Combined effects of dietary fructooligosaccharide and Bacillus licheniformis on growth performance,body composition,intestinal enzymes activities and gut histology of triangular bream (Megalobrama terminalis)

This study was conducted to investigate the effects of fructooligosaccharide (FOS) and Bacillus licheniformis (B. licheniformis) on growth performance, body composition, intestinal enzymes activities and gut histology of Megalobrama terminalis. Nine experimental diets were formulated to contain three FOS levels (0, 3 and 6 g kg^?1) and three B. licheniformis levels (0, 1 and 5 × 10⁷ CFU g^?1) following a 3 × 3 factorial design. Accordingly, diets were named as 0/0, 0/3, 0/6, 1/0, 1/3, 1/6, 5/0, 5/3 and 5/6 (B. licheniformis/FOS). At the end of the 8‐week feeding trial, weight gain (WG) and specific growth rate (SGR) of fish fed 6 g kg^?1 FOS were both significantly (P < 0.01 and P < 0.05) higher than that of the other groups in terms of dietary FOS levels. Besides, WG and SGR of fish fed 1 × 10⁷ CFU g^?1 B. licheniformis were significantly (P < 0.05) higher than that of the control group in terms of dietary B. licheniformis levels. In addition, a significant interaction (P < 0.05) between dietary FOS and B. licheniformis was observed in finial weight, WG, SGR as well as the survival rate with the highest values all observed in fish fed diet 1/3. Hepatosomatic index, carcass lipid content, lipase activities as well as microvilli length increased significantly (P < 0.05) from 0 to 1 × 10⁷ CFU g^?1, but no significant difference (P > 0.05) was observed in terms of dietary FOS levels. In addition, a significant (P < 0.05) interaction of FOS and B. licheniformis was observed in both protease and Na⁺, K⁺‐ATPase activities with the highest value obtained in fish fed diet 1/3. The results indicated that the dietary applications of dietary FOS and B. licheniformis alone or in combination can significantly improve the growth performance, survival rate, intestinal enzymes activities as well as microvilli length of triangular bream. In addition, there is a significant interaction between dietary FOS and B. licheniformis. The best combination for this species is 3 g kg^?1 FOS with 1 × 10⁷ CFU g^?1 B. licheniformis.     

Effects of stocking density on the growth performance,digestive enzyme activities,antioxidant resistance,and intestinal microflora of blunt snout bream (Megalobrama amblycephala) juveniles

In this study, effects of stocking density on the growth performance and physiological responses of blunt snout bream, Megalobrama amblycephala juveniles were evaluated. The fish (average body weight, 25.76 ± 2.25 g) were randomly stocked at densities of 30F (30 fish/m³), 60F, 90F and 120F in 12 cages (1 m × 1 m × 1 m) in a concrete pond, with three cages for each density, for a period of 6 weeks. The higher stocking densities had a negative effect on individual growth performance. The results indicated that serum cortisol, triglyceride, alanine aminotransferase, aspartate transaminase, alkaline phosphatase and malondialdehyde activities; and Acinetobacter, Aeromonas, Pseudomonas and Vibrio numbers in the intestinal microflora increased significantly as the stocking density increased. In contrast, the viscerosomatic index, hepatosomatic index survival rate; serum glucose, total cholesterol, lipase, protease, glutathione peroxidase and superoxide dismutase activities; and Clostridium, Bacteroides, Lactococcus, Lactobacillus and Bacillus numbers in the intestinal microflora decreased significantly. The 90F and 120F groups showed obvious enlargement of the lamina propria and goblet cell damage, indicating that the gut showed inflammatory responses. The specific growth rate and weight gain rate increased significantly as the stocking density increased from 30 to 60 fish/m³, but decreased significantly when the stocking density was over 60 fish/m³.     

