Dietary biotin requirement of juvenile blunt snout bream,Megalobrama amblycephala

This study was conducted to determine the optimal dietary biotin requirement of juvenile Megalobrama amblycephala. Quadruple groups of fish (initial average weight 2.01 ± 0.01 g) were fed thrice daily with six isonitrogenous and isoenergetic purified diets containing 0 (basal diet), 0.015, 0.049, 0.158, 0.624 and 2.49 mg kg^?1 biotin, respectively, for 8 weeks. Results showed that survival rate, final weight, feed intake, specific growth rate, protein efficiency ratio and nitrogen retention efficiency all increased significantly (P < 0.01) as dietary biotin levels increased from 0 to 0.049 mg kg^?1, whereas the opposite was true for feed conversion ratio. Dressout percentage, condition factor, hepatosomatic index, viscera/body ratio all showed no significant difference (P > 0.05) within the biotin range tested. Contrary to moisture content, whole‐body protein and lipid contents showed a positive correlation with dietary biotin levels. In addition, liver biotin content increased significantly (P < 0.05) with increasing dietary biotin levels up to 0.624 mg kg^?1. Hepatic pyruvate carboxylase (PC) and acetyl‐CoA carboxylase (ACC) activities both showed an increasing trend as dietary biotin levels increased. Based on the regression analysis of weight gain, hepatic PC and ACC activities, the optimal dietary biotin requirement of juvenile Megalobrama amblycephala is estimated to be 0.063, 0.071 and 0.075 mg kg^?1, respectively.     

Influence of dietary biotin levels on growth and non‐specific immune response in large yellow croaker,Larimichthys crocea R

A 9‐week feeding experiment was conducted to determine the effect of dietary biotin levels on growth performance and non‐specific immune response of large yellow croaker. Fish (6.16 ± 0.09 g) were fed twice daily to apparent satiation with diets containing 0.00 (as the basal diet), 0.01, 0.05, 0.25, 1.24 and 6.22 mg biotin kg^?1 diet. Results showed that fish fed the basal diet had the lowest survival rate, and fish fed 0.05 mg kg^?1 dietary biotin achieved significantly higher final weight and weight gain. Dietary biotin levels had no significant influence on carcass crude lipid, moisture and ash content, but significantly influenced the carcass crude protein. Liver biotin concentration significantly increased with the supplementation of biotin, but no tissue saturation was found within the supplementation scope of biotin. Broken‐line regression analysis of weight gain showed that juvenile large yellow croaker requires a minimum dietary biotin of 0.039 mg kg^?1 for maximal growth. The analyses of serum parameters showed that the moderate‐ (0.05 mg kg^?1) and high‐dose (6.22 mg kg^?1) dietary biotin significantly improved both lysozyme and alternative complement pathway activities, indicating dietary biotin within a certain range could improve the non‐specific immune response of large yellow croaker.     

Dietary biotin requirement determined for Indian catfish, Heteropneustes fossilis (Bloch), fingerlings

Triplicate groups of Indian catfish, Heteropneustes fossilis (Bloch), fingerlings (average wet weight 3.55 ± 0.03 g) were fed semi-purified diets containing six levels of biotin (0, 0.086, 0.26, 0.86, 2.5 and 4.3 mg kg⁻¹ diet) for 15 weeks. After 42 days of feeding, fish fed the control (no biotin) diet had developed severe deficiency signs characterized by convulsions, heavy mortality, listlessness, poor feed conversion and feed intake, dark skin colour, tetanus and weight loss. None of these signs was seen in fish fed biotin-supplemented diets. Among all the biotin-supplemented diets, percentage weight gain was significantly highest for fish fed the diet supplemented with 0.26 mg of biotin kg⁻¹ and significantly lowest for fish fed the diet supplemented with 0.086 mg of biotin kg⁻¹. Feed conversion ratio (FCR) and protein efficiency ratio (PER) patterns were similar to that of percentage weight gain. The carcass protein and lipid contents were influenced by the dietary biotin up to fish fed 0.26 mg of biotin kg⁻¹. Significantly higher body biotin content, liver pyruvate carboxylase and acetyl CoA carboxylase activities were observed in fish fed biotin-supplemented diets than in fish fed the control diet. Broken-line analyses showed that the optimum dietary requirement for biotin for maximal weight gain, body biotin content, liver pyruvate carboxylase and acetyl CoA carboxylase activities was about 0.25 mg kg⁻¹. Associated liver pyruvate carboxylase and acetyl CoA carboxylase activities for normal growth ranged from 105 to 120 units mg⁻¹ protein and from 9 to 11 units mg⁻¹ protein respectively.     

