Effects of dietary protein, and fat level and rapeseed oil on growth and tissue fatty acid composition and metabolism in Atlantic salmon (Salmo salar L.) reared at low water temperatures

A 12‐week feeding trial was conducted to elucidate the interactive effects of dietary fat, protein contents and oil source on growth, whole body proximate composition, protein productive value (PPV) and fatty acid (FA) composition of muscle and liver in Atlantic salmon (Salmo salar L.)` at low water temperatures (4.2 °C). Triplicate groups of Atlantic salmon (initial weight 1168 g) were fed six isoenergetic diets, formulated to provide either 390 g kg⁻¹ protein and 320 g kg⁻¹ fat (high‐protein diets) or 340 g kg⁻¹ protein and 360 g kg⁻¹ fat (low‐protein diets). Within each dietary protein/fat level, crude rapeseed oil (RO) comprised 0, 30 or 60% (R0, R30, R60, respectively) of the added oil. After 12 weeks, the overall growth and feed conversion ratio (FCR) were very good for all treatments [thermal growth coefficient (TGC): 4.76 (±0.23); FCR: 0.85 (±0.02)]. Significant effects were shown owing to the oil source on specific growth rate and TGC only. The liver and muscle FA compositions were highly affected by the graded inclusion of RO. The PPV was significantly affected by the dietary protein level. The results of this study suggest that more sustainable, lower protein diets with moderate RO inclusion can be used in Atlantic salmon culture at low water temperatures with no negative effects on growth and feed conversion, no major detrimental effects on lipid and FA metabolism and a positive effect on protein sparing.     

The minimum dietary lysine requirement,maintenance requirement and efficiency of lysine utilization for growth of Atlantic salmon smolts

This study was conducted to evaluate and refine the dietary lysine requirement of Atlantic salmon (Salmo salar) smolts using both the dose–response technique to determine the minimum dietary requirement and different ration levels to determine the maintenance lysine requirement and the efficiency of lysine utilization for growth. Diets containing 1.97–3.16% lysine (presented in protein‐bound form of lysine) were fed to satiation or at 25–75% ration levels to smolts during the last week in fresh water and the following 9 weeks in seawater. Based on the exponential lysine gain response to increasing dietary lysine concentrations, the minimum lysine requirement was found to be 2.67% of diet (dry matter basis; protein‐bound form or 3.05% free form). The smolts fed the unsupplemented diet maximally utilized the digestible lysine intake for growth. The efficiency of digestible lysine utilization for growth was 0.85 in the diet containing 2.88% lysine and the maintenance requirement was 21.7 mg digestible lysine kg^?0.7 day^?1.     

Establishing an upper level of intake for vitamin A in Atlantic salmon (Salmo salar L.) postsmolts

The vitamin A (VA) concentration in salmon aquaculture feeds is varying and may lead to sublethal adverse effects. In this study, 135 g Atlantic salmon postsmolts were given eight diets in duplicates with 6, 12, 26, 55, 82, 112, 360 and 749 mg retinol (ROL) kg⁻¹ for 116 days. Subsequently, fish given 6, 82 and 749 mg ROL kg⁻¹ were transferred to a common net pen and given a standard commercial diet for further 28 weeks. Feed conversion rate, liver functionality and markers of VA homoeostasis were not negatively affected by dietary VA level, but chronic high VA intakes led to adverse effects on growth and bone health. In plasma, there was an antagonistic effect of dietary ROL on circulating 1,25 (OH)₂ vitamin D₃ (calcitriol). Moreover, a dose–response of VA on craniofacial deformities, condition factor and vertebral morphometry and mechanical strength was observed. Vertebral deformities were observed after 28 weeks on a standard diet and not immediately after the 116 days on the experimental diet. Elevated VA is a risk factor for bone deformities, and the dietary intake of VA should not exceed 37 mg ROL kg⁻¹ body weight day⁻¹ in Atlantic salmon postsmolts.     

