whether there is a negative effect of ARA in sea bass as reported for other species.     

The effect of tissue docosahexaenoic and arachidonic acids levels on hypersaline tolerance and leucocyte composition in striped bass (Morone saxatilis) larvae

Larval striped bass (M. saxatilis), tissue docosahexaenoic (DHA) and arachidonic (AA) acids levels were modulated through dietary enrichments and the effect on growth and survivorship examined. Mean growth was significantly greater in larvae enriched with AA than in larvae enriched with DHA (F-value for AA treatment was 20.5 versus only 5.1 for the DHA treatment). Dietary treatment did not have a significant effect on larval survivorship (56.0±2.4%, p>0.05). When challenged with hypersaline (25 psu) immersion, DHA enriched larvae survived better than AA enriched larvae, but larvae with body tissue levels of 15.4 mg AA g^–1 and 7.2–15.4 mg DHA g^–1 dry weight provided maximal survivorship to the challenge. Elevated levels of body tissue AA was generally associated with elevated levels of whole body cortisol. On the other hand, increasing levels of DHA mainly affected the kinetics of cortisol increase to hypersaline exposure. Larvae injected intraperitoneally with formalin fixed Staphylococcus aureus responded by altering the proportion of lymphocytes, monocytes and neutrophils in perpherial blood. Lymphocytes, which accounted for the largest percentage of white blood cells (over 70%), decreased in all challenged larvae during the first 6 hours post injection then returned to pre-challenge levels after 44 hours. Conversely, the relative proportion of monocytes and neutrophils rose from 14% and 2% up to 28% and 6% of the total circulating leucocytes, respectively. The largest increase occurred in larvae fed a moderate level of both DHA and AA.     

Temperature-dependent pharmacokinetics and tissue distribution of oxolinic acid in sea bass, Dicentrarchus labrax L., after a single intravascular injection

The pharmacokinetics and tissue distribution of oxolinic acid following an intravascular administration (15 mg kg^?1 fish) were determined in sea bass, Dicentrarchus labrax L. (110 g), at 13 °C and 22 °C water temperature. The kinetic profile of the drug was found to be temperature dependent, with increased temperature having a greater effect on distribution after equilibrium and the elimination phase than on the distribution process. The distibution half‐life of oxolinic acid was 1.15 and 2.76 h at 22 °C and 13 °C respectively, whereas the elimination half‐life of the drug was 55 h at 22 °C and 315 h at 13 °C. The values of the apparent volume of distribution (1.44 L kg^?1 at 22 °C and 3.31 L kg^?1 at 13 °C) and the volume of distribution at steady state (5.2 and 14.7 L kg^?1 at the high and low temperature respectively) were considerably different between the two tested temperatures. The total body clearance of the antibiotic was found to be low (1.47 L kg^?1 day^?1 at 22 °C and 0.80 L kg^?1 day^?1 at 13 °C). Lower rates of elimination were found for the liver compared with muscle, the difference increasing with increasing temperature, while elimination rates from the serum were higher than those of other tissues, especially at the high temperature.     

Effects of dietary docosahexaenoic acid (22:6n‐3) and arachidonic acid (20:4n‐6) on the growth,survival, stress resistance and fatty acid composition in black sea bass Centropristis striata (Linnaeus 1758) larvae

The objectives of this study were to determine the effects of the dietary docosahexaenoic acid (DHA) to arachidonic acid (ARA) ratio on the survival, growth, hypersaline stress resistance and tissue composition of black sea bass larvae raised from first feeding to metamorphic stages. Larvae were fed enriched rotifers Brachionus rotundiformis and Artemia nauplii containing two levels of DHA (0% and 10% total fatty acids=TFA) in conjunction with three levels of ARA (0%, 3% and 6% TFA). On d24ph, larvae fed the 10:6 (DHA:ARA) treatment showed significantly (P<0.05) higher survival (62.3%) than larvae fed 0:0 (DHA:ARA) (27.4%). Notochord length and dry weight were also significantly (P<0.05) greater in the 10:6 (DHA:ARA) treatment (8.65 mm, 2.14 mg) than in the 0:0 (DHA:ARA) (7.7 mm, 1.65 mg) treatment. During hypersaline (65 g L⁻¹) challenge, no significant differences (P>0.05) were observed in the median survival time (ST50) between larvae fed 10% DHA (ST50=25.6 min) and larvae fed 0% DHA (ST50=18.2 min). The results suggested that black sea bass larvae fed prey containing 10% DHA with increasing ARA within the range of 0–6% showed improved growth and survival from first feeding through metamorphic stages.     

