Estimating the quantitative essential amino acid requirements of striped bass Morone saxatilis, using fillet A/E ratios

In order to determine the essential amino acid requirements (EAA) of striped bass Morone saxatilis , fillets were analysed to ascertain the relative amino acid concentrations for determining A/E ratios ((EAA/total EAA) × 1000)). Analysis of the striped bass fillets yielded the following concentrations of essential amino acids (g kg^–1) and A/E ratios, respectively: arginine, 12.5, 115; histidine, 5.1, 47; isoleucine, 8.0, 74; leucine, 17.1, 157; lysine, 20.2, 186; methionine + cysteine, 9.2, 85; phenylalanine + tyrosine, 16.0, 147; threonine, 9.8, 90; tryptophan, 1.9, 18; and valine, 9.1, 84. In two experiments, diets with graded levels of EAA were fed to striped bass weighing 111 ± 3 g and 790 ± 122 g per fish, respectively. In both experiments, the dietary A/E ratios were maintained in the same relative concentrations as determined in the striped bass fillets. Statistical analysis of weight gains, feed conversions and nitrogen balance indicated significant differences ( P  < 0.05) between treatments. Non-linear regression analysis of the response criteria pooled from both experiments yielded the following estimates of dietary EAA requirements (g kg^–1 dry diet) when digestible energy equalled 13.39 MJ kg^–1 diet: arginine, 14; histidine, 6; isoleucine, 9; leucine, 19; lysine, 22; methionine + cysteine, 10; phenylalanine + tyrosine, 17; threonine, 11; tryptophan, 3; and valine, 10. The use of fillet A/E ratios allows for the rapid estimation of quantitative EAA requirements and the development of species specific diets for new aquaculture species. The data presented here are the first to simultaneously describe all the dietary EAA requirements for M. saxatilis.     

Re-evaluation of total sulphur amino acid requirement and determination of replacement value of cystine for methionine in semi-purified diets of juvenile Nile tilapia, Oreochromis niloticus

Two feeding experiments were conducted to re-evaluate the total sulphur amino acid (TSAA; methionine and cystine) requirement and determine the replacement value of cystine for methionine of juvenile Nile tilapia ( Oreochromis niloticus ). Semi-purified diets used in both experiments contained 3510 kcal gross energy and 280 g of protein per kilogram diet from casein, gelatin and crystalline amino acids. The basal diet of the first experiment contained 3.1 g methionine and 0.4 g cystine per kilogram. l -methionine was added to the seven remaining diets at 1.0 g kg⁻¹ increment to produce methionine levels ranging from 3.1 to 10.1 g kg⁻¹ diet. Each diet was fed to four replicate groups of juvenile Nile tilapia (1.28 g mean weight) in a recirculation system for 8 weeks. Broken-line regression analysis of weight gain data indicated that the TSAA requirement of juvenile Nile tilapia was 8.5 g kg⁻¹ of the diet or 30.4 g kg⁻¹ of dietary protein. In the second experiment, TSAA level was set at 95% of the requirement value determined in the first experiment. Seven diets were made with different ratios of l -methionine and l -cystine (20 : 80, 30 : 70, 40 : 60, 50 : 50, 60 : 40, 70 : 30 and 80 : 20, based on an equimolar sulphur basis). Each diet was also fed to four replicate groups of juvenile Nile tilapia (4.14 g mean weight) in a recirculation system for 8 weeks. Regression analysis of weight gain data using broken-line model indicated that cystine (on a molar sulphur basis) could replace up to 49% of methionine requirement in semi-purified diets for juvenile Nile tilapia.     

