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.     

The dietary linoleic and linolenic fatty acids requirements of the prawn Penaeus monodon

The dietary requirement of the prawn Penaeus monodon for linoleic (LOA) and linolenic (LNA) fatty acids was examined in the absence of other long-chain polyunsaturated and highly unsaturated fatty acids (PUFA-20:2, 20:3, 22:2, 22:3 and HUFA-18:4, 20:4, 20:5, 22:4, 22:5, 22:6, respectively). Incremented dietary amounts of LOA (7, 14, 21, 28 and 35% of total fatty acids) and LNA (0, 7, 14, 21 and 28% of total fatty acids) were examined in a 5 × 5 factorial growth experiment lasting 50 days. An additional diet containing both PUFA and HUFA (cod-liver oil) was provided as a reference. The total lipid content (excluding sterols) of each of the 26 diets was maintained at 70 g kg⁻¹ of dry diet. The fatty acid composition of the neutral lipid was manipulated by blending different plant oils and supplementing with purified free fatty acids to provide the desired fatty acid composition upon addition to the total diet. At the end of the 50-day growth experiment, the prawn digestive gland (DG) was quantitatively analysed for lipid and fatty acid content. Prawns fed the reference diet increased in weight (mean ± SEM) by 214 ± 6%. Growth was generally greater when combinations of LOA and LNA were used. The best growth (213 ± 17%) was obtained with the diet containing a fatty acid content of 14% LOA and 21% LNA. This growth was comparable to that of the reference diet. The digestibility of the total lipid in the diet was usually higher when both fatty acids were present. The lipid content of the DG was highest in prawns fed diets containing both LOA and LNA, similar to the growth response. The fatty acid composition of the prawn's DG lipid reflected the fatty acid composition of the diet. However, the maximum assimilation of LNA in the DG lipid (14.2% of DG lipid fatty acids) was about half that of LOA (32.5% of DG lipid fatty acids).     

Nutritional evaluation of fatty acids for the open thelycum shrimp, Litopenaeus vannamei: II. Effect of dietary n-3 and n-6 polyunsaturated and highly unsaturated fatty acids on juvenile shrimp growth, survival, and fatty acid composition

This study evaluated the nutritional value of dietary n‐3 and n‐6 polyunsaturated fatty acids (PUFA) such as linoleic (LOA) and linolenic (LNA) acids, and highly unsaturated fatty acids (HUFA) such as arachidonic (AA), eicosapentaenoic (EPA), and docosahexaenoic (DHA) acids for juvenile Litopenaeus vannamei, based on their effects on growth, survival, and fatty acid composition of hepatopancreas and muscle tissue. Diets contained 5% total lipid. A basal diet contained palmitic and stearic acids each at 2.5% of diet. Five diets contained 0.5% dry weight of LOA, LNA, AA, EPA, or DHA. An additional diet evaluated HUFA in combination by supplementing at 0.5% of diet, a mixture of n‐3 HUFA. All HUFA showed higher nutritional value than PUFA for shrimp and produced significantly (P < 0.05) higher final weight, weight gain, and total lipid in shrimp muscle. Fatty acid profiles of shrimp tissues reflected the composition of the dietary lipids. In general, saturated fatty acids were more abundant in the neutral factions, while PUFA and HUFA were more abundant in the polar fractions of tissues. Under these experimental conditions, HUFA had much greater nutritional value than PUFA for juvenile L. vannamei; moreover, dietary requirements for PUFA were not demonstrated.     

Nutritional evaluation of fatty acids for the open thelycum shrimp, Litopenaeus vannamei: I. Effect of dietary linoleic and linolenic acids at different concentrations and ratios on juvenile shrimp growth, survival and fatty acid composition

A 6‐week feeding trial was conducted to evaluate the nutritional value of dietary linoleic (18:2n‐6, LOA) and linolenic (18:3n‐3, LNA) acids for juvenile Litopenaeus vannamei by determining their effects on growth, survival and fatty acid composition of hepatopancreas and muscle tissue. Diets were formulated to contain 5% total lipid. A basal diet contained only palmitic and stearic acids, each at 2.5% of diet. Six diets contained one of three levels (0.25, 0.5 and 1%) of either LOA or LNA, and three diets had different ratios of LNA/LOA (1, 3, 9) at a combined inclusion level of 0.5% of diet. An additional diet contained 0.5% of a mixture of n‐3 highly unsaturated fatty acids (HUFA). The fatty acid profile of hepatopancreas and muscle of shrimp reflected the profile of the diets. HUFA of the n‐3 family showed higher nutritional value than LOA or LNA for juvenile L. vannamei by producing significantly (P < 0.05) higher final weight and weight gain. Neither LOA nor LNA, alone or in combination, improved growth significantly compared with shrimp fed the basal diet.Thus, dietary requirements for LOA and LNA were not demonstrated under these experimental conditions.     

