The role of dietary methionine concentrations on growth,metabolism and N‐retention in cobia (Rachycentron canadum) at elevated water temperatures

This study determined impacts of dietary methionine concentrations at two temperatures on growth, feeding efficiency and N‐metabolites in juvenile cobia. Methionine concentrations of the experimental diets were deficient (M9; 9 g/kg), sufficient (M12; 12 g/kg) and surplus (M16, 16 g/kg). Water temperature was normal (30°C) or elevated (34°C). Twenty cobia in triplicate tanks were fed the experimental diets for 6 weeks. Both methionine and temperature affected cobia's growth and feeding efficiency. Cobia fed M9 performed lower than the fish fed M12 and M16 diets. Additionally, cobia reared at 34°C performed poorer than at 30°C, probably due to lower voluntary feed intake in the fish reared at 34°C. Protein efficiency ratio and protein productive value in cobia fed M9 diet were less than M12 or M16 diets. This was confirmed with the improved retentions of indispensable amino acids (AAs). No interactions between methionine and temperature were observed in growth and protein accretion. At 30°C, CF improved, while HSI and VSI declined upon methionine supplementation levels. Of which an interaction between temperature and methionine was present. Plasma, muscle and liver free AA and N‐metabolites were affected by methionine and temperature. Furthermore, temperature affected cobia's lipid class composition, resulting in increased phospholipids and cholesterol at 34°C.     

Influences of dietary soy protein concentrate with taurine on growth and biochemical status of Acanthopagrus schlegelii juveniles

Taurine is a component that tends to be deficient in fish diets that contain higher plant‐derivative proteins. Acanthopagrus schlegelii juveniles (IBW ? 11.28 g) were fed nine diets containing SPC levels (SPC40, SPC60, SPC80 and SPC100), or supplemented with 5 g/kg of taurine at each replacement level. Our results showed higher WGR and SGR in SPC60 + TAU (WGR; 224.67, SGR; 2.10) followed by SPC40 + TAU (WGR; 221.75, SGR; 2.09). Phosphorus retention efficiency was significantly raised as the SPC increased when compared with the control diet. Taurine addition showed an incline in the nitrogen retention efficiency. An effect was observed within FR between fish and the SPC content. ADCs of dry matter, crude lipid and phosphorous were affected by SPC, while taurine addition affected only crude lipid. A significant effect was seen in ash and phosphorus by the SPC and taurine. TAU supplementation showed an increased T‐AOC activity in the SPC80 + TAU (1.90), while malondialdehyde and catalase activity were reduced except in completely replaced fish meal groups. It could be concluded that black sea bream can accept the substitution of FM with 60% of SPC while at the inclusion of dietary TAU (5 g/kg) growth could be improved.     

Dietary protein requirement for large‐size Songpu mirror carp (Cyprinus carpio Songpu)

Dietary protein requirements of large‐size Songpu mirror carp (Cyprinus carpio Songpu) were investigated through an an 82‐day feeding trial. Six practical diets were formulated to contain increasing crude protein levels of 240, 260, 280, 300, 320 and 340 g/kg. Six triplicate groups of fish (25 individuals per replicate with initial mean weights of 246.00 ± 17.36 g) were reared in the floating net cages. Results indicated that the highest weight gain rate (WGR), specific growth rate (SGR), protein efficiency ratio (PER) and lowest feed conversion ratio (FCR) were observed in the 260 g/kg treatment. The whole‐body crude protein content was significantly higher in the 260 and 280 g/kg treatments (p < .05). The expression of growth hormone (GH) in dorsal white muscle, Rh a glycoprotein (Rhag), Rh b glycoprotein (Rhbg) and Rh c glycoprotein 1 (Rhcg1) in gills increased significantly with increasing protein levels, while the expression of target of rapamycin (TOR), eukaryotic translation initiation factor 4E binding protein 1 (4EBP1) and S6 kinase (S6K) in muscle and intestine increased first and then decreased significantly. Based on these findings, 260 g/kg seems to be the proper level of protein in a practical diet for large‐size Songpu mirror carp (Cyprinus carpio Songpu).     