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

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

Methionine requirement of juvenile Japanese flounder Paralichthys olivaceus estimated by the oxidation of radioactive methionine

Growth and amino acid oxidation studies were conducted to estimate methionine requirement of juvenile Japanese flounder, Paralichthys olivaceus , by using the purified diets containing 500 g kg^–1 crude protein from casein, gelatine and crystalline amino acids (CAA). Diets with six graded levels of methionine (5.3, 8.3, 11.3, 14.3, 17.3 and 20.3 g kg^–1 diet) were fed to triplicate groups of the juvenile (initial weight 2.8 ± 0.05 g) twice a day for 40 days. To prevent leaching losses, CAA were precoated using carboxymethylcellulose (CMC), and further diets were bound by CMC and κ-carrageenan. Based on broken-line analysis of percentage weight gain and feed conversion efficiency, the methionine requirements of Japanese flounder in the presence of 0.6 g kg^–1 of cystine were 14.9 and 14.4 g kg^–1 dry diet, respectively. After the growth study was finished, a direct estimate of methionine requirement was made by examining the influence of dietary methionine level on ¹⁴C-methionine oxidation by determining radioactive carbon dioxide, protein and nonprotein fractions of the whole body. The dose–response curve between expired radioactive CO₂ and dietary methionine levels showed that the optimum methionine level for the flounder was estimated to be within the range of 14.3–17.3 g kg^–1 of diet in high agreement with values obtained from the growth study.     

Effects of dietary protein to metabolizable energy ratios and total protein concentrations on the performance of yellow perch Perca flavescens

Juvenile yellow perch Perca flavescens were fed semipurified diets with varying protein to metabolizable energy ratios (PME, g protein MJ⁻¹ metabolizable energy) and nutrient densities in three experiments to determine recommended dietary protein and energy concentrations. Experiment 1 fish (18.6 g) were fed diets containing 450 g crude protein kg⁻¹ dry diet and 14.5–18.8 MJ ME kg⁻¹ dry diet for 10 weeks. No differences were found in the growth of experiment 1 fish fed the different diets. Experiment 2 fish (21.9 g) were fed diets containing 15.7 MJ ME kg⁻¹ dry diet and 210–420 g crude protein kg⁻¹ dry diet for 8 weeks. Fish fed the diet containing 340 g kg⁻¹ protein (diet PME = 22) exhibited the greatest weight gain. Experiment 3 fish (27.1 g) were fed diets with a PME of 22 and varying nutrient density (yielding 205–380 g crude protein kg⁻¹ dry diet) for 8 weeks. No differences were found in the growth of experiment 3 fish. Yellow perch fed the semipurified diets exhibited increased liver fat content, liver size and degree of liver discoloration compared with fish fed a commercial fish meal-based diet. Liver changes may have resulted from high dietary carbohydrate levels. We conclude that a protein level of 210–270 g kg⁻¹ dry diet is suitable for juvenile yellow perch provided that the dietary amino acid profile and carbohydrate content are appropriate for yellow perch.     

Dietary methionine requirement of juvenile Arctic charr Salvelinus alpinus (L.)

The dietary methionine requirement of juvenile Arctic charr Salvelinus alpinus (L.) was assessed by feeding diets supplemented with graded levels of DL-methionine (9, 12, 15, 18, 21, and 24 g kg⁻¹dietary protein) for 16 weeks at 12°C. All diets contained 400 g kg⁻¹ protein, 170 g kg⁻¹ lipid, 66 g kg⁻¹ ash and an estimated 17.5 MJ digestible energy (DE) kg⁻¹. When live-weight gain was examined using quadratic regression, the estimate of methionine requirement for optimal growth was 17.6 g kg⁻¹ of dietary protein (DP) or 7 g kg⁻¹ of the diet. Requirements estimated on the basis of carcass protein and energy gains were 18.8 and 17.9 g kg⁻¹ DP, respectively. Plasma methionine concentrations and ocular focal length variability measurements did not provide a sensitive measure of requirement, because each responded in a linear fashion to increasing dietary methionine levels. Based on the prevalence of cataracts, the methionine level required to prevent lens pathology (26.7 g kg⁻¹ DP) appears to be higher than that required for maximum growth.     

