Shewanella putrefaciens Pdp11 probiotic supplementation as enhancer of Artemia n‐3 HUFA contents and growth performance in Senegalese sole larviculture

Adequate enrichment of live prey like Artemia, naturally deficient of essential highly unsaturated fatty acids (HUFA), such as docosahexaenoic acid (22:6n‐3, DHA), is critical for the rapidly developing tissues, survival, normal development and production of good‐quality fingerlings. The aim of the study was to evaluate the effects of a pulse (10–30 dah) of Shewanella putrefaciens Pdp11 (2.5*10⁷ cfu/ml) using Artemia metanauplii as live vector, on its proper lipid profiles and resultant Solea senegalensis body composition and performance. Probiotic administration significantly increased total lipids and specifically n‐3 HUFA levels in Pdp11‐enriched Artemia. The live prey lipid modulation was also reflected in the total lipid contents and fatty acid profiles of Pdp11 sole specimens, which achieved a higher growth performance. A fatty acid multivariate principal component analysis confirmed a neat separation of two groups corresponding to Control and probiotic fish for each age sampled (23, 56, 87 and 119 dah). In addition, a further SIMPER analysis highlighted that the Pdp11 Artemia effect on sole lipid profile was different for each fatty acid and was gradually diluted with age. Results suggest an ability of Pdp11 strain to produce n‐3 HUFA as an effective tool for fish marine larviculture optimization.     

Effects of enriched Artemia by fish and soybean oils supplemented with vitamin E on growth performance,lipid peroxidation,lipase activity and fatty acid composition of Persian sturgeon (Acipenser persicus) larvae

This study aimed to evaluate the effects of enriched Artemia by fish and soybean oils supplemented with vitamin E on growth performance, lipid peroxidation, lipase activity and fatty acid composition of Persian sturgeon (Acipenser persicus) larvae. For this purpose, five experimental diets including non‐enriched Artemia (control diet), Artemia enriched with soybean oil supplemented with 15% and 30% vitamin E (S15 and S30 diets) and fish oil supplemented with 15% and 30% vitamin E (F15 and F30 diets) were used. The larvae were fed to apparent satiation four times per day for 22 days. The results indicated that fish fed enriched Artemia had no significant differences compared with those fed non‐enriched Artemia in terms of growth and survival, but increase in vitamin E levels from 15 to 30% improved growth performance of larvae. Vitamin E content in fish fed S15 and S30 diets was significantly higher. Fish fed non‐enriched Artemia had significantly higher thiobarbituric acid and lower lipase activity. The highest HUFA and n‐3/n‐6 ratio were observed in fish fed F15 and F30 diets. Our results demonstrated that fish oil can completely replace with soybean oil in larval diets. Therefore, using S30 diet is recommended for feeding of Persian sturgeon larvae.     

Effect of dietary n‐3 HUFA on growth performance and tissue fatty acid composition of gibel carp Carassius auratus gibelio

A 12‐week growth trial was conducted with gibel carp Carassius auratus gibelio (initial weight: 2.69 g) to evaluate the effects of dietary n‐3 highly unsaturated fatty acids (n‐3 HUFA) on growth performance and tissue fatty acid composition. Five diets of different n‐3 HUFA levels from 0 to 17 g kg^?1 diet were supplemented at 80 g kg^?1 dietary lipid by including fish oil (FO) at 0, 25, 50, 75 and 100% of supplemental lipid. The remainder was coconut oil. The results showed that fish fed FO₂₅ and FO₅₀ obtained highest specific growth rate and lowest with FO₀. Feed efficiency was highest at FO₁₀₀ and lowest at FO₀. Apparent digestibility coefficient of lipid increased with increasing dietary n‐3 HUFA. The fish fed FO₀ diet had the lowest thiobarbituric acid reactive substance in serum and muscle and highest moisture and lowest lipid content in viscera. Fatty acid compositions of muscle and liver were correlated with dietary fatty acids. Fish muscle concentration of 20:5n‐3 increased with increasing dietary n‐3 HUFA while the concentration of 22:6n‐3 was distinctly reduced in FO₀ group. It suggested that 4 g kg^?1 n‐3 HUFA in diet could permit gibel carp normal growth performance and provide considerable n‐3 HUFA in fish muscle. Excessive n‐3 HUFA showed impact on growth performance of gibel carp.     

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.     