Effects of dietary lipid levels on growth,feed utilization,digestive tract enzyme activity and lipid deposition of juvenile Manchurian trout,Brachymystax lenok (Pallas)

Six isoproteic diets were designated to evaluate the effects of dietary lipid levels (from 70 to 270 g/kg) on the growth performance, feed utilization, digestive tract enzyme activity and lipid deposition of juvenile Brachymystax lenok (average initial weight 0.54 ± 0.04 g). Each diet was fed to triplicate tanks (30 fish per tank) in an indoor closed recirculating system for 9 weeks. Final body weight and weight gain were highest in fish fed 190 g/kg diet and lowest in fish fed the 70 g/kg diet. Specific growth rate of fish fed with 190 g/kg diet was significantly higher than those fed with 70 and 270 g/kg diets (p < .05). Protein efficiency ratio of fish fed with 70 g/kg diet was significantly lower than the 110–230 g/kg treatments and was not significantly different from the 270 g/kg treatment. Fish fed with 270 g/kg diet had significantly higher hepatosomatic index and viscerosomatic index than those fed with 70–190 g/kg diets (p < .05). Intraperitoneal fat ratio and the whole‐body lipid content had a trend to increase with increase in dietary lipid level. Muscle crude lipid content increased up to 190 g/kg with increase in dietary lipid level. Lipid retention decreased with increase in dietary lipid level, while no significant differences in protein intake and retention levels were observed in fish among all treatments. Lipase activity of the mixture of pyloric caeca and foregut in fish fed 190 and 230 g/kg diets was significantly higher than those fed 70 and 110 g/kg diets. Midgut and hindgut lipase activities of fish were significantly higher than those fed the 190 and 230 g/kg diets. In conclusion, based on the second‐order polynomial model of WG and FCR, this study suggested that 173.8–195.0 g/kg dietary lipid levels were appropriated for B. lenok.     

Effects of dietary linolenic acid on growth,fatty acid composition,immune function and antioxidant status of juvenile blunt snout bream,Megalobrama amblycephala

A nine‐week feeding trial was performed to determine the dietary linolenic acid (LNA; 18:3n–3) requirements of juvenile blunt snout bream. Six iso‐nitrogenous, semi‐purified diets were prepared with different concentrations of LNA (0–25 g/kg). Dietary LNA had no significant effects on survival rate. However, final fish weight, weight gain (WG), specific growth rate (SGR) and feed efficiency ratio (FER) increased with increasing dietary LNA concentrations up to 20 g/kg. Dietary LNA increased muscle LNA and total n‐3 polyunsaturated fatty acid (PUFA) contents, but decreased total saturated fatty acid content. Fish fed 20 g/kg LNA had the highest plasma alkaline phosphatase activity, total protein, albumin and white blood cell count levels. Additionally, fish fed 20 g/kg LNA had higher triglyceride levels than control fish. Plasma glucose increased with increasing dietary LNA concentrations. Superoxide dismutase and glutathione peroxidase activities significantly increased with increasing dietary LNA concentrations up to 15 g/kg. Based on SGR and FER, the optimal dietary LNA requirements of juvenile blunt snout bream were 17.5 and 15.6 g/kg respectively.     

Lipid metabolism in juveniles of Yellowtail,Seriola dorsalis,fed different levels of dietary methionine containing a low level of cholesterol: Implication in feed formulation

The involvement of dietary methionine in the lipid metabolism of Seriola dorsalis, at different levels of dietary methionine (Met), and low content of cholesterol was investigated. Four diets containing different Met levels and a low amount of cholesterol (0.23 g/kg diet) were prepared. One hundred and eighty juveniles (16.2 ± 1.5 g) were randomly distributed into each 12 500‐L ponds. After 60 days, the growth in weight (%) significantly increased following the Met supplementation, whereas SGR of all the experimental treatments was higher than the Basal Diet. The expression of the insulin‐like growth factor (IGF‐1) increased significantly (0.2‐fold) and was complemented by significant changes in Mat, BHMT and HMG‐CoA, for fish that were in the low and higher Met levels. The cholesterol content in the whole body and liver increased following methionine supplementation, similarly to that observed for crude lipids, in particular in the whole body and liver tissues. However, the relative amount of most fatty acids remained unchanged. Only the oleic acid increased at a higher amount of Met. Therefore, once the dietary requirement of methionine is met, the methionine is used to synthesize cholesterol. It is recommended to supply a higher amount of methionine to spare energy for growth.     