厚颌鲂仔稚鱼消化酶活性变化研究

测定了厚颌鲂(Megalobrama pellegrini)仔稚鱼从出膜到30日龄的生长、可溶性蛋白含量及消化酶活性。结果表明:初孵仔鱼即能检测到胰蛋白酶、淀粉酶、脂肪酶活性,2日龄检测到碱性磷酸酶活性。投喂组可溶性蛋白含量随日龄的增加显著增加(P<0.05),饥饿组可溶性蛋白含量6日龄前增加不显著,6日龄后开始下降。所测消化酶的全活力与比活力呈现出不同的变化模式。各消化酶的全活力基本是随日龄的增加而增加。投喂组胰蛋白酶比活力在10日龄达最大值,随后显著下降;饥饿组胰蛋白酶比活力在4日龄达最大值,随后显著下降。投喂组淀粉酶在5日龄达到最大值,从6日龄到12日龄维持在一个较低水平,随后增加至15日龄的(0.912±0.090)mU/mg,在试验后期均维持在一个相对较低的水平;饥饿组淀粉酶比活力的最大值出现在1日龄,从2日龄到8日龄变化不显著,8日龄后显著下降。投喂组脂肪酶比活力在18日龄达最大值,随后显著下降;饥饿组脂肪酶比活力1日龄到6日龄变化不显著,6日龄后显著下降。投喂组碱性磷酸酶比活力从2日龄到12日龄显著上升,12日龄达到最大值,随后显著下降;饥饿组碱性磷酸酶比活力从2日龄到8日龄显著上升,8日龄达到最大值,随后显著下降。     

Molecular cloning of Insulin-like Growth Factor-I from triangular bream (Megalobrama terminalis)

Triangular bream (Megalobrama terminalis) IGF-I DNA and gene were cloned from triangular bream liver for the first time by RT-PCR. Sequence analysis indicated that the IGF-I cDNA consisted of 486 nucleotides encoding 117 amino acids which spanned the complete signal peptide, B, C, A, D and E domains. Analysis of the E domain indicated that triangular bream IGF-I belongs to the IGF-I Ea-2 subtype. Compared to bluntnose bream (Megalobrama amblycephala), another member of the same Megalobrama genus, triangular bream IGF-I shared 99.8% identity in cDNA sequence and 99.4% in predicted amino acid sequence. However, considerable differences were found in comparison to with common carp (Cyprinus carpio) and grass carp (Ctenopharyngodon idellus), which are members of the same family but of a different genus. This revised version was published online in July 2006 with corrections to the Cover Date.     

黑龙江三角鲂的胚胎和早期仔鱼发育观察

以产自黑龙江的三角鲂（Megalobrama terminalis）为研究对象,对其胚胎和早期仔鱼发育的形态特征和时间进行观察和描述。将受精卵去膜,在Holtfreter氏液中培育,着重观察去膜卵在原肠胚及后期发育的形态特征。在水温（23.5±1.0）℃条件下,黑龙江三角鲂胚胎从受精到出膜历时30 h 10 min,历经7个主要发育阶段（分为卵裂前期、卵裂期、囊胚期、原肠胚期、神经胚期、器官形成期、出膜期）,25个发育时期。初孵仔鱼体长为（4.33±0.16）mm,在（25.5±1.0）℃条件下,经144 h,卵黄囊被完全吸收,此时全长为（7.73±0.20）mm。早期仔鱼可根据眼色素沉积、体色素沉积、鳔和卵黄囊的变化分为眼色素增多期、体色素增多期、鳔出现期、鳔充气期和卵黄囊完全吸收期5个时期。黑龙江三角鲂在鳔充气膨大（出膜后96 h）前无摄食行为。在出膜后96~144 h为混合营养期,此时适宜将仔鱼转入池塘进行夏花培育。本研究旨为黑龙江三角鲂的资源保护和苗种培育提供科学依据。     

Expression of recombinant triangular bream (Megalobrama terminalis) pre-IGF-I in Escherichia coli

The insulin-like growth factor I (IGF-I) cDNA (GenBank Accession No. AY247412) of triangular bream (Megalobrama terminalis) was expressed for the first time in Escherichia coli. To construct the expression plasmid, the IGF-I cDNA was subcloned into prokaryotic-expressing vector pGEX-4T-1. The E.coli JM109 was transformed with the recombinant plasmid pGEX-4T-1-IGF-I, and the transgene expression was observed after being induced with isopropyl-β-D-thiogalactoside (IPTG). The results of SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting indicated that the recombinant fusion protein had immune activity and the molecular weight was about 47 kDa. The results of SDS-PAGE and thin layer scanning showed that the yield of fusion protein had been enlarged with prolonging time. When the time of induced expression was 1, 2, 3, 4, 5, and 6 h, the expression amount was approximately 1.4, 4.3, 8.1, 11.3, 16.3 and 18.8% of total bacterial protein respectively.     