Modelling of threonine and methionine requirements of Oreochromis niloticus due to principles of the diet dilution technique

Two growth studies (56 days) were conducted with juvenile all male Oreochromis niloticus, using six mixed diets with threonine and five mixed diets, with methionine as the first limiting amino acid (LAA), respectively. Diets were randomly assigned to four repetition tanks with 25 fish per tank. Growth results and body nutrient analyses yielded protein deposition data depending on the uptake of the LAA. For further modelling, a nonlinear N‐utilization model was utilized for assessing the dietary efficiency of threonine and methionine utilization, respectively. By use of the observed dietary AA efficiency, threonine and methionine requirement data were established, depending on the daily protein deposition. The optimal dietary AA concentrations (8.3 g kg⁻¹ versus 9.9 g kg⁻¹ threonine; 7.3 g kg⁻¹ versus 8.8 g kg⁻¹ methionine) were derived for 70% of the potential for protein deposition (ND_maxT) and predicted daily feed intake (3% versus 2.5% of BW). Observed methionine requirements are imposing for the upper limit for methionine replacement by cystine and minimal catabolism of methionine to yield cystine. Generally, the modelling provided requirement data in line with current recommendations. Variation of the dietary AA efficiency by modelling procedure ranks this factor as very important for the conclusion of amino acid requirement data.     

Energy and nutrient utilization of Atlantic cod, Atlantic salmon and rainbow trout fed diets differing in energy content

The growth and feed utilization of Atlantic cod (Gadus morhua) (437 g), Atlantic salmon (Salmo salar) (485 g) and rainbow trout (Oncorhynchus mykiss) (413 g) fed a diet (170 g kg⁻¹ fat, 600 g kg⁻¹ crude protein; LE) similar to that used in commercial cod production or one that was top dressed with additional fat (280 g kg⁻¹ fat, 530 g kg⁻¹ crude protein; HE), were compared in an 11‐week trial. In the cod, relative feed intake was 41–58% and thermal growth coefficient 63% of that in the salmonids, but the feed efficiency ratio (FER) was 38% better (P ≤ 0.05). In contrast to the cod where there was no effect of diet on feed intake, growth or FER, both the salmon and trout fed the HE diet had greater feed intake than those fed the LE diet, but the effect of this was only positive for growth in the salmon. The cod retained more of the digested nitrogen (44.9 ± 2.7%) than the salmon (39.4 ± 0.8%), and both of these species retained more than the trout (33.6 ± 1.1%) (P ≤ 0.05). The retention of digested energy was significantly higher in the salmon (52.2 ± 0.9%) than in the trout (44.8 ± 1.1%), with the cod (44.9 ± 4.9%) not different from either of the other species. There were no differences between the species in the retention of absorbed phosphorus (65.9 ± 3.6%). There were very few dietary effects on nutrient utilization in this trial and, for the cod, this indicates that higher energy diets may be feasible for use in production.     

Dietary protein requirement of zebra sea bream (Diplodus cervinus,Lowe 1838) juveniles

A 14 weeks growth trial was performed to estimate the protein requirement for growth and maintenance of zebra sea bream (Diplodus cervinus) juveniles. For that purpose, nine isolipidic diets were formulated to contain increasing protein levels (from 50 to 550 g kg^?1) at the expense of carbohydrate. Each diet was assigned to duplicate groups of 20 fish, with an average body weight of 7.7 g. Feed efficiency improved with dietary protein up to 400 g kg^?1, no further differences being noticed at higher protein levels. Fish fed the 50 g kg^?1 protein diet lost weight during the trial. In the other groups, weight gain improved as dietary protein increased up to 350–400 g kg^?1. Fish fed diets with 250 g kg^?1 protein or lower had lower whole‐body protein content than the other groups. A curvilinear‐plateau model was used to adjust weight gain and protein gain (g kg ABW^?1 day^?1) to dietary protein levels. Based on that model, the optimum dietary protein requirement for maximum weight gain was estimated to be 437.6 g kg^?1 and for maximum protein gain 461.9 g kg^?1, corresponding to a protein intake of 7.63 g kg ABW^?1 day^?1. Protein requirement for maintenance was estimated to be 1.01 g kg ABW^?1 day^?1.     