Response of gibel carp (Carassius auratus gibelio) to increasing levels of dietary lysine in zero fish meal diets

To investigate the response of gibel carp to dietary lysine levels in zero fish meal (FM) diets, 8 experimental diets were designed. Among them, diet 1 and diet 2 with 150 g/kg FM, methionine, lysine and threonine were supplemented in diet 2. Diet 3 was without FM but matched the amino acid profile of diet 2 except for lysine. Diets 4–8 were based on diet 3, with graded levels of lysine. After 8‐week feeding trail, final body weight, weight gain (WG), specific growth rate, feed conversion ratio (FCR) and feed intake were significantly improved by diets 1–2 and 6–8 compared with diet 3. Plasma glucose, triglyceride, immunoglobulin G and aspartate aminotransferase levels in diets 4–8 were higher than those in diet 1. Apparent digestibility coefficients (ADCs) of protein, energy, methionine, lysine, valine, histidine and phenylalanine in diet 1 were significantly lower than those in diet 6. The highest ADC of isoleucine was found in diet 3. TOR, IGF‐1, JAK3, STAT4, STAT6 and PepT2 mRNA levels were significantly increased with increasing dietary lysine up to a certain level in zero FM diet and lower than diets 1–2. Based on WG and FCR, the optimal lysine requirement of juvenile gibel carp was determined to be 24.4 g/kg and 24.2 g/kg (65.9 g/kg and 65.4 g/kg of dietary protein) in zero FM diet, respectively, using quadratic regression analysis.     

Effects of dietary inulin and Jerusalem artichoke (Helianthus tuberosus) on intestinal microbiota community and morphology of Nile tilapia (Oreochromis niloticus) fingerlings

This study investigated the effects of dietary inulin and Jerusalem artichoke (JA) on intestinal microbiota and morphometry of Nile tilapia fingerlings. Five treatment diets were designed to supplement inulin at 0 (basal diet), 2.5 and 5.0 g/kg, and JA at 5.0 and 10.0 g/kg. Nile tilapia larvae were fed experimental diets from the first feeding through the fingerling stage (84 days). The cultivation‐dependent technique showed that dietary inulin at 5.0 g/kg and JA (at both levels) increased lactic acid bacteria and Bifidobacterium spp., but decreased Vibrio spp. (p < .05). PCR‐DGGE targeting 16S ribosomal RNA gene revealed that dietary inulin and JA generated different profiles of microbial community compared with fish fed a basal diet. Compared with fish fed the basal diet, a greater intestinal villi height was observed in fish fed 5.0 g/kg inulin and JA at both levels (p < .05). A larger relative goblet cell number were observed in the anterior intestine of fish fed 5.0 g/kg inulin or JA (p < .05). Overall, dietary inulin (5.0 g/kg) and JA (5 and 10.0 g/kg) since the first feeding had effects on modulating the intestinal microbiota and morphology of Nile tilapia fingerlings.     

The optimal dietary protein level of large‐size grouper Epinephelus coioides

An 8‐week feeding trial was conducted to determine the requirement of protein for large‐size grouper Epinephelus coioides (initial body weight: 275.07 ± 1.56 g). Six iso‐lipidic (124 g/kg) diets were formulated containing graded levels of protein (350, 400, 450, 500, 550 and 600 g/kg). Grouper was hand‐fed twice daily to apparent satiation with triplicate. The results showed that significantly high weight gain, specific growth rate and significantly low feed conversion ratio were observed in fish fed 450 g/kg protein group. High‐protein level diets significantly increased protein content and significantly decreased lipid content of fish body and muscle. Total protein and cholesterol content in serum of 600 g/kg group were significantly higher than those of 350 g/kg group. However, serum glucose and triglyceride contents of fish fed low‐protein diets were significantly higher than those of fish fed high‐protein diets. Meanwhile, liver glutamic‐pyruvic transaminase and glutamic‐oxaloacetic transaminase in high‐protein diet groups were significantly higher than those of low‐protein diet groups. The intestinal protease activity in high‐protein diet groups was significantly higher that of low‐protein diet groups, but lipase and amylase showed opposite trend. With the increasing of dietary protein level, the activities of alkaline phosphatase, superoxide dismutase and lysozyme in liver of grouper increased significantly compared with 350 g/kg group, while the activities of acid phosphatase decreased significantly. With specific growth rate as the evaluation index, the optimum dietary protein level of large‐size grouper Epinephelus coioides was 438.39 g/kg by fitting the broken‐line regression analysis.     