Genetic parameter estimates for growth‐related traits of blunt snout bream (Megalobrama amblycephala) using microsatellite‐based pedigree

Blunt snout bream (Megalobrama amblycephala) is an herbivorous freshwater fish species native to China and has been recognized as a main aquaculture species in the Chinese freshwater polyculture system with high economic value. The genetic parameter estimates for important economic traits are needed for its selective breeding. The aim of this study was to estimate the heritabilities for its growth‐related traits and explore the genetic and phenotypic correlations among the traits using microsatellite‐based pedigree approach. Offspring from a mass‐spawning of 92 broodstocks (42 sires and 50 dams) were reared in a communal pond and nine microsatellites were used to identify the parents of each sampled offspring. Of 749 offspring randomly selected, 708 (94.53%) could be assigned directly to a single parental pair, which was used for heritability estimates. Data were analysed using the method of restricted maximum likelihood (REML) using animal model and the results showed that the heritabilities of body weight, body length, total length and body height were 0.65, 0.53, 0.53 and 0.50, respectively. High genetic correlations were found among these four traits. According to these results, selection for growth seems to be feasible in M. amblycephala and the other growth traits will be heightened accordingly with the selection based on body length.     

Dietary choline requirement for juvenile blunt snout bream,Megalobrama amblycephala

A 12‐week feeding trial was conducted to determine dietary choline requirement for juvenile Megalobrama amblycephala. The basal diet was formulated to contain 310 g kg^?1 diet from vitamin‐free casein and gelatine. Choline chloride was supplemented to the basal diet to formulate six purified diets containing 0, 250, 500, 1000, 2000 and 4000 mg kg^?1, respectively. Each diet was randomly fed to quadrupled groups of Megalobrama amblycephala with initial average weight 1.84 ± 0.04 g in a flow‐through system. Results showed weight gain was increased significantly with increasing dietary choline levels (P < 0.01). Lipid content of liver decreased significantly as dietary choline concentration increased (P < 0.01), whereas lipid content of dressed carcass showed opposite trend (P < 0.01), and lipid content of whole‐body was unaffected by dietary choline supplementation. Broken‐ line regression of weight gain, liver and muscle choline concentration showed choline requirements of Megalobrama amblycephala of 1198, 1525 and 1365 mg kg^?1, respectively. In addition, dietary choline supplementation significantly improved lipid content of dressed carcass but not the content of whole body of blunt snout bream.     

Ontogenetic development of digestive tract and digestive enzymatic activities in Squaliobarbus curriculus larvae

Squaliobarbus curriculus is an economically important freshwater fish. The ontogenetic development of the digestive system of S. curriculus larvae was studied histologically and enzymatically from hatching to 30 days posthatching (DPH). Amylase, lipase, alkaline phosphatase and pepsin activities were detected from the hatching stage, indicating that these enzymes were genetically preprogrammed. Marked increases in intestinal amylase, trypsin and alkaline phosphatase activities between 10 and 20 DPH corresponded to feed acquisition and transformation. Larval development in S. curriculus could be divided into three phases: phase I (endotrophic period): 1–3 DPH; phase II (endo‐exotrophic period): 4–5 DPH; and phase III (exclusively exotrophic period): from 6 DPH onward. At hatching, the digestive tract of the larvae was an undifferentiated straight tube. On 3 DPH, the digestive tract differentiated into the mouth cavity, oesophagus and intestine. On 6 DPH, feeding was totally exotrophic and the yolk sac was completely exhausted. During the growth of S. curriculus larvae, the intestinal mucosa formed and the number of goblet cells and microvilli increased, demonstrating maturation of the digestive system. The study about the digestive development of S. curriculus larvae will contribute to better larval‐rearing strategies.     

