Effects of dietary vitamin C on growth,flesh quality and antioxidant capacity of juvenile golden pompano Trachinotus ovatus

To investigate the effects of dietary vitamin C on growth, flesh quality and antioxidant capacity of juvenile golden pompano Trachinotus ovatus, a 56‐day feeding trial with five graded levels of dietary VC (D1: 11.69, D2: 34.89, D3: 59.10, D4: 114.26 and D5: 227.93 mg VC per kg of diet) was performed on 375 fish (triplicate groups of 25 fish per diet, initial weight 13.57 ± 0.09 g). Results showed that fish of D3 group exhibited the maximum specific growth rate (SGR) and the highest liver enzymatic activities of catalase (CAT) and glutathione peroxidase (GSH‐PX), which were consistent with the expression levels of cat and gsh‐px. Besides, the D3 group also showed higher contents of protein and lipid, and lower cooking loss, drip loss and malondialdehyde content in muscle than D1 group. The docosahexaenoic acid proportion in muscle increased with increasing dietary VC levels. Furthermore, the lowest expression levels of carnitine palmitoyltransferase1 (cpt1) and peroxisome proliferator‐activated receptor α (pparα) were detected in livers of D3 group. The optimum dietary VC level was 49.73 mg/kg from the broken‐line analysis based on the SGR, in which better growth performance, antioxidative ability and flesh quality were observed in T. ovatus juveniles.     

Effect of dietary valine levels on the growth performance,feed utilization and immune function of juvenile golden pompano,Trachinotus ovatus

This experiment was designed to investigate the effects of dietary valine on the growth performance, feed utilization, digestive enzymes, serum antioxidant and immune indices of juvenile Trachinotus ovatus and determine its valine requirement. Six diets with different concentrations of L‐valine (15.0, 16.6, 18.6, 20.7, 23.5 and 25.4 g/kg dry diet, defined as diet Val‐1 to Val‐6.), were formulated to contain 430 g/kg crude protein with fish meal, soybean meal, peanut meal and precoated crystalline amino acids. Each diet was randomly assigned to triplicate treatments of 20 fish (the initial body weight was 5.34 ± 0.03 g) for 8 weeks. The results indicated that the final body weight and percent weight gain (PWG) increased with increasing valine concentration up to 18.6 g/kg (diet Val‐3), whereas the diets containing higher valine concentration reduced the growth performance significantly (p < .05). Moreover, the protein efficiency ratio, body protein deposition (BPD), muscle protein content, intestinal amylase and pepsin activities, serum T‐AOC, LZM activities, IgM, complement 3 and complement 4 concentration had a similar trend with PWG, and the trend of feed conversion ratio, serum AST, ALT activities, urea and MDA content was opposite. Meanwhile, the lipid contents of whole fish and muscle in diet Val‐6 were particularly lower than other diets (p < .05). The survival rate of diet Val‐1 was lowest in this study and was significantly lower than diet Val‐2 (p < .05). The results of polynomial regression based on PWG and BPD indicated that the optimal dietary valine requirement for Trachinotus ovatus reared in seawater‐floating net cages was 19.87–20.17 g/kg valine of dry diet, correspondingly 46.22–46.91 g/kg of dietary protein.     

Effect of dietary sodium butyrate on growth performance,enzyme activities and intestinal proliferation‐related gene expression of juvenile golden pompano Trachinotus ovatus

The present study was conducted to investigate the effects of dietary sodium butyrate on growth performance, intestine enzyme activities and intestinal proliferation‐related gene expression of juvenile golden pompano Trachinotus ovatus. A basal diet was supplemented with sodium butyrate at 0.0 (control), 0.25, 0.5, 1, 2 and 4 g/kg feed for 8 weeks. The final body weight, weight gain (WG), specific growth rate (SGR) and condition factor (CF) increased with increasing dietary sodium butyrate up to 2.0 g/kg, and thereafter declined, while feed conversion ratio exhibited an opposite trend. Compared with the control, the 2.0 g/kg sodium butyrate group had higher condition factor (CF) significantly (p < 0.05). Whole body ash decreased with increasing dietary sodium butyrate level, with the lowest whole body ash content in 4.0 g/kg sodium butyrate (p < 0.05). Compared with the control, the 2.0 g/kg sodium butyrate group had significantly increased plasma glucose, cholesterol, albumin level, A/G ratio, ALT and AST contents (p < 0.05), while significantly decreased plasma ALT/AST ratio (p < 0.05). As for the intestinal digestive and brush border enzymes activities, compared with the control, the 2.0 and 4.0 g/kg sodium butyrate groups had significantly increased intestinal protease, amylase, AKP and Na⁺‐K⁺‐ATPase activities (p < 0.05), respectively. The relative level of intestinal CDX2 mRNA of fish significantly increased with dietary sodium butyrate level. Compared with the control, the 2.0 g/kg sodium butyrate groups had significantly increased the expression of intestinal CDX2 and CREB mRNA (p < 0.05), respectively. In conclusion, these results suggested that the optimum sodium butyrate level for juvenile golden pompano could be 2.0 g/kg of the diet.     

