Biomarkers of bone development and oxidative stress in gilthead sea bream larvae fed microdiets with several levels of polar lipids and α‐tocopherol

Although dietary marine phospholipids are able to improve culture performance of marine fish larvae in a further extend than soybean lecithin, both types of phospholipids (PL) markedly increase oxidative risk. The inclusion of a fat‐soluble antioxidant such as the vitamin E α‐tocopherol could allow a better control of oxidative stress. The objective of this study was to determine the combined effect of graded levels of α‐tocopherol with different levels and sources of krill phospholipids (KPL) and soybean lecithin (SBL) on growth, survival, resistance to stress, oxidative status, bone metabolism‐related genes expression and biochemical composition of sea bream larvae. Sea bream larvae were completely weaned at 16 dph and fed for 30 days seven microdiets with three different levels of PL (0, 40 and 80 g kg^?1 diet) and two of α‐tocopherol 1500 and 3000 mg kg^?1 diet. Sea bream larvae fed diets without PL supplementation showed the lowest survival, growth and stress resistance, whereas increase in PL, particularly KPL, markedly promoted larval survival and growth. However, feeding SBL markedly increased TBARs and GPX gene expression increasing the peroxidation risk in the larvae. Besides, KPL inclusion improved incorporation of n‐3 HUFA and, particularly, EPA into larval tissues, these fatty acids being positively correlated with the expression of BMP‐4, RUNX 2, ALP, OC and OP genes and to bone mineralization for a given larval size class. The increase in dietary α‐tocopherol tends to improve growth in relation to the n‐3 HUFA levels in the diet, denoting the protective role of this vitamin against oxidation. Indeed, dietary α‐tocopherol decreased the oxidative stress in the larvae as denoted by the reduction in larval TBARs contents and gene expression of SOD and CAT, but not GPX. Thus, increase in dietary α‐tocopherol effectively prevented the formation of free radicals from HUFA, particularly EPA, but did not affect the incidence of bone anomalies or the expression of genes related to osteogenetic processes.     

Influences of dietary fatty acid profile on growth, body composition and blood chemistry in juvenile fat cod (Hexagrammos otakii Jordan et Starks)

This study was conducted to investigate the influence of dietary lipid source and n‐3 highly unsaturated fatty acids (n‐3 HUFA) level on growth, body composition and blood chemistry of juvenile fat cod. Triplicate groups of fish (13.2 ± 0.54 g) were fed the diets containing different n‐3 HUFA levels (0–30 g kg^?1) adjusted by either lauric acid or different proportions of corn oil, linseed oil and squid liver oil at 100 g kg^?1 of total lipid level. Survival was not affected by dietary fatty acids composition. Weight gain, feed efficiency and protein efficiency ratio (PER) of fish fed the diets containing squid liver oil were significantly (P < 0.05) higher than those fed the diets containing lauric acid, corn oil or linseed oil as the sole lipid source. Weight gain, feed efficiency and PER of fish increased with increasing dietary n‐3 HUFA level up to 12–16 g kg^?1, but the values decreased in fish fed the diet containing 30 g kg^?1 n‐3 HUFA. The result of second‐order polynomial regression showed that the maximum weight gain and feed efficiency could be attained at 17 g kg^?1 n‐3 HUFA. Plasma protein, glucose and cholesterol contents were not affected by dietary fatty acids composition. However, plasma triglyceride content in fish fed the diet containing lauric acid as the sole lipid source was significantly (P < 0.05) lower than that of fish fed the other diets. Lipid content of fish fed the diets containing each of lauric acid or corn oil was lower than that of fish fed the diets containing linseed oil or squid liver oil only. Fatty acid composition of polar and neutral lipid fractions in the whole body of fat cod fed the diets containing various levels of n‐3 HUFA were reflected by dietary fatty acids compositions. The contents of n‐3 HUFA in polar and neutral lipids of fish increased with an increase in dietary n‐3 HUFA level. These results indicate that dietary n‐3 HUFA are essential and the diet containing 12–17 g kg^?1 n‐3 HUFA is optimal for growth and efficient feed utilization of juvenile fat cod, however, excessive n‐3 HUFA supplement may impair the growth of fish.     