Comparative study on the utilization of crystalline methionine and methionine hydroxy analogue calcium by Pacific white shrimp (Litopenaeus vannamei Boone)

The objective of this study was to compare the supplemental effects of crystalline DL‐methionine (DL‐Met) and methionine hydroxy analogue calcium (MHA‐Ca) on growth performance of Pacific white shrimp. Eight isoproteinic (355.3 ± 2.0 g/kg diet) and isolipidic (70.0 ± 2.2 g/kg diet) diets were prepared as positive diet (20% fish meal), negative diet (15% fish meal) and DL‐Met, MHA‐Ca‐supplemented diets with the supplementation of 0.03%, 0.06%, 0.09% DL‐Met and 0.04%, 0.07%, 0.1% MHA‐Ca in negative diet respectively. Pacific white shrimp (0.92 ± 0.03 g) were fed one of the eight diets for 49 days. The results showed that dietary DL‐Met did not affect weight gain (WG) and feed conversion ratio (FCR) (p > 0.05), but the supplementation of 0.1% MHA‐Ca significantly increased WG, protein and lipid retention, and reduced FCR (p < 0.05) when compared to the negative group, and reached the similar levels as the positive control. The total free amino acids (TFAA) in haemolymph of MHA‐Ca groups and PC, NC group peaked at the 3rd hr after feeding, but the peaking time of DL‐Met groups was advanced to the 2nd hr. Activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in haemolymph of all MHA‐Ca groups peaked at the 2nd hr, but DL‐Met groups peaked at the 1st hr after feeding. The above results indicated that the supplementation of 0.1% MHA‐Ca in a low fish meal diet could improve the growth performance and feed utilization of Pacific white shrimp, but dietary DL‐Met did not significantly affect the growth.     

Effects of replacement of fishmeal with cottonseed protein concentrate on the growth,intestinal microflora,haematological and antioxidant indices of juvenile golden pompano (Trachinotus ovatus)

A 56‐day feeding trial was conducted to investigate the effects of replacing fish meal with cottonseed protein concentrate (CPC) (free gossypol < 7.9 mg/kg) in the diets on the growth, intestinal microflora, haematological and antioxidant indices of juvenile golden pompano (Trachinotus ovatus). Six diets were designed: fishmeal diets (FM) which contained 340 g/kg fishmeal, as well as five CPC diets, each with differing CPC concentrations (120, 240, 360, 480 and 600 g/kg) to replace the fish meal. The weight gain rate (WGR) and specific growth rate (SGR) showed no significant difference among groups (p > .05) with the dietary CPC level ranged from 0 to 360 g/kg. Serum cholesterol (CHO) of C36 and triglyceride (TG) levels of C36 and C12 were significantly higher than the FM (p < .05). Total protein (TP) levels of C12 were significantly lower than the FM (p < .05). Among the treatments, C36 had higher glutathione peroxidase (GSH‐PX) and total superoxide dismutase (T‐SOD) than FM (p < .05). From the data analysis of 16s sequencing, with increasing CPC concentration, the proportion of harmful microbial taxa (Proteobacteria and Vibrio) increased. The results of this study support that CPC products are acceptable in practical diets for golden pompano. And the optimal dietary CPC replacement of golden pompano was estimated to be 259.3 g/kg.     

