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.     

Dietary soybean β‐conglycinin suppresses growth performance and inconsistently triggers apoptosis in the intestine of juvenile grass carp (Ctenopharyngodon idella) in association with ROS‐mediated MAPK signalling

The current study aimed to investigate the effects of dietary soybean β‐conglycinin on growth performance and intestine apoptosis in juvenile grass carp (Ctenopharyngodon idella). For fish fed with the 80 g β‐conglycinin/kg diet for 7 weeks, the specific growth rate and feed intake were decreased. In the proximal intestine, dietary β‐conglycinin did not induce DNA fragmentation, tended to decrease the reactive oxygen species (ROS) content, and decreased ROS‐generating enzyme (NADPH oxidase [NOX]) activity. Subsequently, in the mid‐intestine, dietary β‐conglycinin caused DNA fragmentation, tended to increase the ROS content, increased caspase‐3, caspase‐8 and caspase‐9 activities, upregulated the mRNA levels of proapoptotic molecules (apoptotic protease‐activating factor‐1 [Apaf1] and Bcl‐2‐associated X protein [BAX]) and mitogen‐activated protein kinase (MAPK)‐related signal molecules (Jun N‐terminal kinase (JNK) and p38 MAPK) and increased the protein levels of p38 MAPK and phospho‐p38 MAPK. Moreover, in the distal intestine, dietary β‐conglycinin induced DNA fragmentation, elevated NOX activity and the ROS content and increased caspase‐3, caspase‐8 and caspase‐9 activities, death ligand (TNF‐α) mRNA expression level, and p38 MAPK and phospho‐p38 MAPK protein levels. In summary, dietary soybean β‐conglycinin suppressed fish growth and inconsistently caused apoptosis among the different intestinal segments which was partially associated with ROS‐mediated MAPK signalling.     

Growth,digestive and absorptive abilities and antioxidative capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idellus Val.) fed graded levels of dietary manganese

To study the effects of manganese on growth performance, digestive and absorptive abilities, as well as the antioxidative capacity in the hepatopancreas and intestine, young grass carp (Ctenopharyngodon idellus Val.) (264 ± 1 g) were fed diets containing graded levels of manganese at 3.65 (control), 8.62, 13.48, 18.24, 22.97 and 27.86 mg kg^?1 diet for 8 weeks. Per cent weight gain (PWG) and feed intake were the poorest in fish fed the basal diet (P < 0.05). The activities of trypsin, lipase and alkaline phosphatase in the intestine were significantly enhanced with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05). Additionally, in the hepatopancreas and intestine, the protein carbonyl and malondialdehyde contents were the lowest in fish fed the diet with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05), while the anti‐hydroxyl radical capacities, manganese superoxide dismutase (MnSOD), glutathione peroxidase and glutathione‐S‐transferase activities were significantly enhanced with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05). Moreover, the catalase activity and glutathione content in the intestine were the highest in fish fed the diet with dietary manganese level at 18.24 mg kg^?1 diet (P < 0.05). These results indicated that optimum dietary manganese promoted growth, enhanced the digestive and absorptive abilities, and improved the antioxidative capacity in young grass carp. Based on the quadratic regression analysis for PWG and intestinal MnSOD activity, the manganese requirements for young grass carp with the initial body weight of 264 g were 16.91 and 18.21 mg kg^?1 diet respectively.     

Evaluation of some intestinal cytokines genes expression and serum innate immune parameters in common carp (Cyprinus carpio) fed dietary loquat (Eriobotrya japonica) leaf extract

The aim of this study was to illustrate the effects of dietary loquat leaf extract (LLE) on the expression of some intestinal cytokines as well as serum innate immune parameters in common carp (Cyprinus carpio) fingerlings. The fish were fed with experimental diets supplemented with 0 (control), 0.25% (0.25LLE), 0.5% (0.5LLE) and 1% (1LLE) LLE over a 7‐week period. At the end of the trial, the serum lysozyme (Lys) activity, ACH50, total Ig as well as the expression of IL1B, IL8, TNF‐alpha, LYZ and TGF‐β in intestine were evaluated. The results showed that administration of 0.5% or 1% LLE significantly increased serum total Ig. However, in case of serum Lys activity significant elevation was observed just in fish fed 0.5% LLE. Also, supplementation of diet with LLE significantly increased ACH50 compared to the control group, regardless of inclusion levels. Gene expression studies revealed upregulation of TNF‐alpha, IL1B, IL8 and LYZ in intestine of fish fed LLE. However, the effects varied dependent on LLE levels and the tested immune related gene. Also, in case of TGF‐β significant downregulation was observed just in 1% LLE treatment. In conclusion, dietary LLE supplementation significantly upregulated immune related genes in intestine and improves innate immune responses. Altogether, LLE can be recommended as fish immunostimulant in early stage of carp culture.     