Supplemental citric acid and particle size of fish bone-meal influence the availability of minerals in rainbow trout Oncorhynchus mykiss (Walbaum)

Juvenile rainbow trout Oncorhynchus mykiss (Walbaum) were fed six low-phosphorus (P) diets supplemented with two different sizes of ground fish bone-meals (fine, 68 μm or less; coarse, 250–425 μm) and a coarse bone-meal diet containing four levels of citric acid (0, 4, 8 or 16 g kg⁻¹ diet) to investigate the effects of pH and bone particle size on P bioavailability. The basal diet provided 3.4 g P   kg⁻¹ and bone-meal increased P contents to 5.4–6.0 g P   kg⁻¹. Coarse bone-meal diets supplemented with 0, 4, 8 or 16 g kg⁻¹ of citric acid had pH values of 6.0, 5.7, 5.4 and 5.0, respectively. Weight gain and whole-body water, protein and lipid contents were not influenced by bone-meal supplementation. Supplementing the basal diet with both coarse and fine bone-meal significantly increased whole-body ash content. Fish fed no bone-meal were hypophosphataemic compared with fish fed with either fine or coarse bone-meals. Phosphorus in fine bone-meal had higher availability than P in coarse bone-meal. Bone-meal supplementation significantly decreased whole-body manganese content from 8.9 μg g⁻¹ in fish fed no bone-meal to 2.3 and 4.5 μg g⁻¹ in fish fed with fine and coarse bone-meals, respectively. The concentration of magnesium increased but zinc concentration was not affected by bone-meal supplements. Citric acid increased whole-body ash content but the influence of citric acid on the body P content was not significant ( P  = 0.07). Dietary acidification by citric acid significantly increased whole-body iron in a linear fashion. The bioavailability of dietary P can be improved by fine grinding the bone in fish meals.     

Dietary arginine requirement of juvenile swimming crab,Portunus trituberculatus

An 8‐week feeding trial was conducted to determine the optimal dietary arginine requirement for juvenile swimming crab Portunus trituberculatus. Six isonitrogenous and isolipidic experimental diets were formulated to contain graded arginine levels which ranged from 15.9 to 33.0 g kg^?1. Each diet was randomly assigned to triplicate groups of 60 juvenile swimming crabs (4.72 ± 0.12 g). The results indicated that dietary arginine had significant effects on weight gain (WG), specific growth rate (SGR), protein productive value, feed efficiency and protein efficiency ratio. Weight gain and SGR significantly increased with the dietary arginine increasing from 15.9 to 27.4 g kg^?1, while with the further increasing from 27.4 to 33.0 g kg^?1, WG and SGR did not increase significantly. Maximum arginine, proline and total essential amino acid contents in muscle were observed in 27.4 g kg^?1 group diet. The swimming crab fed the diet with lower dietary arginine level showed higher AST and lower ALT in the serum. Crab fed with the lower dietary arginine level had significantly lower ALT in the serum than the other groups. Haemolymph indexes were significantly affected by the dietary arginine level except for the cholesterol concentration, and the highest values were all found in 27.4 g kg^?1 group diet. The two slope broken‐line model using SGR showed that the optimal dietary arginine requirement was 27.7 g kg^?1 of the dry matter (56.0 g kg^?1 dietary protein) for juvenile swimming crab.     

Effect of methionine on intestinal enzymes activities, microflora and humoral immune of juvenile Jian carp (cyprinus carpio var. Jian)

An 8-week feeding experiment was conducted to determine the effect of dietary methionine supplementation on intestinal microflora and humoral immune of juvenile Jian carp (initial weight of 9.9 ± 0.0 g) reared in indoor flow-through and aerated aquaria. Eight amino acid test diets (350 g kg⁻¹ crude protein, CP), using fish meal, soybean-condensed protein and gelatin as intact protein sources supplemented with crystalline amino acids, were formulated to contain graded levels of methionine (0.6–22.0%) at a constant dietary cystine level of 3 g kg⁻¹. Each diet was randomly assigned to three aquaria. Growth performance and feed utilization were significantly influenced by the dietary methionine levels ( P  < 0.05). Maximum weight gain, feed intake occurred at 12 g kg⁻¹ dietary methionine ( P  < 0.05). Methionine supplementation improved hepatopancreas and intestine weight, hepatosomatic and intestine index, intestinal γ-glutamyltransferase and creatine kinase activity, Lactobacillus count, Bacillus count, lysozyme activities, lectin potency, sim-immunoglobulin M content, addiment C3,C4 contents and serum total iron-binding capacity and declined Escherichia coli and Aeromonas counts. Quadratic regression analysis of weight gain against dietary methionine levels indicated that the optimal dietary methionine requirement for maximum growth of juvenile Jian carp is 12 g kg⁻¹ of the dry diet in the presence of 3 g kg⁻¹ cystine.     

