Optimum Level of Dietary n‐3 Highly Unsaturated Fatty Acids for Juvenile Sea Cucumber,Apostichopus japonicus

A feeding trial was conducted to estimate the optimum level of dietary n‐3 highly unsaturated fatty acids (HUFAs) for juvenile sea cucumber, Apostichopus japonicas, based on growth performance and fatty acid compositions. Diets with five n‐3 HUFAs levels (0.15, 0.22, 0.33, 0.38, and 0.46%) were fed to sea cucumber juveniles (1.97 ± 0.01 g) once a day for 60 d. The sea cucumbers fed diets containing 0.22% n‐3 HUFAs showed significantly (P < 0.05) higher body weight gain, feed efficiency, and protein efficiency ratio than the sea cucumbers fed diets containing 0.15% n‐3 HUFAs, but not significantly different (P > 0.05) from those of sea cucumbers fed diets containing 0.33, 0.38, and 0.46% n‐3 HUFAs. The sea cucumbers fed diets containing 0.46% n‐3 HUFAs showed significantly (P < 0.05) higher eicosapentaenoic acid and saturated fatty acid than the sea cucumber fed diets containing 0.15% n‐3 HUFAs, but not significantly different (P > 0.05) from those of sea cucumbers fed diets containing 0.22, 0.33, and 0.38% n‐3 HUFAs. The results of growth performance and n‐3 HUFA compositions of body wall indicated that the optimum level of dietary n‐3 HUFAs for juvenile sea cucumber is between 0.22 and 0.46%.     

Dietary ratios of n‐3/n‐6 fatty acids do not affect growth of Nile tilapia at optimal temperatures (28°C) nor at temperatures that simulate the onset of winter (22°C)

Nile tilapia (Oreochromis niloticus) juveniles were fed diets containing 13 g/kg total polyunsaturated fatty acids (PUFAs) at different n‐3/n‐6 dietary ratios (0.2, 0.5, 0.8, 1.3 and 2.9) for 56 days, at 28°C. Subsequently, fish were submitted to a winter‐onset simulation (22°C) for 33 days. PUFA n‐3/n‐6 dietary ratios did not affect fish growth at either temperature. At 28°C, tilapia body fat composition increased with decreasing dietary PUFA n‐3/n‐6. Winter‐onset simulation significantly changed feed intake. The lowest dietary n‐3/n‐6 ratio resulted in the highest feed intake. At both temperatures, body concentrations of α‐linolenic acid, docosahexaenoic acid, eicosatrienoic acid and docosapentaenoic acid decreased as dietary n‐3/n‐6 decreased. Body concentrations of eicosapentaenoic acid (EPA, 20:5 n‐3) increased with decreasing concentrations of dietary EPA. The n‐6 fatty acids with the highest concentrations in tilapia bodies were linoleic acid and arachidonic acid (ARA, 20:4 n‐6). At 28°C, SREBP1 gene expression was upregulated in tilapia fed the lowest n‐3/n‐6 diet compared to tilapia fed the highest n‐3/n‐6 ratio diet. Our results demonstrate that a dietary PUFA of 13 g/kg, regardless of the n‐3/n‐6 ratio, can promote weight gains of 2.65 g/fish per day at 28°C and 2.35 g/fish per day at 22°C.     

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.     

Long‐chain polyunsaturated fatty acid metabolism in carnivorous marine teleosts: Insight into the profile of endogenous biosynthesis in golden pompano Trachinotus ovatus

Golden pompano Trachinotus ovatus is an important farmed carnivorous marine teleost. Although some enzymes for long‐chain polyunsaturated fatty acid (LC‐PUFA) biosynthesis have been identified, the ability of T. ovatus for endogenous biosynthesis is unknown. Here, we evaluated in vivo LC‐PUFA synthesis in a 56‐day culture experiment using six diets (D1–D6) formulated with linseed and soybean oils to produce dietary linolenic/linoleic acid (ALA/LA) ratios ranging from 0.14 to 2.20. The control diet (D0) used fish oil as lipid source. The results showed that, compared with the corresponding indices of fish fed D0, the weight gain rate and specific growth rate as well as the contents of eicosapentaenoic (EPA) and docosahexaenoic acids in tissues (liver, muscle, brain and eye) of D1–D6 groups were significantly lower (p < .05). These data suggested that T. ovatus could not synthesize LC‐PUFA from C₁₈ PUFA or such ability was very low. However, tissue levels of 20:4n‐3 in fish fed diets D1–D6 were higher than that of D0 fish (p < .05), and positively correlated with dietary ALA/LA ratio, while levels of EPA showed no difference among the D1–D6 groups. These results indicated that Δ5 desaturation, required for the conversion of 20:4n‐3 to EPA, may be lacking or very low, suggesting incomplete LC‐PUFA biosynthesis ability in T. ovatus.     