Effects of dietary carbohydrate to lipid ratio on growth,feed utilization,body composition and digestive enzyme activities of golden pompano (Trachinotus ovatus)

An 8‐week feeding trial was conducted to investigate the effects of dietary carbohydrate to lipid ratio (CHO: L) on growth, feed utilization, body composition and digestive enzyme activities of golden pompano, Trachinotus ovatus. Five iso‐nitrogenous (450 g/kg protein) and iso‐energetic (19 MJ/kg gross energy) diets with varying CHO: L ratios of 0.68, 1.02, 1.62, 2.61 and 4.35, respectively, were fed to triplicate groups of 30 fish (average 13.8 ± 0.1 g). Results showed that dietary CHO: L ratios did not show any significant influence on survival of golden pompano (p > .05) but significantly affected its growth performance and feed utilization (p < .05). Fish fed diets with CHO: L ratios at 1.62 and 2.61 exhibited the highest final body weight, weight gain ratio, specific growth rate, feed efficiency ratio and protein efficiency ratio. Fish body lipid and liver glycogen contents were also significantly influenced by CHO: L ratio (p < .05). Hepatic amylase activity increased firstly and then decreased as the dietary CHO: L ratio increased, while lipases activity decreased with increasing dietary CHO:L level. The regression model analysis showed that the most suitable dietary CHO: L ratio (protein 450 g/kg) to reach the highest weight gain ratio is 2.38.     

Nutritional regulation of gene expression and enzyme activity of phosphoenolpyruvate carboxykinase in the hepatic gluconeogenesis pathway in golden pompano (Trachinotus ovatus)

Phosphoenolpyruvate carboxykinase (PEPCK) is known to exist as mitochondrial (PEPCK‐M) and cytosolic isoforms (PEPCK‐C). They are involved in glucose production and play vital roles in regulating glucose homeostasis. This study aimed to (1) clone and characterize PEPCK‐C cDNA from golden pompano fish (Trachinotus ovatus), which are known to have poor utilization of dietary carbohydrates, and (2) analyze the regulation of its expression by varying dietary carbohydrate‐to‐lipid ratios and nutritional status. A full length golden pompano PEPCK‐C cDNA fragment of 2,652 bp was cloned, which contains an open reading frame of 1,875 bp encoding 624 amino acids and shows a high homology to cobia (Rachycentron canadum) PEPCK‐C sequence (92% similarity). The analysis of tissue distributions of PEPCK‐C mRNA showed that a high abundance of PEPCK‐C was expressed in golden pompano liver, followed by kidney; while lower expression levels were found in the heart and intestine. In liver, PEPCK is under the regulation of dietary composition and nutritional status at the enzymatic and molecular levels. Our results indicate that PEPCK‐C plays a modulating role in the adaptation of hepatic gluconeogenesis to different nutrient conditions. Thus, the absence of molecular inhibition of gluconeogenic PEPCK gene expression in rainbow trout (Oncorhynchus mykiss) and gilthead sea bream (Sparus aurata), which can at least partially account for their poor utilization of dietary carbohydrate, is specific to these species and is not observed in golden pompano. Further studies that examine, at the enzymatic and molecular levels, other key enzymes involved in hepatic glucose metabolism are necessary to understand the poor utilization of carbohydrates in golden pompano.     

Effects of different dietary lipid sources on growth performance,body composition and lipid metabolism‐related enzymes and genes of juvenile golden pompano,Trachinotus ovatus

Three groups of juvenile golden pompano, Trachinotus ovatus (54.75 ± 0.25 g), were each fed one of three diets containing different lipid sources: fish oil (FO), soybean oil (SO) and lard oil (LO). Fish were reared in sea cages for 8 weeks, and the fish fed the FO diet had significantly higher specific growth rate (SGR) but lower condition factor (CF) than the other treatments. The fatty acid (FA) composition of whole‐body lipids was closely correlated with those in the diets. Although no differences can be found in hepatic fatty acid synthase (fasn) activity, the carnitine palmitoyl transferase 1 (cpt1) activity in fish fed the FO diet was significantly higher compared with other treatments. In addition, the relative gene expression of lipid metabolism‐related enzymes, such as cpt1, fas, apolipoprotein B100 (apoB100), delta‐6 fatty acyl desaturase (fadsd6) and fatty acid‐binding protein 1 (fabp1), was also influenced by the different dietary lipid sources. Serum triglyceride (TG) and glucose content in fish fed the LO and FO diets were significantly higher than those in the SO group. Accordingly, it can be concluded that FO could not be completely replaced by SO or LO in golden pompano diets. The lipid sources of a diet could impose significant influence on body condition factor and hepatic lipid metabolism of golden pompano.     

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.     