Dietary biotin requirement for growth of juvenile zebrafish Danio rerio (Hamilton‐Buchanan)

This study was conducted to estimate the dietary biotin requirement for maximum growth of zebrafish Danio rerio. Six isonitrogenous and isocaloric purified diets containing 0.031 (biotin‐unsupplemented diet), 0.061, 0.263, 0.514, 1.741 and 2.640 mg biotin kg^?1 diet were fed in triplicate tanks for a total of 12 weeks to juvenile zebrafish (initial mean body mass 0.13 ± 0.001 g). From 4 weeks of feeding, fish fed diets with ≤0.061 mg biotin kg^?1 showed biotin deficiency signs, such as retarded growth and skeletal deformity, was observed on some dead fish. At the end of the study, gill disorders were observed on some fish and liver glycogen accumulation were also observed in fish fed these diets. Fish fed the biotin‐unsupplemented diet exhibited a lower final body weight, protein efficiency ratio and feed utilization than the fish fed the biotin‐supplemented diets, whereas the highest values were observed with the diet containing 0.51 mg biotin kg^?1 diet (P < 0.05). A linear relationship (r² = 0.77; P < 0.0001) was observed between whole‐body biotin content and dietary biotin level. A broken‐line analysis indicated that the optimum dietary biotin content for maximal growth expressed as final body weight is 0.51 mg kg^?1 diet.     

Effects of dietary ethoxyquin on growth,feed utilization and residue in the muscle of juvenile Japanese seabass,Lateolabrax japonicus

Ethoxyquin (EQ) is the most common synthetic antioxidant used for preventing rancidity in fish foodstuffs. However, literature related to the effects of dietary EQ on performance of fish was limited. The present study was conducted to investigate the effects of EQ on performance and EQ residue in muscle of juvenile Japanese seabass Lateolabrax japonicus and to estimate the optimal EQ concentration in the diet. Graded levels [0 (control), 50, 150, 450 and 1350 mg EQ kg^?1 diet] of EQ were added to the basal diet, resulting in five dietary treatments in the experiment. Each diet was fed to triplicate groups of seabass (initial body weight 8.01 ± 0.76 g) for 12 weeks in floating sea cages (1.5 × 1.5 × 2.0 m, 30 fish per cage). Survival ranged from 78.9 to 86.7%, and was irrespective of dietary EQ levels. The specific growth rate (SGR) of fish fed diets supplemented with ≤50 mg kg^?1 EQ had significantly (P < 0.05) higher SGR than fish fed diets supplemented with ≥150 mg kg^?1 EQ, the highest SGR was observed in fish fed diet with 50 mg kg^?1 EQ supplementation. Feed intake (FI) and feed efficiency (FE) were not significantly (P > 0.05) different among dietary treatments. Fish fed diets with 50 and 1350 mg kg^?1 EQ had a significant (P < 0.05) lower body lipid content than fish in the control group. Muscle EQ level significantly increased when dietary EQ increased. Optimal EQ concentration estimated by polynomial regression based on maximum growth of juvenile Japanese seabass was 13.78 mg kg^?1 diet.     

