Effects of dietary L‐lysine supplementation on growth,body composition and muscle‐growth‐related gene expression with an estimation of lysine requirement of GIFT tilapia

An 8‐week feeding trial was conducted with extruded practical diets containing 320 g/kg of crude protein from plant‐based sources to determine the effects of L‐lysine on growth performances, muscle‐growth‐related gene (myoD, myogenine and myostatin) expression and haemato‐biochemical responses in juvenile genetically improved farmed tilapia (GIFT). Five isonitrogenous and isoenergetic diets were formulated to contain graded levels of lysine (14.3, 16, 17.5, 19 and 20.5 g/kg of diet). Each diet was randomly assigned to triplicate groups of 30 juvenile fish (5.2 ± 0.1 g), which were fed thrice a day (9:00, 13:00 and 17:00 hr). Maximum growth performances were observed in fish fed with lysine at 19 g/kg of diet. There was no significant (p > .05) effect on whole‐body composition and amino acids profile by dietary lysine supplementation. Significant (p < .05) changes were observed in relative expression of muscle‐growth‐related genes namely myoD and myogenine and in plasma metabolites by dietary lysine supplementation. In contrast, the relative expression of myostatin was not affected by dietary lysine supplementation. Broken‐line regression analysis and second‐order polynomial regression analysis of weight gain and N gain against dietary lysine levels showed that the dietary lysine requirement for juvenile GIFT tilapia was 18–18.3 g/kg of diet and 19.3–19.5 g/kg of diet, respectively.     

Metabolic responses of juvenile GIFT strain of Nile tilapia (Oreochromis niloticus) to dietary L‐tryptophan supplementation

A 60‐day indoor feeding trial was conducted to evaluate the effects of dietary tryptophan supplementation on growth performances, whole‐body chemical composition, expression of muscle growth‐related genes (MyoD, myogenin and myostatin), and haematological and biochemical responses of juvenile genetically improved farmed tilapia (GIFT). Five corn–soy‐based isonitrogenous and isoenergetic diets were formulated to contain graded levels of dietary tryptophan (2.6, 3.2, 3.7, 4.2 and 4.8 g/kg of diet). Each diet was randomly assigned to triplicate groups of 30 fish (5.3 ± 0.1 g) per experimental unit, which were fed thrice a day (9:00, 13:00 and 17:00 hr). Maximum growth performances and feed utilization were observed in fish fed tryptophan at 3.7 g/kg of diet. There was no significant (p > .05) effect on whole‐body composition and amino acid profile by dietary tryptophan supplementation. However, significant (p < .05) differences were observed in plasma metabolites and the mRNA expression of MyoD, myogenin and myostatin. Serum cortisol level was found significantly lowest in fish fed tryptophan at 3.7 g/kg of diet. Second‐order polynomial regression analysis of weight gain and nitrogen gain against dietary tryptophan levels indicated that the optimum dietary tryptophan requirement for maximum growth and feed utilization of juvenile GIFT tilapia was 3.8 g/kg of diet.     

The Modification of Poultry By-product Meal for Use in Hybrid Striped Bass Morone chrysops×M. saxatilis Diets

Abstract— A pair of experiments were performed to assess amino acid supplementation of pet food grade poultry by‐product meal for utilization as the sole protein source for hybrid striped bass Morone chrysops×M. saxatilis. The first experiment determined the available amino acids from menhaden fishmeal and poultry by‐product meal for hybrid striped bass. The second experiment determined the efficacy of supplementing poultry by‐product meal with amino acids based on an ideal amino acid profile of hybrid striped bass muscle. The positive control diet contained 40% digestible protein solely from menhaden fishmeal and the negative control diet contained 40% digestible protein solely from pet food grade poultry by‐product meal. The negative control diet was additively supplemented with lysine, methionine, threonine, and leucine at 1.16,0.57,0.31 and 0.47% of the diet, respectively. Lysine supplementation alone did not improve fish performance based on any measured response. Moreover, the negative control diet and the lysine supplemented diet had lower weight gain and feed efficiency than the positive control diet. Supplementation of the diet containing pet food grade poultry by‐product with lysine and methionine; lysine, methionine, and threonine; or lysine, methionine, threonine, and leucine improved weight gain and feed efficiency above that of the negative control diet. The diet containing poultry by‐product supplemented with lysine, methionine, and threonine produced weight gains statistically indistinguishable from those of the positive control diet. Protein and energy retention efficiencies also improved with supplementation of at least lysine and methionine and were statistically indistinguishable from those observed in fish fed the positive control diet. Supplementation with lysine and methionine reduced the hepatosomatic index to levels similar to those found in fish fed the menhaden fishmeal diet. Intraperitoneal fat levels were similar among treatments (6.1‐6.6%) with the exception that fish fed the diet supplemented with lysine, methionine, and threonine exhibited lower (5.5%) fat levels. Supplementing the poultry by‐product meal diet with only lysine and methionine increased muscle ratio to levels equivalent to those found in fish fed the positive control (fishmeal) diet. In conclusion, amino acid supplementation of pet food grade poultry by‐product meal can be used to replace fishmeal in diets for hybrid striped bass without a reduction in fish performance.     