Growth and fatty acid composition of juvenile Cerastoderma edule (L.) fed live microalgae diets with different fatty acid profiles

The importance of dietary 20:5n‐3 (EPA), 22:6n‐3 (DHA) and 20:4n‐6 (ARA) for growth, survival and fatty acid composition of juvenile cockles (Cerastoderma edule) was investigated. Cockles of 6.24 ± 0.04 mm and 66.14 ± 0.34 mg (live weight) were distributed into three treatments where live microalgae diets were fed constantly below the pseudofaeces production threshold, for three weeks. Diets had distinct fatty acid profiles: high EPA (53% Chaetoceros muelleri + 47% Pyramimonas parkeae), no DHA (47% Brachiomonas submarina + 53% Tetraselmis suecica) and low ARA concentrations (73% P. parkeae + 27% Phaeodactylum tricornutum). Growth was positively affected by high EPA and low ARA diets, whereas no significant growth was observed for the no DHA diet. High mortality of cockles fed no DHA diet raises questions about its suitability for cockles. In balanced diets with EPA and DHA, lower concentrations of ARA do not limit growth. The impact of dietary fatty acids was evident in the fatty acids of neutral and polar lipids of cockles. In polar lipids of all cockles, there was a decrease in EPA, in contrast to an increase in DHA. The combination of EPA and DHA in a live microalgae diet was beneficial for the growth and survival of juvenile cockles.     

Aurantiochytrium sp. meal as DHA source in Nile tilapia diet,part II: Body fatty acid retention and muscle fatty acid profile

Nile tilapia juveniles (8.35 ± 0.80 g) were fed on four levels (0.0%; 0.5%; 1.0%; 2.0%, 4.0%) of Aurantiochytrium sp. meal (ALL‐G‐RICH?), a source of docosahexaenoic acid (DHA). The 1% Aurantiochytrium sp. meal diet was compared to a control diet, which contained the same amount of DHA as cod liver oil (CLO) at 1.7% diet. Groups of 25 fish were stocked in 100 L tanks and fed twice daily until apparent satiation, for 57 days, at 28°C. Increasing dietary Aurantiochytrium sp. meal reduced the body retention of DHA and n‐3 polyunsaturated fatty acids (n‐3 PUFA) but increased the body retention of alpha‐linolenic (α‐LNA), linoleic (LOA) and n‐6 polyunsaturated fatty acids (n‐6 PUFA). Fatty acid profile in tilapia muscle was affected by increasing dietary inclusions of Aurantiochytrium sp. meal, with an increase in DHA, α‐LNA, n‐3 PUFA and n‐3 long chain‐polyunsaturated fatty acids (n‐3 LC‐PUFA) but a decrease in monounsaturated fatty acids (MUFA), n‐6 PUFA and n‐6 long‐chain polyunsaturated fatty acids (n‐6 LC‐PUFA). There was a larger body retention of DHA, α‐LNA, LOA, n‐3 PUFA and n‐6 PUFA fatty acids and a higher percentage of DHA, n‐3 PUFA and n‐3 LC‐PUFA in muscle fatty acid profile in fish fed on CLO diets than in those fed on 1% Aurantiochytrium sp. Therefore, Aurantiochytrium sp. meal is an alternative source of DHA for Nile tilapia diets.     

