Supplementations of dl‐Methionine and Methionine Dipeptide in Diets are Effective for the Development and Growth of Larvae and Juvenile Red Sea Bream,Pagrus major

Growth trials for larvae and juvenile red sea bream, Pagrus major, were conducted to elucidate the efficacy of two molecular forms of methionine; dl ‐methionine (dl ‐Met) and methionine dipeptide (Met‐Met). For the larvae experiment, five experimental diets were formulated and fed to fish (42 mg) for 30 days. A diet which has 15% soy protein isolate served as the control diet. Similarly, test diets supplemented with dl ‐Met and Met‐Met at 0.5%, which were either precoated by zein or intact, were also formulated. For the juvenile experiment, five experimental diets were formulated wherein the control diet contained 25% soy protein isolate. Test diets were supplemented with dl ‐Met and Met‐Met at 0.75%, which were either coated by carboxymethycellulose or intact and fed to juveniles (0.75 g) for 56 days. The results of two feeding trials showed both dl ‐Met and Met‐Met can be equally utilized by red sea bream larvae and juveniles. Coating the amino acid significantly improved both fish larval and juvenile growth performance. The development of digestive protease activity of larvae was significantly influenced by coating the amino acid, but the type of methionine was not a factor in changing the protease activity of larvae.     

Dietary nucleotides improve the growth performance,antioxidative capacity and intestinal morphology of turbot (Scophthalmus maximus)

A growth trial was conducted to evaluate the effects and safety of nucleotides in low fish meal diets on the growth performance, antioxidative capacity and intestinal morphology of turbot (Scophthalmus maximus). High fish meal control diet was formulated with 500 g kg^?1 fish meal. Seven levels (0.075, 0.15, 0.225, 0.300, 1.5 and 3.0 g kg^?1, respectively) of nucleotides were added to a low fish meal basal diet, which was formulated with 400 g kg^?1 fish meal. The eight experimental diets were fed to groups of juvenile turbot (initial weight: 6.0 ± 0.03 g) for 60 days. Results showed that compared with high fish meal control diet, low fish meal basal diet treatment had lower total antioxidative capacity (T‐AOC), glutathione peroxidase activity, fold height of proximal and distal intestine, enterocyte height of all evaluated enteric section and microvillus height of mid‐intestine and distal intestine (P < 0.05). However, supplemented nucleotides in diets could significantly improve growth (specific growth rate, SGR), feed utilization, antioxidative capacity and intestinal morphology of turbot (P < 0.05). Broken‐line regression analysis of SGR and T‐AOC showed that the optimal supplemental levels of dietary nucleotide for juvenile turbot were 0.366 and 0.188 g kg^?1, respectively. In summary, 0.300 g kg^?1 of dietary nucleotides was helpful in improving growth, feed utilization, antioxidative capacity and intestinal morphology of turbot fed with low fish meal diet. Excessive dietary nucleotides (3.0 g kg^?1) might cause oxidative stress and morphological damage in intestine and then reduce the growth of turbot.     

Proteolytic enzyme activity of juvenile Asian sea bass, Lates calcarifer (Bloch), is increased with protein intake

The effect of high dietary protein intake on proteolytic enzyme activity of feeding juvenile Asian sea bass, Lates calcarifer (Bloch) was studied. Ninety fish [mean body weight ± standard error (SE) 304.62 ± 34.84 g] were randomly assigned to two dietary treatments, each with three replicates. In treatment 1, fish were fed by‐catch (Thunnus albacares) and in treatment 2, a formulated diet containing 50% protein. Proteolytic enzyme activity was determined in pyloric caecae and intestine at day 0, 7, 15, and 30. Initial proteolytic enzyme activity in sea bass ranged from 174 to 232 azocasein units (U^AC.) per mg of protein. After 7 days there was no significant difference in proteolytic enzyme activity of fish fed the two diets. However, a marked increase was observed in fish fed the formulated diet at day 15. After 30 days, the proteolytic enzyme activity in fish fed the formulated diet was threefold higher than that in fish fed the by‐catch diet. Fish fed the formulated diet had significantly higher total protein intake at day 7 than did fish fed by‐catch. Thereafter, a twofold weekly increase in the total protein intake was observed in both fish fed the by‐catch and formulated diets until day 30. These results suggest that a high dietary protein intake induces increased proteolytic enzyme activity in Asian sea bass.     