Effect of taurine supplementation on the metabolism and body lipid‐to‐protein ratio in juvenile Atlantic salmon (Salmo salar)

Previously, we reported that methionine intake determined the taurine concentration in the liver of on‐growing Atlantic salmon fed plant protein diets. Further, the methionine intake and/or the increased taurine concentration following increased methionine intake affected the liver lipid metabolism. The following study therefore aimed to test whether taurine affected the growth or the type of growth when added in high plant protein diets naturally low in taurine but equal and adequate in dietary methionine. Juvenile Atlantic salmon [initial body weight (BW) of 2 g] were fed plant protein diets (16.5% fishmeal), which were supplemented with taurine or not for a period of 56 days. As a control for growth and normal metabolism, a fishmeal‐based commercial diet (68% fishmeal) was used. Supplementation with taurine to high plant protein diets had a slightly negative effect on weight gain, but the final body weight was not different. Interestingly, the pool of free amino acids in the liver and muscle was significantly higher in fish fed the supplemented diet as compared with fish fed the plant protein diet without taurine supplementation. Liver polyamine concentration was higher in fish fed the supplemented diet than in fish fed the similar diet without supplementation. Additionally, juvenile salmon fed the plant‐based diet supplemented with taurine had a lower body lipid‐to‐protein ratio due to a reduced whole‐body lipid content, while the whole‐body protein content was similar between treatments. Our study thus indicates that the addition of a low concentration of taurine to high plant protein diets interacts with lipid metabolism and storage, concomitantly affecting the general metabolism as the concentrations of the free amino acids and polyamines in the liver were significantly higher. The possible reasons for these changes are discussed.     

d‐lysine can be effectively utilized for growth by common carp (Cyprinus carpio)

An 8‐week feeding trial was conducted to compare the effects of different lysine isomers on growth, nutrient utilization and metabolic enzyme activity in common carp (Cyprinus carpio). A basic experimental diet (the control group) with 30% of dietary protein from fish meal and the remaining 70% from soybean meal was used either with or without supplementation of d‐ lysine, l‐ lysine or dl‐ lysine, respectively. The total feed intake was significantly improved by supplementation with lysine regardless of isomeric form (P < 0.05). Supplementation with d‐ lysine or l‐ lysine (but not dl‐ lysine) generally increased the weight gain, specific growth rate, feed efficiency ratio and protein efficiency ratio. Dietary lysine supplementation generally increased the apparent digestibility of dry matter, crude protein and crude lipid, but no statistically significant differences were observed between the control group and the lysine‐supplement groups (P > 0.05). The d‐ amino acid oxidase and d‐ aspartate oxidase activities were generally higher in the liver, kidney and intestine of fish fed diet containing d‐ lysine compared to fish fed diet containing l‐ lysine. The lysine, protein and lipid gains in fish were markedly increased by the supplementation with d‐ lysine or l‐ lysine, and the lysine (r = 0.618, P < 0.05) and protein gains (r = 0.671, P < 0.05) were linearly related to dietary lysine intake. Lysine retention efficiency linearly decreased with the increase of dietary lysine intake (r = ?0.579, P < 0.05). These results indicate that common carp are capable of utilizing the supplemental crystalline d‐ lysine·HCl (4 g kg^?1 dry diet) for growth when the rest of dietary amino acids are protein‐bound forms.     