Partial substitution of fish oil with rapeseed, linseed and olive oils in diets for European sea bass (Dicentrarchus labrax L.): effects on flesh fatty acid composition, plasma prostaglandins E2 and F2α, immune function and effectiveness of a fish oil finishing diet

Triplicate groups of European sea bass (Dicentrarchus labrax L.), of initial weight 90 g, were fed four practical‐type diets in which the added oil was 1000 g kg^?1 fish oil (FO) (control diet), 600 g kg^?1 rapeseed oil (RO) and 400 g kg^?1 FO, 600 g kg^?1 linseed oil (LO) and 400 g kg^?1 FO, and 600 g kg^?1 olive oil (OO) and 400 g kg^?1 FO for 34 weeks. After sampling, the remaining fish were switched to the 1000 g kg^?1 FO diet for a further 14 weeks. Fatty acid composition of flesh total lipid was influenced by dietary fatty acid input but specific fatty acids were selectively retained or utilized. There was selective deposition and retention of docosahexaenoic acid (DHA; 22:6n‐3). Eicosapentaenoic acid (EPA; 20:5n‐3) and DHA were significantly reduced and linolenic (LNA; 18:3n‐3), linoleic (LA; 18:2n‐6) and oleic (OA; 18:1n‐9) acids significantly increased in flesh lipids following the inclusion of 600 g kg^?1 RO, LO and OO in the diets. No significant differences were found among different treatments on plasma concentrations of prostaglandin E₂ and prostaglandin F_2α. Evaluation of non‐specific immune function, showed that the number of circulating leucocytes was significantly affected (P < 0.001), as was macrophage respiratory burst activity (P < 0.006) in fish fed vegetable oil diets. Accumulation of large amounts of lipid droplets were observed within the hepatocytes in relation to decreased levels of dietary n‐3 HUFA, although no signs of cellular necrosis was evident. After feeding a FO finishing diet for 14 weeks, DHA and total n‐3 HUFA levels were restored to values in control fish although EPA remained 18% higher in control than in the other treatments. This study suggests that vegetable oils such as RO, LO and OO can potentially be used as partial substitutes for dietary FO in European sea bass culture, during the grow out phase, without compromising growth rates but may alter some immune parameters.     

Interactive effects of dietary vitamin C and phospholipid in micro‐bound diet for growth,survival, and stress resistance of larval red sea bream,Pagrus major

This study was conducted to examine the effects of dietary ascorbic acid (AsA) and phospholipid (PL) and their interaction on growth, survival, and stress resistance in red sea bream larvae. Twenty‐six days old red sea bream were fed nine micro‐bound diets supplemented three levels of AsA (0, 800 and 1600 mg kg^?1 diet) and PL (0, 20 and 40 g kg^?1 diet) for 15 days. Dietary AsA and PL were both significant factors on survival rates. There was also an interaction between dietary AsA and PL on survival rate (P < 0.05). The larvae fed 800 or 1600 mg kg^?1 AsA with 40 g kg^?1 PL diets showed the highest survival rate, with values similar to those of the live‐food supplemented group. Stress resistance against low salinity exposure significantly increased with increased dietary level of AsA and PL. However, significant interaction of AsA and PL was not detected. The larvae fed 1600 mg kg^?1 AsA with 40 g kg^?1 PL diet showed the highest stress resistance among all diets, but it was not significantly different than that of larvae fed 800 mg kg^?1 AsA with 40 g kg^?1 PL diet. This study clearly demonstrated that combined use of AsA and PL can improve survival of 26–40 days posthatching red sea bream larvae. Moreover, the present study suggested that 800 mg kg^?1 AsA with 40 g kg^?1 PL in diet was needed for producing high quality seedling under the stressful conditions.     

