Refined understanding of sulphur amino acid nutrition in hybrid striped bass, Morone chrysops♀×M. saxatilis♂

Previous studies have indicated that the level of total sulphur amino acids (TSAA) (methionine+cystine) is most limiting in practical diet formulations for hybrid striped bass (HSB), especially if animal feedstuffs are replaced with plant feedstuffs. Reduction in costly animal feedstuffs such as fish meal, while maintaining adequate dietary levels of TSAA, may enhance the cost effectiveness of production. Therefore, this study investigated three different aspects of sulphur amino acid nutrition of HSB including: (1) the efficacy of crystalline methionine hydroxy analogue (MHA) and liquid MHA (Alimet?) relative to l‐ methionine in meeting the requirement for TSAA; (2) the cystine‐sparing value for methionine; and (3) the influence of various sulphur amino acid supplements on ammonia excretion. During three separate feeding trials (2, 6 and 10 weeks in duration), juvenile HSB were fed various diets including a basal diet deficient in TSAA (0.33% or 0.51% of diet), and experimental diets supplemented on an equal‐sulphur basis with different levels of either l‐ methionine, Alimet? or crystalline MHA. Diets containing TSAA at 1% of diet and different ratios of cystine to methionine (60:40, 55:45, 50:50 and 45:55) were also fed to re‐evaluate the sparing effects of cystine on methionine. In trial 1, over the course of 10 weeks, Alimet? was 73% as effective in promoting weight gain as l ‐methionine at the same concentration while MHA was 83% as effective. After 6 weeks in trial 2, fish fed Alimet? at 1.25% of diet displayed similar growth performance as those fed TSAA at 1.0% of diet, while weight gain of fish fed Alimet? at 1% was only 58% of that displayed by fish fed TSAA at 1.0%. No significant differences in weight gain, feed utilization or survival were observed among fish fed diets containing various ratios of cystine to methionine, although the diet with 60:40 cystine to methionine had the lowest numerical responses. Inclusion of MHA or Alimet? did not affect TAN excretion of HSB. These findings will aid in refining diet formulations for HSB to ensure adequate sulphur amino acid nutrition.     

Influence of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder, Paralichthys olivaceus

A feeding trial using five semi-purified diets (50% crude protein) was conducted to investigate the effects of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder. The control diet contained casein and gelatin as intact protein sources and four other diets contained 30% casein–gelatin (2:1, w/w) and 20% crystalline amino acids (CAA). CAA were added to the diets to simulate the amino acid pattern found in red sea bream egg protein (REP), Japanese flounder larvae whole body protein (FLP), Japanese flounder juvenile whole body protein (FJP), and brown fish meal protein (BFP), respectively. The test diets were fed to triplicate groups of juveniles (2.75±0.05 g) twice a day for 40 days to evaluate weight gain, survival, feed conversion efficiency (FCE), protein efficiency ratio (PER), and apparent protein utilization (APU). The apparent retention of total dietary amino acids in the whole body and A/E ratios of the whole body were also evaluated. The highest weight gain was observed in fish fed the diet containing the dietary amino acid pattern of BFP followed by fish fed the control, FJP, FLP and the REP diets. Percent survival, FCE, PER and APU were also significantly (P<0.05) affected by the amino acid pattern in the diets, indicating the highest value in fish fed the BFP diet. Except for a few amino acids, the amino acid composition of the whole body did not show marked differences with different dietary amino acid pattern. Results suggest that BFP could be more suitable as a reference amino acid pattern in the diet of juvenile Japanese flounder compared to the amino acid pattern of FLP, FJP or REP.     

Total sulphur amino acid requirement and cystine replacement value for fingerling rohu,Labeo rohita: effects on growth,nutrient retention and body composition

Two feeding experiments were conducted to quantify the total sulphur amino acid (TSAA) requirement and replacement value of cystine for methionine for fingerling Labeo rohita. In Experiment I, isonitrogenous (380 g kg^?1 CP) and isocaloric (17.90 kJ g^?1 GE) amino acid test diets with graded levels of methionine (4, 6, 8, 10, 12, 14 g kg^?1 dry diet) and 0.4 g kg^?1 cystine were fed to fish (4.62 ± 0.2 cm; 0.66 ± 0.1 g) and methionine requirement determined by analysing absolute weight gain (AWG) (5.48), feed conversion ratio (FCR) (1.26), protein retention efficiency (PRE%) (39%) and energy retention efficiency (ERE%) (85%) data which were best at 10 g kg^?1 methionine of dry diet. In Experiment II, six diets with different ratios of L‐cystine and L‐methionine on equimolar sulphur basis were fed to fish (4.71 ± 0.1 cm; 0.69 ± 0.2 g) under identical conditions. Maximum AWG (5.58), best FCR (1.24), PRE (41%) and ERE (86%) in fish fed Diet IV indicated cystine replacement value to be 40%. On the basis of the broken‐line and second‐degree polynomial regression analyses of results obtained in Experiments I and II, it is concluded that inclusion of TSAA in the range of 25.2–31.31 g kg^?1 of protein is optimum of which 33–39% could be spared by cystine.     