Optimizing zinc supplementation levels of rainbow trout (Oncorhynchus mykiss) fed practical type fishmeal‐ and plant‐based diets

Rainbow trout (23.1 ± 0.4 g) were fed either a fishmeal‐ or plant‐based diet supplemented with various levels of zinc (0, 15, 30, 60 or 120 mg kg^?1) for 12 weeks. Trout fed the fishmeal diet had significantly higher weight gain than with the plant‐based diet. Zinc supplementation in the fishmeal diet had no effect on growth performance, suggesting that additional dietary supplementation of zinc is not required. However, in trout fed the plant‐based diet, growth increased significantly up to 30 mg kg^?1 zinc after which growth was not affected. Trout fed the plant‐based diet containing no zinc exhibited severe growth retardation, and in fish fed the 0 and 15 mg kg^?1 zinc diets, cataracts were present. Use of broken‐line quadratic modelling suggests that dietary supplementation of zinc needed to prevent deficiency and promote adequate growth in rainbow trout fed the plant‐based diet in this study was 30.1 mg kg^?1 (80 mg kg^?1 total dietary zinc). This is higher than the NRC (2011, Nutrient Requirements of Fish and Shrimp) dietary recommended level of 15 mg kg^?1 for rainbow trout. Following the NRC recommendation could lead to zinc deficiency in rainbow trout fed a plant‐based diet.     

Effect of fishmeal replacement by poultry by‐product meal on growth performance,proximate composition,digestive enzyme activity,haematological parameters and gene expression of gilthead seabream (Sparus aurata)

Two feeding trials examined the replacement of fishmeal (FM) with poultry by‐product meal (PBM) in the diet of juvenile Sparus aurata. In Feeding trial I (100 days), three diets were formulated, where FM protein was replaced by 50% (PBM50) and 100% (PBM100) PBM, while in Feeding trial II (110 days), four diets were formulated using the same FM control diet, but FM was replaced at lower levels: 25% (PBM25), and 25% (PBM25 + ) and 50% (PBM50 + ) with the supplementation of lysine and methionine amino acids. PBM protein can successfully replace 50% of FM protein in the diet of S. aurata without adverse effects on survival, feed intake, growth performance and feed utilization, given that the diet is balanced with lysine and methionine. The proximate composition of body and muscle was unaffected by the diet, but the total FM replacement resulted in reduced lipid and energy contents in fish. A fifty per cent FM replacement by PBM did not affect haematological parameters indicating a good fish health. Similarities in trypsin and chymotrypsin activities with FM‐fed fish suggest a high digestibility of PBM. High dietary levels of PBM reduced the liver gene expression of GH/IGF axis and of cathepsin D suppressing fish growth and modulating the protein turnover.     

Fishmeal levels can be successfully reduced in white shrimp (Litopenaeus vannamei) if supplemented with DL‐Methionine (DL‐Met) or DL‐Methionyl‐DL‐Methionine (Met‐Met)

The main objective of this study was to evaluate the effect of methionine supplementation when reducing fishmeal levels in diets for white shrimp (Litopenaeus vannamei). Tested diets consisted of a positive control with 260 g/kg fishmeal (D1), two negative controls with 100 g/kg fishmeal and no amino acid (AA) supplementation (D2) or supplemented with lysine but not methionine (D3), and four additional diets with 100g/kg fishmeal supplemented with increasing levels of DL‐Met (1.0, 2.0 or 3.0 g/kg) (D4, D5, D6) or Met‐Met (1.0 g/kg) (D7). Each diet was fed to four groups of 30 shrimp for 8 weeks at a daily rate of 70 g/kg body weight. Reduction in fishmeal from 260 g/kg down to 100 g/kg did not significantly affect survival rate, feed conversion ratio (FCR), protein efficiency ratio (PER) or protein retention efficiency (PR%) of white shrimp. However, growth performance (final body weight, FBW; weight gain, WG; specific growth rate, SGR) was reduced when dietary fishmeal level was reduced from 260 g/kg (D1) to 100 g/kg without methionine supplementation (D2). The growth performance (FBW, WG and SGR) of shrimp was significantly increased by supplementation of the 100 g/kg fishmeal diet with increasing levels of DL‐Met (p < .05). Same performance as positive control (D1) was achieved with diets containing 100 g/kg fishmeal and supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. The highest values of growth performance (FBW, WG and SGR) were found in shrimp fed D6 and D7 diets, which were significantly higher than those of shrimp fed D2 and D3 diets (p < .05) but without statistical differences with shrimp fed D1, D4 and D5 diets (p > .05). The highest values of whole‐body and muscle protein contents were found in shrimp fed D1 diet, which were significantly higher than those of shrimp fed all other diets (p < .05). The highest value of intestinal tract proteolytic enzyme activity was found in shrimp fed Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) and 3 g/kg DL‐Met‐supplemented diet (D6) (p < .05). The highest values of apparent digestibility coefficients (ADCs) of dry matter and crude protein were found in Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) (p < .05). Shrimp fed the D1 diet showed the highest value of total essential amino acid (EAA) and was significantly higher than shrimp fed D2–D3 (p < .05) but without significant difference with shrimp fed D4–D7 (p > .05). In conclusion, results showed that same performance can be achieved with diets containing 260 or 100 g/kg fishmeal supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. Moreover, supplementation of limiting methionine in low‐fishmeal diets seems to improve the digestive proteolytic activity, improving digestibility of dry matter and protein, and eventually to promote growth of juvenile white shrimp in fishmeal reduction diets.     