Protein‐sparing capability of dietary lipid in herbivorous and omnivorous freshwater finfish: a comparative case study on grass carp (Ctenopharyngodon idella) and tilapia (Oreochromis niloticus × O. aureus)

Two, 8‐week feeding trials were conducted to compare protein‐sparing capability of dietary lipid in herbivorous grass carp (Ctenopharyngodon idella) and omnivorous tilapia (Oreochomis niloticus × O. aureus). Utilizing a 2 × 3 factorial design, experimental diets containing two levels of crude protein (380 and 250 g kg⁻¹) and three levels of lipid (0, 40 and 100 g kg⁻¹) were formulated for use in both feeding trials. Growth performances showed better response of both fish fed 380 g kg⁻¹ protein diet than those fed 250 g kg⁻¹ protein diet. Despite the dietary protein level, weight gain (WG), specific growth ratio (SGR), feed conversion ratio (FCR) and protein efficiency ratio were much higher (P < 0.05) for grass carp fed 40 g kg⁻¹ lipid diet than those fed 100 g kg⁻¹ lipid diet; however, there were no significant differences in tilapia fed the two diets. The feed intake of grass carp fed lipid‐free diet was the lowest, but it tended to decrease with increase in dietary lipids in tilapia. Lipid retention (LR) was negatively correlated with dietary lipid concentration of both fish. Viscerosomatic index (VSI), hepatosomatic index (HSI), intraperitoneal fat ratio (IPF) and whole‐body and liver lipid content positively correlated with dietary lipid concentration of both fish. Plasma parameters and liver enzymes activities were also positively correlated with dietary lipid concentration of both fish. Liver lipid contents were higher and enzymes activities were lower in grass carp when compared with tilapia. These data suggested that there was no evidence of a protein‐sparing effect of dietary lipids in grass carp. Tilapia has relatively higher capacity to endure high dietary lipid level compared to grass carp.     

Effects of dietary phenylalanine on growth,digestive and brush border enzyme activities and antioxidant capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idella)

This study investigated the effects of phenylalanine on growth, digestive and absorptive ability and antioxidant status of young grass carp (Ctenopharyngodon idella). Young grass carp were fed diets containing 3.4 (basal diet), 6.1, 9.1, 11.5, 14.0 and 16.8 g phenylalanine kg^?1 diet with a fixed of 10.7 g tyrosine kg^?1 diet for 8 weeks. Percent weight gain (PWG), feed efficiency and feed intake of fish were the lowest in fish fed the basal diet (P < 0.05). Trypsin, lipase and amylase activities in the hepatopancreas, and antioxidants including glutathione contents and glutathione reducase activities in the hepatopancreas and intestine were all the highest in fish fed 11.5 g phenylalanine kg^?1 diet (P < 0.05). Trypsin, chymotrypsin and amylase activities in whole intestine, and creatine kinase, Na⁺, K⁺‐ATPase and alkaline phosphatase activities in the proximal intestine, and superoxide dismutase activities in the hepatopancreas and intestine were all the highest when phenylalanine at level of 9.1 g kg^?1 diet (P < 0.05). In conclusion, phenylalanine improved growth, digestive and absorptive ability, and antioxidant capacity of young grass carp. The phenylalanine requirement of young grass carp (256–629 g) based on PWG was 10.4 g kg^?1 diet or 3.44 g 100 g^?1 protein.     