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.     

Improvement of quality of sesame Seasamum indicum seed meal protein with supplemental amino acids in feeds for rohu Labeo rohita (Hamilton) fingerlings

An 8-week feeding trial was conducted in a static indoor rearing system to examine the effects of partial substitution of fish meal (FM) protein with sesame seed meal protein with and without supplemental amino acids in diets for rohu Labeo rohita fingerlings (average weight 3.82 ± 0.05 g). Before incorporation into diets, sesame Seasamum indicum seed meal was fermented with lactic acid bacteria Lactobacillus acidophilus in order to reduce/eliminate the antinutritional factors tannin and phytic acid present in it. Twelve experimental diets (diets D1 to D12) were formulated replacing the FM protein from a reference diet with sesame seed meal protein at different levels (four sets of diets, of which each set of three diets contained 30%, 40% and 50% replacement of FM protein by sesame seed meal protein respectively). Diets D1 to D3 were not supplemented with any amino acid. Lysine was supplemented to diets D4 to D6. Diets D7 to D9 were supplemented with methionine–cystine (together), and diets D10 to D12 contained lysine and methionine–cystine (together). Lysine and methionine–cystine were added to the diets at 5.7% and 3.1% of dietary protein respectively. The groups of fish fed diets without any supplemental amino acids had significantly lower percentage weight gain, specific growth rate (SGR) and higher feed : gain ratio (FGR) than the groups of fish fed on other experimental diets. The addition of lysine and methionine–cystine to the diet in which 50% of FM protein was replaced by sesame meal protein (diet D12) significantly improved fish weight gain and FGR. The percentage live weight gain and SGR values differed significantly (P < 0.01) from each other in the fish fed diets D10 to D12, which were supplemented with all three amino acids. The results of the present study suggest that rohu fingerlings can effectively utilize the supplemented amino acids and that sesame seed meal protein can replace up to 50% of FM protein in the diets for rohu if the sesame seed meal is properly processed (fermented) and supplemented with deficient amino acids.     

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.     

Effect of fish-meal replacement with poultry by-product meal on the growth, tissue composition and hematological parameters of largemouth bass (Micropterus salmoides) fed diets containing different lipids

We conducted a 12-week feeding trial with largemouth bass (LMB) to examine the interactive effects of non-fish dietary proteins with different lipid sources on the performance of LMB (3.4 g ± 0.05 g). Practical diets contained poultry by-product meal (PBM) in place of fish meal on a digestible-protein basis, and 12% blood meal (BM). Diets were supplemented with 10% lipid as canola (CAN), chicken (CHK), CHK + MFO (50 : 50), menhaden fish oil (MFO) or CHK + CAN (50:50). A commercial trout diet (SC) was included as a positive control. Growth, survival, feed intake (FI), feed conversion ratio (FCR), protein efficiency ratio (PER), body composition, fatty acid composition of liver and muscle, hematological parameters and lysozyme and alternative complement activity (ACH50) were measured to assess diet effects. Weight gain (23.8 ± 1.8 g), survival (92 ± 4%), FI (44.3 ± 4.1 g), FCR (2.1 ± 0.1) and PER (1.3 ± 0.04) of LMB did not differ among the replacement diets. However, weight gain (38.5 ± 1.5), FI (53.9 ± 1.1), and PER (1.6 ± 0.1) of LMB fed the SC diet was higher and FCR (1.4 ± 0.1) was lower than that of fish fed the test diets. No signs of essential fatty acid deficiency were observed, although tissue concentrations of n-3 highly unsaturated fatty acids declined markedly in LMB fed diets without fish oil. Lymphocytes were elevated in fish fed diets with ≥ 7% n-3 fatty acids. Fish fed diets with 10% MFO or CHK + MFO had higher ACH50 than fish fed the diet with 10% CHK. There were no differences in serum lysozyme activity among fish fed the test diets. Hematocrit, hemoglobin, ACH50, and lysozyme activity were higher in fish fed the SC diet than in those fed the test diets. Regardless of lipid source, the ACH50 and lysozyme activity were greatly reduced in LMB fed PBM diets compared to LMB fed diets with 30% fish meal in a previous trial. The amino acid composition of the PBM was similar to that of fish meal. However, the availability of some essential amino acids from PBM to LMB may have been limited, and poor diet palatability caused by BM also may have reduced feed intake and performance of LMB.     