Evaluation of Poultry By‐product Meal in Commercial Diets for Hybrid Striped Bass,Morone chrysops ♀ × Morone saxatilis ♂, in Pond Production

The efficacy of replacing fish meal with petfood‐grade poultry by‐product meal (PBM) on an ideal protein basis in commercial diets for hybrid striped bass (HSB) was evaluated under production conditions in pond culture. A generic production diet (GEN) for HSB was formulated to contain 45% protein, 12% lipid, and 3.7 kcal/kg. Protein in the generic diet was supplied by a mix of animal and plant sources typically used by the industry that included more than 20% select menhaden fish meal and less than 10% PBM. A positive control diet (GEN + AA) was formulated by supplementing the generic diet with feed‐grade Met and Lys to match the level of those amino acids in HSB muscle at 40% digestible protein. Substitution diets were formulated by replacing 35, 70, or 100% of fish meal in the GEN diet with PBM on a digestible protein basis and then supplementing with Met and Lys (designated 35PBM, 70PBM, and 100PBM, respectively) as needed to maintain concentrations equal to those in the GEN + AA diet. Diet formulation and extrusion were conducted by a commercial mill, and all diets met or exceeded known nutritional requirements for HSB. Twenty 0.10‐ha ponds (4 ponds/diet) were randomly stocked with juvenile HSB (76 ± 10 g; mean ± SD) at a density of 7400/ha and fed for 600 d (October 2004 to May 2006). Diets were fed once daily to apparent satiation to a maximum of 95 kg feed/ha. Total weight and number of fish in each pond were determined at harvest. Weight distributions in each pond were estimated by selecting every 15th fish during harvest. Subsets of ten fish from each of these samples were selected randomly for the determination of body composition and nutrient and energy retention. The availability of indispensable amino acids as well as ammonia production from the commercial test diets were determined in separate tank trials. Most production characteristics were not statistically different (P > 0.10) among dietary treatments. Distributions of individual fish weights from each of the ponds were not affected by poultry by‐product level in the diet. Multivariate analysis of body compositional indices grouped diets into two clusters composed of GEN, GEN + AA, 35PBM vs. 70PBM, and 100PBM mainly because fish fed the 70PBM and 100PBM diets had greater (P = 0.001) body fat (visceral somatic indices) than fish fed the other diets. Ammonia production in tanks was not different among diets and peaked 6–8 h after feeding when fish were fed at 1.5% of body weight; ammonia‐N excretion ranged from 197 to 212 mg/kg/d and 18.5–21.5% of nitrogen intake. Some imbalances in the levels and ratios of selected amino acids to Lys were found in the diets containing higher amounts of PBM and were attributed to a lack of accurate availability coefficients during formulation for some dietary proteins. These imbalances in essential amino acids may have been the predominant factor in the somewhat fattier fish observed fed diets containing the two highest levels of PBM. Nevertheless, these results from fish stocked at commercial densities and raised to market size in ponds suggest that formulating diets on an available amino acid basis for all protein sources while balancing limiting amino acids, particularly Met, Lys, Thr, and Trp, on an ideal protein basis will yield significant improvements in HSB performance when fed commercial diets in which all fish meal is replaced with PBM.     