Combination effects of dietary EPA and DHA plus alpha‐tocopherol: effects on performance and physiological status of Caspian brown trout (Salmo trutta caspius) fry

The effects of dietary n‐3 highly unsaturated fatty acids [eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA)] with α‐tocopherol on growth, non‐specific immune response and oxidative status were investigated in Caspian brown trout, Salmo trutta caspius, fry. Six experimental diets containing three different dietary levels of n‐3 HUFAs (low: 1 + 0.5% of total fatty acids, DHA+ EPA, medium: 2 + 1%, DHA + EPA, high: 4 + 2%, DHA + EPA) with two different levels of α‐tocopherol (low: 300 and high: 1000 mg kg^?1 diet) were prepared and named: LL, LH, ML, MH, HL and HH (HUFA/α‐tocopherol) groups, respectively. Diets were fed to triplicate groups of 60 fry with an initial weight of 600 ± 25 mg for 10 weeks. Results showed that increase in dietary DHA and EPA up to high level improved fry growth in terms of the body weight and specific growth rate, particularly when dietary α‐tocopherol levels were high, suggesting a higher antioxidant protection value when these fatty acids are high. At all dietary n‐3 HUFA levels, increase in α‐tocopherol from low to high level enhanced the alternative complement (ACH50) activity. Fry fed diets medium and high n‐3 HUFA displaying significantly higher lysozyme activity (P < 0.05). Moreover, fish fed medium or high levels of n‐3 HUFA had significantly lower prostaglandin E2 (PGE2) than those fed low n‐3 HUFA (P < 0.05). Significant differences in antioxidant enzymes (catalase, glutathione peroxidase, glutathione S‐transferase, glutathione reductase and superoxide dismutase) activity were also observed between groups, with higher activity in high levels of n‐3 HUFA (P < 0.05). Results of this study suggest that the effect of dietary n‐3 HUFA on examined non‐specific immunity parameters are not uniform; however, these impacts are closely related to the α‐tocopherol supplement and their interaction. In conclusion, increased dietary levels of n‐3 HUFA and α‐tocopherol would enhance growth performance and welfare of this species.     

Lipid enrichment for Senegalese sole (Solea senegalensis) larvae: effect on larval growth, survival and fatty acid profile

Results from three larval Senegalese sole (Solea senegalensis) feeding trials using non-enriched Artemia and Artemia enriched with Super HUFA®, Arasco®, sunflower oil and microalgae are presented and the effects on larval survival, growth and fatty acid (FA) composition are reported. The FA profile of Senegalese sole eggs was analysed to gather information about the nutritional requirements of the early larval stages and a quite high DHA/EPA ratio (4.3) was found. However, there was no evidence of a high dietary demand for DHA or EPA, given that no relationship was found between dietary HUFA concentration and larval growth and survival. When larvae were fed non-enriched Artemia a significantly better growth and comparable survival were obtained than with Artemia enriched with Super HUFA® (containing the highest HUFA level and DHA/EPA ratio). The FA profiles of the larvae generally reflected those of their diets. DHA was an exception, as it was present in high proportions, even in larvae fed DHA-deficient prey. Total FAME concentration decreased during larval development, with SFA, MUFA and PUFA being equally consumed; HUFA appeared to be less used, with its relative concentration being either kept constant (particularly EPA and ARA) or increased (DHA). A specific requirement for ARA in the first larval stages could not be confirmed but it was always present in considerable amounts, even in larvae fed an ARA poor diet.     

The effects of dietary long‐chain essential fatty acids on growth and stress tolerance in pikeperch larvae (Sander lucioperca L.)

The nutritional requirements of pikeperch larvae have been sparsely examined. Dietary polyunsaturated fatty acids, arachidonic acid (ARA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) may affect growth and physiological stress response in marine fish larvae, but these mechanisms have not received as much attention in freshwater fish. Pikeperch larvae were reared on Artemia from day 3 until 21 days posthatch. Artemia were enriched with six formulated emulsions, with inclusion of either fish oil, pure olive oil (POO) or olive oil supplemented with various combinations of ARA, EPA and DHA. Larval tissue FA was significantly related to the content in the diets, but larval growth was similar for all treatments. When exposed to stress by confinement in small tanks with culture tank water or saline water (15 g L^?1.), mortality in larvae treated with POO was significantly higher than in the remaining treatments while tissue cortisol contents in these fish seemed lower. The findings of a lower stress response in larvae fed POO may be related to the lower tissue content in these larvae of essential fatty acids especially DHA but also EPA and ARA.     

