Requirements of vitamin C (l-ascorbyl-2-sulphate and l-ascorbyl-2-polyphosphate) and its effects on non-specific immune responses of grouper, Epinephelus malabaricus

An 8‐week feeding trial was conducted to determine two vitamin C derivatives, l ‐ascorbyl‐2‐sulphate (C2S) and l ‐ascorbyl‐2‐polyphosphate (C2PP), to satisfy vitamin C requirement and to test their effects on the non‐specific humoral immune responses of juvenile grouper, Epinephelus malabaricus. C2S and C2PP were each supplemented at 20, 50, 80, 150, 250 and 400 mg kg^?1 diet in the semi‐purified basal diet providing of 7, 16, 28, 55, 86, 142 mg ascorbic acid (AA) equivalent of C2S kg^?1 diet and 4, 9, 15, 31, 49, 75 mg AA equivalent of C2PP kg^?1 diet, respectively. Basal diet without AA supplemented was included as a control. Each diet was fed to triplicate groups of grouper (mean initial weight: 6.69 ± 0.07 g). Fish fed diets with ≥28 mg AA equivalent of C2S or ≥4 mg AA equivalent of C2PP kg^?1 had significantly (P < 0.05) greater weight gain (WG) than fish fed the unsupplemented control diet. Liver AA concentrations were higher in fish fed diets with ≥16 mg AA equivalent of C2S or ≥9 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Alternative pathway of complement activation (ACP) was higher in fish fed diets with ≥55 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the control diet. Lysozyme activity was higher in fish fed ≥86 mg AA equivalent of C2S or ≥15 mg AA equivalent of C2PP kg^?1 than fish fed the 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 46.2 mg AA equivalent of C2S kg^?1 diet and 17.8 mg AA equivalent of C2PP kg^?1 diet, and it also indicated that C2S is approximately 39% as effective as C2PP in meeting the vitamin C requirement for grouper. The data suggest that both C2S and C2PP supplementation support non‐specific immune responses of grouper.     

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.     

Effects of dietary L‐carnitine level on growth performance,body composition and antioxidant status in beluga (Huso huso L. 1758)

A 17‐week feeding trial was carried out to evaluate the effects of dietary L‐carnitine level in beluga, Huso huso. A total of fish averaging 1247 ± 15.6 g (mean ± SD) were randomly distributed into 18 fibreglass tanks, and each tank holding 10 fish was then randomly assigned to one of three replicates of six diets with 50, 150, 350, 650, 950 and 1250 mg L‐carnitine kg^?1 diet. At the end of 17 weeks of feeding trial, average weight gain (WG), feed efficiency (FE), protein efficiency ratio (PER) and condition factor (CF) of fish fed 350 mg kg^?1 diet were significantly (P < 0.05) higher than those of fish fed 50, 150, 950 and 1250 mg kg^?1 diets. WG, FE, PER and CF of beluga fed 650 mg kg^?1 diet were also significantly higher than those of fish fed 50, 950 and 1250 mg kg^?1 diets. Whole body and muscle protein were significantly improved by the elevation of dietary L‐carnitine level up to 350 mg kg^?1. Liver superoxide dismutase and glutathione peroxidase activities of fish fed 350 and 650 mg kg^?1 diets were significantly higher than those of fish fed 50, 950 and 1250 mg kg^?1 diets. The dietary L‐carnitine level of 350–650 mg kg^?1 diet could improve growth performance, feed utilization, protein‐sparing effects of lipid, antioxidant defence system and reproductive success. Polynomial regression of WG suggested that the optimum dietary L‐carnitine level was 480 mg kg^?1 diet. Therefore, these results may indicate that the optimum dietary L‐carnitine could be higher than 350 but <650 mg kg^?1 diet in beluga reared in intensive culture conditions.     

Synergistic effects of dietary vitamin E and selenomethionine on growth performance and tissue methylmercury accumulation on mercury‐induced toxicity in juvenile olive flounder,Paralichthys olivaceus (Temminck et Schlegel)

An 8‐week feeding trial was conducted to evaluate the synergistic effects of dietary vitamin E and selenomethionine (SeMet) on induced methylmercury (MeHg) toxicity in juvenile olive flounder Paralichthys olivaceus. Nine semi‐purified diets were formulated to contain three different vitamin E levels as DL‐α‐tocopheryl acetate (0, 100 and 200 mg TAkg^?1 diet) and three different selenium (Se) levels (0, 2 and 4 SeMet mg kg^?1 diet) on the constant mercury toxicity level (20 mg MeHgkg^?1 diet). Nine experimental diets, in a 3² factorial design (E₀Se₀, E₀Se₂, E₀Se₄, E₁₀₀Se₀, E₁₀₀Se₂, E₁₀₀Se₄, E₂₀₀Se₀, E₂₀₀Se₂ and E₂₀₀Se₄), were fed to triplicate groups of fish averaging 2.3 ± 0.04 g (mean ± SD) in the semi‐recirculation system. After 8 weeks of feeding trial, vitamin E and Se showed significant effects on weight gain (WG) of fish (P < 0.05). We found that there was a clear trend of increasing WG with elevating vitamin E and Se levels in the diets. Feed efficiency (FE), specific growth rate (SGR), protein efficiency ratio (PER) and survivability exhibited a similar trend with WG. Both antioxidants had significant interaction effects on FE and PER (P < 0.05). Methylmercury concentrations in fish muscle, liver and kidney decreases in a dose‐dependent manner as dietary vitamin E and Se levels increase. Interestingly, the most significant interactive effects of vitamin E and Se were found in liver tissue for depleting Hg concentrations (P < 0.05). These findings suggest that dietary vitamin E more than 100 mg TA kg^?1 diet with 2 or 4 mg SeMet kg^?1‐supplemented diets could have synergistic effects on growth and liver mercury bioaccumulation on MeHg‐induced toxicity in juvenile olive flounder.     