Predicting dietary essential amino acid requirements for hybrid striped bass

Three experiments were conducted that were designed to evaluate our ability to predict essential amino acid (EAA) needs of hybrid striped bass using the quantified lysine requirement and whole‐body amino acid concentrations. In the first experiment, six diets containing various amino acid profiles were fed to triplicate groups of fish initially weighing 7.7 g per fish. At the end of the 8‐week experiment, no significant differences were detected in growth rates or feed efficiencies (FE) between fish fed a practical diet containing 510 g kg^?1 herring fish meal (FM) and fish fed a purified diet containing the amino acid profile of herring fish meal (CAA‐FM). Growth responses of fish fed purified diets containing 100 (HSB), 110 (HSB110), 120 (HSB120) or 140 g 100 g^?1 (HSB140) of the amino acid profile of hybrid striped bass whole‐bodies were significantly lower than those of fish fed diet FM. In the second experiment, triplicate groups of fish (5.6 g per fish) were fed diets containing various energy : protein (E : P) ratios (34.8, 41.2, 47.5 and 53.9 kJ g^?1 protein) and one of two amino acid profiles (CAA‐FM and HSB120) in a 4 × 2 factorial design. Carbohydrate concentration was varied to achieve the desired energy concentrations. At the end of the 8‐week experiment, weight gain and FE were significantly higher in fish fed diets formulated to simulate the amino acid profile of herring fish meal (CAA‐FM) compared with fish fed diets formulated to contain 120 g 100 g^?1 of the amino acid profile of hybrid striped bass whole‐bodies (HSB120). Weight gain, FE and survival data indicated the optimum dietary E : P was 41.2 kJ g^?1 protein. Dietary treatments in the final experiment included three amino acid profiles and four levels of lipid in a 3 × 4 incomplete factorial design. Dietary amino acid treatments included the amino acid profile of herring fish meal (CAA‐FM) or 120 g 100 g^?1 of the predicted EAA requirement profile for hybrid striped bass (HSB120). The amino acid profile of the remaining dietary treatment (PRED+) was similar to that of the HSB120 treatment, but contained additional threonine, isoleucine and tryptophan. Diets CAA‐FM and HSB120 contained either 90, 130, 170 or 210 g kg^?1 lipid, whereas diet PRED+ contained 130 g kg^?1 lipid. Dietary treatments were fed for 10 weeks to triplicate groups of fish initially weighing 81.0 g per fish. Weight gain and FE were not significantly affected by dietary amino acid profile. Feed efficiency was significantly reduced in fish fed diets containing 210 g kg^?1 lipid compared with fish fed diets containing 90–170 g kg^?1 lipid. Intraperitoneal fat (IPF) ratio and hepatosomatic index (HSI) values generally increased as dietary lipid concentrations increased. Total liver lipid concentrations were significantly reduced in fish fed diets containing 210 g kg^?1 lipid compared with those of fish fed 90–130 g kg^?1 lipid. Results of this study indicate an appropriate dietary amino acid profile can be predicted for hybrid striped bass using the quantified lysine requirement and whole‐body amino acid concentrations. Further, the optimum E : P appears to be 40 kJ g^?1 protein.     

Feeding of juvenile cobia,Rachycentron canadum: Evaluation of practical feeds,comparison of commercial fishmeal replacers,and estimation of essential amino acid requirements

Three studies were conducted with juvenile cobia, Rachycentron canadum: (Study 1) a 10‐week feeding trial within floating net cages to test the nutritional efficacy of different dietary feeding regimes (trash‐fish control diet, a semimoist diet, an in‐house dry formulated diet, and a commercial cobia feed); (Study 2) a 10‐week feeding trial within an indoor water‐recirculating tank‐based system to test the nutritional efficiency of different potential dietary fishmeal replacers (poultry byproduct meal, soy protein concentrate, feather meal), a diet without taurine supplementation, and a commercial cobia feed; and (Study 3) estimation of the essential amino acid (EAA) requirements of cobia based on EAA whole‐body composition in fast‐growing cobia fed a trash‐fish‐based diet. Fish performance in terms of growth and feed efficiency was the greatest within the outdoor net‐cage feeding trial, with fish fed the control trash‐fish‐based diet exhibiting the best performance. Although fish growth was poorer within the indoor feeding trials, fish performance was similar for most diets, with apparent crude protein digestibility coefficients of over 75% being obtained in all experimental diets in both feeding trials. The estimated EAA requirements of cobia obtained during this study were similar to those reported for other similar marine carnivorous fish species.     