Whole-body amino acid pattern of juvenile,preadult, and adult pacu,Piaractus mesopotamicus,with an estimation of its dietary essential amino acid requirements

In the present study, juvenile (live body weight, 54.3 ± 8.2 g), preadult (live body weight, 822.5 ± 33.9 g), and adult (live body weight, 1,562.8 ± 41.8 g) pacu, Piaractus mesopotamicus, were used to estimate their dietary essential amino acid (EAA) requirements using the whole-body amino acid (AA) pattern. The results showed that whole-body moisture, crude protein, total lipid, and ash contents expressed on a wet weight basis (%) were significantly different among the studied growth phases. No significant differences were observed in the dietary EAA requirements estimated for the three growth phases of pacu. These dietary EAA requirements were found to be different than those previously estimated for the same fish through its muscle AA pattern. Based on whole-body EAA to total EAA ratios {A/E ratios; [(each EAA/total EAA) × 1,000]}, EAA requirements were estimated to be histidine (0.42%), arginine (1.36%), threonine (0.82%), valine (0.90%), methionine (0.45%), isoleucine (0.83%), leucine (1.29%), phenylalanine (0.74%), lysine (1.64%), and tryptophan (0.14%) for pacu. These estimated requirements may serve as a reference line in the formulation of practical and experimental diets until dose–response-based optimum EAA requirements are available for pacu.     

Optimum dietary soybean meal level for maximizing growth and nutrient utilization of on-growing gilthead sea bream (Sparus aurata)

Six isonitrogenous [450 g kg⁻¹ crude protein (CP)] and isoenergetic diets (23 kJ g⁻¹) with six levels of defatted soybean meal inclusion (0, 132, 263, 395, 526 and 658 g kg⁻¹) in substitution of fish meal were evaluated in gilthead sea bream of 242 g initial weight for 134 days. Fish fed diets S0, S13, S26 and S39 had a similar live weight (422, 422, 438 and 422 g, respectively) but fish fed diets S53 and S66 obtained the lowest final weight (385 and 333g, respectively), and similar results were presented in specific growth rate (SGR). Fish fed diets S53 and S66 also obtained the highest feed conversion ratio (FCR). Quadratic multiple regression equations were developed for SGR and FCR which were closely related to dietary soybean level. The optimum dietary soybean levels were 205 g kg⁻¹ for maximum SGR and 10 g kg⁻¹ for minimum FCR. Sensorial differences were appreciated by judges between fish fed S0 and S39 soybean level, but after a re-feeding period of 28 days with diet S0, these differences disappeared.     

Dietary arginine requirement of juvenile grass carp Ctenopharyngodon idella (Valenciennes in Cuvier & Valenciennes, 1844)

This experiment was conducted to estimate the optimum requirement of arginine for juvenile grass carp Ctenopharyngodon idella. Six isonitrogenous (38%) and isoenergetics (16 MJ kg^?1) semi‐purified diets containing casein and gelatine with graded level of arginine (0.93, 1.20, 1.51, 1.84, 2.10 and 2.41 g 100 g^?1 DM) were formulated. Each diet was randomly assigned to triplicate groups of 25 fish each tank (initial weight: 3.84 ± 0.01) for 10 weeks. The highest weight gain (WG, %) was recorded when arginine level was 2.10% of the diet. Dietary arginine level higher than 1.84% significantly increased the protein contents of whole body. Whole body amino acid composition of juvenile grass carp was not significantly affected by the dietary arginine level. Plasma‐free arginine level was increased linearly with increasing of arginine level in the diets, and the plasma‐free ornithine level was significantly higher when the dietary arginine level was 2.41% compared with other groups. Quadratic model analysis of SGR data indicated that the minimum recommended dietary arginine requirement for grass carp was 2.17% of the diet, corresponding to 5.71% of dietary protein.     