Dietary isoleucine requirement of juvenile blunt snout bream,Megalobrama amblycephala

A 9‐week feeding trial was conducted to estimate the dietary isoleucine requirement of juvenile blunt snout bream. Six isonitrogenous and isoenergetic experimental diets were formulated to contain graded isoleucine levels ranging from 5.3 to 20.1 g kg^?1 dry diet. At the end of the experiment, weight gain (WG), specific growth rate (SGR), feed efficiency ratio (FER) and protein efficiency ratio (PER) increased with increasing dietary isoleucine level up to 11.1 g kg^?1 dry diet, and dietary isoleucine level above 14.2 g kg^?1 dry diet declined these performances. Dietary isoleucine levels (14.2 and 17.3 g kg^?1 dry diet) significantly improved whole‐body protein content, but decreased whole‐body lipid, plasma triglyceride and cholesterol contents. Significantly lower visceral fat index (VFI) in fish fed with 14.2 g kg^?1 dietary isoleucine was observed compared to those fed with deficient or excessive isoleucine. Dietary isoleucine supplementation significantly increased plasma isoleucine concentration, while plasma valine and leucine concentrations showed a reversed trend. Dietary isoleucine levels regulated the target of rapamycin (TOR) gene expression and improved plasma superoxide dismutase (SOD) activity in juvenile blunt snout bream. Based on second‐order polynomial regression model analysis of SGR and FER, the optimum dietary isoleucine requirement was estimated to be 13.8 g kg^?1 dry diet (40.6 g kg^?1 dietary protein) and 14.0 g kg^?1 dry diet (41.2 g kg^?1 dietary protein), respectively.     

The responses of digestive enzymes in juvenile and adult Rapana venosa (Valenciennes, 1846) to different temperatures

The veined rapa whelk, Rapana venosa (Valenciennes, 1846), is an important and valuable fishery species; however, it has not been routinely artificially cultured on a large scale. Here, we studied the effect of temperature (6, 16, 22 and 28°C) on the digestive enzymes in R. venosa of different sizes (Juvenile (small, S: 1.2–1.8 cm) and adult (medium, M: 4.1–5.2 cm; large, L: 7.6–8.9 cm)) by detected the enzyme activity in different digestive glands (the salivary gland, Leiblein's gland (oesophageal gland) and liver) using spectrophotometric method, to determine the optimal temperature for artificial cultivation and to explore the feeding habits of this whelk. The optimal temperature was determined to be approximately 22°C. The food intake was significantly higher at 22°C than at other temperatures, and the activity of the digestive enzyme, trypsin, was at the highest in the main digestive gland of R. venosa, the liver, at 22°C. In addition, the observed presence of cellulase and α‐amylase may suggest that carbohydrates are necessary during R. venosa development. The results of this study will help in the formulation of technical procedures for R. venosa aquaculture and in preventing its overexploitation, and explained its invasion mechanism to some extent.     

Ontogenic development of enzymatic activity and digestive system in Jullien's golden carp (Probarbus jullieni Sauvage, 1880)

Ontogenic development of the main enzymes and histological structure of digestive organs were studied in Jullien's golden carp (Probarbus jullieni) from hatching until 50 days after hatching (DAH). The larval fish were produced by artificial insemination and fed only Moina sp. till end of experiment. Body weight (mg) and total length (cm) of Jullien's golden carp increased exponentially and linearly. The results indicate the fish weight grew fast with increasing rate, while length increased at a constant rate over the studied period. Up‐regulation of acid protease was observed in newly hatched larvae and the specific activity gradually decreased with time. Trypsin specific activity was relatively stable within the first 35 DAH, while fluctuations in chymotrypsin were observed. Among these three proteolytic enzymes, acid proteases exihibited relatively high specific activity in newly hatched larvae, suggesting a role in yolk protein degradation. Alkaline proteases became more prominent with age and correlated with an abrupt decrease in acid proteases. Increased lipase‐specific activity appeared within 3 DAH and then gradually decreased with time, indicating the capacity to digest yolk lipid reserve. Amylase and cellulase‐specific activities changed in a similar manner, and the sensitivity to time was higher in amylase than in cellulase. The digestive organs and accessory organs developed around 3–5 DAH. However, intestinal histology was almost fully developed around 18 DAH. These findings should be useful for deciding the preferred timing for weaning, as well as on developing artificial diets referenced to the physiological changes of digestive enzymes.     

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.     