Prebiotics effect on growth performance,hepatic intermediary metabolism,gut microbiota and digestive enzymes of white sea bream (Diplodus sargus)

This study aimed at evaluating the effects of short‐chain fructooligosaccharides (scFOS), xylooligosaccharides (XOS) and galactooligosaccharides (GOS) on growth performance, hepatic metabolism, gut microbiota and digestive enzymes activities of white sea bream juveniles. Four diets were formulated: a control diet with fish meal (FM) and plant feedstuffs (PF) (30FM:70PF) and three test diets similar to control but supplemented with 10 g of scFOS, XOS or GOS per kilo diet, which were fed to fish during 12 weeks. Prebiotics had no effect on growth, feed efficiency or gut microbiota. Plasmatic triglycerides were lower in fish fed XOS than FOS and GOS diets. Malic enzyme activity was lower in fish fed XOS than FOS diet. Fish fed XOS diet had lower fatty acid synthetase (FAS), a key lipogenic enzyme and higher alanine aminotransferase activities. Fifteen days after the start of the trial, an enhancement of total alkaline protease, trypsin and lipase activities was observed in fish fed prebiotics, but such effect disappeared at 12 weeks. In conclusion, scFOS, XOS or GOS seem to have limited applicability in white sea bream feed.     

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

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

Inosine supplementation effectively provokes the growth,immune response,oxidative stress resistance and intestinal morphology of juvenile red sea bream,Pagrus major

A feeding trial was conducted to determine effects of dietary inosine on growth, immune responses, oxidative stress resistance and intestinal morphology of juvenile red sea bream. A semi‐purified basal diet supplemented with 0 (D1, control), 2 g kg^?1 (D2), 4 g kg^?1 (D3), 6 g kg^?1 (D4) and 8 g kg^?1 (D5) dietary inosine, respectively, to formulate five experimental diets. Each diet was randomly allocated to triplicate groups of fish (initial weight: 8 g). After 50 days of feeding trial, fish fed diets with 4 g kg^?1 inosine had the highest (p < .05) final weight, weight gain and specific growth rate. Similarly, improved feed intake, feed conversion ratio and protein efficiency ratio were also found at 4 g kg^?1 inosine supplemented group. Some non‐specific immune parameters such as total serum protein, lysozyme activity and bactericidal activity tended to be higher for fish fed diets supplemented with inosine. Peroxidase and catalase activity also influenced (p < .05) by dietary inosine supplementation. Fish fed 2 g kg^?1 and 4 g kg^?1 inosine supplemented diets showed the least oxidative stress condition. Inosine supplementation significantly increased (p < .05) anterior enterocyte height and posterior fold height, enterocyte height and microvillus height compared with control. We concluded that 4 g kg^?1 dietary inosine supplementation effectively provokes growth and health performance of red sea bream by increasing growth, immune response, oxidative stress resistance and intestinal health condition.     

Effect of fermented Enteromopha prolifera on the growth performance,digestive enzyme activities and serum non‐specific immunity of red tilapia (Oreochromis mossambicus × Oreochromis niloticus)