Dietary lipid sources for seabream and seabass: growth performance, tissue composition and flesh quality

Due to its traditionally good availability, digestibility and high content of n ? 3 HUFA, fish oil is the main lipid source in fish feeds. However, world demand for this product has grown significantly in recent years, whereas its production, based on fisheries landings, is static. The purpose of the present study was to assess the effect of partial replacement of fish oil in compound diets for gilthead seabream and seabass, by several vegetable oil sources, on growth, dietary fatty acid utilization and flesh quality. Five iso‐energetic and isoproteic experimental diets were formulated (25% lipid content). Fish oil was the only added lipid source in the control (FO) diet, and it was included in the other experimental diets at a level high enough (40% of FO diet) to keep the n ? 3 HUFA levels well over 3% in order to cover the essential fatty acid requirements of these species. Fish oil was replaced by soyabean oil (SO), rapeseed oil (RO) and linseed oil (LO) or a mixture (Mix) of them. Feed intake in all dietary groups was in the range of results obtained for commercial diets in both species, and growth and feed utilization were very good. The results show that, providing a minimum content of essential fatty acids in the diet, it is possible to replace up to 60% of the fish oil by SO, LO and RO or a mixture of them in diets for seabream and seabass, without compromising fish growth. Fatty acid composition of liver and muscle reflected that of the diet, but utilization of dietary lipids differed between these two tissues and was also different for the different fatty acids. Despite reduction in dietary saturated fatty acids by the inclusion of vegetable oils, their levels in fish liver were as high as in fish fed the fish oil diet, whereas, in muscle, levels were reduced according to that in the diet. Linoleic and linolenic acids were accumulated in the liver proportionally to their levels in the diet, suggesting a lower oxidation of these fatty acids in comparison to other 18C fatty acids. Regarding eicosapentaenoic acid (20 : 5n ? 3; EPA), docosahexaenoic acid (22 : 6n ? 3; DHA) and arachidonic acid (20 : 4n ? 6; ARA), these essential fatty acids were reduced in the liver at a similar rate, whereas DHA was preferentially retained in the muscle in comparison with the other fatty acids, denoting a higher oxidation particularly of EPA, in the muscle. Some other PUFA increased despite their low dietary levels in seabream fed LO diets and in seabass fed SO diet, suggesting the stimulation of delta‐6 and delta‐5 desaturase activity in marine fish. Despite differences in fatty acid composition, fillet of fish fed vegetable oils was very well accepted by trained judges when assessed cooked.     

Growth, survival, lipid composition and pigmentation of turbot (Scophthalmus maximus) larvae fed live-prey enriched in Arachidonic and Eicosapentaenoic acids

Turbot larvae were fed live-prey enriched with different levels of arachidonic (ARA) and eicosapentaenoic (EPA) acids to study the effects of these fatty acids on body composition and pigmentation success. Significantly reduced pigmentation was obtained in those fish fed medium and high ARA diets for 43 days. Growth and survival were the same for all groups. The incorporation of ARA and EPA in fish eyes, brains, livers and carcasses reflected the percentage of these fatty acids in the diets. ARA accumulation was similar in all tissues, but brain accumulated EPA was less efficient than the other tissues examined. A highly significant, negative correlation was found between the %ARA in turbot juvenile brain total lipids and pigmentation success. A weaker, positive correlation was found between brain EPA and pigmentation. Increasing dietary ARA affected the fatty acid composition of turbot brain phosphoglycerides more than increasing dietary EPA, especially in phosphatidylinositol (PI) and phosphatidylethanolamine (PE). A negative relationship was found between percentage normal pigmentation and ARA levels in brain phosphatidylcholine (PC), PE and phosphatidylserine (PS). Elevated levels of ARA in PI also resulted in malpigmented juveniles, but EPA:ARA ratios ≥1 in PI were associated with normal pigmentation. We conclude that, given a sufficiency of dietary docosahexaenoic acid (DHA), the optimum dietary level of EPA is not a function of DHA, but of dietary ARA.     

Requirement of n-3 long chain polyunsaturated fatty acids for European sea bass (Dicentrarchus labrax) juveniles: growth and fatty acid composition

European sea bass juveniles (14.4±0.1 g mean weight) were fed diets containing different levels of fish oil then of n-3 highly unsaturated fatty acids (n-3 HUFA) for 12 weeks. The fish performance as well as fatty acid (FA) composition of neutral and polar lipids from whole body after 7 and 12 weeks feeding were studied. The requirements of juvenile sea bass for n-3 highly unsaturated fatty acids (n-3 HUFA) were studied by feeding fish diets containing six different levels of n-3 HUFA ranging from 0.2% to 1.9% of the diet, with approximately the same DHA/EPA ratio (1.5:1).