Effect of dietary hydrolysate supplementation on growth performance,non‐specific immune response and disease resistance of olive flounder (Paralichthys olivaceus) challenged with Edwardsiella tarda

A 9‐week feeding trial was conducted to investigate the effects of dietary supplementation with protein hydrolysates on growth, innate immunity and disease resistance of olive flounder. A fishmeal (FM)‐based diet was regarded as a control, and three diets were prepared by partial replacement of FM with krill hydrolysate, shrimp hydrolysate or tilapia hydrolysate (designated as Con, KH, SH and TH, respectively). Triplicate groups of fish (24.5 ± 0.3 g) were fed one of the diets to apparent satiation twice daily for 9 weeks and then challenged by Edwarsellia tarda. Fish‐fed KH diet showed significantly (P < 0.05) higher growth performance and feed utilization compared with the Con diet. Dry matter digestibility of the diets was significantly increased by KH and TH supplementation. All the examined innate immune responses were significantly increased in fish fed KH diet. Significantly, higher respiratory burst and superoxide dismutase (SOD) activities were found in fish‐fed SH diet. Lysozyme and SOD activities were significantly increased in fish‐fed TH diet. However, no significant effect was found on fish disease resistance. This study indicates that dietary supplementation of the hydrolysates, particularly KH, can improve growth performance, feed utilization and innate immunity of olive flounder.     

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.     

Effects of dietary essential amino acid deficiencies on the growth performance and humoral immune response in silvery‐black porgy (Sparidentex hasta) juveniles

A 6‐week feeding trial was conducted for determining the effects of dietary essential amino acids (EAA) deficiencies on growth performance and non‐specific immune responses in silvery‐black porgy juveniles (4.7 ± 0.1 g initial weight). Eleven isoproteic (ca. 47%) diets were formulated including a control diet containing the optimum quantity of EAA, and ten EAA‐deficient diets. All diets contained 36% fish meal and 18.5% crystalline EAA and non‐essential amino acids (NEAA) as the main source of dietary proteins. All the EAA and NEAA incorporated in the crystalline amino acids mixture of the control diet simulated the amino acids profile of the fish meal. The other 10 EAA‐deficient diets were formulated by the deletion of each of the 10 EAA (crystalline form) from the control diet and replaced by a mixture of NEAA for the adjustment of dietary nitrogen contents. At the end of the experiment, fish fed with threonine‐deficient diet showed the lowest survival rate (p < .05), whereas growth performance decreased in fish fed all EAA‐deficient diets, although the reduction in body growth varied depending on the EAA considered. Plasma total protein decreased in all experimental groups except for fish fed the phenylalanine‐deficient diet. Fish fed with arginine‐ and lysine‐deficient diets had the lowest plasma C3, C4, lysozyme, total immunoglobulin and total superoxide dismutase activity (p < .05). Present results indicated that lysine, methionine and threonine were the most limiting EAA in terms of growth performance; however, arginine, threonine and lysine were the most limiting EAA for innate immunity responses in silvery‐black porgy juveniles.     

Assessing feasibility of replacement of fishmeal with enzyme‐treated feather meal in the diet of juvenile turbot (Scophthalmus maximus L.)

A feeding trial was conducted to examine the potential of enzyme‐treated feather meal (EFM) as a fishmeal substitute in the diet of turbot. Fishmeal (650 g/kg) was incrementally replaced (0, 160, 320, 480, 640 and 800 g/kg) by EFM (0, 80, 160, 240, 320, 400 g/kg) to prepare six experimental diets (Control, EFM8, EFM16, EFM24, EFM32 and EFM40). Diet EFM24 was supplemented with lysine and methionine to form seventh diet (EFM24 + AA). The eighth diet was formulated with 240 g/kg steam‐processed feather meal to replace 480 g/kg fishmeal (SFM24). Eight diets were each fed to triplicate groups of juvenile turbot (37.47 g) in a recirculating aquaculture system for 7 weeks. The results showed that fish fed diet EFM24 grew better than those fed SFM24 (p < .05). The growth performance, body composition and serum biochemical indices of fish fed diet EFM8 were not statistically different compared to the control treatment (p > .05). However, further increase in fishmeal replacement (32%–80%) affected negatively growth, nutrients digestibility, serum biochemical indices and body composition (p < .05). Supplementation of lysine and methionine (EFM24 + AA) significantly improved growth performance and diet utilization compared to EFM24 (p < .05). Altogether results suggested that the EFM could comprise 80 g/kg in a diet for juvenile turbot by concurrent reduction of the fishmeal content by 160 g/kg without lysine and methionine supplementation.     