Dietary calcium requirement of red swamp crayfish (Procambarus clarkia)

To investigate dietary calcium requirement of red swamp crayfish (Procambarus clarkia), six semi‐purified diets were formulated to contain different concentrations of calcium (2.7(control group), 6.1, 11.9, 17.6, 23.5 and 29.1 g/kg calcium). Each diet was hand‐fed to triplicate of 15 crayfish with average initial body weight (6.22 ± 0.87) g for 8 weeks. The results showed that weight gain rate (WGR) significantly increased and feed conversion ratio (FCR) significantly decreased from 11.9 to 23.5 g/kg groups (p < .05). Protease activities in intestine and hepatopancreas and parathyroid hormone concentrations in serum significantly decreased with increasing dietary calcium levels (p < .05), while calcium and phosphorus contents in exoskeleton, calcium content in muscle and calcitonin concentrations in serum significantly increased (p < .05). The activities of lipase and amylase in intestine and hepatopancreas, serum alkaline phosphatase and total vitamin D concentrations in serum had significant increase as dietary calcium content increased up to 11.9–17.6 g/kg (p < .05). The inorganic phosphorus content in 29.1 g/kg group was significantly lower than those in other groups (p < .05). Broken‐line model analysis based on WGR and quadratic curve model analysis based on FCR showed that optimal dietary calcium requirement of red swamp crayfish ranged from 12.7 to 17.1 g/kg.     

The optimum dietary isoleucine requirement of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)

A six‐week growth trial was conducted to evaluate the optimum dietary isoleucine requirement of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). Seven isoenergetic (3,400 kcal/kg of dry matter), isoproteic (496 g/kg of dry matter) and isolipidic (70 g/kg of dry matter) diets were formulated to contain graded Ile levels (7.3, 11.3, 15.7, 19.6, 23.5, 26.8 and 30.8 g/kg, dry‐matter basis). Each experimental diet was fed to triplicate groups of 12 hybrid grouper juveniles (average initial body weight: 6.00 ± 0.01 g/fish). Experimental fish were randomly distributed into 21 glass tanks (L 60 × W 45 × H 50 cm) connected to mechanical and biological water filters as a recycling system. Fish were fed twice daily (08:00 and 16:00) to apparent satiation. After the sampling of the growth trial, the remaining fish in each group were fed their corresponding diets for 2 d and then exposed to 4 mg Cu²⁺ · L^?1 water for 24 hr. Results showed that growth performance and feed utilization were significantly affected by different dietary Ile levels (p < .05). Weight gain percentage (WG%), protein productive value (PPV), protein efficiency ratio (PER) and feed efficiency (FE) were increased as dietary Ile level increased, reaching a peak value at 19.6 g/kg dietary Ile, and thereafter, these four parameters declined as dietary Ile level continued to increase. Daily feed intake (DFI) showed an opposite tendency of variations as FE. The quadratic regression analysis of WG%, PPV, PER and FE against dietary Ile levels indicated that the optimum dietary Ile requirement for hybrid grouper was estimated to be 19.8, 20.8, 19.4 and 19.1 g/kg dry matter, respectively. Among all experimental treatments, fish fed 19.6 g/kg dietary Ile had the highest expression of growth and protein synthesis‐related genes, including growth hormone (GH) in pituitary, insulin‐like growth factor 1 (IGF‐1), growth hormone receptor 1 (GHR1), target of rapamycin (TOR) and S6‐kinase 1 (S6K1) in liver. Gut micromorphology was significantly influenced by dietary Ile levels. After the exposure to 4 mg Cu²⁺ · L^?1 water for 24 hr, fish fed 19.6 g/kg dietary Ile had the highest survival and the best immunologic manifestation among all experimental treatments. Generally, the optimum dietary Ile requirement for maximum growth of hybrid grouper was estimated to be 19.8 g/kg dry matter, corresponding to 39.9 g/kg dietary protein.     

Dietary histidine affects intestinal antioxidant enzyme activities,antioxidant gene expressions and inflammatory factors in juvenile blunt snout bream (Megalobrama amblycephala)