The growth rate at the end of the trial showed significant differences. Fish fed low dietary n-3 HUFA (0.2% DM of the diet) showed significantly lower growth than the diet 3 (0.7%), then no further improvement (P>0.05) of growth performance was seen by elevating the n-3 HUFA level in the diet up to 1.9% (diet 6). No difference in feed efficiency, protein efficiency ratio or protein retention was observed among treatments, nor in protein and total lipid content. However, the n-3 HUFA levels in diets highly influenced fish fatty acid composition in neutral lipid, while polar lipid composition was less affected. Comparison of polar lipid content after 7 or 12 weeks indicated that DHA remained stable at the requirement level, while arachidonic acid decreased with time. Results of this experiment suggest that the requirement for growth of n-3 HUFA of juvenile sea bass of 14 g weight is at least 0.7% of the dry diet.     

Development of a broodstock diet to improve developmental competence of embryos in European eel,Anguilla anguilla

We examined the effect of dietary arachidonic acid (ARA) and eicosapentaenoic acid (EPA) on the production of embryos and hatched larvae in the European eel, Anguilla anguilla. Two diets with high and intermediate levels of ARA and low and intermediate levels of EPA (Feed 1: ARA 1.9%, EPA 4.2%; Feed 2: ARA 1.2%, EPA 5.1% of total fatty acids) were tested against a commercial diet (DE: ARA: 0.5%, EPA: 8.2% of total fatty acids). After 24 weeks of feeding, ARA levels in the muscles and ovaries increased to 0.9% and 1.3% of total fatty acids, respectively, in Feed 1 and were significantly higher than in Feed 2 and DE. Female broodstock was not fed during hormonal treatment to induce vitellogenesis and ovulation. EPA levels in females fed the test diets decreased in the both muscle and ovary and were significantly lower in eggs from females fed Feed 1. The highest percentage of stripped females, producing viable eggs and larvae, were those females fed the highest dietary ARA levels (Feed 1). The level of lipid peroxidation products in eggs was similar among treatment, indicating that the lowest dietary levels of vitamin C and vitamin E were sufficient. In the unfertilized eggs, ARA levels were also highest (1.1% of total fatty acids) in the diet with highest ARA levels (Feed 1).     

Effects of varying dietary docosahexaenoic acid levels on growth,proximate composition and tissue fatty acid profile of juvenile silver pomfrets,Pampus argenteus (Euphrasen, 1788)

This study investigated the effects of varying dietary levels of decosahexaenoic acid (DHA) on growth performance, proximate composition and whole body fatty acid profiles of juvenile silver pomfret, Pampus argenteus. Triplicate groups of fish (30.55 ± 0.08 g) were fed diets containing 5.2%, 9.31% and 13.38% DHA (% of total fatty acids) or 0.85%, 1.52% and 2.18% DHA on dry diet weight for diets 1, 2 and 3 respectively. Survival was not affected by dietary DHA levels. The growth performance and feed utilization parameters of fish fed diets 2 and 3 were significantly (P < 0.05) higher than those fed diet 1, although these parameters in diets 2 and 3 did not differ significantly (P > 0.05). Whole body lipid and fatty acid profiles were influenced by dietary DHA levels. Significantly higher n‐3 fatty acids particularly DHA, DHA:EPA(eicosapentaenoic acid) ratios and n‐3:n‐6 ratios were observed in fish fed diets 2 and 3 compared to those fed diet 1. Better growth performance and higher whole body DHA:EPA (2.31, 2.29) ratios and n‐3:n‐6 ratios (2.17, 2.12) observed in fish fed diets 2 and 3, respectively, suggests that silver pomfret juveniles have a higher requirement for n‐3 fatty acids, notably DHA for optimum growth and survival.     