The Effect of Diet on the Biochemical Composition of Juvenile Artemia: Potential Formulations for Rock Lobster Aquaculture

The lipid class and fatty acid (FA) composition of juvenile Artemia fed continuously on four diets—the microalga Tetraselmis suecica , a mix of oat bran-wheat germ-lecithin (OWL), OwL-eicosapentaenoic acid (EPA), and OWL-EPA-arachidonic acid (AA)—were examined over a 9-d experiment in an attempt to approximate the FA profile of phyllosoma larvae of wild southern rock lobster Jasus edwardrii . The main difference in lipid class composition of Artemia fed the four diets was the relative level of polar lipid (PL) and triacylglycerol (TAG). By day 9, the algal-fed Artemia were highest in PL (95% of total lipid) and lowest in TAG (2%), whereas the remaining diets resulted in Artemia with 16–30% PL and 41–82% TAG. After 2 d, the relative FA composition of all Artemia treatments closely reflected those of the diets, with no marked change after further feeding (to day 9). In terms of the content of essential polyunsaturated fatty acids (PUFA), by day 5 Artemia fed: 1) with the algal diet contained 7 mg/g FA dry mass (0.3% DHA, 6.3% EPA, 3.4% AA of total FA); 2) with the OWL diet contained 3 mg/g (0.3% DHA, 0.9% EPA, 0.7% AA); 3) with the OWL-EPA diet contained 55 mg/g (6.2% DHA, 11.6% EPA, 1.1% AA); and 4) with the OWL-EPA-AA contained 83 mg/g (3.8% DHA, 7.5% EPA, 17.4% AA). The PUFA profiles of Artemia using the OWL-oil diets were similar to wild rock lobster phyllmmata, although levels of doco-sahexaenoic acid (DHA) were lower (10% DHA) than in J. edwardsii larvae. On the basis of PUFA composition data alone, the results suggest the suitability of the OWL-oil mixed diets for consideration for feeding to Artemia used in the culture of southern rock lobster larvae, particularly if the level of DHA can be further enhanced.     

Utilization of different dietary lipid sources at high level in herbivorous grass carp (Ctenopharyngodon idella): mechanism related to hepatic fatty acid oxidation

Herbivorous grass carp (Ctenopharyngodon idella) has been reported to exhibit low capacity to utilize high dietary lipid, but different lipid sources might affect this limited capacity. In order to compare the effects of different lipid sources with different lipid levels, juvenile grass carp were fed one of nine diets containing three oils [lard, plant oil mixed by maize and linseed oil, and n‐3 high unsaturated fatty acid‐enriched (HUFA‐enriched) fish oil] at three lipid levels (20, 60 and 100 g kg^?1 dry diet) for 8 weeks. Decreased feed intake, poor growth performance, hepatic pathology and higher blood lipid peroxidation were found in 60 and 100 g kg^?1 fish oil groups. Conversely, in lard and plant oil groups, even at 100 g kg^?1 dietary lipid level, feed intake and growth performance did not decrease, despite histological observation revealed hepatic pathology in these groups. Plasma triglyceride and cholesterol contents increased significantly in all 100 g kg^?1 dietary lipid groups. In the comparison of hepatic FA β‐oxidation among three oil groups at 60 g kg^?1 dietary lipid level, impaired mitochondrial and peroxisomal FA oxidation capacity was observed in fish oil group. The results confirmed the relatively low capacity of grass carp to utilize high dietary lipid, and furthermore excess HUFA intake will result in more serious adverse effects than other FA.     