Six semipurified diets of graded histidine levels (from 4.1 to 14.2 g/kg) were fed to fish for 8 weeks. The results showed that the intestinal content of malondialdehyde (MDA), activities of total superoxide dismutase (T‐SOD) and glutathione peroxidase (GPx) were noticeably lower in 8.0 g/kg diet compared to control group. While the lowest activities of copper–zinc superoxide dismutase (Cu/Zn‐SOD) and manganese superoxide dismutase (Mn‐SOD) were observed in 9.9 g/kg diet, the intestinal activities of total antioxidant capacity (T‐AOC) and catalase (CAT) in 14.2 g/kg diet were noticeably higher than those in 8.0 and 9.9 g/kg diets. Plasma biochemical indexes were not significantly affected by dietary histidine levels. In the intestine, the Kelch‐like ECH‐associated protein 1 (Keap1) mRNA levels were increased in 8.0 g/kg diet, which suppressed the nuclear translocation of nuclear factor E2‐related factor 2 (Nrf2), and subsequently decreased CAT, GPx1, Cu/Zn‐SOD and Mn‐SOD expression levels. The lowest mRNA levels of interleukin 8 (IL‐8) and tumour necrosis factor‐α (TNF‐α) were observed in 8.0 g/kg diet, whereas the highest mRNA levels of transforming growth factor‐β (TGF‐β) and interleukin 10 (IL‐10) were observed in 8.0 g/kg diet. These results indicated that dietary histidine plays a major role in maintaining intestinal health in juvenile blunt snout bream.     

Evaluation of replacing fish meal with corn protein concentrate in Nile tilapia Oreochromis niloticus fingerlings commercial diet

Four isocaloric‐isonitrogenous diets containing 0, 50, 100 and 190 g/kg corn protein concentrate (CPC) as replacement for dietary fish meal were fed to Nile tilapia Oreochromis niloticus fingerlings for 8 weeks. Tilapia growth parameters were not significantly (p > .05) different in fish fed diets with 0, 50 and 100 g/kg CPC and found to be superior compared to those fed on 190 g/kg CPC. Fish dressing ratios and body composition were similar among all treatments. The electron microscope indicated that the stomach size of control fish was slightly smaller and the wall was thinner while the stomach of the fish fed all other levels of CPC undergone a remarkable size increase and their walls were thicker after feeding diets with CPC. Total aerobic bacterial and coliform counts were significantly decreased in fish intestine when fed diets with 100 and 190 g/kg CPC compared with fish fed diets with 0 g/kg or 50 g/kg CPC. This study indicates that it is possible to replace up to 534 g/kg of dietary fish meal in tilapia fingerlings using 100 g/kg of CPC without any negative effect on fish growth and proximate body composition.     

Effects of dietary tryptophan levels on growth performance,whole body composition and gene expression levels related to glycometabolism for juvenile blunt snout bream,Megalobrama amblycephala

To investigate the effects of dietary tryptophan on growth and glycometabolism in juvenile blunt snout bream, 450 fish (initial weight 23.33 ± 0.03 g) were fed six practical diets with graded levels of tryptophan (from 0.79 g/kg to 5.96 g/kg dry matter) for 8 weeks. Results showed that final weight, per cent weight gain (PWG), protein efficiency rate, feed intake and feed conversion ratio (FCR) were significantly improved by 2.80 g/kg diet. The maximum values of protein and ash were observed in 2.80 g/kg diet, while moisture was minimum. Lipid content of fish fed 3.95 g/kg diet was significantly higher than other diets. The highest plasma insulin‐like growth factor‐1 (IGF‐1) content was observed in 0.79 g/kg diet. In the liver, IGF‐1 mRNA levels were significantly downregulated by 2.80 g/kg dietary tryptophan, while glucokinase levels were by 3.95 g/kg, while glucose‐6‐phosphatase and phosphoenolpyruvate carboxykinase mRNA levels showed a converse trend compared with IGF‐1. Based on PWG and FCR, the optimal dietary tryptophan level was determined to be 1.99 g/kg (6.20 g/kg of dietary protein) and 1.96 g/kg (6.11 g/kg of dietary protein), respectively, using broken‐line regression analysis.     