Comparison of dietary essential amino acid requirements determined from group‐housed versus individually‐housed juvenile bluegill,Lepomis macrochirus

Two 60‐day experiments were conducted sequentially to determine (i) lysine requirement of juvenile bluegill, Lepomis macrochirus based on the dose–response method, (ii) requirements for other essential amino acids (EAAs) using whole‐body amino acid profile and (iii) whether differences in growth rates of group‐housed versus individually‐housed bluegills lead to different lysine requirement levels because of the presence and absence, respectively, of social hierarchies. Seven, semi‐purified, experimental diets (isonitrogenous, isocaloric) were prepared to contain graded levels of digestible lysine (10–31 g kg⁻¹). Experiment‐1 involved group‐housed bluegills (approximately 27 g, n = 10 fish/chamber, 4 chambers/diet) whereas experiment‐2 involved individually‐housed bluegills (approximately 30 g, n = 1 fish/chamber, 14 chambers/diet). Fish were fed twice daily to apparent satiation. Bluegill growth responses in both experiments generally improved (P < 0.05, anova ) with increasing dietary lysine levels from 10 to 16 g kg⁻¹, and then levelled off with further increase in lysine level (P > 0.05). Optimal dietary lysine level (digestible basis) was estimated to be 15 g kg⁻¹ based on broken‐line regression analyses of relative growth rate and feed conversion ratio with no differences being observed between the two rearing methods. Determined dietary requirement levels for other EAAs ranged from 2.4 g kg⁻¹ (tryptophan) to 15.3 g kg⁻¹ (leucine).     

Effect of replacing fish meal with soybean meal and of DL‐methionine or lysine supplementation in pelleted diets on growth and nutrient utilization of juvenile golden pompano (Trachinotus ovatus)

The experiment was conducted to determine the effect of dietary DL‐methionine addition in fish meal reduction diet for juvenile golden pompano (Trachinotus ovatus). The trial comprises the following of 11 diet treatments. (i) D1 diet as the standard was formulated with fish meal as the main protein source. (ii) D2–D5 diets were formulated to replace fish meal with soybean meal at 60, 120, 180 and 300 g kg^?1, respectively, and the amino acid profiles (including methionine and lysine) of D2–D5 were referred to the amino acid profile of D1. (iii) D6‐D9 diets were the same as D2–D5, respectively, but without methionine supplementation. (iv) D10 and D11 diets were the same as D3 and D5 diets, respectively, but without lysine supplementation. Each diet was randomly fed to groups of 20 fish (initial average weight about 18 g) per net cage (1.0 × 1.0 × 1.5 m) in triplicate and the feeding experiment lasted for 56 days. Weight gain (WG, %) and final body weight (FBW, g) of fish fed D3 diet were significantly higher than those of fish fed D8 and D9 diets (P < 0.05), but without significant differences with fish fed other diets (P > 0.05). Feed conversion ratio (FCR) of D3, D4 and D5 diet treatments were significantly lower than that of D11 diet treatment (P < 0.05), but without significant difference with other diet treatments (P > 0.05). Whole‐body protein contents of fish fed D3–D5 were significantly higher than those of fish fed D7–D9 (P < 0.05), but without significant differences with fish fed other diets (P > 0.05). In conclusion, the replacement of fish meal by soybean meal could reach 300 g kg^?1 without detrimental but profitable effect on growth and survival when enough methionine and lysine were supplemented in the diet; moreover, lysine is not the first‐limiting amino acid relative to methionine for juvenile T. ovatus from the present growth and proximate composition results.     