Effect of krill phospholipids versus soybean lecithin in microdiets for gilthead seabream (Sparus aurata) larvae on molecular markers of antioxidative metabolism and bone development

The objective of the present study was to compare the effectiveness of dietary marine phospholipids (MPL) obtained from krill and soybean lecithin (SBL) on the rearing performance and development of seabream (Sparus aurata) larvae. Larvae were fed from 16 to 44 day posthatching (dph) five formulated microdiets with three different levels (50, 70 and 90 g kg^–1) of phospholipids (PL) obtained either from an MPL or from a SBL source. Larvae‐fed MPL show a higher survival, stress resistance and growth than those‐fed SBL, regardless the dietary PL level. Overall, the increase in MPL up to 70 g kg^–1 total PL in diet was enough to improve larval gilthead seabream performance, whereas even the highest SBL inclusion level (90 g kg^–1 PL) was not able to provide a similar success in larval growth or survival. Inclusion of SBL markedly increased the peroxidation risk as denoted by the higher TBARs in larvae, as well as a higher expression of CAT, GPX and SOD genes. Moreover, SBL tends to produce larvae with a lower number of mineralized vertebrae and a lower expression of osteocalcin, osteopontin and BMP4 genes. Finally, increasing dietary MPL or SBL lead to a better assimilation of polyunsaturated fatty acids in the larvae, n‐3HUFA (especially 20:5n‐3) or n‐6 fatty acids (especially 18:2n‐6), respectively. In conclusion, MPL had a higher effectiveness in promoting survival, growth and skeletal mineralization of gilthead seabream larvae in comparison with SBL.     

The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) and docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on growth, survival and osmotic stress tolerance of Eriocheir sinensis zoea larvae was studied in two separate experiments. In experiment I, larvae were fed rotifers and Artemia enriched with ICES emulsions with 0, 30 and 50% total n-3 HUFA levels but with the same DHA/EPA ratio of 0.6. In experiment II, larvae were fed different combinations of enriched rotifers and Artemia, in which, rotifers were enriched with emulsions containing 30% total n-3 HUFA, but different DHA/EPA ratio of 0.6, 2 and 4; while Artemia were enriched with the same emulsions, but DHA/EPA ratio of 0.6 and 4. In both experiments, un-enriched rotifers cultured on baker's yeast and newly-hatched Artemia nauplii were used as control diets. Larvae were fed rotifers at zoea 1 and zoea 2 stages; upon reaching zoea 3 stage, Artemia was introduced.Experiment I revealed no significant effect of prey enrichment on the survival of megalopa among treatments, but higher total n-3 HUFA levels significantly enhanced larval development (larval stage index, LSI) and resulted in higher individual dry body weight of megalopa. Furthermore higher dietary n-3 HUFA levels also resulted in better tolerance to salinity stress. Experiment II indicated that at the same total n-3 HUFA level, larvae continuously receiving a low dietary DHA/EPA ratio had significantly lower survival at the megalopa stage and inferior individual body weight at the megalopa stage, but no negative effect was observed on larval development (LSI). The ability to endure salinity stress of zoea 3, zoea 5 and megalopa fed diets with higher DHA/EPA ratio was also improved.     

Enhancement of gilthead seabream (Sparus aurata) larval growth by dietary vitamin E in relation to two different levels of essential fatty acids

The objective of this study was to determine the effect of dietary vitamin E on gilthead seabream (Sparus aurata) growth and survival, at two different highly unsaturated fatty acids (HUFAs) levels. Eighteen days old gilthead seabream larvae were fed four formulated experimental diets combining two different dietary levels of HUFAs (M: medium 2.5 + 1.5, DHA + EPA, H: high 5 + 2.5 DHA + EPA g per 100 g) with two different levels of vitamin E (M: medium 540 mg kg^?1, H: high 2900 mg kg^?1): MM, MH, HM, HH (HUFA/vitamin E). After 2‐week feeding trial, the average survival rate was 52.6% and there were no significant differences found among treatments. Increase in vitamin E up to high level markedly improved larval growth, particularly when dietary HUFA levels were lower, suggesting a higher protection value when these fatty acids are more limiting. At medium dietary HUFA levels, increase in vitamin E from medium to high level enhanced larval growth performance in terms of total length. Moreover, increase in vit E enhanced HUFAs content in the larval polar lipids denoting the anti‐oxidative effect of vitamin E.     