Evaluation of silver perch (Bidyanus bidyanus, Mitchell) nutritional requirements during grow-out with high and low protein diets at two feeding levels

Silver perch (Bidyanus bidyanus, Mitchell) of 65 g average weight were reared at high density under controlled conditions on diets containing 24.8% and 40.6% protein. Diets were fed at 2% or 4% of the fish biomass day^?1. Both protein concentrations and feeding level of the diet influenced growth and proximate composition of the fish at the end of the 50‐day growth trial. Energy requirement for maintenance for these fish was found to be 77 cal BW^?0.8 and for each unit of energy retained 3.78 cal BW^?0.8 needed to be supplied, and for each gram of protein retained per metabolic body weight 5.04 g were required. The results indicate that it is possible to obtain the same weight increment when feeding a 24.8% protein diet at a level of 4% body weight day^?1 as compared with only 2% body weight day^?1 of a 40.6% protein diet. This study indicates that the nutritional requirements for early grow‐out are similar to those found in a previous study with juvenile silver perch and adds information concerning the required relationships between dietary amino acids. The results provide information required for further refinement of nutrition for this species.     

The effect of feeding rate on the growth performance of green sturgeon (Acipenser medirostris) fry

Four one‐week growth trials were conducted on green sturgeon fry to determine the effect of feeding rate on their growth performance at 18 °C when they were fed a salmonid soft moist feeds containing 445–457 g kg^?1 of crude protein and 201–207 g kg^?1 of lipid. The fry used in Trials I‐IV were 5–8 weeks after their initiation of exogenous feeding. Their average initial body weights were 1.63 ± 0.01, 2.63 ± 0.03, 5.08 ± 0.08 and 7.49 ± 0.05 g, respectively. Six feeding rates used were as follows: 2.5–15.0% body weight per day (% BW day^?1) with a 2.5% increment in Trial I; 1.25–7.50% BW day^?1 with a 1.25% increment in Trial II; and 2.0–7.0% BW day^?1 with a 1.0% increment in Trials III and IV. Four replicates with 50 fry per tank in Trials I‐III and 30 fry per tank in Trial IV were assigned randomly to each feeding rates. The final body weight, specific growth rate, feed efficiency, protein retention, and whole‐body moisture, lipid, and energy contents were significantly (P < 0.05) affected by the feeding rates. The optimum feeding rates determined by the broken‐line model were 7.1, 5.7 and 5.3% BW day^?1 for Trials I, II and IV, when the fry were 5, 6 and 8 weeks after their initiation of exogenous feeding, respectively.     

Protein and lysine requirements for maintenance and for tissue accretion in Atlantic salmon (Salmo salar) fry

Available data on the quantitative requirement for lysine (Lys) in different salmonids show much variability. So far, there are very limited data on the maintenance requirements of indispensable (I) amino acids (AA) in fish. In the present study, we determined simultaneously the Lys requirements for maintenance and for protein accretion in Atlantic salmon fry by adapting a protocol established for the piglet. Groups of fish having an initial body weight of 1.5 g were fed for 28 days on isoenergetic diets with increasing nitrogen (N) content supplied by cod meal and a mixture of crystalline AAs (50% of total dietary N). Except the protein-free diet (PF; 0.2% dry matter (DM)), the N content of the other diets was either low (2.9% DM), medium (6.2% DM) or high (8.5% DM). Two types of diets with the same N content were formulated. The AA pattern of three control diets was based on the AA pattern of the cod meal protein. For the other three diets, Lys·HCl was totally omitted from the low-N diet (LPD), and 50% of Lys·HCl was removed from the medium- and high-N diets. After a 28-day feeding trial, carcass N and Lys gains were estimated. N and Lys requirements for maintenance and for growth were calculated regressing daily N or Lys gain against N or Lys intakes. The daily N requirement for growth above maintenance was 3.05 g per g protein gain and for maintenance it was 54 mg kg body weight^− 0.75. From the regression between protein and Lys intake, we calculated that for the accretion of 1 g body protein, the dietary Lys requirement was 152 mg, and that the Lys maintenance requirement for zero N gain was 20 mg kg body weight^− 0.75/day. This last value is higher than the previous estimations obtained for rainbow trout and could be explained by the lower body weight or age of the fish used here.     