Effects of dietary 60 g kg−1 dried distiller's grains in least‐cost practical diets on production and gut allochthonous bacterial composition of cage‐cultured fish: comparison among fish species with different natural food habits

Dried distiller's grain (DDG) is considered as an alternative ingredient of dietary feed due to its high contents of protein, fibre and fat. In this study, 60 g kg^‐1 of DDG was used to feed grass carp (Ctenopharyngodon idella), bluntnose black bream (Megalobrama amblycephala), gibel carp (Carassius gibelio) and black carp (Mylopharyngodon piceus) for 8 weeks, and its effect on fish production and gut allochthonous microbiota was investigated for the development of a suitable fish feed high in nutrients and low in cost for polyculture freshwater fish. DDG supplementation resulted in the less weight gain and higher feed conversion ratio of black carp (P < 0.05), but had no significant effects on other fish or parameters. PCR–denaturing gradient gel electrophoresis (DGGE) analysis indicated that all four fish species had some common and unique bacteria in their digestive tracts, and the gut microbiota of bluntnose black and gibel carp fed the control diet and DDG diets were very similar (Cs > 91%); of them, the total counts of intestinal bacteria studied by qPCR increased in grass carp (P < 0.05) and depressed in black carp (P < 0.05) when fed dietary DDG. Thus, we assumed that dietary DDG modulated production and gut microbiota of fish in a host‐specific way.     

Ammonia nitrogen excretion in Mandarin Fish (Siniperca chuatsi) and Grass Carp (Ctenopharyngodon idellus) fed practical diets: the effects of water temperature

To understand the actual production of fish culture about the utilization of dietary protein and excreta impact on the environment between mandarin fish (Siniperca chuatsi) and grass carp (Ctenopharyngodon idellus), the study to investigate the effect of temperatures (19 ± 0.5°C, 24 ± 0.5°C and 29 ± 0.5°C) on ammonia‐N excretion in mandarin fish and grass carp under fed and fasted states was conducted. These two species were fed a practical diet containing 325.2 g kg^?1 crude protein at 3% body weight per day. The ammonia‐N excretion rate was significantly increased when temperature increased from 19 to 29°C, and a linear relationship between ammonia‐N excretion rate and temperature. The maximum ammonia‐N excretion levels of mandarin fish and grass carp were observed at 4–8 h and 2–4 h after feeding, respectively, and the minimum values for both species were observed at 24 h after feeding. Under the feeding condition, mandarin fish had a lower ammonia‐N excretion level compared to grass carp at 24°C and 29°C. The average amount of ammonia‐N excreted by mandarin fish at 24 h is significantly higher than grass carp under fasting conditions, except 19 ± 0.5°C. These results indicated that mandarin fish might make better use of protein at higher temperature than grass carp when fed practical diets in commercial production. These results of this study suggested that mandarin fish had a lower ammonia‐N excretion level compared with grass carp, making a less contribution to environmental loading in an intensive fish culture.     

Effect of dietary chitosan on growth performance,haematology, immune response,intestine morphology,intestine microbiota and disease resistance in gibel carp (Carassius auratus gibelio)

A 75‐day experiment was conducted with juvenile gibel carp (Carassius auratus gibelio) (4.80 ± 0.01 g) to evaluate effects of dietary chitosan on fish growth performance, haematology, intestine morphology and immune response. Six isonitrogenous (crude protein: 383 g kg^?1), isolipid (97.5 g kg^?1) and isocaloric (gross energy: 16.7 kJ g^?1) diets were formulated to contain 0, 1800, 4000, 7500, 10 000, 20 000 mg kg^?1 chitosan, respectively. The results showed that the growth was depressed when the fish fed with 10 000 mg kg^?1 chitosan. Serum cholesterol, triglyceride and low‐density lipoprotein decreased in 10 000 and 20 000 mg kg^?1 chitosan. On day 75, blood leucocyte phagocytic activity respiratory burst and alternative pathway of complement haemolytic activity were enhanced in 4000 mg kg^?1 chitosan. The number of goblet cell, intraepithelial lymphocyte of mid‐intestine and microvilli height of distal intestine increased at 4000 mg kg^?1 dietary chitosan. Dietary chitosan modulated intestine microbiota, depressed pathogen bacteria Aeromonas veronii‐like and improved Cellulomonas hominis‐like, Bacillus oceanisediminis‐like and two uncultured bacterium‐like species on day 75. Dietary 7500 and 10 000 mg kg^?1 chitosan enhanced the protection against Aeromonas hydrophila infection. In conclusion, oral administration of dietary 7500 mg kg^?1 chitosan for 75 days is recommended for the survival of gibel carp.     