Effect of dietary microbial phytase supplementation on growth performance, nutrient utilization, body compositions and haemato-biochemical profiles of Cyprinus carpio (L.) fingerlings fed soyprotein-based diet

An 8‐week feeding trial was conducted in flow through system to examine the effects of dietary supplementation of microbial phytase on growth, nutrient utilization, haemato‐biochemical status and body compositions in common carp, Cyprinus carpio (L.) fingerlings (average weight 6.66 ± 0.08 g). The aim of experiment was also to determine whether dicalcium phosphates (DCP), tracemineral premix and lysine and methionine supplemental levels in carp diet could be reduced if microbial phytase was supplemented. Control diet (diet 1) supplemented with DCP, tracemineral premix and lysine and methionine without microbial phytase supplementation. Four experimental diets were formulated with reduction of either dietary DCP or trace minerals or lysine and methionine or all four supplementation levels by 100% of control diet without microbial phytase supplementation (diet 2, 3, 4 and 5 respectively). Another two experimental diets were formulated with reduction of all dietary DCP, trace minerals, lysine and methionine supplementation levels by 0 and 100% of control diet with dietary microbial phytase supplementation at 500 FTU kg⁻¹ diet (diet 6 and 7 respectively). After acclimation of fish for 2 weeks, 20 fish were randomly stocked into triplicate tanks for each of seven treatments (T₀, T₁, T₂, T₃, T₄, T₅ and T₆ for diet 1, 2, 3, 4, 5, 6 and 7 respectively) and fed with respective diet to apparent satiation. Results indicated that phytase was effective in releasing most of the phytate bound proteins, amino acids and minerals for optimum utilization and performance. Results also suggested that DCP, trace minerals and lysine and methionine supplementation level could be replaced by microbial phytase supplementation at 500 FTU kg⁻¹ in soybean‐based common carp diets without altering the optimum performance of fish. Optimum replacement level of dietary DCP, trace minerals and lysine and methionine with phytase supplementation at 500 FTU kg⁻¹ soybean‐based carp diet needs to be standardized in further investigations.     

A study on dietary l‐lysine requirement of juvenile yellow catfish Pelteobagrus fulvidraco

An 8‐week feeding trial was conducted to determine lysine requirement of juvenile yellow catfish (Pelteobagrus fulvidraco) by feeding formulated diets containing crystalline l ‐lysine. Six isonitrogenous and isoenergetic diets (405 g kg^?1 protein, 18 kJ g^?1 gloss energy) containing fish meal together with soybean protein concentrate as protein sources and fish oil together with soybean oil as lipid sources were formulated. Crystalline l ‐lysine was added into the six diets to acquire lysine concentrations of 17.3, 21.8, 26.0, 31.3, 35.5 and 41.9 g kg^?1 dry diets, respectively. Mixture of crystalline amino acid was supplemented to simulate the amino acid profile in muscle of yellow catfish. The results indicated that final body weight (FBW), weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and protein efficiency (PE) increased with the increase in dietary lysine level from 17.3 to 31.3 g kg^?1 of diet and then decreased as the dietary lysine levels further increased. No significant difference in survival rate was found among all the dietary treatments. One‐slope, quadratic broken‐line analysis on the basis of SGR showed that the dietary l ‐lysine requirement of juvenile yellow catfish was 33.1 g kg^?1 of dry diet (83.2 g kg^?1 of dietary protein).     

Dietary lysine requirement of juvenile dusky kob,Argyrosomus japonicus

To determine dietary lysine requirement of dusky kob, Argyrosomus japonicus, six isonitrogenous and isoenergetic diets (431 g/kg crude protein, 141 g/kg lipid and 20 kJ/kg) were formulated with graded levels of crystalline L‐lysine (18–42 g/kg of the dry diet). The protein source in the basal diet comprised fishmeal and soya, where a combination of L‐aspartic and L‐glutamic acids was maintained at a ratio of 1:1, and all diets were supplemented with a mixture of crystalline essential amino acids to simulate the amino acid profile in dusky kob. Dietary treatments were randomly assigned to triplicate groups of 12 fish (4.5 ± 0.2 g, mean weight; 66.5 ± 1.1 mm, total length ± SD), which were fed to apparent satiation three times daily for 12 weeks. The fish fed dietary L‐lysine at 21, 29 and 33 g/kg dry diet showed the highest specific growth rates (SGR) and the lowest feed conversion ratio. For most amino acids, retention in the body of the fish increased with an increase in dietary lysine from 18 to 21 g/kg, and it reached a maximum somewhere between 21 and 33 g/kg, where after amino acid retention decreased with increasing dietary lysine. Based on SGR and using segmented broken‐line analysis, the dietary L‐lysine requirement of juvenile dusky kob was estimated at 31.7 ± 1.6 g/kg dry diet corresponding to 73.5 g lysine per kg protein.