Hypervitaminosis A in first‐feeding fry of the Atlantic salmon (Salmo salar L.)

Atlantic salmon fry were reared on a fishmeal based diet with increasing levels of vitamin A (VA) (6, 122 and 938 mg retinol kg^–1 dry feed) from startfeeding and for 14 weeks. Signs of VA stress, such as reduced fat stores, liver size and growth, were found for groups receiving 122 and 938 mg retinol kg^–1. Signs of vitamin A toxicity, such as increased mortality, abnormal vertebral growth, and reduced growth, were found for groups receiving 938 mg retinol kg^–1. These results suggest that excess VA in the early life stages of Atlantic salmon is deleterious for normal development.     

Protein-sparing effect of carbohydrate in silver barb, Puntius gonionotus fry

A 30‐day study was undertaken to examine the protein‐sparing effect of carbohydrate in diets for silver barb, Puntius gonionotus fry. Six semi‐purified experimental diets were formulated with two levels of protein (200 and 250 g kg⁻¹ diet) and three levels of carbohydrate (300, 340 and 380 g kg⁻¹ diet). In addition to the six experimental diets, a diet containing the protein and carbohydrate requirement levels of 300 and 260 g kg⁻¹ diet, respectively, as reported earlier for this species, was used as a reference diet. For each dietary treatment, 30 healthy fry of 20 days age (0.12 ± 0.01 g) were stocked in triplicate tanks using a flow‐through system. The fish were fed ad libitum four times a day to a level close to apparent satiation. Batch weighing of fish was done after 15 days of stocking to measure growth and general health status of the fish. The fish fed 250 g protein and 340 g carbohydrate kg⁻¹ diet with a protein to energy ratio of 17.86 g protein MJ⁻¹ performed equally well in terms of growth and nutrient utilization as the reference diet group. The study indicates that dietary protein can be reduced from 300 to 250 g kg⁻¹ diet by increasing carbohydrate from 260 to 340 g kg⁻¹ diet without sacrificing the growth of silver barb fry.     

Methionine and lysine requirements for maintenance and efficiency of utilization for growth of two sizes of tilapia (Oreochromis niloticus)

Different ration levels were used to determine the digestible methionine (DMet) and lysine (DLys) maintenance requirements and the utilization efficiencies for gain above maintenance for two different sizes of tilapia (20.7 and 165 g), by feeding a soybean meal–based diet. Protein gain and amino acid (AA) gain (e.g. methionine, Met; lysine, Lys; R² = 0.98) were best‐fit linear functions of DMet and DLys intake in both fish size classes. Slopes of these regression lines showed that the DMet utilization efficiencies for growth were 0.76 and 0.55 for juvenile and adult fish, respectively. The DMet maintenance requirements were 3.12 and 16.5 mg BW(kg)^−0.7 day⁻¹ for juvenile and adult fish, respectively. The DLys utilization efficiencies for gain were 0.72 and 0.52, whereas the DLys maintenance requirements were 16.9 and 68.8 mg BW (kg)^−0.7 day⁻¹, for juvenile and adult fish, respectively. These results suggested that there was an obvious difference in the maintenance requirements and utilization efficiencies for gain above maintenance for DMet and DLys in two different sizes of tilapia. The AA maintenance needs increased as fish increased in size, being greater in adult fish than in juvenile; however, the AA utilization efficiencies for gain above maintenance decreased with the increment of fish size.     