Effects of dietary glutamate supplementation on growth performance,digestive enzyme activities and antioxidant capacity in intestine of grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was carried out to investigate the effect of dietary glutamate (Glu) supplementation on growth performance, digestive enzyme activities and intestinal antioxidant capacity of grass carp (Ctenopharyngodon idella). A total of 540 grass carp with average initial weight of 370 ± 3.8 g were fed three diets supplemented with 0, 8 and 16 g kg^?1 Glu. The per cent weight gain, specific growth rate, feed conversion ratio, and protein and lipid productive value were significantly improved by dietary Glu supplementation. Feed intake, condition factor, visceral somatic index and perivisceral fat ratio were not significantly different between dietary groups. Hepatosomatic index, intestinal somatic index, intestine length index, intestine protein content, trypsin activity in hepatopancreas and intestine, lipase activity in proximal and mid‐intestine (MI) and hepatopancreas, alkaline phosphatase and gamma‐glutamyl transpeptidase activities in intestine, creatine kinase activities in the mid‐ and distal intestine, intestinal folds height in the proximal and MI were significantly increased with increased Glu. Malondialdehyde and protein carbonyl contents in the intestine decreased with increased Glu. Superoxide anion and hydroxyl radical scavenging ability, glutathione reducase, catalase and total superoxide dismutase activities, and glutathione content in the intestine were increased with increased Glu. These results suggested that dietary Glu supplementation increased intestinal antioxidant capacity, digestive and absorptive ability and improved fish growth.     

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.     

The effect of ultrafiltered fish protein hydrolysate level on growth performance,protein digestibility and mRNA expression of PepT1 in juvenile turbot (Scophthalmus maximus L.)

The intention of the study was to investigate the effect of ultrafiltered fish protein hydrolysate (UF) level on growth, feed utilization, apparent digestibility coefficients and proximal intestine peptide transporter 1 (PepT1) mRNA level for juvenile turbot (Scophthalmus maximus L.). Experimental diets (UF‐0, UF‐5, UF‐10, UF‐15 and UF‐20) were prepared containing about 68% plant protein, and fish meal protein was, respectively, replaced by 0%, 5%, 10%, 15% and 20% UF of dietary protein. Diet PP contained about 78% plant protein, and diet CAA contained about 10% crystalline amino acid mixture. All diets were fed to seven triplicate groups of turbot (initial weight 16.05 ± 0.03 g) for 68 days. Fish fed diet UF‐10 had an increasing tendency in growth compared with diets contained UF, while dietary UF level was not significantly correlated with specific growth rate and feed intake. Feed efficiency, protein efficiency ratio and protein productive value significantly correlated with dietary UF level, and fish fed diets contained low‐level UF had higher digestibility than that diets UF‐0, PP and CAA. There was a decreasing tendency in PepT1 expression level with dietary UF level. The results indicated that low‐level UF showed a positive effect on growth and feed utilization in juvenile turbot.     

Total aromatic amino acid requirement of juvenile grass carp (Ctenopharyngodon idella)

This experiment was conducted to investigate total aromatic amino acid requirement of juvenile grass carp Ctenopharyngodon idella. Six isonitrogenous and isoenergetic semipurified diets containing casein and gelatin with graded level of phenylalanine (7.8, 11.1, 14.4, 17.6, 21.7, 24.9 g kg^?1 DM) were formulated. Each diet was randomly assigned to triplicate group of 30 fish (3.58 ± 0.002 g, mean ± SEM) each tank for 8 weeks. The highest weight gain (WG, %), final body weight (g) and specific growth rate were recorded when phenylalanine level was 17.6 g kg^?1 of the diet. Fish muscle protein content, protein efficiency ratio (PER), feed conversion ratio and alanine aminotransferase were significantly affected by dietary phenylalanine level. The polynomial regression calculated using WG and PER indicated that the optimal dietary total aromatic amino acid (phenylalanine + tyrosine) requirement for juvenile grass carp was 24.4 g kg^?1 of the diet, corresponding to 65.9 g kg^?1 of dietary protein.     