Effect of Lactobacillus acidophilussupplementation on growth performances,digestive enzyme activities and gut histomorphology of striped catfish (Pangasianodon hypophthalmusSauvage, 1878) juveniles

The influence of dietary supplementation of Lactobacillus acidophilus, on growth performance, digestive enzyme activities, gut histomorphology and gut microflora were evaluated in juveniles striped catfish (Pangasianodon hypophthalmus). Five experimental diets were formulated by supplementing 0, 10³, 10⁵, 10⁷ and 10⁹ CFU/g L. acidophilus in fishmeal and casein‐based semi‐purified diet. Triplicate groups of striped catfish (21.69 ± 0.18 g) were stocked in 15 fiberglass tanks with stocking density of 25 individuals per tank and fed twice daily at 2.5% of the fish body weight for 12 weeks. Weight gain, specific growth rate, feed conversion ratio and the protein efficiency ratio were significantly higher in fish fed with 10⁵ and 10⁷CFU/g L. acidophilussupplemented diets compared with the other treatment groups. Compared with the control and fish fed low (10³ CFU/g) L. acidophilus supplementation, those fed with 10⁵ and 10⁷ CFU/g had significantly higher (p < 0.05) apparent protein digestibility. Inclusion of L. acidophilus at 10⁷ CFU/g diet significantly increased amylase, protease and lipase activities. Microscopic analysis showed that the villi length in both the anterior and posterior gut and microvilli length in the posterior gut increased significantly in fish fed L. acidophilus supplementation at 10⁵ and 10⁷ CFU/g of diet. The fish fed L. acidophilus supplemented diets significantly increased the total lactic acid bacteria counts in the gut of striped catfish compared with the control‐fed group. Based on gut histomorphology and growth performance, inclusion of L. acidophilus at 10⁵ CFU/g appears to have the most positive effect on striped catfish farming.     

Morphological changes during the transition from freshwater to sea water in an amphidromous goby,Sicyopterus lagocephalus (Pallas 1770) (Teleostei)

The widely distributed amphidromous goby Sicyopterus lagocephalus needs drastic change of habitat to fulfil its life cycle: adults live and spawn in rivers, where eggs hatch into prolarvae that have to reach the sea to acquire characteristics of planktonic larvae. Postlarvae return to rivers where they recruit and grow to the adult reproductive stage. Here, we describe the prolarval stages, namely from hatching to first contact with sea water, as well as the first marine larval stages. The observations were made under experimental conditions. We described 3 prolarval substages in freshwater (L1a–L1c). Prolarvae present a slight but visible ontogenetic development in freshwater, during which the yolk sac begins to reduce, the pigmentation increases on the body and in the eyes, and the lenses appear, although the eyes are not functional. Prolarvae need to reach the sea in a maximum of 96 h to pursue their development. Their transfer in sea water at a salinity of 36.5 induces important morphological modifications (i.e. yolk sac full absorption, appearance of pectoral fins, migration of the eyes in anterolateral position of the head, opening of mouth and anus), enabling the organisms to adapt to their new environment. This marine stage is divided into two substages: L2a corresponding to the organisation of the morphological structures adapted to the marine environment and L2b during which these morphological structures become functional. Whether it is in freshwater or sea water, the duration of the substages depends on the water temperature, but is similar for all individuals for a given temperature.     

Effect of cortisol and triiodothyronine bath treatments on the digestive enzyme profile and growth of Catla catla larvae during ontogenic development

Digestive enzyme profile is a good indicator of the nutritional and health status of the fish. The present investigation aims to evaluate the effect of exogenous bath treatment of hormones, cortisol and triiodothyronine, on the digestive enzyme activities and growth of carp Catla catla (Ham.) during ontogenic development. Catla larvae (4 days old) were given bath treatment with cortisol (hydrocortisone, 0.2 mg L^?1), 3,5,3′‐triiodothyronine (T3, 2.5 mg L^?1) and a combination of cortisol and T3 for 30 min. Digestive enzyme profile was recorded on every third day and was continued for 30 days. Larvae were fed with live food for initial 14 days and then weaned to mix feeding of live food and prepared diet. Significantly (P < 0.05) higher amylase, total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were found in the hormone‐treated groups compared to the control one during ontogenic development. Among the treated groups, amylase activity was highest in cortisol‐treated larvae. Total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were significantly (P < 0.05) higher in larvae exposed at combined treatment of cortisol and T3 compared to the other two groups in most sampling days. Average length, weight and specific growth rate of treated larvae were higher compared to the control one. The combined bath treatment of cortisol and triiodothyronine influenced the digestive enzyme activities of catla larvae and thereby enhanced the growth at early developmental stage. This helps the larvae to overcome the problems associated with early developmental stage.     