Dietary methionine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

Indian major carp, Cirrhinus mrigala fingerling (3.85 ± 0.50 cm, 0.50 ± 0.02 g) were fed isonitrogenous and isocaloric diets (40% CP, 4.28 kcal g⁻¹, GE) containing casein, gelatin and crystalline amino acids with graded levels of L- methionine (0.50, 0.75, 1.00, 1.25, 1.50 and 2.00 g/ 100 g, dry diet) with 1.00% cystine fixed, to determine its dietary methionine requirement. A feeding trial was conducted in triplicate for six weeks. Diets were fed twice a day at 0800 and 1600 h at 5% of body weight/day. The ration size and feeding regime were worked out prior to the start of the feeding trial. Weight gain (158%) and food conversion ratio (1.45) were significantly (P < 0.05) higher in fish fed diet containing 1.00% methionine with 1.00% cystine fixed. Second degree polynomial regression analysis of the weight gain data indicated the dietary methionine requirement to be 1.20 g/100 g of dry diet, corresponding to 3.00% of dietary protein. Second degree polynomial regression analysis was also employed to determine the relationship between food conversion ratio (FCR) and dietary methionine levels which indicated that the best FCR occurred at approximately 1.20% dietary methionine level. Carcass composition of fish fed diet containing graded levels of methionine varied significantly (P < 0.05) except carcass ash content which showed insignificant (P > 0.05) differences among the dietary methionine levels. This revised version was published online in August 2006 with corrections to the Cover Date.     

Influence of multiple amino acid supplementation on the performance of rainbow trout, Oncorhynchus mykiss (Walbaum), fed soya based diets

Using rainbow trout, Oncorhynchus mykiss (Walbaum), of approximately 50 g, a 63 day feeding trial was undertaken to evaluate the efficacy of individual and multiple amino acid supplements in diets where soyabean meal was used as the principle protein source. Iso-nitrogenous diets (45% crude protein) were formulated where the control diet (F) contained fishmeal as the reference protein and a solvent extracted soyabean meal (S) replaced approximately 66% of this protein source. The soyabean containing diets were then supplemented with crystalline amino acids thus; methionine only, dual supplemented with two methionine and lysine levels and finally, a supplement comprising methionine, lysine, tryptophan, threonine, arginine and histidine. The results showed that soyabean meal (SBM) was inferior to the reference protein when SBM was used to replace 66% of the fishmeal and that no significant restoration in growth, feed efficiency and apparent net protein utilization was obtained by either methionine only or dual methionine and lysine supplementation. However, by comparison with the fish fed the unsupplemented, single and dual supplemented diets, multiple amino acid incorporation was associated with significantly improved percentage weight gain, specific growth rate and marginal improvements in apparent net protein utilization. However, performance was not equal to that of the fish fed the fishmeal based control diet. The results are discussed with respect to the level of each of the essential amino acids (EAA) as a proportion of the total EAAs (A/E index) of test diets by reference to the whole body tissue amino acid profile of rainbow trout.     

Juvenile Atlantic salmon decrease white trunk muscle IGF‐1 expression and reduce muscle and plasma free sulphur amino acids when methionine availability is low while liver sulphur metabolites mostly is unaffected by treatment