Effects of dietary biotin supplement on growth,body composition,intestinal enzyme activities and microbiota of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1400 juvenile Jian carp (Cyprinus carpio var. Jian) (7.72 ± 0.02 g) were fed seven purified diets containing 0.010 (basal diet), 0.028, 0.054, 0.151, 0.330, 1.540 and 2.680 mg biotin kg^?1 for 63 days to investigate the effects of biotin on growth, body composition, intestinal enzyme activities and microbiota. Specific growth rate (SGR), feed intake, feed efficiency and protein retention value were the highest when dietary biotin level was 0.151 mg kg^?1 diet. Crude protein, lipid and ash content of fish carcass improved with increasing dietary biotin levels up to 0.054, 0.151 and 0.028 mg kg^?1 diet, respectively (P < 0.05). Intestinal folds height, trypsin, chymotrypsin, lipase, amylase, alkaline phosphatase, Na⁺, K⁺‐ATPase, γ‐glutamyl transpeptidase and creatinekinase activities increased with dietary biotin levels up to 0.151–0.330 mg kg^?1 diet (P < 0.05). Intestinal Aeromonas and Escherichia coli significantly decreased with increasing dietary biotin up to 0.151 mg kg^?1 diet, while Lactobacillus and Bacillus significantly increased with dietary biotin levels up to 0.054 and 0.151 mg kg^?1 diet, respectively. In conclusion, biotin could improve digestive and absorptive ability of fish, and the dietary biotin requirement for SGR of juvenile Jian carp (7.72–32.67 g) was 0.15 mg kg^?1 diet.     

Dietary myo‐inositol requirement for juvenile gibel carp (Carassius auratus gibelio)

An 11‐week growth trial was conducted to determine dietary myo‐inositol (MI) requirement for juvenile gibel carp (Carassius auratus gibelio). Myo‐inositol was supplemented to the basal diet to formulate six purified diets containing 1, 56, 107, 146, 194 and 247 mg MI kg^?1 diet, respectively. Each diet was fed to triplicate groups of juvenile gibel carp (initial body weight 3.38 ± 0.27 g, mean ± SD) in a flow‐through system. The diets were randomly assigned to different fish tanks. Fish fed ≥ 107 mg MI kg^?1 diet had significantly higher weight gain (WG), feed efficiency (FE) and protein efficiency ratio than those fed 1 mg MI kg^?1 diet. Fish fed ≥ 56 mg MI kg^?1 diet had higher feeding rate and survival compared with fish fed 1 mg MI kg^?1 diet. Dietary supplemental inositol did not affect fish liver inositol concentration. Fish fed ≥ 56 mg MI kg^?1 diet had higher body dry matter, crude protein and gross energy and lower hepatosomatic index than fish fed 1 mg MI kg^?1 diet. Dietary inositol supplementation decreased fish body ash. Quadratic regression of weight gain indicated that the myo‐inositol requirement to maximum growth for juvenile gibel carp was 165.3 mg MI kg^?1 diet.     

Replacement of dietary fish meal by soybean meal supplemented with crystalline methionine for Japanese seabass (Lateolabrax japonicus)

Two 8‐week feeding trials were conducted to evaluate soybean meal (SBM) as a fish meal substitute in diets for Japanese seabass, Lateolabrax japonicas. In trial I, a control diet (C) contained 400 g kg^?1 fish meal, and 20%, 40%, 60% and 80% of the fish meal were replaced with SBM, supplied with 3 g kg^?1 DL‐methionine and 2 g kg^?1 L‐lysine (S20, S40, S60 and S80). In trial II, 60% and 80% of the fish meal in diet C were replaced with SBM, supplied with DL‐methionine at 3 g kg^?1 (S60, S80) or 6 to 7 g kg^?1 (RS60, RS80). The feed intake was lower in fish fed diet C than in fish fed diets S20, S40, S60 and S80 (trial I). No significant differences were found in the weight gain, nitrogen retention efficiency and body composition between fish fed diets C, S20, S40 and S60 (trial I), between fish fed diets S60 and RS60 or between fish fed diets S80 and RS80 (trial II). This study indicates that dietary fish meal level for Japanese seabass can be reduced to 160 g kg^?1 by using SBM as a fish meal substitute.     