Short-term dietary supplementation with the microalga Parietochloris incisa enhances stress resistance in guppies Poecilia reticulata

Two trials were conducted to determine the effects of dietary enrichments with the microalga Parietochloris incisa , rich in arachidonic acid (ARA), on stress resistance in guppies Poecilia reticulata . The microalga was added to commercial diets as a neutral lipid (NL) extract and its fractions or as broken cells. Experimental diets were applied for a period of 14 days. In trial 1, commercial diets were supplemented with NL (containing 25 mg ARA and 0.11 mg β-carotene g⁻¹ feed), its triacylglycerol (TAG) fraction (containing 25 mg ARA g⁻¹ feed and no β-carotene) and the β-carotene fraction (containing 0.11 mg carotenoid g⁻¹ feed and minute amounts of ARA). Neutral lipid-fed fish demonstrated the highest resistance ( P <0.05) to osmotic stress (32-ppt NaCl), followed by fish fed with diets supplemented with TAG and β-carotene alone, which were more resistant than control ( P <0.05). In trial 2, fish fed diets supplemented with higher levels of broken alga (26.1 mg ARA g⁻¹ feed) were more resistant ( P <0.05) to stress as compared with fish fed lower ARA (16.3 mg g g⁻¹) or an unsupplemented control diet. We suggest a dietary supplementation with broken P. incisa cells to enhance stress resistance in guppies before a stressful event.     

Quantitative dietary threonine requirement of juvenile Pacific white shrimp, Litopenaeus vannamei (Boone) reared in low-salinity water

An 8-week feeding trial was conducted to determine the threonine requirement of juvenile Pacific white shrimp Litopenaeus vannamei (Boone) in low-salinity water (0.50–1.50 g L⁻¹). Diets 1–6 were formulated to contain 360 g kg⁻¹ crude protein with fish meal, wheat gluten and pre-coated crystalline amino acids with six graded levels of l -threonine (9.9–19.0 g kg⁻¹ dry diet). Diet 7, which was served as a reference, contained only intact proteins (fish meal and wheat gluten). Each diet was randomly assigned to triplicate groups of 30 shrimps (0.48±0.01 g), each four times daily. Shrimps fed the reference diet had similar growth performance and feed utilization efficiency compared with shrimps fed the diets containing 13.3 g kg⁻¹ or higher threonine. Maximum specific growth rate (SGR) and protein efficiency ratio were obtained at 14.6 g kg⁻¹ dietary threonine, and increasing threonine beyond this level did not result in a better performance. Body compositions, triacyglycerol and total protein concentrations in haemolymph were significantly affected by the threonine level; however, the threonine contents in muscle, aspartate aminotransferase and alanine aminotransferase activities in haemolymph were not influenced by the dietary threonine levels. Broken-line regression analysis on SGR indicated that optimal dietary threonine requirement for L. vannamei was 13.6 g kg⁻¹ dry diet (37.8 g kg⁻¹ dietary protein).     