We previously reported that juvenile Atlantic salmon with mean initial BW 11.5 g offed a methionine deficient diet had lower weight gain due to a reduced protein accretion, while lipid gain was unaffected. Muscle of the fish fed the methionine deficient diet was depleted for sulphur amino acids, while in liver, the concentration of these metabolites was maintained within narrow limits. We speculated whether this could be due to an increased muscle proteolysis to support a prioritized liver metabolism in fish fed the low methionine diets. In this study, we assessed whether genes associated with muscle proteolysis increased under methionine deficiency. The composition of the diets was similar to those used previously containing 1.6 or 2.1 g Met/16 g N. We confirmed that the fish fed the low methionine diet gained less protein compared to fish fed the DL‐methionine enriched diet (P = 0.014), but growth did not reduce significantly. Also the deficient fish maintained the concentrations of liver sulphur amino acids and reduced muscle free methionine. Several of the other free amino acids within muscle increased. Further, methylation capacity was maintained in liver but reduced in the muscle (P = 0.78 and 0.04, respectively). Gene expression of muscle IGF‐1 was lower (P = 0.008) and myosin light chain 2 tended (MLC2, P = 0.06) to be reduced in fish fed low methionine diet, concurrently the activity of cathepsins B+L increased (P = 0.047) in muscle of fish fed the low methionine diet. Gene expression of the muscle‐specific E3 ubiquitine ligases (Murf and MaFbx) was not affected by treatment. Thus, the lower protein gain observed in fish fed the low methionine diet may be caused by reduced protein synthesis in line with the reduced IGF‐1 gene expression in the white trunk muscle. Thus, to support metabolism, the dietary protein needs to be balanced in amino acids to support metabolism in all compartments of the body and secure maximal protein gain.     

Methionine Requirement in Practical Diets of Juvenile Nile Tilapia,Oreochromis niloticus

Two feeding experiments were conducted to confirm methionine requirement in practical diets of juvenile Nile tilapia, Oreochromis niloticus. Test diets used in both experiments contained 414 kcal gross energy, 28 g protein, and 5 g lipid per 100 g diet. In the first experiment, seven diets were made using cottonseed meal (CSM), dehulled solvent‐extracted soybean meal (DSESM), and gelatin as intact protein sources. Methionine was added to five of these diets at 0.03 or 0.06% increments to produce methionine levels ranging from 0.33 to 0.57% of the diet. Each diet was fed to four replicate groups of male juvenile Nile tilapia (5.62 ± 0.13 g) in a recirculation system for 8 wk. Broken‐line regression analysis of weight gain indicated that methionine requirement of juvenile Nile tilapia was 0.49% of the diet or 1.75% of dietary protein at cystine level of 0.45% of the diet. The second experiment was designed based on methionine requirement determined in the first experiment and also contained seven test diets. The first six diets contained CSM and DSESM as protein sources. Methionine was added to five of these diets at an increasing rate of 0.06% to produce methionine levels ranging from 0.49 to 0.79% of the diet. In the last diet (Diet 7), a portion of DSESM was replaced by gelatin to reduce methionine level to 0.33% of the diet in order to test whether methionine is limited. Each diet was also fed to four replicate groups of male juvenile Nile tilapia (2.32 ± 0.06 g) in a recirculation system for 9 wk. At the termination of the second experiment, there were no significant differences in terms of weight gain, survival, and feed efficiency ratio (FER) among the fish fed the first six diets. However, weight gain and FER of the fish fed these diets were significantly better than those fed Diet 7, confirming the methionine requirement value as has been determined in the first experiment.     

Effect of dietary protein reduction with synthetic amino acids supplementation on growth performance,digestibility, and body composition of juvenile Pacific white shrimp, <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

Two experiments were conducted to evaluate the effect of lowering crude protein level and fish-meal inclusion rate by using commercially available synthetic amino acid supplements in practical diets on the growth performance of Litopenaeus vannamei. In experiment 1, three diets were formulated to assess whether 50% of fish meal could be replaced by soybean meal with synthetic amino acid supplementation. Diet 1 was formulated as the normal control with 20% fish meal and 36% crude protein; diet 2 was the negative control with 34% crude protein and half of the fish meal was replaced with soybean meal; and diet 3 was similar to diet 2 but was supplemented with amino acids to ensure the level of lysine, methionine plus cystine, and threonine similar to that in the diet 1. After a 70-day feeding trial, weight gain and specific growth rate of shrimps fed diet 2 were significantly lower than those fed diet 3, and numerically lower than those fed diet 1. Feed intake of shrimps fed diet 3 was significantly higher than those fed diets 1 and 2. There were no significant differences in feed conversion ratio among shrimps fed different diets. In experiment 2, four diets were prepared with diet 1 as the normal control with 41.26% crude protein, diets 2–4 were formulated to contain 39.81, 38.40, and 35.52% of crude protein with synthetic amino acids were added to simulate the amino acid levels of the diet 1. After a 70-day feeding trial, it was found that reducing dietary crude protein from 41.26 to 35.52% did not affect weight gain or feed conversion ratio. The survival of crude protein 35.52% treatment was significantly lower than other treatments. No difference was observed in body protein, lipid composition, and apparent digestibility coefficient among dietary treatments. Results of this study suggested that dietary crude protein could be reduced from 41.26 to 35.52% in the diets of L. vannamei as long as synthetic amino acids were supplemented.     