This study was conducted to evaluate the effect of dietary inclusion of different levels of fermented macroalga, Enteromorpha prolifera, on the growth performance, digestive enzyme activities and serum non‐specific immunity of red tilapia (Oreochromis mossambicus × Oreochromis niloticus). Fish were fed the control diet or the five diets supplemented with fermented E. prolifera for 7 weeks (diets 1–5 containing 10, 20, 30, 40 and 50 g kg^?1 of fermented E. prolifera respectively). The results showed that the percentage weight gain and specific growth rate (SGR) improved significantly in fish fed diets 3, 4 and 5, and regression analysis showed SGR reached the maximum in fish fed diet containing 37 g kg^?1 fermented E. prolifera, while the feed conversion ratio reduced significantly in fish fed diets 2, 3, 4 and 5, and regression analysis showed it reached its minimum in fish fed diet containing 41 g kg^?1 fermented E. prolifera. The pepsin, erepsin, gastric amylase, intestinal amylase, gastric lipase and intestinal lipase activities in fish fed diets 2, 3 and 4 were higher significantly than the control, with the exception of the pepsin activity in fish fed diet 4. The serum total superoxide dismutase, lysozyme, acid phosphatase and alkaline phosphatase activities enhanced significantly in fish fed diets 3 and 4. In conclusion, fermented E. prolifera had positive effects on the growth performance, digestive enzyme activities and serum non‐specific immunity of red tilapia and the recommended dose of fermented E. prolifera is 37–41 g kg^?1 in the diet of red tilapia.     

Dietary organic acids blend alone or in combination with an essential oil on the survival,growth, gut/liver structure and de hemato‐immunological in Nile tilapia Oreochromis niloticus

This study evaluated the effects of dietary supplementation of organic acids blend (OAB) alone or in combination with essential oil, Lippia origanoides (OAE) for Nile tilapia fed supplemented diets for 30 days. Fish (1.1 ± 0.04 g) were fed control (Control), or OAB 0.5% or OAB 0.5% + essential oil 0.125% (OAE) respectively. At the end of the experiment, samples were collected for de hemato‐immunological, histological analysis of the intestine and liver, as well as microbiology of the intestine. The pH of the diets supplemented with OAB and OAE reduced 0.92 and 0.19 respectively. The growth and FCR were unaffected by the treatments, but survival was significantly higher in the OAB treatment. Fish fed the OAB diet showed reduced concentration of total heterotrophic bacteria and Pseudomonas sp. in the intestine. Increased glucose in fish fed OAB and high number of circulating monocytes in fish fed OAE diet were observed. The anterior intestine of fish fed OAE diet showed larger number of goblet cells and increased villi height. The diet supplemented with OAB, mainly, improved the intestinal health and survival of tilapia juveniles and can be used in juvenile production.     

Growth,tissue metabolites and digestive functionality in Sparus aurata juveniles fed different levels of macroalgae,Gracilaria cornea and Ulva rigida

The aim of this study was to evaluate the inclusion of two macroalgae as dietary ingredients on growth, body composition, intermediate metabolism, intestinal enzymes and gut histology of Sparus aurata juveniles. Six isocaloric and isonitrogenous diets incorporating 5%, 15% or 25% of Gracilaria cornea or Ulva rigida meal were tested against a control diet. Duplicate groups of fifteen fish (14 g) were fed the experimental diets for 70 days. Growth performance showed an inverse linear relationship with the inclusion level of Gracilaria. Feed conversion ratio increased significantly in fish fed 25% Gracilaria. Ulva presented a quadratic relationship with the lowest SGR at 15% of inclusion. Fish fed 25% Ulva exhibited significantly lower lipid content than those fed control diet. Overall, metabolites did not show differences with specimens fed control diet, except for plasmatic glucose and muscle lactate. Intestinal proteolytic activity was reduced with the macroalgae inclusion level, but all fish showed the same enzyme pattern. The histological study revealed no negative effect on liver and intestine structure by the macroalgae inclusion. The present work demonstrated that these ingredients can be used in practical diets for sea bream, but a more comprehensive research is needed to determine the optimum dietary level in a longer feeding trial.     