Use of salt to control ichthyophthiriosis and prevent saprolegniosis in silver perch, Bidyanus bidyanus

The ciliate protozoan, Ichthyophthirius multifiliis , and the fungus, Saprolegnia parasitica , cause the diseases ichthyophthiriosis and saprolegniosis respectively. Both diseases are difficult to control and can cause high mortalities of freshwater fish, including the Australian silver perch, Bidyanus bidyanus (Mitchell). The efficacy of salt (NaCl) in controlling and preventing these diseases in silver perch was evaluated in aquaria and tanks. Low pH levels were also evaluated as a control for ichthyophthiriosis. Concentrations of 2 or 3 g L⁻¹ salt controlled infestations of I. multifiliis , and fish were free of both theronts and trophonts by day 8 at temperatures of 17.3–21.3 °C and by day 6 at 19.2–23.5 °C. Fish treated with 1 g L⁻¹ salt remained infested and all fish in a control treatment (0 g L⁻¹ salt) died. Although the mean survival rates of infested fish at pH levels of 5 or 6 were only 13.9% and 7.6%, respectively, there were no theronts or trophonts on surviving fish after 12 days. Silver perch harvested from a pond and treated with 2 or 3 g L⁻¹ salt did not become infected with S. parasitica and survival was 100%, whereas 16.6% of untreated (0 g L⁻¹ salt) fish became infected and survival was only 66.7%. A concentration of 2 g L⁻¹ NaCl is recommended for the control of ichthyophthiriosis and the prevention of saprolegniosis in silver perch held in tanks, aquaria and re-circulating aquaculture systems.     

Predicting the optimal dietary essential amino acid profile for growth of juvenile yellow perch with whole body amino acid concentrations

Rapid methods of estimating dietary essential amino acid (EAA) requirements might facilitate increases in aquaculture production, particularly for new or emerging industries. We conducted a 12‐week feeding study to test the hypothesis that whole body EAA concentrations and the quantified methionine requirement could be used to predict the remaining dietary EAA requirements for juvenile all‐female yellow perch. Six purified diets were developed and fed to triplicate groups of fish for 12 weeks. The diets contained the EAA profile of fishmeal (FM), the profile as predicted by whole body analysis, the quantified methionine requirement and resulting A/E ratios (PRED), PRED + 20% of all EAA (PRED20), PRED + 40% of all EAA (PRED40), PRED + 20% threonine, isoleucine and tryptophan (PRED320), and PRED + 40% threonine, isoleucine and typtophan (PRED340). Mean weight gain and feed consumption were significantly higher in fish fed PRED20 (35.7 ± 3.2 and 55.0 ± 5.3 g, respectively) than in fish fed FSM (25.1 ± 0.4 g wt gain, 41.0 ± 1.9 g cons), PRED (23.4 ± 2.3 g wt gain, 40.1 ± 4.2 g cons) and PRED340 (22.9 ± 3.3 g wt gain, 35.0 ± 3.8 g cons). There was no significant difference in feed efficiency among treatments. We recommend an EAA profile similar to PRED20 for feeding all‐female juvenile yellow perch.     

A dietary energy level of 14.6 MJ kg−1 and protein-to-energy ratio of 20.2 g MJ−1 results in best growth performance and nutrient accretion in silver barb Puntius gonionotus fingerlings

Five iso-nitrogenous (300 g protein kg⁻¹ diet) and iso-lipidic (80 g kg⁻¹ diet) semi-purified experimental diets with variable energy levels of 10.5 (D-1), 12.5 (D-2), 14.6 (D-3), 16.7 (D-4) and 18.8 (D-5) MJ kg⁻¹ diets were fed to Puntius gonionotus fingerlings (average weight 1.79 ± 0.02 g) in triplicate groups (15 healthy fishes per replicate) for a period of 90 days to assess the optimum dietary energy level and protein-to-energy ratio (P/E). Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min⁻¹ were used for rearing the fish. Maximum specific growth rate, protein efficiency ratio, protein productive value, RNA : DNA ratio, whole body protein content, digestive enzyme activity and minimum feed conversion ratio was found in fish-fed diet D-3 with 14.6 MJ kg⁻¹ energy level. There were no improvements in all these parameters with the further rise in dietary energy level. Hence, it may be concluded that the optimum dietary gross energy level for maximum growth and nutrient utilization of silver barb is 14.6 MJ kg⁻¹ diet with a resultant P/E ratio of 20.2 g protein MJ⁻¹ diet, when the dietary protein and lipid are maintained at optimum requirement levels of 300 and 80 g kg⁻¹ diet, respectively, for this species.     