Total sulphur amino acid requirement and maximum cysteine replacement value for methionine for fingerling Catla catla (Hamilton)

Two separate 12 weeks feeding trials were performed to quantify the total sulphur amino acid (TSAA) requirement (experiment I) and cysteine replacement value for methionine (experiment II) of fingerling Catla catla. In experiment I, six casein–gelatin based (33% crude protein; 16.72 kJ g^?1 gross energy) diets with graded levels of TSAA (0.56%, 0.81%, 1.06%, 1.31%, 1.56%, 1.81% dry diet) were fed to triplicate groups of fish (3.55 ± 0.06 cm; 0.65 ± 0.02 g) near to satiation. The TSAA requirement was determined by quadratic regression analysis of absolute weight gain (AWG), protein efficiency ratio (PER), feed efficiency (FE), protein gain (PG) and TSAA gain (TSAAG) against dietary TSAA concentrations at 95% maximum response. Above analysis revealed that inclusion of TSAA at 1.28% dry diet (1.22% methionine + 0.06% cysteine), corresponding to 3.87% of dietary protein is optimum. In experiment II, to determine the replacement value of cysteine for methionine, six diets containing 1.28% TSAA determined in experiment I with different ratios of l ‐methionine and l ‐cysteine (80:20, 70:30, 60:40, 50:50, 40:60, 30:70) on equimolar sulphur basis were fed to fish (3.65 ± 0.08 cm; 0.67 ± 0.04 g). Broken‐line regression analysis of AWG, PER, FE, PG and TSAAG against varying methionine to cysteine ratios yielded the optimum cysteine replacement value at 40.3%. Based on above analysis, it is recommended that inclusion of 1.28% dietary TSAA, corresponding to 3.87% of dietary protein is optimum of which 40.3% could be spared by cysteine. Data generated during this study would be useful in formulating TSAA balanced, cost‐effective feeds for the intensive culture of this fish.     

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.     

Dietary Threonine Requirement of Juvenile Red Drum Sciaenops ocellatus

Threonine is an indispensable amino acid required by all animals for normal growth and metabolic functions. An experiment was conducted in a brackish water (5 ± 1 ppt) recirculating system to quantify the minimum dietary threonine requirement of juvenile red drum Sciaenops ocellatus . The experimental diets contained 350/0 crude protein from red drum muscle and crystalline amino acids and 3.2 kcal available energy/g diet. Incremental levels of L-threonine were added to the diets and fed to juvenile red drum initially averaging 2.8 g/fish for 8 wk. Weight gain and feed efficiency of fish fed the various diets were significantly ( P < 0.05) affected and increased linearly as dietary threonine increased until plateauing around 0.8% of dry diet. Protein efficiency ratio and protein conversion efficiency values for fish fed the different diets also were significantly affected by threonine level and indicated requirement values of 0.8–0.9% of dry diet. Free threonine in plasma also significantly responded to increasing dietary threonine but indicated a slightly higher requirement value of approximately 1.0% of dry diet. Based on these data, the minimum threonine requirement of juvenile red drum was determined to be approximately 0.8% of dry diet (2.28% of dietary protein). This requirement level is similar to values reported for some other fish species. Based on this information diets may be formulated more precisely for aquacultural production of red drum.     

Dietary vitamin E requirement, fish performance and carcass composition of rohu (Labeo rohita) fry

A 90-day feeding experiment was conducted using flow through system to evaluate the requirement of vitamin E for rohu fry in relation to growth performance and to assess the relationship between the dietary vitamin E levels and carcass composition. Five semi-purified diets supplemented with 0, 50, 100, 150, 200 mg vitamin E/kg dry diet as --tocopheryl acetate were fed to rohu fry (av. wt.±S.E.=0.58±0.01 g) in triplicate groups. At the end of the experiment, growth and dietary performance were evaluated and vitamin E deposition in the tissue, erythrocyte fragility, thiobarbituric acid reactive substances (TBARs) were analyzed. The average net weight gains were 6.82±0.09, 7.38±0.03, 8.20±0.03, 8.07±0.09 and 8.12±0.08 (g/90 days), respectively, for fish fed diets 0, 50, 100, 150, 200 mg vitamin E/kg. The fish fed diets containing less than 100 mg supplemental vitamin E/kg had significantly (P<0.05) reduced weight gain, feed efficiency and other nutritional indices compared to those fed diets supplemented with vitamin E at 100–200 mg/kg. With the increasing level of vitamin E, both TBARs and erythrocyte fragility values were reduced. Regression analysis of weight gain data using broken-line model indicated a minimum vitamin E requirement of 131.91 mg/kg dry diet. The erythrocyte fragility and TBARs were maximum in fish fed the vitamin E-deficient diet. From the present experiment, it may be concluded that the vitamin E requirement of Labeo rohita fry for optimum growth and other dietary performances is 131.91 mg/kg of dry diet.     