Tributyrin‐supplemented high‐soya bean meal diets of juvenile black sea bream,Acanthopagrus schlegelii: Study on growth performance and intestinal morphology and structure

An 8‐week growth trial was conducted to evaluate the effects of different levels of tributyrin supplementation in a high‐soya bean meal diet on juvenile black sea bream (11.30 ± 0.16 g). The positive control (PC) diet contained 45% fishmeal and 20% soya bean meal, while the negative control (NC) contained 12% fishmeal and 45% soya bean meal. Graded levels of tributyrin were added to the NC diet at 0.05% (TB 0.05), 0.1% (TB 0.1), 0.2% (TB 0.2), 0.4% (TB 0.4) and 0.8% (TB 0.8). Ultimately, the fish fed the PC diet had a higher weight gain and specific growth rate than the fish fed other diets. The fish fed the NC diet had the lowest growth, and TB 0.05–TB 0.2 diets increased growth performance while TB 0.4–TB 0.8 diets caused reduction in growth. Dietary tributyrin supplementation improved protease activity and enhanced antioxidant capacity. Compared with the fish fed the NC diet, the fish fed the tributyrin‐supplemented diets had improved gut morphology and structure, and the results were similar to those of the fish fed the PC diet. Furthermore, the analysis of the dose response with second‐order polynomial regression indicated that the optimum tributyrin supplementation for juvenile black sea bream is 2.24 g/kg in the 45% soya bean meal diet.     

Effects of Dietary Vitamin C on Blood Chemistry and Nonspecific Immune Response of Juvenile Red Sea Bream,Pagrus major

A trial was conducted to determine the effect of ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) on blood chemistry and nonspecific immune response of red sea bream juveniles. Test diets with three levels of AsA (free, 107, and 325 mg/kg diet) were fed to juvenile red sea bream (36.0 ± 1.3 g) two times a day for 3 wk. There were no significant differences in hematocrit, glucose, and blood urea nitrogen. Total cholesterol and triglyceride in plasma of fish fed AsA‐free diet was significantly (P < 0.05) higher than that of fish fed two other diets. There were no significant differences in serum albumin, total bilirubin, and total serum protein. Glutamyl oxaloacetic transaminase in serum of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) lower than that of fish fed AsA‐free diet. Serum lysozyme activity (LA) of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) higher than that of fish fed AsA‐free diet. There was no significant difference in mucus LA. The results mentioned above demonstrated that AMP‐Na/Ca is a bioavailable AsA source for red sea bream juveniles. Supplement of more than 107 mg AsA/kg in diets improved blood chemistry and nonspecific immune function of red sea bream juveniles.     

Soybean meal as a protein source in gilthead sea bream (Sparus aurata L.) diets: effects on growth and nutrient utilization

The value of defatted soybean meal as a protein source for sea bream fingerlings (15.2±4.4 g on average) growing to market size (300–350 g) was evaluated by feeding extruded isonitrogenous and isoenergetic diets (46% protein and 22 MJ kg⁻¹) containing 20%, 30%, 40% and 50% soybean meal considering two phases. On day 87, the fish weight ranged between 66 and 81 g. The specific growth rate (SGR) of sea bream fed 50% soybean was lower (1.73% day⁻¹) than that of fish fed 20% (1.87% day⁻¹) and 30% (1.93% day⁻¹), but the food conversion ratio (FCR) was not significantly affected, and a quadratic significant trend was observed for the feed intake (FI) in relation to the dietary soybean level. At the end of the second phase on day 309, fish weight was between 303 and 349 g, but SGR and FCR were similar for all diets, and ranged between 0.64 and 0.69% day⁻¹, and 1.95 and 2.10% day⁻¹ respectively. The final biometric parameters were not affected by the diets, although the levels of some free amino acids in the muscle were affected. Sensory differences were detected by panellists in fish fed diet 20% as compared those fed diet 50%, which had a less marine flavour and was less juicy. The global growth results suggest the possibility of feeding sea bream weighing less than 80 g with 30% soybean meal, and for fish weighing more than 80 g, a 50% dietary soybean meal can be used until the fish reach commercial weight, with no negative effects on growth or feed efficiency. Nevertheless, when sensory analysis and economic aspects are considered, the maximum inclusion level of soybean was 20–22%.     