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.     

The use of clove oil, metomidate, tricaine methanesulphonate and 2-phenoxyethanol for inducing anaesthesia and their effect on the cortisol stress response in black sea bass (Centropristis striata L.)

Juvenile and adult black sea bass (Centropristis striata L.) were exposed to various concentrations of four anaesthetics to determine practical dosages for handling as well as for procedures such as bleeding, ovarian biopsy or tag implantation. In experiment 1, juveniles exposed to either 2.0 mg L^?1 metomidate, 15 mg L^?1 clove oil, 70 mg L^?1 tricaine methanesulphonate (TMS) or 200 mg L^?1 2‐phenoxyethanol (2‐PE) reached stage II of anaesthesia in 3–5 min and could be handled for weighing and measuring. All fish had completed recovery to stage III within 6 min. In experiment 2, the established concentrations of each anaesthetic were tested on juveniles to determine their ability to prevent a reflex to a subcutaneous needle puncture. All of the fish exposed to clove oil (20 mg L^?1) and 40% of the TMS‐treated (70 mg L^?1) fish reacted while none of the fish anaesthetized in metomidate (2.0 mg L^?1) or 2‐PE (200 mg L^?1) responded to the needle puncture. In experiment 3, metomidate (5.0 mg L^?1), clove oil (30 mg L^?1) TMS (125 mg L^?1) or 2‐PE (300 mg L^?1) were all effective for performing an ovarian biopsy or tag implantation on adults. In experiment 4, TMS (125 mg L^?1) exacerbated the cortisol response to a short handling stressor during a 30 min exposure. Fish anaesthetized in 2‐PE (300 mg L^?1), metomidate (5.0 mg L^?1) or clove oil (40 mg L^?1) had increased cortisol levels associated with the handling stressor but there were no further increases during the remainder of the experimental period. The results demonstrate that these anaesthetics are effective for sedation and anaesthesia of black sea bass and that the best choice is dependant upon the procedures to be performed.     

High n‐3 HUFA levels in the diet of Atlantic salmon affect muscle and mitochondrial membrane lipids and their susceptibility to oxidative stress

Atlantic salmon were fed one of four diets with increasing levels (11–58%of total fatty acids (FAs)) of n‐3 highly unsaturated FAs (HUFAs) in order to investigate the effect on muscle and mitochondrial membrane lipids and their susceptibility to oxidative stress. The high n‐3 eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) groups contained lower percentages of sphingomyelin and cardiolipin in total muscle than the intermediate n‐3 HUFA group. Cardiolipin and sphingomyelin are particularly susceptible to peroxidation, and a reduced percentage of these lipids showed that mitochondrial membranes had been damaged by oxidation. The intermediate n‐3 HUFA group had the highest level of mitochondrial integrity and tendencies of lower thiobarbituric acid reactive substances (TBARS) level than the other dietary groups. The high caspase‐3 activity, in addition to casp3a and bax gene expression levels, in the n‐3 DHA group also suggests that some degree of oxidative stress had occurred. Electron microscopy images showed a higher degree of myofibre–myofibre detachment in fish fed the high HUFA diets than in fish fed the intermediate n‐3 HUFA diet. Our findings show that intermediate levels of n‐3 HUFAs in salmon diets gave the best protection against oxidative damage of mitochondrial membranes and muscle structure.     