Effects of dietary calcium levels on growth and tissue mineralization in Japanese seabass,Lateolabrax japonicus

A feeding trial was conducted to investigate the effects of different levels of calcium (Ca) on growth and tissue mineralization in Japanese seabass, Lateolabrax japonicus. Six experimental diets were formulated to contain different levels of Ca (2.9, 4.2, 6.5, 7.9, 10.2 and 31.0 g kg^?1) from dietary ingredients and Ca‐lactate·5H₂O. The diets were fed to three triplicate groups of Japanese seabass (initial weight, 12.5 ± 0.0 g) for 56 days. Dietary Ca had no significant effect on survival or feed efficiency; however, the highest Ca (31.0 g kg^?1) diet significantly reduced weight gain, feeding rate and whole‐body and muscle protein and lipid contents, as well as serum Ca concentration and alkaline phosphatase activity. A significant reduction in vertebral Ca, P, Zn, Fe and Mn contents and scale Ca, P, Mg and Mn contents was observed in Japanese seabass as dietary Ca level increased. Deformed fish were primarily found in the 2.9 and 31.0 g Ca kg^?1 groups, indicating that these fish had poor bone mineralization.     

Effects of dietary squid viscera meal on growth and cadmium accumulation in tissues of Japanese seabass, Lateolabrax japonicus (Cuvier 1828)

Cadmium (Cd) is a toxic environmental pollutant with a long biological half‐life and can produce both hepatic and renal injuries in mammals and fish. Squid viscera meal (SVM), an effective attractant for aquatic animals, is widely used as an ingredient in aquafeeds. However, SVM is rich in Cd and its complexes. A study was conducted to evaluate the effects of dietary SVM on the growth and Cd deposition in tissues of Japanese seabass, Lateolabrax japonicus. Three practical diets were formulated to contain 0, 50 and 100 g SVM kg^?1 diet, respectively, containing 0.21, 7.26 and 12.08 mg Cd kg^?1 diet. Each diet was randomly assigned to triplicate groups of 80 Japanese seabass (mean initial weight, 10.89±0.21 g) in floating sea cages (1.5 × 1.5 × 2.0 m). Fish were fed twice daily (06:30 and 16:30 hours) to satiation for 8 weeks. The results showed that there were no significant differences in fish survival among three dietary treatments, but significant higher specific growth rates (SGR) were observed in the fish fed diets with 50 or 100 g SVM kg^?1 diet than that from the control group (P<0.05). The Cd concentrations in the kidney, liver and gill were found in a decreasing order at each treatment, and positively correlated with dietary Cd levels. Fish fed diets with 50 and 100 g SVM kg^?1 diet had significantly higher Cd accumulations in the kidney (3.25, 5.85 mg kg^?1), liver (0.76, 1.26 mg kg^?1) and gill (0.42, 0.58 mg kg^?1) compared with the control group (0.82, 0.34 and 0.32 mg kg^?1 respectively) (P<0.05). The Cd concentration in fish muscle; however, was undetectable in any treatment. Therefore, based on these results, accumulation of Cd in edible tissue (muscle) of farmed Japanese seabass is not a food safety issue. However, long‐term feeding of diets with SVM may result in accumulation of Cd in the kidneys, liver and gills of fish.     

The effects of dietary lipid levels on performance and heat‐shock protein response of juvenile white seabass,Atractoscion nobilis

A feeding trial was conducted to study the effect of dietary lipid on growth performance and heat‐shock protein (HSP70 and HSP60) response of white seabass (WSB), Atractoscion nobilis. Five diets were formulated to contain 440 g kg^?1 protein from 300 g kg^?1 fish meal, 240 g kg^?1 soybean meal and 100 g kg^?1 soy protein concentrate with different levels of lipid: 100, 120, 140, 160 or 180 g kg^?1. At the end of the trial, heat shock response based on HSP70 and HSP60 was measured in liver and white muscle from fish at ambient temperature and temperature shock conditions. Final weight and percent gain were significantly higher for fish fed the 100 g kg^?1 lipid diet than for fish fed the rest of the diets (P ≤ 0.05). Feed conversion ratio was lowest for fish fed the 100 g kg^?1 lipid diet. The HSP70 and HSP60 responses were positively correlated to dietary lipid levels following temperature shock. At ambient temperature, HSP60 and HSP70 responses in muscle and HSP60 response in liver increased with dietary lipid level. Temperature shock significantly increased the HSP response of fish in all treatments. Results of this study demonstrated that a moderate (110–120 g kg^?1) level of dietary lipids would be recommended for production diets but a higher dietary lipid level may be required for optimal stress tolerance.     

