Effects of dietary linolenic acid on growth,fatty acid composition,immune function and antioxidant status of juvenile blunt snout bream,Megalobrama amblycephala

A nine‐week feeding trial was performed to determine the dietary linolenic acid (LNA; 18:3n–3) requirements of juvenile blunt snout bream. Six iso‐nitrogenous, semi‐purified diets were prepared with different concentrations of LNA (0–25 g/kg). Dietary LNA had no significant effects on survival rate. However, final fish weight, weight gain (WG), specific growth rate (SGR) and feed efficiency ratio (FER) increased with increasing dietary LNA concentrations up to 20 g/kg. Dietary LNA increased muscle LNA and total n‐3 polyunsaturated fatty acid (PUFA) contents, but decreased total saturated fatty acid content. Fish fed 20 g/kg LNA had the highest plasma alkaline phosphatase activity, total protein, albumin and white blood cell count levels. Additionally, fish fed 20 g/kg LNA had higher triglyceride levels than control fish. Plasma glucose increased with increasing dietary LNA concentrations. Superoxide dismutase and glutathione peroxidase activities significantly increased with increasing dietary LNA concentrations up to 15 g/kg. Based on SGR and FER, the optimal dietary LNA requirements of juvenile blunt snout bream were 17.5 and 15.6 g/kg respectively.     

Molecular cloning,characterization and expression analysis of the fatty acid‐binding protein (MnFABP), involved in dietary lipid sources response in oriental river prawn,Macrobrachium nipponense

A fatty acid‐binding protein (FABP) gene designated as MnFABP10 was cloned and characterized from the freshwater prawn Macrobrachium nipponense. The full‐length cDNA of MnFABP10 was 646 bp encoding a 130 amino acid. Real‐time quantitative RT‐PCR showed that the MnFABP10 gene was expressed in various tissues with the highest expression in the hepatopancreas. The MnFABP10 mRNA levels in the hepatopancreas and ovary of M. nipponense were dependent on the stages of ovarian development. Western blot results revealed a single immunoreactive band with an estimated molecular mass of approximate 14 kDa in the developmental ovary. Then, M. nipponense with an initial body weight of 0.090 ± 0.0010 g were fed with four isonitrogenous and isocaloric diets with different oils, that is, beef tallow (BT), soybean oil (SO), pollack fish oil (FO) and a mixture of fish oil and soybean oil (FO/SO 2 : 1 w/w) for 52 days. The mRNA levels of MnFABP10 in the hepatopancreas were influenced by different lipid sources, with a peak expression observed in prawns fed SO. This study suggests that MnFABP10 may have a putative function in ovary maturation, and its mRNA expression in the hepatopancreas can be regulated by the source of dietary lipids in M. nipponense.     

Effects of dietary supplementation with coconut oil on the growth,fatty acid profiles and some lipid metabolism relative gene expressions of orange‐spotted grouper Epinephelus coioides

This study investigated the effects of coconut oil as a dietary supplement on the growth, lipid metabolism and related gene expressions of juvenile orange‐spotted grouper Epinephelus coioides. Coconut oil at concentrations of 0, 10, 30 and 50 g/kg was used to replace dietary lipids in a basal diet containing 150 g/kg lipids. The four experimental diets were, respectively, fed to triplicate groups of juvenile groupers (initial weight: 8.53 ± 0.13 g) in a recirculating system for 8 weeks. Fish fed the diet containing 50 g/kg coconut oil exhibited lower (p < .05) weight gain than did fish fed the diet containing 30 g/kg coconut oil; however, no significant differences in weight gain were observed between fish fed diets containing 0 and 10 g/kg coconut oil. Hepatic carnitine palmitoyltransferase‐1, fatty acid synthase, fatty acid elongase, fatty acid desaturase and peroxisome proliferator‐activated receptor gamma gene expressions were all the highest in fish fed the diet containing 10 g/kg coconut oil. Fish fed the coconut oil‐free basal diet demonstrated upregulated gene expression of neuropeptide Y. The results suggest that dietary supplementation with 10 g/kg coconut oil exerted beneficial effects on lipid metabolism by E. coioides.     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.     