Effects of Starvation and Feeding on Lipid Class and Fatty Acid Profile of Late Stage Mud Crab,Scylla serrata,larvae

Lipid class and fatty acid (FA) analysis were conducted on newly molted, fed, and starved zoea V and megalopa of the mud crab, Scylla serrata (S. serrata). Larvae starved for 4 d showed a substantial decrease in total FA content, from 49.67 μg/mg to 13.94 μg/mg ash‐free dry weight (AFDW) at the zoea V stage, and from 38.47 μg/mg to 10.40 μg/mg AFDW at the megalopa stage. This depletion indicates that S. serrata larvae effectively utilize stored lipid reserves for energy during periods of food deprivation. Megalopa subjected to longer starvation periods, however, did not utilize lipid as the major energy source after day 4, suggesting increased reliance on protein catabolism during prolonged starvation. At both larvae stages the major FAs were 18:1n‐9, 16:0, 20:5n‐3 (eicosapentaenoic acids, EPA), 18:3n‐3 (linolenic acid, LNA), 18:0 and 22:6n‐3 (docosahexaenoic acid, DHA) and this FA profile persisted in both fed and starved larvae. The highly unsaturated fatty acids (HUFA), EPA, DHA, and arachidonic acid (20:4n‐6, AA) were not conserved in tissue during starvation, indicating that HUFA requirements might be lower for S. serrata larvae than shown for other crustaceans. Similarly, a high level of LNA in newly molted zoea V and megalopa were rapidly depleted in unfed larvae, indicating that this FA had an important role as an energy reserve. Throughout the study, FAs from the polar lipid fraction dominated larvae tissues, while FAs from the neutral lipid constituted the largest accessible energy reserve during starvation (depleted from 23.05 to 1.23 μg/mg AFDW in zoea V, and from 19.00 to 1.27 μg/mg AFDW in megalopa). The results of this study provide new insight into lipid utilization of S. serrata larvae during development, an important step toward development of formulated diets for use in mud crab hatcheries.     

Optimizing the essential fatty acids, eicosapentaenoic and docosahexaenoic acid, in the diet of the prawn, Penaeus monodon

The dietary requirements of Penaeus monodon for eicosapentaenoic (20:5n‐3; EPA) and docosahexaenoic (22:6n‐3; DHA) acids were examined. These requirements were examined when dietary levels of linoleic (18:2n‐6; LOA) and linolenic acids (18:3n‐3; LNA) were also provided at previously established optimal levels of 14 and 21% respectively of the total lipid fatty acids. A 5 × 5 factorial design was used with incremental amounts (0, 4, 8, 12 and 16% of total fatty acids) of EPA and/or DHA. An additional diet containing cod‐liver oil was provided as a reference diet. The total lipid content of all of the 25 treatments and reference diets was maintained at the same level of 75 g kg^?1. Growth of prawns fed with the reference diet after 50 days was 244 ± 21%. The greatest response to singular additions of EPA or DHA was with a 12% inclusion of either fatty acid, resulting in 287 ± 21 and 293 ± 18% weight gain, respectively. Growth was generally better when combinations of EPA and DHA were used, the optimal combination being EPA 4% and DHA 4%, resulting in 335 ± 25% weight gain. Addition of high levels of either of the highly unsaturated fatty acids (HUFA) in the diet had a negative effect on growth. Digestibilities of the total neutral lipid and specific fatty acids were examined during the growth trials. The digestibility of total neutral lipid was usually higher when either or both HUFA were present, however there were few significant differences between treatments that contained either or both HUFA. Following the growth trials, digestive glands (DG) of prawns fed with the various diets were analysed to determine the total lipid content and fatty acid composition. Total lipid in the digestive gland increased with the inclusion of DHA, but was not significantly affected by the addition of EPA. The fatty acid composition of the digestive gland lipid generally reflected that of the diet. However, the maximum retention of EPA (11.1% of total DG fatty acids) and DHA (10.7% of total DG fatty acids), was not directly proportional to the amount of either fatty acid present in the diet. These results demonstrate that both EPA and DHA have considerable growth promoting capacity. This growth promoting capacity is enhanced when an optimal balance of both fatty acids are incorporated into the diet.     