Comparative study on the effect of high dietary carbohydrate on the growth performance,body composition,serum physiological responses and hepatic antioxidant abilities in Wuchang bream (Megalobrama amblycephala) and black carp (Mylopharyngodon piceus Richardson, 1846)

A 90‐day trial investigated the effect of dietary carbohydrate level on growth performance, body composition, serum physiological responses and hepatic antioxidant abilities of a herbivorous fish, Wuchang bream (Megalobrama amblycephala) and a carnivorous fish, black carp (Mylopharyngodon piceus Richardson, 1846). For each species, two isonitrogenous and isolipidic semi‐purified diets (28.6% crude protein and 5.3% crude lipid for Wuchang bream) were formulated to contain 30.4% and 52.9% carbohydrate (WB‐NCHO and WB‐HCHO); Two isonitrogenous and isolipidic semi‐purified diets (32.6% crude protein and 5.3% crude lipid for black carp) were formulated to contain 20.5% and 40.5% carbohydrate (BC‐NCHO and BC‐HCHO). It was shown that, compared to Group WB‐NCHO, Wuchang bream in Group WB‐HCHO had lower weight gain rate, specific growth rate, serum total protein, hepatic superoxide dismutase activity and total antioxidative capacity, while higher feed conversion ratio, serum triglyceride and cortisol levels (P < 0.05). Compared to Group BC‐NCHO, black carp in Group BC‐HCHO had lower final weight, weight gain rate, specific growth rate, protein efficiency ratio, serum alkaline alkaline phosphatase activity, serum total protein, hepatic superoxide dismutase activity and total antioxidative capacity, while higher feed conversion ratio, whole body crude lipid, serum triglyceride, cholesterol, glutamic‐oxaloacetic transaminase activity and cortisol levels (P < 0.05). The results of this study suggested that Wuchang bream had better capacity of utilizing dietary carbohydrate than that of black carp, while high levels of dietary carbohydrate might not only be detrimental to growth performance and feed utilization but also the immune response and health for both species.     

Essential amino acids supplementation to practical diets affects growth,feed utilization and glucose metabolism‐related signalling molecules of blunt snout bream (Megalobrama amblycephala)

Six diets were formulated to investigate the success of fish meal (FM) replacement by plant proteins; diet 1 reflected a commercial feed (8% FM), diet 3 contained 4% FM, and diet 5 was devoid of FM. Whereas, diet 2, diet 4, and diet 6 reflected diet 1, diet 3, diet 5, respectively, and supplemented with essential amino acid (EAA). At the end of 8‐week trial, there was no significant difference in survival rate. Significantly higher final weight, weight gain rate, and specific growth rate were recorded in the group fed diet 2 compared with the other treatments (except diet 4) (p < 0.05). Feed conversion ratio of fish fed diet 2 was significantly lower than those fed diets 1, 3, and 5 (p < 0.05). The lowest feed intake and highest protein efficiency rate were found in fish fed diet 2 (p < 0.05). There was no significant difference in whole body compositions between treatments. Plasma aspartate transaminases, alanine aminotransferase, and glucose were significantly affected by dietary treatments (p < 0.05), while plasma protein and albumin contents were not influenced by the treatments. The relative expression of target of rapamycin (TOR) and phosphatidylinositol 3‐kinase, regulatory subunit 1 (alpha) (PIK3R1) in fish fed diet 3 (4% FM) were significantly down‐regulated compared with those fed diet 6 for TOR and diets 4 and 6 for PIK3R1 (p < 0.05). Insulin receptor substrate 1 (IRS‐1) and janus kinase 3 (JAK3) expressions were fluctuated, with the higher levels in fish fed diets 4 and 6. In conclusion, the findings of this study indicate that plant protein mixture supplemented with EAA could be used to substitute FM in practical diet for Megalobrama amblycephala.