n‐3 and n‐6 fatty acid bioconversion abilities in Eurasian perch (Perca fluviatilis) at two developmental stages

The aim of this work was to study the fatty acid (FA) bioconversion ability in Eurasian perch fed with diets differing in their polyunsaturated fatty acids (PUFA) from n‐3 and n‐6 series content at two development stages: adults in exogenous vitellogenesis, and juveniles during the on‐growing phase. Duplicate groups of adults and juveniles were fed for 12 weeks with four diets: D1 and D2, two diets prepared with fish oil partially or totally as the lipid source, and so containing long‐chain PUFA (LC‐PUFA). Those two diets differed by their n‐3/n‐6 FA dietary ratio (0.2 and 7.0, respectively), D1 being characterized by a high n‐6 LC‐PUFA level, while D2 had a high level of n‐3 LC‐PUFA. D3 and D4 were constituted only with vegetable oils, and were therefore devoid of LC‐PUFA. D3 was characterized by a high level of 18:2 n‐6 (n‐6/n‐3 ratio of 0.3), while D4 was characterized by a high level of 18:3 n‐3 (n‐3/n‐6 ratio of 1.9). Both groups of fish were able to elongate and desaturate the 18:3 n‐3 precursor into eicosapentaenoic acid and docosahexaenoic acid, regarding the FA profile of livers. Furthermore, total elongation/desaturation from [1‐¹⁴C]18:3 n‐3 of LC‐PUFA was higher in fish fed with the high dietary 18:3 n‐3 level compared to the diet rich in n‐3 LC‐PUFA. By opposition, the bioconversion of 18:2 n‐6 into LC‐PUFA was limited, regarding the elongation/desaturation activity of LC‐PUFA from [1‐¹⁴C]18:2 n‐6. In view of the great ability for bioconversion of n‐3 FA, linseed oil is a promising alternative to fish oil in formulating feed for juveniles perch as there were no differences in terms of specific growth rate between the treatments, but adults undergoing maturation should have at least partially LC‐PUFA in their diet, particularly arachidonic acid (ARA) which is important during maturation, as breeders are not able to bioconvert 18:2 n‐6 into ARA.     

Effects of dietary fish oil replacement by microalgae raw materials on growth performance,body composition and fatty acid profile of juvenile olive flounder,Paralichthys olivaceus

Replacing dietary fish oil with DHA‐rich microalgae Schizochytrium sp. and EPA‐rich microalgae Nannochloropsis sp. for olive flounder (Paralichthys olivaceus) was examined. Three experimental isonitrogenous and isolipidic diets with lipid source provided by 50% fish oil (F50S50), 50% (M50F25S25) and 100% microalgae raw material (M100) respectively were compared with a soybean oil (S100) diet as control. Triplicate groups of olive flounder juveniles (16.5 ± 0.91 g) were fed the experimental diets, and a group was fed the control diets for 8 weeks in a recirculation system. Results showed feed efficiency and growth performance were not significantly changed when fish oil (FO) was totally substituted by soybean oil (SO) or microalgae raw material (MRM). The whole‐body composition, lipid content of liver and muscle, and lipid composition of plasma were not significantly influenced by the total substitution of FO by MRM. The polyunsaturated fatty acids (PUFA) content of muscle and liver declined in fish fed S100 diet, whereas it was not significantly reduced in fish fed M50F25S25 and M100 diets. The total substitution of FO by MRM not only maintained the levels of arachidonic acid, EPA or DHA but also increased n‐3/n‐6 ratio. In conclusion, MRM as the sole lipid source is sufficient to obtain good feed efficiency, growth performance and human health value in olive flounder juveniles.     