Dietary ascorbic acid requirement of cobia,Rachycentron canadum Linneaus

A 10‐week feeding trial was conducted to determine the optimal requirement of cobia (Rachycentron canadum Linneaus) for dietary ascorbic acid (AA). Graded levels of L‐ascorbyl‐2‐polyphosphate (LAPP) were supplemented in basal diet to formulate six semi‐purified diets containing 2.70 (the control diet), 8.47, 28.3, 80.6, 241 and 733 mg AA equivalent kg^?1 diet, respectively. Each diet was randomly fed to triplicate groups of fish in flow‐through plastic tanks (300 L), and each tank was stocked with 25 fish with average initial weight of 4.59 ± 0.36 g. Observed deficiency signs included poor growth, higher mortality and lower feeding rate (FR) in the fish of the control group. Fish fed the control diet had significantly lower weight gain (WG), lower feed efficiency ratio (FER) and lower tissue AA concentrations in fish liver and muscle. With the increase of dietary AA, the survival, WG, FER, hepatic and muscular AA concentrations of cobia significantly increased and then levelled off. The dietary AA requirement of cobia was estimated to be 44.7 mg kg^?1 based on WG, 53.9 mg kg^?1 or 104 mg kg^?1 based on either hepatic or muscular AA concentration, respectively.     

Quantitative dietary lysine requirement of juvenile striped bass Morone saxatilis

Two feeding trials of 8 and 10 weeks each were conducted to quantify the dietary lysine requirement of juvenile striped bass, Morone saxatilis. Diets in both experiments contained approximately 420 g crude protein kg^–1 and 13.4 MJ digestible energy (DE) kg^?1. L ‐Lysine‐HCl was added to the basal diet to yield five and six treatments in the two experiments. Diets in the first experiment were determined to contain 9.2, 14.1, 14.6, 19.9 and 21.0 g available lysine kg^?1 on a dry‐matter basis. Diets in the second experiment were determined to contain 14.8, 18.1, 21.3, 24.5, 27.6 and 30.9 g available lysine kg^?1 on a dry‐matter basis. Weight gain, specific growth rate (SGR), feed conversion ratio (FCR), and apparent nitrogen utilization (ANU) were significantly (P < 0.05) improved by increasing dietary lysine concentrations to approximately 20 g kg^?1 of diet. Least‐squares regression analysis of weight gain and SGR in the first experiment indicated a minimum dietary lysine requirement of 20.1 ± 2 g kg^?1 dry diet. Least‐squares regression analysis of the same criteria measured in the second experiment yielded the following estimates of dietary lysine requirements (g kg^?1 dry diet): 19.8 ± 2.3 for weight gain, 21.7 ± 1.5 for SGR, 23.7 ± 3.5 for FCR and 18.6 ± 1.3 for ANU. From these results the minimum recommended dietary lysine requirement for optimal growth of juvenile striped bass is approximately 21 g kg^?1 dry diet which equates to 49 g kg^?1 dietary protein or 1.57 mg kJ^?1 DE. Although higher than that reported for hybrid striped bass, this requirement level is similar to those reported for many other fish species.     

The potential of land animal protein ingredients to replace fish meal in diets for cuneate drum, Nibea miichthioides, is affected by dietary protein level

A net pen experiment was carried out to examine the effect of dietary protein level on the potential of land animal protein ingredients as fish meal substitutes in practical diets for cuneate drum Nibea miichthioides. Two isocaloric basal (control) diets were formulated to contain 400 g kg^?1 herring meal but two different digestible protein (DP) levels (400 versus 350 g kg^?1). At each DP level, dietary fish meal level was reduced from 400 to 280, 200, 80 and 0 g kg^?1 by incorporating a blend that comprised of 600 g kg^?1 poultry by‐products meal (PBM), 200 g kg^?1 meat and bone meal (MBM), 100 g kg^?1 feather meal (FEM) and 100 g kg^?1 blood meal (BLM). Cuneate drum fingerling (initial weight 42 g fish^?1) were fed the test diets for 8 weeks. Fish fed the test diets exhibited similar feed intake. Final body weight, feed conversion ratio and nitrogen retention efficiency was not significantly different between fish fed the basal diets containing 350 and 400 g kg^?1 DP. Weight gain decreased linearly with the reduction of dietary fish meal level at the 350 g kg^?1 DP level, but did not decrease with the reduction of dietary fish meal level at the 400 g kg^?1 DP level. Results of the present study suggest that fish meal in cuneate drum diets can be completely replaced with the blend of PBM, MBM, FEM and BLM at the 400 g kg^?1 DP level, based on a mechanism that excessive dietary protein compensate lower contents of bio‐available essential amino acid in the land animal protein ingredients relative to fish meal.     