Ascorbate dynamics in embryos and larvae of sea bass and sea bream, originating from broodstocks fed supplements of ascorbic acid

The transfer of L-ascorbic acid (AA) from broodstock to the fertilised eggs and it's dynamics in embryos and fasting larvae, has been studied in sea bass (Dicentrarchus labrax L.) and sea bream (Sparus aurata L.). Two types of diets were fed to the broodstocks: one containing sufficient ascorbate for normal growth and the other with an extra addition of a very high dose (2,000 mg/kg feed) of ethyl-cellulose-coated L-ascorbic acid.The concentration of AA and total ascorbate (L-ascorbic acid plus dehydroascorbic acid) was detected through several embryo and larval development stages, including larvae just before feeding. In sea bass and sea bream, the mean (SD) concentration of total ascorbate in fertilised eggs originating from broodstock fed AA supplemented diet, was 218.5 (17.7) and 122.4 (5.1) g/g wet weight respectively. This was significantly higher (P < 0.01) than the unsupplemented groups, which contained only 155.9 (6.9) and 103.9 (3.5) g/g wet weight respectively. A diet with a vitamin C content adequate for normal growth, may not be sufficient for broodstock when the goal is transfer of TAA to embryos. © Rapid Science Ltd. 1998     

Effect of increasing docosahexaenoic acid content in weaning diets on survival,growth and skeletal anomalies of longfin yellowtail (Seriola rivoliana,Valenciennes 1833)

Five isoproteic (54.8%) and isolipidic (24.1%) microdiets, which varied in their docosahexaenoic acid (DHA) content (0.25%, 0.75%, 1.64%, 1.99% and 3.17%; dw), were manufactured to determine its effects on longfin yellowtail Seriola rivoliana larvae in terms of fish biological performance, whole body fatty acid profile and incidence of skeletal anomalies from 30 dah (11.31 ± 1.79 Total Length, TL) to 50 dah (19.80 ± 0.58 mm TL). The inclusion of dietary DHA up to 3.17% (dw) improved larval resistance to air exposure, although DHA did not significantly affect fish final growth or final survival. Indeed, high levels of dietary DHA (1.99% and 3.17%, dw) tended to increase the incidence of skeletal anomalies in S. rivoliana larvae, albeit no significant differences were observed. Furthermore, the occurrence of severe anomalies such as kyphosis and lordosis, was mainly associated to the larvae fed the highest levels of dietary DHA. In terms of survival, increasing dietary DHA levels did not significantly affect longfin yellowtail survival rate, despite a tendency for enhanced survival. The results of the present study proved that the inclusion of dietary DHA in inert diets up to a 3.17% (dw) and a DHA/EPA ratio above 3.1 increased the final survival and stress resistance in S. rivoliana larvae.     

Dietary docosahexaenoic acid to eicosapentaenoic acid (DHA/EPA) ratio influenced growth performance,immune response,stress resistance and tissue fatty acid composition of juvenile Japanese seabass,Lateolabrax japonicus (Cuvier)

A 10‐week feeding experiment was conducted to investigate the effects of dietary DHA/EPA ratio on juvenile Japanese seabass reared in sea floating cages. Six practical diets were formulated differing only in DHA/EPA ratio: 0.55 (Diet D/E0.55), 1.04 (D/E1.04), 1.53 (D/E1.53), 2.08 (D/E2.08), 2.44 (D/E2.44) and 2.93 (D/E2.93). All diets had the same contents of total n‐3 long‐chain polyunsaturated fatty acid (LC‐PUFA) and arachidonic acid (ARA). The results showed that the final weight and specific growth rate significantly increased with increasing dietary DHA/EPA ratio from 0.55 to 2.08 and thereafter declined. Activities of lysozyme and superoxide dismutase in serum in groups with DHA/EPA of 1.53–2.93 was significantly higher compared to group D/E0.55 while the activity of serum alternative complement pathway in group D/E2.93 was significantly lower compared with group D/E1.53. However, no difference was observed in activities of both respiratory burst of head kidney macrophage and serum catalase among dietary treatments. The per cent survival after air exposure in group D/E1.53, D/E2.08 and D/E2.93 was significantly higher compared with group D/E0.55. The fatty acid composition of whole body and tissues reflected closely those of diets, while fish accumulated more DHA than EPA in fish tissues, especially in muscle. These results suggested that at the same dietary contents of n‐3 LC‐PUFA (appr. 18% of TFA) and ARA (appr. 0.7% of TFA), moderate dietary DHA/EPA ratios of 1.53–2.08 significantly enhanced growth performances, certain innate immune responses, and the stress tolerance of Japanese seabass, in accordance with the preferential incorporation of DHA into fish tissues.