牙鲆幼鱼对EPA和DHA的营养需求

研究了EPA和DHA水平对牙鲆生长的影响,饲料中含0.5%EPA和1.0%～1.5%DHA能保证牙鲆幼鱼最适生长,鱼体水分最低,肝体指数最小,脂肪含量有较大幅度提高,肝脏极性脂中EPA和DHA达到最大积累;在肝脏和肌肉的非极性脂部分,各组间的脂肪酸组成没有显著变化,而极性脂部分能体现出饲料中n-3 HUFA含量对鱼体脂肪酸组成的影响,极性脂中的EPA和DHA含量远高于非极性脂;在肌肉和肝脏的极性脂和非极性脂中都含有较高的16:0和18:1n-9; 18:1n-9/n-3HUFA可以作为必需脂肪酸满足程度的一个判据,18:1n-9值的升高往往是缺乏必需脂肪酸的表现,在生长最佳时18:1n-9/n-3HUFA比值下降,为0.62和0.74.     

Comparison of effects of vegetable oils blended with southern hemisphere fish oil and decontaminated northern hemisphere fish oil on growth performance, composition and gene expression in Atlantic salmon (Salmo salar L.)

Replacement of fish oil with sustainable alternatives, such as vegetable oil, in aquaculture diets has to be achieved without compromising the nutritional quality, in terms of n-3 highly unsaturated fatty acid (HUFA) content, of the product. This may be possible if the level of replacement is not too high and oil blends are chosen carefully but, if high levels of fish oil are substituted, a fish oil finishing diet prior to harvest would be required to restore n-3HUFA. However, a decontaminated fish oil would be required to avoid increasing undesirable contaminants. Here we test the hypotheses that blending of rapeseed and soybean oils with southern hemisphere fish oil will have a low impact upon tissue n-3HUFA levels, and that decontamination of fish oil will have no major effect on the nutritional quality of fish oil as a feed ingredient for Atlantic salmon. Salmon (initial weight ~ 0.8 kg) were fed for 10 weeks with diets in which 60% of fish oil was replaced with blends of soybean, rapeseed and southern hemisphere fish oil (SVO) or 100% decontaminated northern fish oil (DFO) in comparison with a standard northern fish oil diet (FO). Decontamination of the oil was a two-step procedure that included treatment with activated carbon followed by thin film deodorisation. Growth performance and feed efficiency were unaffected by either the SVO or DFO diets despite these having lower gross nutrient and fatty acid digestibilities than the FO diet. There were also no effects on the gross composition of the fish. Liver and, to a lesser extent flesh, lipid levels were lower in fish fed the SVO blends, due to lower proportions of neutral lipids, specifically triacylglycerol. Tissue lipid levels were not affected in fish fed the DFO diet. Reflecting the diet, flesh eicosapentaenoic acid (EPA) and total n-3 fatty acids were higher, and 18:1n-9 lower, in fish fed DFO than FO, whereas there were no differences in liver fatty acid compositions. Flesh EPA levels were only slightly reduced from about 6% to 5% although docosahexaenoic acid (DHA) was reduced more severely from around 13% to about 7% in fish fed the SVO diets. In contrast, the liver fatty acid compositions showed higher levels of n-3 HUFA, with DHA only reduced from 21% to about 18% and EPA increased from under 8% to 9–10% in fish fed the SVO diets. The evidence suggested that increased liver EPA (and arachidonic acid) was not simply retention, but also conversion of dietary 18:3n-3 and 18:2n-6. Increased HUFA synthesis was supported by increased hepatic expression of fatty acyl desaturases in fish fed the SVO diets. Flesh n-3HUFA levels and desaturase expression was significantly higher in fish fed soybean oil than in fish fed rapeseed oil. In conclusion, partial replacement of fish oil with blends of vegetable oils and southern hemisphere fish oil had minimal impact on HUFA levels in liver, but a greater effect on flesh HUFA levels. Despite lower apparent digestibility, decontamination of fish oil did not significantly impact its nutritional quality for salmon.     

Effects of docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limanda ferruginea): a live food enrichment experiment