Efficacy of Three Methionine Sources in Diets for Pacific White Shrimp, Litopenaeus vannamei

Abstract.— To examine the need for supplemental dietary methionine and to determine the ability of different methionine sources to meet this need for Pacific white shrimp, Litopenaeus vannamei, a growth trial was conducted. A control diet (30% crude protein) was formulated to be deficient in methionine by inclusion of high levels of dehulled, solvent‐extracted soybean meal and with no fish meal (methionine level was 0.45% of diet as fed, 1.5% of total amino acids). Three test diets were manufactured based on the control diet but supplemented with l ‐methionine, dl ‐methionine, or an analog of methionine (2‐hydroxy‐4‐methylthio butanoic acid [HMTBA], calcium salt) at the level of 0.5% as fed (equivalent to 1.5% of total amino acids). Cystine was constant among these diets at 0.5% of diet as fed. A commercial feed (30% crude protein) was included as a reference. The culture vessels were flat‐bottomed, cylindrical, black fiberglass tanks (1.5‐m diameter) initially filled with 1000 L seawater and supplied with aeration and seawater from a well at a rate of 2 L/min. At the end of the trial, all surviving shrimp were counted and weighed. The trial was run outdoors, with a mean water temperature of 25.9 C. The mean survival, final weight, growth rate, and feed efficiency of shrimp in the control group were 95.9%, 10.8 g, 0.93 g/wk, and 0.44 g/g, respectively. The mean values of all the test treatments were 96.9%, 11.6 g, 1.00 g/wk, and 0.47 g/g, respectively. The final weight and growth rate of shrimp fed the control diet were significantly lower than those of shrimp fed the methionine‐supplemented diets, indicating that this species has a requirement for dietary methionine. There were no significant differences (P > 0.05) in any of the parameters among the test treatments, indicating that l ‐methionine, dl ‐methionine, and HMTBA can be used to meet the methionine requirement of this species under these experimental conditions. It is concluded that the three sources of methionine examined in this trial, l ‐methionine, dl ‐methionine, and HMTBA, are similar in their ability to meet the methionine requirement of this species.     

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.     

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.     

Optimal dietary methionine requirement of juvenile Chinese sucker,Myxocyprinus asiaticus

A total of 630 juvenile Chinese sucker, with an average initial weight of 1.72 ± 0.05 g, were fed seven diets for 56 days to study the effect of dietary methionine levels on growth, feed utilization, body composition and haematological parameters on juvenile Chinese sucker. Diet 1 using fish meal as the sole protein source and diets 2–7 using fish meal and fermented soybean meal as intact protein sources supplemented with crystalline amino acids contained six levels of l ‐methionine ranging from 6.4 to 18.9 g kg^?1 of dry diet at a constant dietary cystine level of 3.7 g kg^?1. Each diet was randomly assigned to three aquaria. Results indicated that the highest weight gain, specific growth rate (SGR), feed efficiency ratio, protein efficiency ratio and protein productive value occurred at 13.9 g methionine kg^?1 diet among the methionine supplemented dietary groups, beyond which they showed declining tendency. The whole body and muscle protein contents of juvenile Chinese sucker were positively correlated with dietary methionine level, while muscle lipid content was negatively correlated with it. The total essential amino acids content of muscle was increased significantly with increasing dietary methionine level from 6.4 to 13.9 g kg^?1 (P < 0.05). Apparent digestibility coefficients of dietary protein were significantly affected by dietary treatments. Serum protein, cholesterol and triacylglycerol increased with increasing dietary methionine levels, but showed a relatively lower value for fish fed the 18.9 g methionine kg^?1 diet. Quadratic regression analysis of SGR against dietary methionine level indicated that optimal dietary methionine requirement for juvenile Chinese sucker was 14.1 g kg^?1 of the diet in the presence of 3.7 g kg^?1 cystine (corresponding to 32.0 g kg^?1 of dietary protein on a dry‐weight basis).