Effect of dietary vitamin C on the growth performance,antioxidant ability and innate immunity of juvenile yellow catfish (Pelteobagrus fulvidraco Richardson)

A 12‐week feeding trial was conducted to evaluate the effects of dietary vitamin C on growth performance, antioxidant status and innate immune responses in juvenile yellow catfish, Pelteobagrus fulvidraco. Six isonitrogenous and isolipidic diets (44% crude protein and 7% lipid) were formulated to contain six graded dietary vitamin C (ascorbate‐2‐poly‐ phosphate, ROVIMIX^® STAY‐C^® 35) levels ranging from 1.9 to 316.0 mg kg^?1 diet. The results of present study indicated that fish fed the lowest vitamin C diet had lower weight gain (WG) and specific growth rate (SGR) than those fed the diets supplemented vitamin C. WG and SGR did significantly increase with dietary vitamin C levels increasing from 1.9 to 156.5 mg kg^?1. However, no significant increase was observed with further dietary vitamin C levels increasing from 156.5 to 316 mg kg^?1. Survival, protein efficiency ratio and feed efficiency were not significantly affected by the dietary vitamin C levels. The activities of serum superoxide dismutase, catalase and glutathione peroxidase significantly increased when dietary vitamin C levels increased from 1.9 to 156.5 mg kg^?1, fish fed the lowest vitamin C diet had higher serum malondialdehyde content than those fed the diets supplemented with vitamin C. Fish fed the diet containing 156.5 mg kg^?1 vitamin C had the highest lysozyme, total complement activity, phagocytosis index and respiratory burst of head kidney among all treatments. The challenge test with Aeromonas hydrophila indicated that lower cumulative survival was observed in fish fed the lowest vitamin C diet. Analysis by broken‐line regression of SGR and lysozyme activity indicated that the dietary vitamin C requirement of juvenile yellow catfish was estimated to be 114.5 and 102.5 mg kg^?1 diet, respectively.     

Effects of diet supplementation of soya‐saponins,isoflavones and phytosterols on Atlantic salmon (Salmo salar,L) fry fed from start‐feeding

A 14‐week trial was conducted to investigate the effects of antinutritional factors (ANFs) commonly present in soybean ingredients, singly and in combination, on Atlantic salmon (Salmo salar L.) fed from start‐feeding. The experimental diets consisted of a negative control fish meal diet (FM), and a positive control diet with 167 g kg^?1 soybean meal inclusion (SBM) and four diets based on the FM diet supplemented with 2 g kg^?1 soya‐saponins (SAP), 1.5 g kg^?1 isoflavones (IFL), 0.3 g kg^?1 phytosterols (PHS) or a mixture of these (MIX). Fish fed the SAP diet showed significantly higher growth performance than those fed FM, while the IFL treatment significantly decreased growth performance of salmon fry. Fish fed the IFL diet had significantly lower maltase activity and higher trypsin activity in proximal intestine than fish fed the FM diet. Histological differences were observed in the liver of fish fed the IFL diet, characterized by reduced size of the hepatocytes. Fish fed the PHS and IFL diets showed the highest frequencies of skeletal deformities among the six treatments. In conclusion, the results indicate that purified isoflavones may negatively affect growth performance, intestinal function, liver metabolism and bone formation of salmon fry.     

Necessity of dietary taurine supplementation for preventing green liver symptom and improving growth performance in yearling red sea bream Pagrus major fed nonfishmeal diets based on soy protein concentrate

The necessity of dietary taurine supplementation for preventing green liver symptom and improving growth performance of red sea bream Pagrus major fed nonfishmeal (non-FM) diets was investigated. Yearling red sea bream (initial body weight, 580 g) were fed for 36 weeks on non-FM diets based on soy protein concentrate (SPC) supplemented with taurine at levels of 0%, 0.5%, 1.0%, 1.5%, and 2.0%. Specific growth rate (SGR) and feed conversion ratio (FCR) of fish fed the taurine-unsupplemented SPC diet were markedly inferior. In these fish, incidence of green liver was markedly higher and was accompanied by a decrease of tissue taurine concentration and an increase of hepatopancreatic bile pigment content. The green liver symptom was mainly caused by an increase of hemolysis since the erythrocytes became osmotically fragile due to taurine deficiency. Physiological abnormality and growth performance (SGR and FCR) were markedly improved by taurine supplementation to the SPC diets. These results indicate that dietary taurine supplementation is necessary for yearling red sea bream fed non-FM diet based on SPC to maintain normal physiological condition and growth performance.     