The role of dietary ratios of docosahexaenoic acid (DHA, 22:6n−3), eicosapentaenoic acid (EPA, 20:5n−3) and arachidonic acid (AA, 20:4n−6) on early growth, survival, lipid composition, and pigmentation of yellowtail flounder was studied. Rotifers were enriched with lipid emulsions containing high DHA (43.3% of total fatty acids), DHA+EPA (37.4% and 14.2%, respectively), DHA+AA (36.0% and 8.9%), or a control emulsion containing only olive oil (no DHA, EPA, or AA). Larvae were fed differently enriched rotifers for 4 weeks post-hatch. At week 4, yellowtail larvae fed the high DHA diet were significantly larger (9.7±0.2 mm, P<0.05) and had higher survival (22.1±0.4%), while larvae fed the control diet were significantly smaller (7.3±0.2 mm, P<0.05) and showed lower survival (5.2±1.9%). Larval lipid class and fatty acid profiles differed significantly among treatments with larvae fed high polyunsaturated fatty acid (PUFA) diets having higher relative amounts of triacylglycerols (18–21% of total lipid) than larvae in the control diet (11%). Larval fatty acids reflected dietary levels of DHA, EPA and AA while larvae fed the control diet had reduced amounts of monounsaturated fatty acids (MUFA) and increased levels of PUFA relative to dietary levels. A strong relationship was observed between the DHA/EPA ratio in the diet and larval size (r²=0.75, P=0.005) and survival (r²=0.86, P=0.001). Following metamorphosis, the incidence of malpigmentation was higher in the DHA+AA diet (92%) than in all other treatments (50%). Results suggest that yellowtail larvae require a high level of dietary DHA for maximal growth and survival while diets containing elevated AA exert negative effects on larval pigmentation.     

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.     

Influence of highly unsaturated fatty acids in live food on larviculture of mud crab Scylla paramamosain (Estampador 1949)

Two experiments were carried out to investigate the effects of docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and arachidonic acid (ARA) levels in rotifers (Brachionus plicatilis) and Artemia on the survival, development and metamorphosis of mud crab Scylla paramamosain larvae. Five different lipid emulsions, varying in the level of total n‐3 and n‐6 highly unsaturated fatty acids (HUFA), DHA, EPA and ARA were used to manipulate the fatty acid profile of the live food. Fatty acid profiles of the live food and crab larvae at zoea one, three and five stages were analysed to study the HUFA uptake by the larvae. The fatty acid content of the live food affected the fatty acid profiles of the crab larvae. In both experiments, the survival rate in the zoeal stages was not statistically different among treatments. However, larval development rate and metamorphosis success were affected by the dietary treatments. In this respect, the DHA/EPA ratio in the live food seems to be a key factor. Enrichment emulsions with a very high (50%) total HUFA content but a low DHA/EPA ratio (0.6), or zero total HUFA content caused developmental retardation and/or metamorphosis failure. An emulsion with a moderate total HUFA (30%) and a high DHA/EPA ratio (4) was the best in terms of larval development during the zoeal stages and resulted in improved metamorphosis. Dietary ARA seemed to improve first metamorphosis, but its exact role needs further clarification. For the larval rearing of S. paramamosain, an enrichment medium containing about 30% total n‐3 HUFA with a minimum DHA/EPA ratio of 1 is recommended. Further investigation is needed on the total HUFA and optimum DHA/EPA ratio requirements for each crab larval stage.     

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.     

Dietary effects of n-3 highly unsaturated fatty acid levels on egg and larval quality, and the fatty acid composition of the eggs of Chilean flounder Paralichthys adspersus broodstock

The effects of dietary n-3 highly unsaturated fatty acid (n-3 HUFA) on eggs and larval quality were investigated in the Chilean flounder Paralichthys adspersus . Broodstock were fed with three formulated diets with similar proximate compositions but different n-3 HUFA (2.1%, 3.1% or 4.1%) estimated levels from 5 months before and during the spawning period. The diet with an intermediate n-3 HUFA level resulted in a significantly higher ( P <0.05) percentage of buoyant eggs (68.2 ± 2.9%), fertilization (92.8 ± 3.9%), normal cell cleavages (93.5 ± 1.9%), hatching rate (87.7 ± 4.1%) and normal larvae (76.3 ± 3.7%) compared with the other two diets. In contrast, high levels of n-3 HUFA produced larvae with a higher survival capacity when subjected to fasting. The diet with the lowest content of n-3 HUFA produces lower quality eggs and larvae. The n-3 HUFA level in eggs increased with an increase in the dietary level, and the n-3/n-6 ratios were 1:1, 2:1 and 3:1. The DHA/EPA and EPA/ARA ratios of 2 and 4 in eggs, respectively, were associated with improved egg and larval quality and were similar to the ratios found in eggs from wild broodstock. Attainment of optimal fatty acid contents in broodstock diets is one of the key factors for producing the high-quality spawning required for managed culture of this flounder.