Assessment of dietary lipid sources in tropical gar,Atractosteus tropicus larvae: Growth parameters and intermediary lipogenic gene expression

Five experimental diets containing different lipid sources, fish oil (D1), soybean lecithin (D2), corn oil (D3), canola oil (D4) and olive oil (D5), were evaluated in Atractosteus tropicus larvae for the relative gene expression of the enzymes fatty acid synthase (fas), acetyl‐CoA carboxylase 1 (acc1) and carnitine palmitoyltransferase 1C (cpt1c), in addition to their effects on larval growth, survival and cannibalism during a 30‐day feeding trial. Higher growth and survival were obtained in treatments D1 and D2, and lower performance in diets D3, D4 and D5. The highest levels of expression of fas and acc1 occurred in larvae fed with D1, which contained high amounts of n‐3 long‐chain polyunsaturated fatty acids (LC‐PUFA), mainly DHA and EPA FA are regulators of lipogenesis. The higher cpt1c expression in plant‐based diets is attributed to the fact that these diets are rich in α‐linolenic acid (ALA) and low DHA, EPA and ARA levels that favour ß‐oxidation. In conclusion, the diets with fish oil (D1) and soybean lecithin (D2) were the best treatments for larval growth, survival and cannibalism and thus appear to meet both lipid and energy requirements of A. tropicus larvae, meanwhile the use of vegetable oils influences the expression of intermediary lipogenic genes.     

Effects of dietary n‐3 to n‐6 fatty acid ratios on spawning performance and larval quality in tongue sole Cynoglossus semilaevis

This study investigated the effect of n‐3 to n‐6 fatty acid ratios in broodstock diets on reproduction performance, fatty acid composition of eggs and gonads of tongue sole Cynoglossus semilaevis. Broodstock were fed five isonitrogenous and isoenergetic diets for 60 days. The supplemented lipids were prepared by a combination of fish oil and soybean oil inclusion FO (fish oil); FSO1 (fish oil: soybean oil = 7:1); FSO2 (fish oil: soybean oil = 2.2:1); FSO3 (fish oil: soybean oil = 1:1); FSO4 (fish oil: soybean oil = 1:4.3) as lipid sources with different n‐3 to n‐6 fatty acid ratios 10.40, 5.21, 2.81, 1.71 and 0.87. Results showed that relative fecundity, fertilization rate and survival rate of larvae at 7 days posthatching were all higher in broodstock fed FSO1 and FSO2 diet and significantly (P < 0.05) decreased in groups fed FSO3 and FSO4 diets. The best result in starvation tolerance test was obtained in FSO2 diet. The present study suggests that n‐3 and n‐6 PUFA ratio in broodstock diet has a considerable effect on spawning performance, egg and larval quality for C. semilaevis.     

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.     

Replacement of fish oil with a DHA‐rich Schizochytrium meal on growth performance,activities of digestive enzyme and fatty acid profile of Pacific white shrimp (Litopenaeus vannamei) larvae

This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.     

Influence of 18:2n‐6/20:5n‐3 ratio in diets on growth and fatty acid composition of juvenile abalone,Haliotis discus hannai Ino

A 120‐day feeding trial was conducted to examine the effects of the ratio of dietary linoleic acid (LA, 18:2n‐6) to eicosapentaenoic acid (EPA, 20:5n‐3) on the growth and fatty acid composition of juvenile Haliotis discus hannai (initial shell length 10.23 ± 1.48 mm; initial body weight 0.13 ± 0.05 g) in a recirculation water system. Five semipurified diets with 35 g kg^?1 total lipid were formulated to contain graded LA/EPA ratios (1 : 0, 0.75 : 0.25, 0.5 : 0.5, 0.25 : 0.75, and 0 : 1, respectively). Twenty‐five juveniles were stocked in a rearing unit, a plastic basket (20 × 20 × 10 cm), as a replicate, and there were three replicates for each dietary treatment. The results showed that abalone survival rates were generally high (90.1–98.3%) and independent of the dietary treatments. However, abalone growth was significantly affected by LA/EPA ratio (P < 0.05). The LA/EPA ratio of 0.25 : 0.75 (Diet 4) produced the highest weight gain rate (WGR, 416.3%), closely followed by the ratio of 0 : 1 (Diet 5, 412.9%), the ratio of 0.5 : 0.5 (Diet 3, 399.7%) and the ratio of 0.75 : 0.25 (Diet 2, 372.1%), but no significant differences were observed among these treatments. The abalone fed the diet without 20:5n‐3 (Diet 1) had the lowest WGR (Diet 1, 363.8%), which was significantly lower than that of Diet 4. Fatty acid profiles in abalone body reflected those of dietary lipids, especially for the polyunsaturated fatty acids. The contents of arachidonic acid (AA; 20:4n‐6) in abalone tissues were positively correlated with dietary level of 18:2n‐6 (P < 0.05). Similar correlation was also observed between the level of docosahexaenoic acid (DHA, 22:6n‐3) in abalone tissues and the level of dietary EPA. It is suggested that abalone, H. discus hannai, have the capacity to synthesize 20:4n‐6 from 18:2n‐6, and maybe 22:6n‐3 from 20:5n‐3.     