Dietary copper requirement of juvenile yellow catfish Pelteobagrus fulvidraco

The present experiment was conducted to quantify dietary copper (Cu) requirement for juvenile yellow catfish Pelteobagrus fulvidraco. The six experimental diets were formulated to contain the graded levels of CuSO₄·5H₂O (0, 0.005, 0.01, 0.02, 0.04 and 0.08 g kg^?1 diet respectively) providing the actual dietary copper values of 2.14 (control), 3.24, 4.57, 7.06, 12.22 and 22.25 mg Cu kg^?1 diet respectively. Each diet was fed to triplicate groups of yellow catfish (initial body weight: 3.13 ± 0.09 g, means ± SD) in an indoor static rearing system for 7 weeks. Fish fed the diet containing 3.24 mg Cu kg^?1 diet had the highest weight gain and specific growth rate, but they were not significantly different from that of fish fed the 4.57 and 7.06 mg Cu kg^?1 diets (P > 0.05). The poorest feed conversion rate, the lowest protein efficiency ratio, the lowest hepatosomatic index and viscerosomatic index were observed in fish fed the diet containing the highest Cu content diet (P < 0.05). Condition factor showed no significant differences among the treatments (P > 0.05). Proximate composition of fish body was significantly affected by dietary copper level (P < 0.05). Cu contents of whole body and liver increased with dietary Cu levels (P < 0.05), but muscle Cu content remained relatively stable (P > 0.05). Analysis by the second‐order regression of SGR and linear regression of whole‐body Cu retention of the fish indicated that dietary Cu requirements in juvenile yellow catfish were 3.13–4.24 mg Cu kg^?1 diet.     

The effects of dietary selenium on growth performances,oxidative stress and tissue selenium concentration of gibel carp (Carassius auratus gibelio)

A 100‐day growth trial was executed to determine the dietary selenium (Se) requirement of juvenile gibel carp (Carassius auratus gibelio). Selenomethionine was supplemented to casein‐gelatin diets at 0, 0.1, 0.25, 0.5, 1.0, 2.5 and 5 mg Se kg^?1, respectively. Each of these seven semi‐purified diets containing 0.34, 0.47, 0.66, 0.82, 1.23, 2.77 and 5.13 mg Se kg^?1 was fed to triplicate groups of gibel carp (2.74 ± 0.02 g) in a flow‐through system. No behaviour abnormalities and no mortality were found in fish exposed to dietary Se concentrations. With the increasing dietary Se supplemented concentrations, weight gain of fish remarkably increased at the levels of ≤1 mg Se kg^?1 diet and then showed no significant difference above 1 mg Se kg^?1 levels. Although growth performances (weight gain, hepatosomatic index, condition factor and survival) were not impaired in gibel carp fed at above the levels of 2.5 mg Se kg^?1, indicators of oxidative stress were changed significantly. Serum superoxide dismutase (SOD) activities significantly declined, hepatic glutathione peroxidase (GPx) activities significantly increased and the tissue Se concentrations significantly raised at the highest supplemented level of 5 mg Se kg^?1. A clear linear relationship between Se‐depended GPx activities and hepatic Se concentrations was observed. The present results indicated that the dietary Se requirement for gibel carp is 1.18 mg Se kg^?1 diet based on weight gain, GPx activities and tissue accumulation.     