The effect of non‐marine HUFA supplementation with fish oil removal on growth and survival of the Pacific white shrimp,Litopenaeus vannamei

The use of non‐marine arachidonic acid (ArA) and docosahexaenoic acid (DHA) as highly unsaturated fatty acid (HUFA) enrichments was evaluated as complete replacements for marine fish oil in practical diets formulated with solvent‐extracted soybean meal (SESM). Litopenaeus vannamei juveniles (0.59 g) were reared over 84 days in an outdoor tank system with no water discharge. Fishmeal was replaced with SESM, while fish oil was replaced with HUFA‐rich algal cells, alternative oil and/or fermentation products. Spray‐dried Schizochytrium algal cells (Schizomeal‐Hi DHA) served as the DHA enrichment source. Oil extracted from Mortierella sp. was used as the ArA enrichment (AquaGrow^® ArA). DHA and ArA sources (Advanced BioNutrition Corp., Columbia, MD, USA) were non‐marine products obtained from a commercial supplier. Five diets were formulated with ArA inclusion levels of 0, 0.65, 1.3, 2.6 and 5.2 g kg^?1. In addition, one diet was formulated to be DHA deficient and another was formulated with menhaden fish oil (control). Different inclusion levels of non‐marine ArA had no effect on survival or growth. Shrimp fed the non‐marine HUFA‐supplemented diets had lower average weight compared to shrimp offered the diet containing fish oil. No differences were detected in average weights of shrimp offered the ArA‐deficient and ArA‐supplemented diets.     

Siberian sturgeon (Acipenser baerii) larvae fed Artemia nauplii enriched with poly‐β‐hydroxybutyrate (PHB): effect on growth performance,body composition,digestive enzymes,gut microbial community,gut histology and stress tests

The effects of feeding Artemia nauplii enriched with or without poly‐β‐hydroxybutyrate (PHB) and/or highly unsaturated fatty acids (HUFA) on Siberian sturgeon (Acipenser baerii) larvae were investigated. Feeding larvae with PHB‐enriched nauplii (PHB, PHB + HUFA) decreased the growth performance of the larvae. PHB affected the body composition by increasing the lipid content of the whole body and decreasing total saturated, monoenoic, n3, n6 and decosahexanoeic acid (DHA) in the larvae, indicating that the PHB addition affected lipid metabolism. A high activity of pepsin was observed in the digestive extracts of PHB treatments (PHB, PHB + HUFA), while PHB suppressed amylase activity in the intestine of the larvae. Based on molecular analysis, PHB changed the microbial community in the distal intestine of the larvae. The highest counts of goblet cells were observed in the HUFA‐containing treatments (HUFA, PHB + HUFA), indicating that HUFA addition may improve the mucosal barrier defence system. The overall quality of the larvae was evaluated by exposing them to different salinities and ammonia stress levels. PHB decreased survival rates in these challenges. Our results show that optimal PHB doses for bio‐encapsulation into Artemia remain to be determined for further application at the earliest larval stages of sturgeon.     

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 .     

Effects of dietary n‐3HUFA on different growth stages of the white shrimp,Litopenaeus vannamei: Growth,haematological characteristics,enzyme activities and fatty acid profiles

This study investigated the effects of n‐3 high unsaturated fatty acid (n‐3HUFA) levels on the growth performance, antioxidant enzyme activities and fatty acid profiles of both subadult and adult Litopenaeus vannamei (L. vannamei). Seven iso‐nitrogenous and iso‐lipidic diets were used, containing n‐3HUFA concentrations of 1.6 (control), 4.8, 7.4, 13.9, 23.9, 29.2 and 34.4 g/kg, respectively. Two 8‐week feeding trials were conducted to determine the dietary n‐3HUFA requirements of L. vannamei with an initial body weight of 4.25 ± 0.00 g (subadults) and 8.50 ± 0.01 g (adults). The results showed that the dietary n‐3HUFA level significantly affected the weight gain rate (WGR), specific growth rate, the feed conversion ratio and the hepatosomatic index (HSI) (p < 0.05), but did not significantly affect the survival rate (p > 0.05). At appropriate level, dietary n‐3HUFA improved growth performance and HSI of both subadult and adult L. vannamei. Both subadults and adults showed significant differences in body composition (p < 0.05), except for moisture and crude ash (p > 0.05). Cholesterol and low‐density lipoprotein significantly decreased with increasing dietary n‐3HUFA both in subadults and adults (p < 0.05); however, triglyceride showed no significant change (p > 0.05). High‐density lipoprotein (HDL) in subadults was significantly affected by dietary n‐3HUFA (p < 0.05), but followed no apparent regularity; HDL significantly changed in adults and showed an upward trend followed by a downward trend (p < 0.05). There was no significant effect on aspartate transaminase (AST) activity in subadults, but AST in adults and alanine transaminase (ALT) in subadults and adults were significantly affected (p < 0.05). Dietary n‐3HUFA significantly affected serum polyphend oxidase, malic dehydrogenase, alkaline phosphatase, superoxide dismutase and sodium‐potassium adenosine triphosphatase enzyme activities in gills (p < 0.05). The fatty acid composition of the shrimp tissue was associated with the fatty acid composition of the diet. Dietary n‐3HUFA supplementation significantly improved the contents of tissue ∑HUFA and n‐3HUFA, increased the n‐3/n‐6 ratio in the tail muscle and decreased the contents of tissue polyunsaturated fatty acid and saturated fatty acid (p < 0.05). Based on the WGR, the broken‐line equations indicated that the optimum requirements of dietary n‐3HUFA were determined to be 9.0 and 5.1 g/kg for subadult and adult L. vannamei, respectively.     