Effects of feeding Atlantic salmon (Salmo salar L.) imbalanced levels of lysine and arginine

Two experiments were performed to study the effects of imbalanced levels of arginine and lysine in diets of Atlantic salmon ( Salmo salar L.) with Zein as the major dietary protein source. In the first experiment, the effects of high levels of lysine on marginal levels of arginine and high levels of arginine on marginal levels of lysine were compared in fish fed these amino acids at either marginal or high levels. Dietary contents of arginine/lysine were 30.0/29.4, 17.0/29.4, 30.0/14.4 and 17.2/14.5 g kg^–1, respectively. In the second experiment, fish were fed increasing levels of lysine in a diet containing arginine at the requirement level. Dietary levels of arginine/lysine in the second experiment were 23.0/17.0, 23.0/34.3, and 23.0/46.4 g kg^–1.
Atlantic salmon fed marginal levels of lysine showed better growth and feed utilization when fed diets supplemented with high levels of arginine. Fish fed diets marginal in arginine exhibited increased growth and feed utilization when fed diets supplemented with high levels of dietary lysine. Increasing dietary lysine did not significantly affect growth in fish fed arginine at the requirement level. Increased growth in fish fed high dietary arginine levels might be explained by increased tissue concentrations of ornithine acting as a precursor for in situ synthesis of polyamines. A slight correlation between specific growth rate (SGR) and concentration of spermidine in muscle and between condition factor and concentration of total polyamines in muscle was seen. Abdominal injection of U-C¹⁴ arginine may indicate lysine inhibition of arginine availability in muscle tissue.     

Total sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton)

The quantitative dietary sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton), was determined by conducting a growth study. The experimental diets contained 400 g crude protein kg⁻¹ from casein, gelatine and supplemental crystalline amino acids. Diets containing six graded levels of methionine (3.2, 6.5, 9.0, 11.5, 14 and 16.5 g kg⁻¹) with a constant level of cystine (1.4 g kg⁻¹) were formulated and fed to triplicate groups of Labeo rohita fingerlings twice a day to satiation for 60 days. The optimum dietary requirement for methionine was estimated using the break-point regression analysis at 11.5 g kg⁻¹ of diet or 28.8 g kg⁻¹ of dietary protein. Thus the total sulphur amino acid (Met + Cys) requirement was determined to be 12.9 g kg⁻¹ of diet or 32.3 g kg⁻¹ of protein. Higher survival, specific growth rate and food conversion efficiency values were observed for fish fed the diet containing optimum levels of sulphur amino acids.     

Dietary threonine requirement to optimize protein retention and fillet production of fast‐growing Nile tilapia

Threonine is the third‐limiting essential amino acid in diets based on cereal ingredients. A 4‐week trial was conducted to determine the threonine requirement of large Nile tilapia based on fish growth, feed efficiency, body composition, protein and amino acid retention. Six hundred fish (563.3 ± 15.1 g) were distributed into twenty 1.2‐m³ cages. Five diets containing 288 g kg^?1 of crude protein, 12.7 MJ kg^?1 of digestible energy and 8.9, 10.5, 12.2, 13.7 and 15.4 g kg^?1 of threonine were elaborated. Fish were hand‐fed five times a day to extruded diets. Significantly, differences in growth performance and amino acids retention among the treatments were observed. Fish fed 10.5 g kg^?1 of threonine showed higher daily weight gain, gutted weight and fillet weight (P < 0.05) compared to fish fed with other experimental diets. Diets containing 8.9–15.4 g kg^?1 of threonine did not affect whole body and muscle proximate composition. Based on second‐order regression analysis, the dietary threonine requirement estimated based on final gain, fillet weight and fillet yield was 12, 12.1 and 11.5 g kg^?1 diet, respectively. The dietary threonine requirement for maximum fillet yield of Nile tilapia was estimated to be 11.5 g kg^?1.