Dietary vitamin C requirement of juvenile Chinese sucker (Myxocyprinus asiaticus)

An 8‐week growth experiment was conducted to quantify the appropriate dietary vitamin C requirement of juvenile Chinese sucker (Myxocyprinus asiaticus). Triplicate groups of 30 experimental fish [initial body weight: (7.1 ± 0.3) g] were cultured in 500 L aquaria and fed with semi‐purified diets containing six levels [10.1 (unsupplemented diet), 37.4, 64.9, 125.2, 244.2 and 482.0 mg kg^?1 diet, respectively] of vitamin C (supplied as L‐ascorbyl‐2‐polyphosphate). Results showed that weight gain of Chinese sucker was significantly increased with increasing dietary vitamin C levels, but there was no significant difference of weight gain among fish fed the diets containing more than 125.2 mg kg^?1 vitamin C. As dietary vitamin C increased, the liver vitamin C content of fish showed the increasing trend firstly and then stabled, while the muscle vitamin C content significantly increased without reaching a constant level. Lower liver malondialdehyde content was observed in 125.2, 244.2 and 482.0 mg kg^?1 vitamin C groups, and higher antioxidant capacity and superoxide dismutase activities were observed in supplemented groups when compared to the unsupplemented group. Liver aspartate aminotransferase and alkaline phosphatase activities were also significant affected by dietary vitamin C. Ash content of fish fed the diet with 244.2 or 482.0 mg kg^?1 vitamin C was significantly higher than that of fish fed the other diets. However, dietary vitamin C had no significant effects on whole‐body crude protein, lipid and moisture contents. The vitamin C requirement of juvenile Chinese sucker was estimated to be 84.6 and 126.1 mg kg^?1 based on weight gain and liver vitamin C concentration respectively.     

Requirements of vitamin C (L-ascorbyl-2-monophosphate-Mg and L-ascorbyl-2-monophosphate-Na) and its effects on immune responses of grouper, Epinephelus malabaricus

An 8‐week feeding trial was conducted to evaluate two vitamin C derivatives, L‐ascorbyl‐2‐monophosphate‐Mg (C2MP‐Mg) and L‐ascorbyl‐2‐monophosphate‐Na (C2MP‐Na), to satisfy the vitamin C requirement and to test their effects on the immune responses of juvenile grouper, Epinephelus malabaricus. C2MP‐Mg and C2MP‐Na were each supplemented at 20, 50, 80, 150, 250, and 400 mg kg^?1 diet in the basal diet providing of 7, 18, 31, 51, 93, 145 mg ascorbic acid (AA) equivalent of C2MP‐Mg kg^?1 diet and 4, 10, 17, 31, 47, 77 mg ascorbic acid (AA) equivalent of C2MP‐Na kg^?1 diet, respectively. Basal diet without AA supplementation was included as control. Each diet was fed to triplicate groups of grouper (mean initial weight 3.20 ± 0.05 g). Fish fed diets supplemented with either C2MP‐Mg or C2MP‐Na had significantly (P < 0.05) greater weight gain (WG), feed efficiency and survival than those fed the unsupplemented control diet. Liver ascorbate concentrations in fish generally increased as dietary C2MP‐Mg or C2MP‐Na supplementation level increased. Haemolytic complement activity was higher in fish fed diets supplemented with 92 mg AA equivalent of C2MP‐Mg kg^?1 or 10–17 mg AA equivalent of C2MP‐Na kg^?1 than fish fed the unsupplemented control diet. Lysozyme activity was higher in fish fed ≥51 mg AA equivalent of C2MP‐Mg kg^?1 or ≥47 mg AA equivalent of C2MP‐Na kg^?1 than fish fed the unsupplemented control diet. Analysis by broken‐line regression of WG indicated that the adequate dietary vitamin C concentration from each vitamin C derivative in growing grouper is 17.9 mg AA equivalent of 2MP‐Mg kg^?1 and 8.3 mg AA equivalent of C2MP‐Na kg^?1, and it also indicated that C2MP‐Mg is about 46% as effective as C2MP‐Na in meeting the vitamin C requirement of grouper.     