Effects of four feeding stimulants in high plant‐based diets on feed intake,growth performance,serum biochemical parameters,digestive enzyme activities and appetite‐related genes expression of juvenile GIFT tilapia (Oreochromis sp.)

The present research evaluated the effects of four feeding stimulants on feed intake (FI), growth performance, body composition, serum biochemical parameters, digestive enzyme activities and appetite‐related gene expression for juvenile GIFT tilapia. Five experimental high plant‐based diets (with 0, 0.4 g kg^?1 dimethyl‐β‐propiothetin (DMPT), 0.6 g kg^?1 dimethylthetin (DMT), 1.8 g kg^?1 tryptophan (Trp) and 6 g kg^?1 betaine (Bet), respectively supplemented) were fed to juvenile GIFT tilapia (Oreochromis sp.) (3.34 ± 0.01 g) for 8 weeks. The results showed that FI was significantly increased by Trp and DMT. The highest specific growth rate (SGR) and protein efficiency ratio (PER) was observed for fish fed Bet diet, followed by DMT. Serum triglyceride was significantly increased by DMPT. Fish fed Bet diet showed significantly higher intestinal protease, liver protease and intestinal amylase activities. The neuropeptide Y (NPY) mRNA expression in brain was significantly improved by Trp, and ghrelin mRNA expression in stomach was significantly increased by DMT. It is concluded that supplementation of Trp or DMT could act as an effective feeding stimulants (FS) in high plant‐based diets for juvenile GIFT tilapia, which might be associated with the relatively higher orexigenic (NPY or ghrelin) genes expression, and supplementation of Bet could improve SGR, PER and reduce feed conversion ratio (FCR) of juvenile GIFT tilapia.     

Effects of partial substitution of dietary fish meal by fermented soybean meal on growth performance,amino acid and biochemical parameters of juvenile black sea bream Acanthopagrus schlegeli

A feeding trial was conducted to evaluate the effect of replacing fish meal protein with fermented soybean meal (FSM) on the growth performance, feed utilization, amino acid profile, body composition, morphological parameters, activity of antioxidant and digestive enzymes of black sea bream (Acanthopagrus schlegeli) juvenile. Five isonitrogenic and isolipidic diets were prepared with levels of 0 (control), 80, 160, 240 and 320 g kg^?1 FSM. Triplicate groups (40 fish per tank) of juvenile black sea bream with initial weight of 1.17 ± 0.04 g were hand‐fed to visual satiation at three meals per day for 8 weeks. The fish fed diets containing different levels of FSM had no significant differences regarding survival and specific growth rate compared with control group. Feed and protein efficiency ratios of fish fed diet containing 320 g kg^?1 FSM were significantly lower than those of control group. Daily feed intake and daily protein intake of fish fed diet containing 240–320 g kg^?1 were significantly higher than those of control group. Hepatosomatic index and condition factor of fish were not affected by different dietary FSM level. Fish fed diets containing 240–320 g kg^?1 FSM had significantly higher visceral somatic index than control group. Whole body proximate and amino acid compositions of fish were not affected by dietary FSM level. The activity of digestive enzymes in the intestine was not affected by dietary FSM level. The activity of glutathione peroxidase in liver was significantly higher for fish fed the diet containing 160 g kg^?1 FSM compared with control group. This study showed that up to 40% fish meal in the diets of juvenile black sea bream could be replaced by fermented soybean meal with supplementation of methionine, lysine and taurine.     