Effects of dietary docosahexaenoic acid on growth performance,fatty acid profile and lipogenesis of blunt snout bream (Megalobrama amblycephala)

Six diets were designed to investigate the effects of dietary docosahexaenoic acid (22:6n‐3; DHA) levels (0.5, 1.3, 2.3, 4.2, 8.1 and 15.9 g/kg diets) on growth performance, fatty acid profile and expression of some lipogenesis‐related genes of blunt snout bream (Megalobrama amblycephala). Fish (average weight: 26.40 ± 0.11 g) were randomly fed one of six diets for 8 weeks. Results indicated that the final body weight (FBW) and specific growth rate (SGR) of fish fed 1.3 g/kg DHA were significantly higher than other groups except for the 2.3 g/kg DHA (p < .05). Compared with other groups, the number of lipid droplet clusters of the liver stained with oil red O in the 2.3 g/kg DHA group was the highest, which was consistent with the lipid contents of whole body and liver. The DHA proportion in liver and muscle significantly increased with the increasing dietary DHA levels (p < .05), which reflected fatty acid profiles of diets. The highest mRNA expressions of acetyl‐CoA carboxylase α (ACCα), fatty acid synthase (FAS) and sterol regulatory element‐binding protein‐1 (SREBP‐1) occurred in the 1.3 g/kg DHA group, followed by 2.3 g/kg DHA. In summary, the supplementation of 1.3–2.3 g/kg DHA could improve growth performance and lipogenesis, and the dietary DHA could improve DHA and PUFA proportion in liver and muscle.     

Influence of dietary linoleic acid (18:2n‐6) and α‐linolenic acid (18:3n‐3) ratio on fatty acid composition of different tissues in freshwater fish Songpu mirror carp,Cyprinus Carpio

Six isonitrogenous and isoenergetic purified diets were formulated to feed Songpu mirror carp for 60 days. The control diet (CD) was only supplemented with soybean oil. The other five experimental diets contained soybean oil, linseed oil and lard oil blended at various inclusion levels to attain different linoleic acid (LA)/α‐linolenic acid (LNA) ratios (0.53, 1.04, 2.09, 3.95, 6.82) with a constant total C₁₈ polyunsaturated fatty acids (PUFA; LA+LNA, 2% dry weight) content. The fatty acid (FA) profiles of hepatopancreas, dorsal muscle, intestine, intraperitoneal fat (IPF), spleen and kidney reflected those of the diets, but with some differences. The spleen showed the lowest correlation with diet compared with other tissues, followed by the hepatopancreas (P < 0.05). The intestine and IPF showed relatively higher correlation. On the other hand, the control group had the lowest tissue‐diet correlation, followed by the LA/LNA0.53 group (P < 0.05), whereas the LA/LNA2.09 showed the highest. The LA/LNA ratios in the tissues were up‐regulated in the LA/LNA0.53, 1.04 groups and down‐regulated in the LA/LNA3.95, 6.82 groups. This was due to when LA (or LNA) was highly added in diet, the decrease in this FA was huge in tissue. The contents of saturated fatty acids and monounsaturated fatty acids increased in the control group, but seemed not influenced by dietary LA/LNA ratios. These results demonstrated that the FA deposition was tissue‐specific, and also influenced by the dietary FA composition in the experimental fish. Finally, we suggest that 2.09 is the optimal LA/LNA ratio (2% C₁₈ PUFA) of Songpu mirror carp for fillet FA composition.     

Effects of dietary alpha‐linolenic acids on growth performance,lipid metabolism and antioxidant responses of juvenile Russian sturgeon Acipenser gueldenstaedtii

Five isonitrogenous diets were formulated with graded alpha‐linolenic acid (LNA) levels (0, 5, 10, 15 and 20 g/kg) to investigate LNA requirement of juvenile Russian sturgeon Acipenser gueldenstaedtii. Weight gain and specific growth rate of fish fed LNA5 and LNA10 were significantly higher than those in other groups, while the feed conversion ratio of these two groups was lower than others. Dietary LNA increased n‐3 polyunsaturated fatty acid and n‐3/n‐6 ratio, but decreased saturated fatty acid contents in the liver. DHA in the fish tissue also increased with the increased dietary LNA. The superoxide dismutase activity was highest in fish fed LNA5. Fish fed LNA10 showed the highest catalase activity and the highest malondialdehyde content. A 459‐bp fragment of Δ6 fatty acid desaturase and a 474‐bp fragment of elongases of very long chain fatty acids 5 were cloned and analysed. The expressions of these two genes were higher in fish fed LNA15 and LNA20. The highest hepatic lipase activity occurred in fish fed LNA 20, and the malate dehydrogenase activity peaked in the LNA5 group. Based on SGR and FCR, the range of optimum dietary LNA concentration for juvenile Russian sturgeon is recommended at 6.85–10.69 g/kg.     