Effects of α‐lipoic acid on growth performance,body composition,antioxidant profile and lipid metabolism of the GIFT tilapia (Oreochromis niloticus) fed high‐fat diets

The wide use of lipid as a non‐protein energy substitute has led to lipid metabolic problems in cultured tilapia. Therefore, studies that reduce the effects of high‐fat diets in genetically improved farmed tilapia (GIFT) are required. This study evaluated the optimum level and effects of dietary α‐lipoic acid (α‐LA) on growth performance, body composition, antioxidant capacity and lipid metabolism of GIFT tilapia. The basal diet (120 g/kg lipid) was supplemented with six concentrations of α‐LA at 0 (control), L300, L600, L900, L1200 and L2400 mg/kg diet to make the experimental diets, which were fed to GIFT tilapia juveniles (initial body weight: 0.48 ± 0.01 g) for 8 weeks. The weight gain of fish improved significantly in the L300 than other dietary treatments. The intraperitoneal fat index and lipid content of fish fed on the L2400 diet decreased significantly than those fed on the control diet. The activities of superoxide dismutase and glutathione peroxidase (GSH‐Px) in serum and liver were significantly higher in fish fed on the L300 diet than the control. The reduced GSH content of fish fed on the L300 in serum and liver was significantly higher than those fed on control diet. The malondialdehyde content in serum and liver was significantly lower in L300 than in the control. The adipose triglyceride lipase gene was significantly up‐regulated in fish fed on the L2400, but the diacylglycerol acyltransferase 2 gene was down‐regulated in adipose. The liver‐type fatty acid‐binding protein gene in the liver was significantly up‐regulated in fish fed on the L300 and L600 diets. Moreover, the acyl‐coenzyme A oxidase gene in liver was significantly up‐regulated in fish fed on the L300, L600, L900 and L1200 diets. Polynomial regression analysis indicated that 439–528 mg/kg α‐LA is an appropriate dosage in high‐fat diet to improve growth performance and relieve lipid oxidative damage by accelerating lipid catabolism and reducing lipid synthesis in GIFT tilapia.     

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 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.     

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.     

Dietary n‐3 highly unsaturated fatty acids affect the biological and serum biochemical parameters,tissue fatty acid profile,antioxidation status and expression of lipid‐metabolism‐related genes in grass carp,Ctenopharyngodon idellus

A 95‐day feeding trial was conducted to determine the effects of n‐3 highly unsaturated fatty acids (n‐3 HUFA) on the growth, antioxidation and lipid metabolism in grass carp (Ctenopharyngodon idellus). Two isonitrogenous and isolipidic diets were formulated with either lard oil (LO) or fish oil (FO) as the main lipid source. The results showed that the intraperitoneal fat (IPF) ratio was significantly lower (P < 0.05) in FO group. The concentration of n‐3 HUFA in muscle, hepatopancreas and IPF was significantly higher in FO group (P < 0.05). The serum low‐density lipoprotein (LDL) content was significantly lower (P < 0.05), and glucose (GLU) content was significantly higher (P < 0.05) in FO group. The serum total superoxide dismutase (T‐SOD) activity was significantly higher (P < 0.05) in FO group, consistent with the serum malondialdehyde (MDA) content. The gene expression of IPF fatty acid synthase (FAS), acetyl‐CoA carboxylase (ACC), sterol regulatory element‐binding protein (SREBP‐1) and peroxisome proliferator‐activated receptor γ (PPARγ) was significantly lower (P < 0.05) and that of peroxisome proliferator‐activated receptor α (PPARα) was significantly higher (P < 0.05) in FO group compared with LO group. Similar trends were found in the hepatopancreas, except for PPARγ. It is suggested that n‐3 HUFA could inhibit lipid accumulation in grass carp by affecting the expression of lipid‐metabolism‐related genes.     

Replacement of de‐oiled rice bran by soaked and fermented sweet potato leaf meal: Effect on growth performance,body composition and expression of insulin‐like growth factor 1 in Labeo rohita (Hamilton), fingerlings

A 60‐day feeding trial was conducted to determine the growth performance and expression of insulin‐like growth factor 1 gene (IGF‐I gene) in Labeo rohita fingerlings fed with either raw, soaked or fermented sweet potato leaf meal (SPLM) by completely replacing de‐oiled rice bran (DORB), following a completely randomized design. Seven isonitrogenous (30%) and isocaloric (1.8 MJ/100 g) diets were prepared by replacing DORB with 50% and 100% raw, soaked and fermented sweet potato leaf meal, maintaining DORB‐containing diets as a control. Weight gain %, SGR (specific growth rate) and PER (protein efficiency ratio) were significantly (p < 0.05) higher when 100% DORB was replaced by fermented SPLM in comparison to other treatment groups. The fermented and soaked SPLM‐fed groups had registered with lower FCR value. The expression of growth regulating gene IGF‐I mRNA and RNA/DNA ratio was found to be significantly higher (p < 0.05) in soaked and fermented SPLM‐fed groups. In this study, the body protein and lipid composition did not vary significantly (p > 0.05). Hence, the study concludes that the fermented sweet potato leaf meal using Chaetomium globosum can replace 100% DORB in the diet of Labeo rohita without any detrimental effect on growth performance.