Dietary leucine requirement of Juvenile Nile tilapia,Oreochromis niloticus

An 8‐week feeding trial was conducted to evaluate the effects of dietary leucine on growth performance, feed utilization, body composition and non‐specific immune responses of juvenile Nile tilapia. Five isonitrogenous and isoenergetic diets were formulated to contain graded levels of L‐leucine (5.3, 8.1, 10.9, 13.2, 15.6 and 18.1 g kg^?1 diet, respectively) from dietary ingredients and crystalline L‐leucine. Each diet was randomly assigned to triplicate groups of 20 juvenile fish (1.94 ± 0.01 g) three times daily to apparent satiation. Results showed that the weight gain (WG) and specific growth rate (SGR) increased as dietary leucine concentrations increased from 5.3 to 13.2 g kg^?1 and then decreased slightly with further increase in dietary leucine concentrations. Quadratic regression analysis (y = ?522.6x² + 1304.x + 132.6, R² = 0.684) on weight gain against dietary leucine levels indicated that the optimal dietary leucine requirement was estimated to be 12.5 g kg^?1 diet (corresponding to 43.1 g kg^?1 of dietary protein). Leucine supplementation had no impact on the survival and body composition of tilapia. Serum lysozyme activity of fish fed diet containing 13.2 g kg^?1 leucine significantly increased compared to fish fed diet containing 5.3 g kg^?1. Serum superoxide dismutase activity and immunoglobulin M (IgM) concentration were not significantly affected by dietary leucine supplementation.     

Effect of high dietary intakes of vitamin E and n-3 HUFA on immune responses and resistance to Edwardsiella tarda challenge in Japanese flounder (Paralichthys olivaceus, Temminck and Schlegel)

A feeding experiment was conducted to investigate the effects of high dietary intake of vitamin E (supplied as dl ‐α‐tocopheryl acetate) and n‐3 highly unsaturated fatty acid (n‐3 HUFA) on the non‐specific immune response and disease resistance in Japanese flounder Paralichthys olivaceus. Nine practical diets were formulated to contain one of three levels of vitamin E namely, 0, 80 or 200 mg kg^?1 (the total α‐tocopherol contents in the diets were 21, 97 and 213 mg kg^?1 based on analysis), and at each vitamin E level with one of three n‐3 HUFA levels i.e. 0.5%, 1.5% or 2.0%. Each diet was randomly assigned to triplicate groups of Japanese flounder (initial body weight: 40.5±1.0 g, mean±SD) in a re‐circulation rearing system. Fish were fed twice daily to apparent satiation at 07:00 and 18:00 hours for 12 weeks. During the experimental period, water temperature was maintained at 18±1°C, salinity 31–35 g L^?1, and pH 7.8–8.2. Dissolved oxygen was not less than 6 mg L^?1, and there were negligible levels of free ammonia and nitrite. The results showed that the increase in dietary n‐3 HUFA from 0.5% to 1.0% significantly decreased muscle α‐tocopherol contents in fish‐fed diets with 21 and 97 mg α‐tocopherol kg^?1 diet (P<0.05). In 1.0% HUFA groups, alternative complement pathway activity (ACH₅₀) of fish fed the diet containing the 213 mg α‐tocopherol kg^?1 diet was significantly higher than noted for fish fed the diet containing 97 mg α‐tocopherol kg^?1 diet (P<0.05). Fish fed the diet with 213 mg α‐tocopherol kg^?1 and 2.0% n‐3 HUFA had the highest lysozyme activity (131.7 U mL^?1) among all the dietary treatments. Fish fed the diets containing 97 and 213 mg α‐tocopherol kg^?1 with 1.0% n‐3 HUFA had significantly higher respiratory burst activity than those fed the diets containing 21 mg α‐tocopherol kg^?1 with 0.5 and 1.0% n‐3 HUFA (P<0.05). In the disease resistance experiment, high intake of dietary vitamin E with 213 mg α‐tocopherol kg^?1 significantly decreased cumulative mortality and delayed the days to first mortality after a 7‐day Edwardsiella tarda challenge (P<0.05). In addition, under the experimental conditions, dietary vitamin E and n‐3 HUFA had a synergistic effect on the non‐specific immune responses and disease resistance in Japanese flounder (P<0.05).