Effects of dietary lipid level on growth performance,body composition and digestive enzymes activity of juvenile sea cucumber,Apostichopus japonicus

A feeding trial aimed to determine the effects of dietary lipid level on growth performance, body composition and digestive enzymes activity of juvenile sea cucumber, Apostichopus japonicus. Diets with six crude lipid levels (1.9, 13.8, 29.1, 43.6, 59.6 and 71.6 g kg^?1) were fed to sea cucumbers (initial weights 0.65 ± 0.01 g) at a density of 30 juveniles, once a day. After 60 days, body weight gain (BWG), specific growth rate (SGR), feed intake (FI) and protein efficiency ratio (PER) decreased with increasing dietary lipid levels. The sea cucumbers fed 1.9 g kg^?1 crude lipid showed significantly higher (P < 0.05) BWG than those of the sea cucumbers fed 59.6 and 71.6 g kg^?1 crude lipid. Intestinal protease and lipase activities generally increased with increasing dietary lipid levels. Eicosapentaenoic acid (EPA) content of body walls generally increased with increasing dietary lipid levels. Docosahexaenoic acid (DHA) content of body walls reached the maximum value at a dietary lipid level of 13.8 g kg^?1. N‐3 highly unsaturated fatty acid content followed the same pattern of DHA. According to the growth performance and body composition of sea cucumbers, it can be indicated that the optimum dietary lipid level for juvenile sea cucumbers is between 1.9 and 13.8 g kg^?1.     

The importance of dietary HUFA for meagre larvae (Argyrosomus regius; Asso, 1801) and its relation with antioxidant vitamins E and C

Despite the interest of meagre (Argyrosomus regius) as a fast‐growing candidate for Mediterranean aquaculture diversification, there is a lack of information on nutrition along larval development. Importance of highly unsaturated fatty acids (HUFA) and the antioxidant vitamins E and vitamin C has not been investigated yet in this species. Six diets with two levels of HUFA (0.4% and 3% dw), two of vitamin E (1500 and 3000 mg kg^?1) and two of vitamin C (1800 and 3600 mg kg^?1) were fed to 15 dah meagre larvae. Larval growth in total length and dry body weight was significantly lowest in larvae fed diet 0.4/150/180 and showed few lipid droplets in enterocytes and hepatocytes and lower HUFA contents than the initial larvae. Increase in dietary HUFA up to 3%, significantly improved larval growth and lipid absorption and deposition. Besides, among fish fed 3% HUFA, increase in vitamin E and vitamin C significantly improved body weight, as well as total lipid, 22:6n‐3 and n‐3 fatty acids contents in the larvae. Thus, the results showed that 0.4% dietary HUFA is not enough to cover the essential fatty acid requirements of larval meagre and a high HUFA requirement in weaning diets is foreseen for this species. Besides, the results also pointed out the importance of dietary vitamin E and C to protect these essential fatty acids from oxidation, increase their contents in the larvae and promote growth, suggesting high vitamin E and C requirements in meagre larvae (higher than 1500 and 1800 mg kg^?1 for vitamin E and vitamin C respectively).     

A determination of the quantitative requirements for docosahexaenoic acid for juvenile barramundi (Lates calcarifer)

A series of diets with varying docosahexaenoic acid (DHA; 22:6n‐3) inclusion levels (1 g kg^?1 3 g kg^?1, 6 g kg^?1, 10 g kg^?1, 15 g kg^?1 and 18 g kg^?1) were fed to juvenile barramundi (Lates calcarifer) for 6 weeks. Two additional diets examined the addition of arachidonic acid (ARA; 20:4n‐6) or eicosapentaenoic acid (EPA; 20:5n‐3) to the diets at 10 g kg^?1 when DHA was also included at 10 g kg^?1. Fish were fed the diets on a pair‐fed feeding regime to eliminate feed intake variability. Fish were weighed, and blood and tissue samples were collected after 6 weeks. Behavioural parameters were also assessed. Improvement in growth was seen with increasing inclusion of DHA up to a maximum at 10 g kg^?1 inclusion, albeit the response was minor. However, the addition of ARA to the diet reduced the growth response, while the addition of EPA improved the growth response. An improvement in feeding behaviour was also seen with increasing DHA up to a peak at 10 g kg^?1, while those animals fed diets low in DHA showed increasingly cryptic behaviour. With the increasing inclusion of DHA, a range of pathologies were observed, but the addition of an EPA component to the diet limited these pathologies, while the addition of ARA made little improvement and in some cases exacerbated the pathologies.     