Effects of Dietary DHA/EPA Ratios on Fatty Acid Composition,Lipid Metabolism‐related Enzyme Activity,and Gene Expression of Juvenile Grass Carp,Ctenopharyngodon idellus

To determine the effects of docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on grass carp, Ctenopharyngodon idellus, a 38‐d feeding trial was conducted using six isonitrogenous and isoenergetic semi‐purified diets containing constant n‐3 long‐chain polyunsaturated fatty acid (LC‐PUFA) (0.5% of dry matter), but varying ratios of DHA to EPA and a control diet (no n‐3 LC‐PUFA was included). The results revealed higher final weight and specific growth rate in the DHA/EPA 0.21 group. The n‐3 LC‐PUFA content increased in the CK (control) groups compared with that in the control diet. Lipoprotein lipase (LPL) activity increased in the treatment groups. Malate dehydrogenase showed lower activity in the DHA/EPA 1.08 group, as well as to the change in the level of glucose‐6‐phosphate dehydrogenase (G6PDH). The gene expressions of LPL increased in the treatment groups and that of peroxisome proliferator‐activated receptor α gene showed higher expressions in DHA/EPA 1.08, 0.49, and 0.21 groups. However, no remarkable differences were found among the six groups in the peroxisome proliferator‐activated receptor γ gene expression. Our findings indicated that dietary n‐3 LC‐PUFA affected fatty acid composition and lipid metabolism of grass carp. Further, fish achieved the best effect in decreasing the lipid accumulation when dietary DHA/EPA ratio was not greater than 1.     

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.     

Beef tallow is suitable as a primary lipid source in juvenile Florida pompano feeds

It is assumed that Florida pompano have dietary EPA (20:5n‐3) and DHA (22:6n‐3) requirements. However, it is unclear whether both are equally important in meeting demand for n‐3 long‐chain polyunsaturated fatty acids (LC‐PUFAs) or whether the requirement(s) can be influenced by other fatty acids. Accordingly, we assessed production performance and tissue composition of juvenile Florida pompano (41.0 ± 0.5 g) fed diets containing fish oil; beef tallow; or beef tallow partially or fully supplemented with EPA, DHA or both. After 8 weeks, no signs of fatty acid deficiency were observed. Although fish performance did not vary significantly among the dietary treatments, fish fed the DHA‐supplemented feeds exhibited numerically superior growth than those fed the other diets. Fillets of fish fed the beef tallow‐based diets contained reduced levels of n‐3 fatty acids and LC‐PUFAs and elevated levels of MUFAs and n‐6 fatty acids, although dietary supplementation with EPA and/or DHA attenuated these effects somewhat. Our results suggest that beef tallow is suitable as a primary lipid source in Florida pompano feeds and n‐3 LC‐PUFA requirements may be met by as little as 4 g/kg EPA and 4 g/kg DHA. However, there may be value in supplementing tallow‐based diets with DHA to enhance tissue levels and possibly growth.     

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.     

Effect of Dietary n‐3 highly unsaturated fatty acids on Growth,Body Composition and Fatty Acid Profiles of Juvenile Black Seabream,Acanthopagrus schlegeli (Bleeker)