Effects of dietary n‐3HUFA on juvenile white shrimp,Litopenaeus vannamei: Growth,feed utilization,antioxidant enzymes activities and fatty acid compositions

Present study investigates the effects of n‐3 high‐unsaturated fatty acid (n‐3HUFA) levels on growth performance, antioxidant enzymes activities and fatty acid compositions of juvenile Litopenaeus vannamei. These represented seven iso‐nitrogenous and iso‐lipidic diets. Analysed n‐3HUFA concentrations were 0.16% (control), 0.48%, 0.74%, 1.39%, 2.39%, 2.92% and 3.44% respectively. A total of 840 juvenile L. vannamei were randomly stocked into 21 0.5 m³ tanks for 56 days. A significant increase (p < 0.05) was observed from 0.16% to 0.74% n‐3HUFA and a decrease when n‐3HUFA was above these levels in weight gain rate (WGR) and specific growth rate. Total cholesterol, triglyceride and low‐density lipoprotein in serum showed a significant decrease, high‐density lipoprotein showed a significant increase (p < 0.05). Phenoloxidase activity in serum and sodium‐potassium adenosine triphosphatase activity in gill were significantly affected by dietary n‐3HUFA (p < 0.05), both of them showing a downward trend after upward. Malic dehydrogenase and superoxide dismutase activities in serum were also significantly affected by dietary n‐3HUFA (p < 0.05), which rose first and then decreased in general, both of them have a maximum in 2.39% group. No significant differences of the activities of aspartate transaminase and alanine transaminase were observed among all groups (p > 0.05). With dietary n‐3HUFA increase, both ∑HUFA and n‐3HUFA contents gradually increased in hepatopancreas and tail muscles (p < 0.05). Based on broken‐line regression analysis of WGR, the optimal n‐3HUFA requirement is 0.89% for juvenile L. vannamei with initial weight of 0.50 ± 0.01 g.     

Investigation of highly unsaturated fatty acid metabolism in the Asian sea bass, <Emphasis Type="Italic">Lates calcarifer</Emphasis>

Lates calcarifer, commonly known as the Asian sea bass or barramundi, is an interesting species that has great aquaculture potential in Asia including Malaysia and also Australia. We have investigated essential fatty acid metabolism in this species, focusing on the endogenous highly unsaturated fatty acid (HUFA) synthesis pathway using both biochemical and molecular biological approaches. Fatty acyl desaturase (Fad) and elongase (Elovl) cDNAs were cloned and functional characterization identified them as ∆6 Fad and Elovl5 elongase enzymes, respectively. The ∆6 Fad was equally active toward 18:3n-3 and 18:2n-6, and Elovl5 exhibited elongation activity for C_18–20 and C_20–22 elongation and a trace of C_22–24 activity. The tissue profile of gene expression for ∆6 fad and elovl5 genes, showed brain to have the highest expression of both genes compared to all other tissues. The results of tissue fatty acid analysis showed that the brain contained more docosahexaenoic acid (DHA, 22:6n-3) than flesh, liver and intestine. The HUFA synthesis activity in isolated hepatocytes and enterocytes using [1-¹⁴C]18:3n-3 as substrate was very low with the only desaturated product detected being 18:4n-3. These findings indicate that L. calcarifer display an essential fatty acid pattern similar to other marine fish in that they appear unable to synthesize HUFA from C₁₈ substrates. High expression of ∆6 fad and elovl5 genes in brain may indicate a role for these enzymes in maintaining high DHA levels in neural tissues through conversion of 20:5n-3.     