Effect of dietary vitamin E on growth,immunity and regulation of hepatopancreas nutrition in male oriental river prawn,Macrobrachium nipponense

We evaluated the effects of dietary vitamin E on growth, immunity and regulation of the hepatopancreas in male oriental river prawn, Macrobrachium nipponense. Shrimps were fed 0, 40, 80, 160, 320 or 640 mg vitamin E/kg for 60 days. The 80 mg/kg group had the highest weight gain rate, specific growth rate and lowest feed conversion rate while there were no significant differences in survival rate and hepatosomatic index. The highest crude fat and lowest crude protein content were observed in the 160 mg/kg group. In the hepatopancreas, lysozyme, acid phosphatase and alkaline phosphatase activities were highest in the 160 mg/kg group, while superoxide dismutase, catalase and glutathione peroxidase activities decreased with increasing vitamin E levels. Malondialdehyde content initially decreased then increased with vitamin E levels, whereas the reverse was seen with total antioxidant capacity. Linoleic acid, DPA, DHA, total n‐3 polyunsaturated fatty acid and total polyunsaturated fatty acid first increased then decreased, while EPA and total saturated fatty acid rose with vitamin E levels. Total n‐6 polyunsaturated fatty acid content declined while there were no significant differences in linolenic and total monounsaturated fatty acid content. Following a toxicity test with Aeromonas hydrophila, hepatopancreas ultrastructure revealed that appropriate vitamin E levels promote an increase in mitochondria, endoplasmic reticulum and Golgi bodies, but excess vitamin E can damage cell structure. These results provide evidence that 80–160 mg/kg dietary vitamin E has a positive impact on growth, immunity and regulation of the hepatopancreas in male shrimp.     

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.     

Optimal dietary alpha‐linolenic acid/linoleic acid ratio improved digestive and absorptive capacities and target of rapamycin gene expression of juvenile grass carp (Ctenopharyngodon idellus)

Growth performance, digestive and absorptive capacities and target of rapamycin (TOR), ribosomal protein S6 kinase 1 (S6K1) and eIF4E‐binding protein (4E‐BP) gene expression in the hepatopancreas and intestine of juvenile grass carp (Ctenopharyngodon idellus) fed graded ratios of dietary alpha‐linolenic acid/linoleic acid (ALA/LNA) (0.01, 0.34, 0.68, 1.03, 1.41, 1.76 and 2.15) for 60 days were investigated. The results showed that ALA/LNA ratio of 1.03 significantly improved (i) per cent weight gain (PWG) and feed efficiency, (ii) hepatopancreatic trypsin, chymotrypsin, lipase, amylase and intestinal creatine kinase (CK) activities, (iii) hepatopancreatic trypsinogen‐2 and chymotrypsinogen mRNA levels. Meanwhile, fish fed with ALA/LNA ratio of 0.68 significantly enhanced, (iv) Na⁺/K⁺‐ATPase and γ‐glutamyl transpeptidase activities in whole intestine, and alkaline phosphatase activities in the proximal intestine (PI) and distal intestine, (v) amylase, intestinal Na⁺/K⁺‐ATPase alpha‐subunit isoform 1, Na⁺/K⁺‐ATPase alpha‐subunit isoform 8 and CK mRNA abundances, (vi) TOR and S6K1 gene expression in the hepatopancreas and intestine of juvenile grass carp. Based on the quadratic regression analysis of PWG, cholecystokinin and leptin contents in the PI, optimal dietary ALA/LNA ratio of juvenile grass carp (8.78–72.00 g) was estimated to be 1.08, 1.19 and 1.05, respectively.     

Effects of salinity acclimation on the growth performance,osmoregulation and energy metabolism of the oriental river prawn,Macrobrachium nipponense (De Haan)

In this study, we evaluated the growth, osmoregulation and energy metabolism of the oriental river prawn, Macrobrachium nipponense, reared during 6 weeks with different salinities (0, 8, 14 and 22 g/L). The results showed that the haemolymph osmolality of M. nipponense increased with an increase in ambient osmotic pressure; the isosmotic point was 490 mOs/kg H₂O. The prawns showed a higher survival rate, weight gain rate and hepatopancreas index in salinity 14 g/L. Digestive enzymes were all affected by salinity, and the highest activities were observed in the salinity 14 g/L. The mRNA expression of Na⁺‐K⁺‐ATPase in gills and p53 in hepatopancreas were the highest in salinity 22 g/L. The expressions of heat shock protein 90 and glutathione S‐transferase genes in hepatopancreas were significantly higher in the salinity 8 g/L. Lipid metabolism‐related genes in hepatopancreas were significantly expressed in the salinity 14 g/L. The glucose‐6‐phosphatase gene in hepatopancreas was highly expressed in the salinity 8 and 22 g/L, and the expression of the ecdysone receptor gene in hepatopancreas was significantly higher in the salinity 14 g/L. The results showed that salinity 14 g/L could promote the growth of M. nipponense. However, higher salinity conditions may cause physiological damage, which provides a theoretical basis for brackish water culture of M. nipponense.     