Effect of lipid supplementation on reproductive performance of female channel catfish,Ictalurus punctatus,induced and strip‐spawned for hybridization

The influence of different lipid sources and n3:n6 ratios on reproductive performance of female channel catfish, Ictalurus punctatus was evaluated. A commercial catfish feed was top coated with 2% oil and offered to brood stock females fish during 70–85 days before spawning season. Four dietary treatments were formulated using the following top coating ratios: diet 1, soybean oil 9.5 g kg^?1 and linseed oil 10.5 g kg^?1; diet 2, soybean oil 17.5 g kg^?1 and linseed oil 2.5 g kg^?1; diet 3, 20.0 g kg^?1 linseed oil, and diet 4, 10.0 g kg^?1 menhaden fish oil, supplemented with 5.0 g kg^?1 arachidonic acid (ARA), and 5.0 g kg^?1 docosahexaenoic acid (DHA). Fatty acid composition of the eggs reflected the effect of dietary treatment offered during spring season. Supplementation of ARA, EPA and DHA in commercial catfish feed in the form of menhaden fish oil with purified liquid algae extracts of ARA and DHA produced from two to five times the number of fry per female body weight when compared to the effect of fed top coated with vegetable oils. Although, this effect was not statistically significant it may represent an economical improvement for the industry.     

Effect of high dietary intakes of vitamin E and n-3 HUFA on immune responses and resistance to Edwardsiella tarda challenge in Japanese flounder (Paralichthys olivaceus, Temminck and Schlegel)

A feeding experiment was conducted to investigate the effects of high dietary intake of vitamin E (supplied as dl ‐α‐tocopheryl acetate) and n‐3 highly unsaturated fatty acid (n‐3 HUFA) on the non‐specific immune response and disease resistance in Japanese flounder Paralichthys olivaceus. Nine practical diets were formulated to contain one of three levels of vitamin E namely, 0, 80 or 200 mg kg^?1 (the total α‐tocopherol contents in the diets were 21, 97 and 213 mg kg^?1 based on analysis), and at each vitamin E level with one of three n‐3 HUFA levels i.e. 0.5%, 1.5% or 2.0%. Each diet was randomly assigned to triplicate groups of Japanese flounder (initial body weight: 40.5±1.0 g, mean±SD) in a re‐circulation rearing system. Fish were fed twice daily to apparent satiation at 07:00 and 18:00 hours for 12 weeks. During the experimental period, water temperature was maintained at 18±1°C, salinity 31–35 g L^?1, and pH 7.8–8.2. Dissolved oxygen was not less than 6 mg L^?1, and there were negligible levels of free ammonia and nitrite. The results showed that the increase in dietary n‐3 HUFA from 0.5% to 1.0% significantly decreased muscle α‐tocopherol contents in fish‐fed diets with 21 and 97 mg α‐tocopherol kg^?1 diet (P<0.05). In 1.0% HUFA groups, alternative complement pathway activity (ACH₅₀) of fish fed the diet containing the 213 mg α‐tocopherol kg^?1 diet was significantly higher than noted for fish fed the diet containing 97 mg α‐tocopherol kg^?1 diet (P<0.05). Fish fed the diet with 213 mg α‐tocopherol kg^?1 and 2.0% n‐3 HUFA had the highest lysozyme activity (131.7 U mL^?1) among all the dietary treatments. Fish fed the diets containing 97 and 213 mg α‐tocopherol kg^?1 with 1.0% n‐3 HUFA had significantly higher respiratory burst activity than those fed the diets containing 21 mg α‐tocopherol kg^?1 with 0.5 and 1.0% n‐3 HUFA (P<0.05). In the disease resistance experiment, high intake of dietary vitamin E with 213 mg α‐tocopherol kg^?1 significantly decreased cumulative mortality and delayed the days to first mortality after a 7‐day Edwardsiella tarda challenge (P<0.05). In addition, under the experimental conditions, dietary vitamin E and n‐3 HUFA had a synergistic effect on the non‐specific immune responses and disease resistance in Japanese flounder (P<0.05).