A factorial approach to defining the energy and protein requirements of Tra Catfish,Pangasianodon hypothalamus

This study assessed the protein and energy requirements of Tra catfish (Pangasianodon hypothalamus) using a bio‐energetic factorial approach. To examine these parameters, a series of inter‐related studies were undertaken to define the various factorial parameters. Studies included controlled laboratory experiments and field‐data collection from commercial farms in the Mekong Delta region of Vietnam. Key components of the model include parameters for maintenance energy (39.7 kJ kg^?0.844 day^?1 at 32 °C) and protein (0.467 g kg^?0.833 day^?1 at 32 °C) demands, the effect of animal live‐weight and temperature on growth and maintenance energy demands, the efficiencies of energy and protein utilization at various energy and protein intake levels and the variability in whole body composition with varying live‐weight. Interestingly, both the energy [0.510· (Energy intake) ? 11.143] and protein [0.322·(Protein intake) + 0.665] utilization efficiencies at near satiety feed intake levels were substantially lower than that observed from other fish species. Modelling using these equations was used to iteratively derive optimal diet designs and feeding rations for growing Pangasius catfish.     

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.     

Bioavailability of oxytetracycline from medicated feed administered to Atlantic salmon (Salmo salar L.) in seawater

Bioavailability of oxytetracycline HCl was investigated in Atlantic salmon (Salmo salar L.) in seawater at 7–8 °C. A group of 80 fish received an intravascular injection of 20 mg kg⁻¹ body weight. Another group of about 90 fish were force-fed with medicated feed containing a dose of 50 mg kg⁻¹ body weight. The oral bioavailability was found to be only 2%.     

Dietary ascorbic acid requirement of juvenile ayu (Plecoglossus altivelis)

To investigate dietary ascorbic acid (AA) requirement of juvenile ayu (Plecoglossus altivelis) weighing 1.27 ± 0.02 g, eight diets were formulated with graded levels (0, 20, 40, 80, 160, 320, 640 and 1280 mg AA kg^?1) of AA supplied as ascorbyl polyphosphate. Each experimental diet was fed to four‐replicate groups to apparent satiation three times a day for 8 weeks. At the end of the feeding trial, fish fed AA‐deficient diet showed visible AA deficiency signs and low survival. Based on the four‐parameter saturation kinetics model, the calculated AA requirement levels for each dose‐dependent response [weight gain, hepatic AA concentration, hydroxyproline (HyPro) concentration in skin and HyPro concentration in backbone] were 116, 226, 47 and 35 mg kg^?1, respectively. Based on the maximal growth performance, a level of 116 mg AA kg^?1 was recommended for commercial diet of juvenile ayu. To maintain tissue HyPro saturation and avoid AA deficiency symptoms, the minimum required dietary AA level was 47 mg kg^?1. Hepatic AA saturation was considered as the most stringent criterion for determination of AA requirement.     

Dietary arginine requirement of fingerling hybrid Clarias (Clarias gariepinus×Clarias macrocephalus)