Complete repertoire of long‐chain polyunsaturated fatty acids biosynthesis enzymes in a cyprinid,silver barb (Barbonymus gonionotus): Cloning,functional characterization and dietary regulation of Elovl2 and Elovl4

The silver barb (Barbonymus gonionotus) is a valuable cyprinid with good deposition of long‐chain polyunsaturated fatty acids (LC‐PUFA) such as docosahexaenoic acid (DHA). We have previously isolated a fatty acyl desaturase (Fads2) and elongase (Elovl5), which fulfils the production of eicosapentaenoic acid (EPA) and arachidonic acid (ARA) from C18 polyunsaturated fatty acid (PUFA) substrates. The isolation, functional characterization and dietary regulation of two elongases, Elovl2 and Elovl4, are reported here. The Elovl2 displayed the capacity to elongate C20 and C22 PUFA substrates, while the showed low activities towards saturated fatty acids and C22 PUFA substrates. This discovery validates the existence of a complete set of enzymes of LC‐PUFA in silver barb. The elovl2 showed higher expression in liver, in limited dietary LC‐PUFA intake conditions. As for Elovl4, tissue expression showed prominence in eye tissue. We also showed that the expression of both genes was upregulated when fish was fed with diets devoid of LC‐PUFA. Fatty acid composition analysis indicates the utilization of C18 polyunsaturated fatty acids for LC‐PUFA biosynthesis.     

Prey–predator dynamics in seahorses (Hippocampus guttulatus): deciphering fatty acid clues

Understanding the flow of fatty acids between trophic levels can provide important clues on prey–predator dynamics and nutritional requirements of the species. This study investigates the fatty acid flow between enrichment emulsions, Artemia nauplii and Hippocampus guttulatus juveniles, and evaluates the nutritional value of enriched and unenriched Artemia for newborn seahorses. The fatty acid profile of Artemia and seahorses generally reflected the dietary composition, but fatty acids were not linearly transferred between trophic levels. The incorporation of dietary fatty acids showed to be a more complex process involving dietary composition, predator metabolism and nutritional requirements. Artemia composition resulted from a dynamic balance between what was assimilated and metabolized by the nauplii during enrichment. Prey fatty acids were incorporated in seahorses, but HUFA, particularly DHA, were selectively retained to fulfil their high requirements. H. guttulatus newborns were not successfully reared on Artemia nauplii, not even on enriched Artemia, with low survival rates (15.0–26.7%) being observed in all feeding treatments. The high MUFA content and low DHA level of Artemia did not fulfil the high SFA and PUFA requirements of newborn juveniles, particularly their great DHA demands. Higher survivorship was obtained with enriched Artemia, but no differences were detected in juvenile growth.     

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.     

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.     

A Comparison of Growth,Survival, and Fatty Acid Composition of the Lined Seahorse,Hippocampus erectus,Juveniles Fed Enriched Artemia and a Calanoid Copepod,Schmackeria dubia

The purpose of this study was to evaluate the effect of varying dietary levels of highly unsaturated fatty acids (HUFAs) in live prey (Artemia nauplii and a calanoid copepod, Schmackeria dubia) on the growth performance, survival, and fatty acid composition of the lined seahorse, Hippocampus erectus, juveniles. Artemia nauplii were enriched with a commercial product (SS^? 50DE‐microcapsule as HUFA source, 2/3 DHA, 1/3 EPA. Shengsuo Fishery Feed Research Center of Shandong Province, Qingdao, China) at four concentrations of 0.0, 14.0, 28.0, and 56.0. Newly hatched juveniles were cultured for 35 days. The content of docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and n‐3 HUFAs in the Artemia nauplii was positively related to the enrichment concentration. At the end of the trials, growth performance of the juveniles was positively related to the enrichment concentration as well. However, the juveniles fed prey enriched with the highest concentration of enrichment (56.0 μL/L) had the significantly lower (P < 0.05) survival rate. The juveniles fed the copepod had the best growth performance and the highest survival rate, suggesting that the copepod, S. dubia, is suitable for feeding the seahorse juveniles. The comparisons between the growth, survival, and fatty acid profiles of the juveniles fed Artemia and copepods indicate that the seahorse juveniles require dietary levels of DHA beyond those achieved by enriching prey with the HUFA enrichment. Surplus EPA resulted from an imbalance between DHA and EPA in the enriched Artemia nauplii probably caused an adverse effect on the seahorse juveniles. This study suggests that DHA and EPA requirement of the lined seahorse juveniles is roughly 32% of total fatty acid, and the optimal DHA/EPA ratio for the species is circa 4:1. To avoid an adverse effect resulting from excessive EPA, maximum proportion of EPA in enriched Artemia nauplii should not exceed 13% of total fatty acid, and a recommended minimum DHA/EPA ratio in the enriched Artemia nauplii is 1.46. Arachidonic acid (20:4n‐6) might not be an essential fatty acid for the seahorse juveniles.     