Carbohydrate utilization by herbivorous and omnivorous freshwater fish species: a comparative study on gibel carp (Carassius auratus gibelio. var CAS III) and grass carp (Ctenopharyngodon idellus)

Two 8‐week feeding trials were conducted to evaluate dietary carbohydrate utilization by omnivorous gibel carp (Carassius auratus gibelio) (2.4 ± 0.1 g) and herbivorous grass carp (Ctenopharyngodon idellus) (6.5 ± 0.1 g). Five isonitrogenous (370 g kg^?1) and isolipid (70 g kg^?1) diets were formulated with increasing corn starch levels (60, 140, 220, 300 and 380 g kg^?1). Results showed that specific growth rate (SGR), feed efficiency (FE) and protein retention efficiency (PRE) of gibel carp significantly increased from dietary starch of 60 to 300 g kg^?1 and then decreased from 300 to 380 g kg^?1, but those of grass carp showed no significant differences between treatments. Independent of dietary starch levels, grass carp gained significantly higher FE and PRE than gibel carp. Feeding rate (FR) of gibel carp was significantly higher than that of grass carp. In two fish species, high dietary starch (300 and 380 g kg^?1) tended to obtain higher hepatosomatic index (HSI), serum triglyceride, hepatic lipid and body lipid contents. Serum glucose concentration of grass carp was not affected, while that of gibel carp fed the starch of 300 g kg^?1 diet was significantly lower than those of the fish fed other four diets (60, 140, 220 and 380 g kg^?1). Grass carp showed high tolerance to dietary starch while dietary corn starch should be no more than 300 g kg^?1 for gibel carp. High starch contents may cause lipid accumulation in the liver and body.     

Growth performance,intestinal morphology,hepatopancreatic antioxidant capacity and growth‐related gene mRNA expressions of juvenile grass carp (Ctenopharyngodon idellus) as affected by graded levels of dietary arginine

An 8‐week growth trial was conducted to investigate the effects of dietary Arg levels (7.6, 12.3, 17.9, 22.4 and 28.6 g/kg diet) on growth performance, hepatopancreatic antioxidant capacity, intestinal morphology and growth‐related gene expressions of juvenile grass carp (Ctenopharyngodon idellus). The results showed that SGR in Arg22 and Arg28 groups was lower than in Arg12 and Arg17 groups (p < 0.05). Serum NO content in Arg7 group was lower than other groups. Hepatopancreatic GSH‐Px activity was higher in Arg17 group than in Arg7 group, while MDA content showed the opposite trend. Hepatopancreatic IGF‐1 expression tended to increase with Arg from 7.6 to 22.4 g/kg and then decreased in Arg28 group (p < 0.05), while IGFBP‐1 expression increased with Arg level. Muscle mRNA expressions of TOR and S6K1 showed quadratic trends as dietary Arg level increased, which were higher in Arg17 group than in Arg22 and Arg28 groups (p < 0.05). Higher mRNA expression levels of y⁺LAT1, y⁺LAT2 and PepT1, as well as higher villus height and villus width in foregut, were all observed in Arg17 group. The optimal dietary Arg level based on SGR by the quadratic model was 15.3 g/kg diet for juvenile grass carp, corresponding to 54.7 g/kg dietary protein.     

草鱼NK-lysin基因的克隆、原核表达与活性分析

为研究草鱼抗菌肽NK-lysin基因的组成结构、在组织中的表达差异及其抑菌活性,实验根据已知鱼类NK-lysin基因保守区序列设计上下游引物,采用RT-PCR和RACEPCR技术克隆草鱼NK-lysin基因(Cinkl),RT-PCR进行各组织间的表达分析,并构建Cinkl成熟肽的原核表达载体,采用琼脂糖孔穴扩散法检测重组蛋白的抑菌活性。结果显示,Cinkl c DNA全长768 bp,编码121个氨基酸;基因组DNA全长3 361 bp,包含4个外显子和3个内含子。经多序列比对分析和结构域预测表明,Ci Nkl氨基酸序列具有SAPLIP家族的特征:含有6个保守的半胱氨酸和saposin B蛋白结构域,与斑马鱼Nklc、Nkld的相似性分别为57.98%和63.03%。构建NK-lysin氨基酸序列系统进化树,发现Ci Nkl与斑马鱼Nklc、Nkld聚为一支。RT-PCR检测结果显示,Cinkl m RNA在所有检测的草鱼组织中均有表达,在脾脏中表达量最高,心脏、鳃、中肾和头肾的表达量次之,皮肤、脑、肝脏和肠有微量表达。p ET-28b-MBP与Ci Nkl成熟肽序列构建的原核表达质粒在宿主菌Rosetta(DE3)表达出约57 ku的可溶性融合蛋白,利用Ni-NTA His·Bind树脂进行纯化,Ci Nkl重组蛋白对G–细菌如大肠杆菌M15株、嗜水气单胞菌,以及G+细菌如金黄色葡萄球菌均具有抑菌活性。基于草鱼NK-lysin的组织表达特征和重组蛋白的抑菌活性,推测NK-lysin在草鱼的先天性免疫防御中发挥调节作用。     