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.     

Effects of dietary lipid source on fatty acid composition,expression of genes involved in lipid metabolism and antioxidant status of grass carp (Ctenopharyngodon idellus)

This study determined the effect of different lipid sources on growth, feed use, lipid metabolism and antioxidant status of grass carp (Ctenopharyngodon idellus). Juvenile fish (56.9 ± 4.7 g) were divided into four triplicate groups and fed diets containing 30 g/Kg of fish oil (FO), olive oil (OO), peanut oil (PO) and linseed oil (LO), respectively, for 60 days. Weight gain and feed conversion ratio were not significantly different between the dietary groups, but we observed changes in the fatty acid composition of muscle and intraperitoneal fat reflecting the fatty acid profile of the dietary lipid source. In the hepatopancreas, the highest mRNA level of fatty acid translocase CD36 (FAT/CD36) and carnitine palmitoyl transferase (CPT‐1A) was both observed in the FO group. In muscle, the expression of FAT/CD36 and CPT‐1A in the LO group was significantly higher than that in other groups, except for CPT‐1A in the PO group. In addition, the lowest and highest content of malondialdehyde in serum was observed in OO and FO groups, respectively. In summary, dietary lipid source altered the fatty acid composition, potential uptake (FAT/CD36) and oxidation (CPT‐1A) of fatty acids, and antioxidant status of grass carp, which should be considered when selecting a lipid source.     

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.     

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.     

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.     

Effects of different dietary lipid sources on tissue fatty acid composition,serum biochemical parameters and fatty acid synthase of juvenile mud crab Scylla paramamosain (Estampador 1949)

A 6‐week study was conducted to determine the effects of different lipid sources in pelleted diets on juvenile mud crab Scylla paramamosain. Five isonitrogenous and isolipidic diets containing 8% level of fish oil (FO), lard (LD), safflower oil (SO), perilla seed oil (PO) or mixture oil (MO; V_FO:V_SO:V_PO = 1:1:1), and a live food of marine bivalve Potamocorbula rubromuscula as the control diet (CF), were fed to groups of 25 juvenile crabs (average initial weight 7.4 g, carapace width 3.5 cm) in triplicate. The results showed that crabs fed MO had the highest survival (P < 0.05). The moisture content was significantly higher in crabs fed LD, SO and PO (P < 0.05). Crabs fed SO exhibited the lowest crude protein and lipid (P < 0.05). Ash contents were obviously lower in LD group (P < 0.05). Highest total lipid in the hepatopancreas and muscle was in LD and FO group respectively. Glucose, total cholesterol and low‐density lipoprotein were higher while high‐density lipoprotein was lower (P < 0.05) in LD group. Tissue fatty acid compositions were consistent with those in diets. FO and MO diets had the same depression effect like CF on fatty acid synthase activity and mRNA expression in the hepatopancreas. The results of this study indicated that FO and mixed oil are suitable for preparation of pelleted diets with better effects for juvenile S. paramamosain compared with live food, and the ratio of n‐6/n‐3 fatty acids in pelleted diets must be <1.     

Dietary fulvic acid effects on survival and expression of immune‐related genes in Litopenaeus vannamei challenged with Vibrio parahaemolyticus

The effects of fulvic acid (FA) on survival and immune‐related gene expression were investigated in Litopenaeus vannamei challenged with Vibrio parahaemolyticus by immersion. Shrimp were fed with different dietary FA concentrations (1, 2, 4 and 6 g/kg feed) for 20 days (first bioassay) or 8 days (second bioassay, 2 g/kg feed of FA added every 2 days) and then challenged with V. parahaemolyticus. In a third bioassay, the expression of three immune‐related genes (translationally controlled tumour protein [TCTP], superoxide dismutase [SOD] and heat‐shock protein 70 [HSP70]) in haemocytes or hepatopancreas of experimental shrimp was measured by real‐time quantitative PCR at 0, 6, 12, 24, 48, 72 and 96 hr after FA (2 g/kg feed) administration. Fulvic acid increased survival at a concentration of 2 g/kg feed supplied every two days. Interestingly, TCTP gene expression was upregulated, whereas gene expression of SOD and HSP70 was downregulated. In conclusion, dietary fulvic acid improves survival in white shrimp challenged with V. parahaemolyticus and modulates the immune response. Therefore, FA merits further evaluation as prophylactic treatment in commercial shrimp farms.