Effect of dietary valine levels on the growth performance,feed utilization and immune function of juvenile golden pompano,Trachinotus ovatus

This experiment was designed to investigate the effects of dietary valine on the growth performance, feed utilization, digestive enzymes, serum antioxidant and immune indices of juvenile Trachinotus ovatus and determine its valine requirement. Six diets with different concentrations of L‐valine (15.0, 16.6, 18.6, 20.7, 23.5 and 25.4 g/kg dry diet, defined as diet Val‐1 to Val‐6.), were formulated to contain 430 g/kg crude protein with fish meal, soybean meal, peanut meal and precoated crystalline amino acids. Each diet was randomly assigned to triplicate treatments of 20 fish (the initial body weight was 5.34 ± 0.03 g) for 8 weeks. The results indicated that the final body weight and percent weight gain (PWG) increased with increasing valine concentration up to 18.6 g/kg (diet Val‐3), whereas the diets containing higher valine concentration reduced the growth performance significantly (p < .05). Moreover, the protein efficiency ratio, body protein deposition (BPD), muscle protein content, intestinal amylase and pepsin activities, serum T‐AOC, LZM activities, IgM, complement 3 and complement 4 concentration had a similar trend with PWG, and the trend of feed conversion ratio, serum AST, ALT activities, urea and MDA content was opposite. Meanwhile, the lipid contents of whole fish and muscle in diet Val‐6 were particularly lower than other diets (p < .05). The survival rate of diet Val‐1 was lowest in this study and was significantly lower than diet Val‐2 (p < .05). The results of polynomial regression based on PWG and BPD indicated that the optimal dietary valine requirement for Trachinotus ovatus reared in seawater‐floating net cages was 19.87–20.17 g/kg valine of dry diet, correspondingly 46.22–46.91 g/kg of dietary protein.     

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

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

Dietary lysine requirement of juvenile Pseudobagrus ussuriensis

An 8‐week feeding trial was conducted to determine dietary lysine requirement of juvenile Pseudobagrus ussuriensis (initial body weight: 0.60 g). Six isonitrogenous (crude protein, 400 g/kg) and isolipidic (crude lipid, 50 g/kg) diets were formulated to contain graded levels of dietary lysine (12.8, 19.9, 26.5, 34.0, 40.8 and 44.1 g/kg dry diets, respectively). The results indicated that weight gain, specific growth rate, productive protein value and protein efficiency ratio increased, while feed conversion ratio decreased with increasing dietary lysine level up to 34.0 g/kg dry diet and then levelled off. Fish fed diet with 12.8 g/kg lysine had the lowest lysine content (58.6 g/kg dry matter) in muscle, while fish fed diet with 34.0 g/kg lysine had the highest value (61.6 g/kg dry matter; p < .05). Broken‐line analysis on the basis of weight gain showed that the optimal dietary lysine requirement for maximum growth of juvenile Pseudobagras ussuriensis is 33.5 g/kg dry diet (82.4 g/kg dietary protein). Quadratic regression analysis of protein efficiency ratio against dietary lysine levels indicated that the optimal dietary lysine requirement of juvenile Pseudobagras ussuriensis is 36.4 g/kg dry diet (89.5 g/kg dietary protein).     

Determination of dietary phosphorus requirement of stinging catfish Heteropneustes fossilis based on feed conversion,growth, vertebrae phosphorus,whole body phosphorus,haematology and antioxidant status

A 12‐week feeding trial was conducted to determine the dietary phosphorus requirement of Heteropneustes fossilis fingerlings (7.7 ± 0.04 g). Fish were fed casein–gelatine‐based purified diets in triplicate groups near satiation with seven different levels of dietary phosphorus (3.2, 5.2, 7.2, 9.2, 11.2, 13.2 and 15.2 g/kg dry diet). All diets were formulated to be isoproteic (400 g/kg) and isoenergetic (17.89 kJ/g). Highest absolute weight gain (68.38 g/fish), best feed conversion ratio (1.48), protein retention efficiency (30.74%), protein gain (12.44 g/fish), haemoglobin (11.19 g/dL), RBCs (3.12 x10⁶/mm³), haematocrit (33.44%) and serum phosphate (2.82 mg/L) were found at 9.2 g/kg phosphorus. Hepatic superoxide dismutase and catalase activity were also significantly influenced by the dietary phosphorus levels. Whole body and vertebrae phosphorus concentrations increased significantly as the amount of dietary phosphorus increased from 3.2 to 11.2 g/kg dry diet and then plateaued. More accurate information on dietary phosphorus requirement was obtained by subjecting the AWG, FCR, vertebrae phosphorus and whole body phosphorus concentrations data against various levels of dietary phosphorus to broken‐line analysis, which yielded the requirement in the range of 9.0–11.0 g/kg for optimum growth and mineralization of H. fossilis.