Dietary krill hydrolysates affect the expression of growth‐related genes in juvenile turbot (Scophthalmus maximus L.)

A 10‐week feeding experiment in indoor flow‐through seawater system was conducted to investigate the effects of dietary krill hydrolysate on the expression of growth‐related genes in juvenile turbot (Scophthalmus maximus L.; initial body weight 9.45 ± 0.01 g). Three isonitrogenous and isolipidic experimental diets containing high plant protein were formulated to contain 0 (control), 50 g/kg (LKH) and 100 g/kg (HKH) krill protein hydrolysate (KH) to replace fishmeal, respectively. Triplicate groups of 30 fish were fed for 10 weeks to apparent satiation twice daily. At the end of the feeding trial, the mRNA expressions of insulin‐like growth factor (IGF‐1) gene in liver, peptide transporters (PepT1) gene in pyloric caeca and proximal intestine and neuropeptide Y (NPY) gene in brain in all groups were determined. IGF‐1, PepT1 and NPY expression levels in HKH group were significantly increased compared with those of LKH and control (p < 0.05), which was consistent with the SGR, feed efficiency, PER and PPV. These results indicated that dietary 100 g/kg krill hydrolysate could improve growth performance and upregulate the mRNA expression of IGF‐1, PepT1 and NPY genes in juvenile turbot.     

Effects of supplemental phytic acid on the apparent digestibility and utilization of dietary amino acids and minerals in juvenile grass carp (Ctenopharyngodon idellus)

A feeding trial was conducted for 8 weeks to evaluate the effects of supplemental phytic acid (PA) on the apparent digestibility and utilization of dietary amino acids (AAs) and minerals in juvenile grass carp. Five experimental diets consisted of graded levels of PA (0.2, 4.7, 9.5, 19.1 and 38.3 g kg^?1, named as P0, P5, P10, P20 and P40). Triplicate groups of fish (initial weight, 22.37 ± 0.16 g) were fed twice daily (08:00 and 16:00 h). The crude protein content in whole body significantly (p < .05) decreased in fish fed with 19.1 and 38.3 g PA kg^?1 diet. Supplemental PA (>4.7 g kg^?1) significantly reduced the apparent digestibility coefficient (ADC) of AAs (Asp, Thr, Ser, Glu, Gly, Ala, Cys, Val, Met, Ile, Leu, Phe, Lys, Pro, His and Arg) and the ADC of minerals (P, Ca, Mg, Zn, Cu, Fe and Mn) in grass carp. The contents of minerals (P, Ca, Mg and Zn) in whole body and bone were also found to be significantly reduced in dietary PA > 4.7 g kg^?1, while the bone ash, serum Alkp and Zn contents were found to be significantly decreased when the PA supplementation level was above 9.5 g kg^?1, and the contents of serum Ca and Mg were found to be markedly altered in higher PA‐supplemented groups. The results indicated that supplemental PA decreased the apparent digestibility and utilization of AAs and minerals, and thus reduced the feed utilization of grass carp, suggesting that the level of total PA should be below 4.7 g kg^?1 in grass carp diet.     

Molecular cloning and expression of kidney‐type glutaminase from common carp (Cyprinus carpio) and its up‐regulation by glutamine in primary culture enterocyte

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney‐type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real‐time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up‐regulated quickly and effectively by Gln.