First step of non‐fish meal,non‐fish oil diet development for red seabream, (Pagrus major), with plant protein sources and microalgae Schizochytrium sp

A feeding experiment was conducted to develop non‐fish meal and non‐fish oil diet for red seabream by using plant protein source and Schizochytrium meal which is rich in 22:6n‐3 (DHA). Three iso‐nitrogenous and iso‐lipidic experimental diets were prepared (CP 41.2% ± 0.4%, CL 16.4% ± 1%). Control diet contained both fish meal (40%) and fish oil (6%). In the second diet, fish meal was replaced by plant meals (soy protein concentrate, soybean meal, corn gluten meal) [FO]. In the third diet, fish meal and fish oil were replaced by algae meal (Schizochytrium sp. powder) and plant proteins [AO]. Duplicated groups of juvenile red seabream (8.8 g ± 1.5) were fed the experimental diets for 12 weeks to near satiation. There was no statistical difference among treatment in specific growth rate. Feed conversion ratio of AO diet group was higher than that of control. In wet basis, whole body protein level was significantly higher in AO diet than FO group while lipid content was lower than control group. In fatty acid profile, AO group had significantly lower 18:4n‐3, 20:4n‐3, 22:5n‐3 and 20:5n‐3 (EPA) level, but significantly higher 18:3n‐3 and DHA level than the other two diet fed fish. The results might suggest that further developments in microalgae diet offer a promising lipid source of n‐3 PUFA as essential fatty acid on marine fish. And it showed possibility to develop non‐fish meal and non‐fish oil feed for marine aquaculture fish by using microalgae.     

The effect of soybean oil containing stearidonic acid on growth performance,n‐3 fatty acid deposition and sensory characteristics of pacific white shrimp (Litopenaeus vannamei)

The ability of shrimp Litopenaeus vannamei to utilize soy oil (SO) modified to contain stearidonic acid (SDA) in replacement of fish oil (FO) by converting SDA to highly unsaturated fatty acids (HUFA) was examined. Six diets with either supplemental modified SO or FO and three levels of fishmeal (FM) replacement (0%, 50% and 100%) by soybean meal (SBM) were fed to shrimp (1.7 g) for 12 weeks. The effect of oil source at the three SBM levels on growth and fatty acid profiles was examined by contrast analysis and sensory attributes by t‐tests (5% error rate). At 0% SBM inclusion, there was no effect of dietary oil source, while at the highest SBM inclusion level, shrimp fed the FO diet outperformed those fed the corresponding SO diet. Oil source had no effect on sensory attributes. The fatty acid profiles of the shrimp reflected that of the diets. SDA SO can replace supplemental FO in diets for shrimp with no reduction in growth when there is sufficient oil present from FM. At low FM, however, replacing FO with SDA SO reduces shrimp performance and tissue n‐3 HUFA levels. It is concluded that SDA is unable to meet the essential fatty acid needs of shrimp.     

Effects of dietary n‐3 PUFA supplements on composition of n‐3 PUFA and expression of fatty acid elongase 5 (AJELOVL5) in sea cucumber,Apostichopus japonicus

This is the first comprehensive study on the effect of dietary polyunsaturated fatty acid (PUFA) levels on the expression of fatty acid elongase 5 (AJELOVL5), PUFA composition, and growth in juvenile sea cucumbers. The specific growth rate (SGR_w) was improved in n‐3 PUFA‐rich diets compared to low n‐3 PUFA diets. AJELOVL5 expression was apparently upregulated in juveniles fed lower PUFA diets relative to higher PUFA diets, with higher expression in the body wall and respiratory tree of juveniles fed diets without ɑ‐linolenic acid (ALA, 18:3n‐3) compared to juveniles fed higher ALA level diets; similar results were also detected in juveniles fed diets with lower eicosapentaenoic acid (EPA, 20:5n‐3), docosahexaenoic acid (DHA, 22:6n‐3), and none of ALA, EPA, or DHA respectively. The concentrations of ALA, EPA, and DHA in tissues were positively related to the content of dietary corresponding PUFA, with higher ALA content in juveniles fed diet ALA_12.71 than in the ALA_7.46 and ALA₀ groups. Similar results were also obtained in sea cucumber fed diets enriched with either EPA or DHA. Interestingly, considerable levels of EPA and DHA were found in the tissues of juveniles fed diets of CK₀ and DHA₀, with no specific input of EPA or DHA, showing that the sea cucumber was capable of biosynthesizing EPA and DHA from their corresponding precursors as ALA and linoleic acid (LA, 18:2n‐6).     

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.