Effect of dietary protein reduction with lysine and methionnine supplementation on growth performance,body composition and total ammonia nitrogen excretion of juvenile grass carp,Ctenopharyngodon idella

A 63‐day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant‐based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day^?1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP‐ and 28CP‐diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P < 0.05), and the interaction between dietary protein level and amino acid supplementation was noted between WG and SGR (P < 0.05). Feed intake (FI) was significantly increased with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05), but feed conversion ratio (FCR) showed a significant decreasing trend (P < 0.05). Two days after total ammonia nitrogen (TAN) concentration test, the values of TAN discharged by the fish 8 h after feeding were 207.1, 187.5, 170.6, 157.3, 141.3 and 128.9 mg kg^?1 body weight for fish fed 32CP, 32AA, 30CP, 30AA, 28CP and 28AA diets, respectively. TAN excretion by grass carp was reduced in plant‐based practical diets with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05). The results indicated that lysine and methionine supplementation to the plant protein sources‐based practical diets can improve growth performance and feed utilization of grass carp, and the dietary crude protein can be reduced from 320 g kg^?1 to 300 g kg^?1 through balancing amino acids profile. The positive effect was not observed at 280 g kg^?1 crude protein level.     

Beneficial effects of dietary exogenous protease on the growth,intestinal health and immunity of GIFT (Oreochromis niloticus) fed plant‐based diets

This study investigated the effects of dietary exogenous protease on the growth performance, intestinal health, immune parameters and disease resistance of genetically improved farmed tilapia (GIFT, Oreochromis niloticus). Five test diets with commercial protease at the levels of 0, 1.38, 2.76, 5.52 and 11.04 U/g (named PE0, PE1, PE2, PE5 and PE11, respectively) were administered to triplicate tanks with 30 fish for 60 days, and then, the fish were challenged with Streptococcus agalactiae for 14 days. The results indicated that weight gain increased as exogenous protease increased from 0 to 5.52 U protease/g diet and then decreased significantly (p < .05) with a further increase in exogenous protease supplementation (p < .05). The height of the villi in the proximal intestine and distal intestine, the width of the villi in three segments of the intestine, and the thickness of the muscle layer in the proximal intestine and mid‐intestine (p < .05) were increased in the fish fed the PE5 diet. Immune and antioxidant indices (except malondialdehyde), and survival after challenged with S. agalactiae were higher in fish fed PE5 diets than in those fed other diets (p < .05). In conclusion, 5.52 U/g protease supplementation in a plant‐based diet could promote the growth performance, intestinal physical barrier function, innate immunity and S. agalactiae resistance of GIFT.     

Effect of dietary administration of Psychrobacter sp. on the growth,feed utilization,digestive enzymes and immune responses of grouper Epinephelus coioides

A 60‐day feeding trial was conducted to assess the effect of incorporating dietary Psychrobacter sp., one predominant bacterium with antagonistic activity in the gut of fast‐growing grouper Epinephelus coioides, on the growth, feed utilization, digestive enzymes and immune responses of E. coioides at regular monthly intervals. The fish were fed with control diet (without probiont) or probiotic diet supplemented with 1.0 × 10⁸ CFU g^?1Psychrobacter sp. No significant improvement in weight gain (WG) and specific growth ratio (SGR) were observed in the probiotic group, but a significant improvement in feed conversion ratio (FCR) was observed in fish fed the probiotic diet for 60 days. Although not statistically significant, enhanced hepatopancreatic protease and lipase activities and intestinal amylase activity were observed in probiotic group at day 60. A slightly increase in phagocytic activity and phagocytic index was observed in fish fed probiotic diet for 60 days. Serum lysozyme activity showed no significant differences between the probiotic group and the control group during the whole experiment period, while an increasing trend of superoxide dismutase (SOD) activity in the probiotic group was observed. Serum complement component 3 (C3) level of the probiotic group was higher than that of control in the whole feeding period, while complement component 4 (C4) level of the probiotic group was significantly higher than that of the control at day 60. The results of the current study firstly demonstrated potential for Psychrobacter sp. as a novel probiont to improve feed utilization, digestive enzymes and innate immunity in E. coioides.