An 8‐wk feeding trial was conducted to investigate the effects of dietary n‐3 highly unsaturated fatty acids (n‐3 HUFA) on growth, body composition, and fatty acid profiles of juvenile Acanthopagrus schlegeli. Three replicate groups of fish (initial mean weight: 8.08 ± 0.09 g, mean ± SD) were fed diets with different levels of n‐3 HUFA (0.76%, HUFA_0.76; 0.83%, HUFA_0.83; 0.90%, HUFA_0.90; 0.97%, HUFA_0.97; 1.04%, HUFA_1.04; 1.12%, HUFA_1.12) at 12.9% of total lipid, with a constant eicosapentaenoic acid (EPA, 20:5n‐3) to docosahexaenoic acid (DHA, 22:6n‐3) ratio of about 2.1. Hepatosomatic index (HSI) and intraperitoneal fat (IPF) ratio were all linearly depressed by dietary n‐3 HUFA levels (P < 0.05), and condition factor (CF) was not affected. Adipocyte diameter in IPF decreased with the elevation of dietary n‐3 HUFA and significance occurred between group HUFA_0.90 and HUFA_1.12. Lipid content in dorsal muscle was significantly lowered by dietary n‐3 HUFA compared to fish fed diet HUFA_0.76. No significance was found in whole fish proximate composition. In liver, dorsal muscle and IPF, ∑SFA, 16:0 or ∑n‐3 HUFA were all positively correlated with dietary n‐3 HUFA, while DHA to EPA ratios remained constant in 2.68, 2.86, and 3.60, respectively. Fatty acid synthase (FAS, EC 2.3.1.85) activities of all treatments remained constant at first and then were significantly elevated by dietary n‐3 HUFA higher than 0.97% (P < 0.05). In contrast, hormone sensitive lipase (HSL, EC 3.1.1.3) changed following an opposite tendency. Quadratic analysis based on weight gain rate (WGR) indicated that dietary n‐3 HUFA requirement for juvenile black seabream was 0.94% of the diet in 12.9% lipid diets .     

Transgenic modification of the n‐3 HUFA biosynthetic pathway in nibe croaker larvae: improved DPA (docosapentaenoic acid; 22:5n‐3) production

Marine fish are generally unable to produce sufficient quantities of n‐3 highly unsaturated fatty acid (n‐3 HUFA) such as eicosapentaenoic acid (EPA; 20:5n‐3) and docosahexaenoic acid (DHA; 22:6n‐3). Consequently, the seed production of marine fish requires careful nutritional enrichment of live feeds such as rotifers and brine shrimp Artemia to meet n‐3 HUFA requirements for normal growth. Another strategy for improving n‐3 HUFA availability is modifying the biosynthetic pathway of marine fish using transgenic technology. In this study, we conducted a feeding trial with non‐transgenic and transgenic nibe croaker Nibea mitsukurii carrying the elongation of very long‐chain fatty acids protein 2 (Elovl2) gene isolated from masu salmon Oncorhynchus masou and three groups of Artemia (non‐enriched and enriched with two products). For all Artemia groups, docosapentaenoic acid (DPA, 22:5n‐3), which is a direct product of Elovl2, was significantly higher in the transgenic fish than that in non‐transgenic fish, despite the absence of DPA in all diets. Thus, applying transgenic techniques to marine fish at the larval stage are a powerful strategy for modifying n‐3 HUFA biosynthetic pathways.     

Long‐chain PUFA enrichment in microalgae and metabolic dynamics in Tapes philippinarum larvae

The possibility of increasing n‐3 and n‐6 long‐chain polyunsaturated fatty acids (PUFA) content in microalgal mixtures used to feed Tapes philippinarum larvae was explored by lowering culture temperature from 26 to 14 °C. Although fatty acid composition of different microalgal species has a genetic basis, the algal cultures grown at 14 °C significantly increased the content of long‐chain n‐3 PUFA in Isocrysis galbana and in Thalassiosira pseudonana, while in Tetraselmis tetrathelo, the PUFA increase only involved shorter chain PUFA, namely 16:4n‐3 and 18:4n‐3. However, larvae fed on the PUFA enriched microalgal mixture did not show improvements in growth and survival performances with respect to the control group fed the microalgal mixture grown at 26 °C. From a biochemical perspective, two key aspects emerged from the results: (i) clam larvae have adequate biotransformation and selection skills to adjust fatty acid profile to their requirements as they can even modulate the incorporation of essential long‐chain PUFA as 20:5n‐3 and 22:6n‐3 when the dietary supply exceeds the physiological requirements; (ii) bivalve can biosynthesize non‐methylene‐interrupted dienoic (NMID) fatty acids as confirmed by the constancy of relative proportion with larvae growth in spite of the NMID fatty acid absence in the diet.