The dietary arginine requirement of fingerling hybrid Clarias (Clarias gariepinus×Clarias macrocephalus) (4.2±0.03 cm, 0.56±0.04 g) was determined by feeding six isonitrogenous (400 g kg⁻¹ crude protein) and isocaloric (17.9 kJ g⁻¹) amino acid test diets containing casein, gelatin and l ‐crystalline amino acids with graded levels of arginine (10.0, 12.5, 15.0, 17.5, 20.0 and 22.5 g kg⁻¹) for 4 weeks to triplicate groups. Diets were fed twice a day at 09:00 and 16:00 hours at 8% body weight day⁻¹. Maximum weight gain (523%), best feed conversion ratio (FCR, 1.41), protein efficiency ratio (1.78) and specific growth rate (6.53%) were recorded in fish fed the diet containing arginine at 20.0gkg⁻¹ of the diet. Second‐degree polynomial regression analysis of live weight gain and FCR values indicated the dietary arginine requirement at 17.8 and 20.0 g kg⁻¹ of dry diet respectively. Significantly higher carcass protein and protein deposition values were recorded at the requirement level (20.0 g kg⁻¹). Higher fat and lower moisture values were obtained in carcass of fish fed the diet with 15.0g kg⁻¹ arginine. The maximum carcass ash value was noticed in the fish fed at 20.0 g kg⁻¹ dietary arginine. We recommend that the diet for hybrid Clarias (C. gariepinus×C. macrocephalus) should contain arginine in the range of 17.8–20.0 g kg⁻¹ of the dry diet, corresponding to 44.5 and 50 g kg⁻¹ of dietary protein respectively.     

Dietary lysine requirement of large yellow croaker (Pseudosciaena crocea,Richardson 1846) larvae

A 30‐day feeding experiment was conducted to estimate the lysine requirement of large yellow croaker larvae (2.75 ± 0.11 mg). Six isonitrogenous (509.5–519.7 g kg^?1 crude protein) and isoenergetic (22.3–22.5 kJ g^?1 energy) microdiets containing graded levels of lysine·HCl ranging from 24.8 to 41.0 g kg^?1 diet in placement of glycine and glutamic acid were formulated. Mixture of crystalline amino acids (MAA) was supplemented to simulate the amino acid (AA) profiles of whole body of this larva, except for lysine. The MAA and supplemented lysine for each diet were coated with tripalmitin. Triplicate groups of 3000 fish were fed to apparent satiation by hand eight times per day. The results showed that specific growth rate (SGR), survival, body composition and the specific activity of digestive enzymes were significantly affected by dietary lysine levels (P<0.05). The optimal dietary lysine requirements estimated by second‐order polynomial model based on SGR and survival were 33.7 (65.5 g kg^?1 dietary protein) and 33.4 (64.9 g kg^?1 dietary protein) g kg^?1 dry diet respectively. The estimated requirements for the other essential AAs were calculated by A/E ratios of whole body AA profile of this larva based on lysine requirement.     

Dietary arginine requirement of Heteropneustes fossilis fry (Bloch) based on growth,nutrient retention and haematological parameters

Dietary arginine requirement of Heteropneustes fossilis fry (3.0 ± 0.5 cm; 5.1 ± 0.3 g) was determined by feeding casein‐gelatin‐based isonitrogenous (400 g kg^?1 crude protein) and isocaloric (17.97 kJ g^?1) amino acid test diets containing graded levels of l ‐arginine (15, 17, 19, 21, 23 and 25 g kg^?1 dry diet) for 12 weeks. Maximum absolute weight gain (AWG) (44.4), best feed conversion ratio (FCR) (1.22), highest protein retention efficiency (PRE%) (41%), energy retention efficiency (ERE%) (75%), best condition factor, hepatosomatic index and viscerosomatic index were noted at 21 g kg^?1 arginine of the dry diet. Maximum body protein (189.8 g kg^?1) was also obtained in fish fed above diet. Highest haematocrit value (35%), Hb concentration (9.54 g dL^?1), RBC count (3.44 × 10⁹ mL^?1) and lowest Erythrocyte sedimentation rate (ESR) (1.93 mm h^?1) were obtained at the above level of arginine in the diet. AWG, FCR, PRE% and ERE% data were analysed using broken‐line and an exponential fit to obtain more precise dietary arginine requirement. On the basis of broken‐line and exponential analyses of AWG, FCR, PRE and ERE data, inclusion of dietary arginine in the range of 20.4–22.6 g kg^?1 dry diet, corresponding to 51–56.5 g kg^?1 dietary protein, is recommended for formulating arginine‐balanced feeds for rearing H. fossilis fry.     

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).