Effect of dietary vitamin C on the growth performance,antioxidant ability and innate immunity of juvenile yellow catfish (Pelteobagrus fulvidraco Richardson)

A 12‐week feeding trial was conducted to evaluate the effects of dietary vitamin C on growth performance, antioxidant status and innate immune responses in juvenile yellow catfish, Pelteobagrus fulvidraco. Six isonitrogenous and isolipidic diets (44% crude protein and 7% lipid) were formulated to contain six graded dietary vitamin C (ascorbate‐2‐poly‐ phosphate, ROVIMIX^® STAY‐C^® 35) levels ranging from 1.9 to 316.0 mg kg^?1 diet. The results of present study indicated that fish fed the lowest vitamin C diet had lower weight gain (WG) and specific growth rate (SGR) than those fed the diets supplemented vitamin C. WG and SGR did significantly increase with dietary vitamin C levels increasing from 1.9 to 156.5 mg kg^?1. However, no significant increase was observed with further dietary vitamin C levels increasing from 156.5 to 316 mg kg^?1. Survival, protein efficiency ratio and feed efficiency were not significantly affected by the dietary vitamin C levels. The activities of serum superoxide dismutase, catalase and glutathione peroxidase significantly increased when dietary vitamin C levels increased from 1.9 to 156.5 mg kg^?1, fish fed the lowest vitamin C diet had higher serum malondialdehyde content than those fed the diets supplemented with vitamin C. Fish fed the diet containing 156.5 mg kg^?1 vitamin C had the highest lysozyme, total complement activity, phagocytosis index and respiratory burst of head kidney among all treatments. The challenge test with Aeromonas hydrophila indicated that lower cumulative survival was observed in fish fed the lowest vitamin C diet. Analysis by broken‐line regression of SGR and lysozyme activity indicated that the dietary vitamin C requirement of juvenile yellow catfish was estimated to be 114.5 and 102.5 mg kg^?1 diet, respectively.     

Evaluation of l-ascorbyl-2-glucose as the source of vitamin C for juvenile Korean rockfish Sebastes schlegeli (Hilgendorf)

This study was conducted to compare l ‐ascorbyl‐2‐glucose (AA2G) with l ‐ascorbyl‐2‐monophosphate‐Na/Ca (AMP‐Na/Ca) for supplying the dietary vitamin C for juvenile Korean rockfish Sebastes schlegeli (Hilgendorf). Fish were fed one of seven semi‐purified diets containing equivalents of 0, 50, 100 and 200 mg ascorbic acid (AA) kg^?1 diet in the form of AA2G or AMP‐Na/Ca for 12 weeks. After 12 weeks of feeding, weight gain, feed efficiency ratio and survival of fish fed the vitamin C‐free diet were significantly lower than those of fish fed the vitamin C‐supplemented diets in either form. The hepatosomatic index, condition factor and survival of fish fed AMP‐Na/Ca₁₀₀, AMP‐Na/Ca₂₀₀, AA2G₁₀₀ and AA2G₂₀₀ diets were significantly higher than fish fed the vitamin C‐free diet. After 9 weeks of feeding, fish fed the vitamin C‐free diet began to show vitamin C deficiency signs such as anorexia and lethargy. At the end of the 12‐week feeding trial, fish fed the vitamin C‐free diet exhibited vitamin C deficiency signs, e.g., anorexia, scoliosis, exophthalmia and fin haemorrhage. Vitamin C retention in the muscle and liver was similar to those of fish fed AA2G‐ or AMP‐Na/Ca‐supplemented diets. In general, there was no significant difference in the muscle and liver vitamin C concen‐tration in fish fed the AA2G and AMP‐Na/Ca diets at the same supplementation levels.     

Dietary nucleotide supplementation enhances growth and immune responses of grouper,Epinephelus malabaricus

Basal diet containing 0.5, 1.0, 1.5 and 2.0 g kg^?1 mixture of inosine monophosphate (IMP), adenosine monophosphate (AMP), guanosine monophosphate (GMP), uridine monophosphate (UMP) and cytidine monophosphate (CMP) (1 : 1 : 1 : 1 : 1) (mixed‐NT; Experiment 1) and 1.5 g kg^?1 from each nucleotides and mixed‐nucleotides (NT; Experiment 2) were fed to triplicate groups of grouper for 8 weeks. Basal diet without NT was used as control in both Experiments. In Experiment 1, fish fed the diet with 1.5 g mixed‐NT kg^?1 had higher (P < 0.05) weight gain (WG) than the control group. The superoxide anion (O₂^?) production ratio was higher in fish fed diets with 1.0–1.5 g mixed‐NT kg^?1 than the fish fed diets with ≤0.5 g mixed‐NT kg^?1. In Experiment 2, fish fed diets with nucleotides had higher WG than the control group. The O₂^? production ratio was higher in fish fed the diet with 1.5 g AMP kg^?1, followed by fish fed diets with 1.5 g UMP and mixed‐NT kg^?1, and lowest in the control group. These results suggest that growth and immune responses were enhanced in grouper fed diet with 1.5 g mixed‐NT kg^?1 diet. Diet with 1.5 g kg^?1 of AMP seems to be more beneficial on the immune responses in fish than other nucleotides.     

Effect of dietary protein reduction with lysine and methionnine supplementation on growth performance,body composition and total ammonia nitrogen excretion of juvenile grass carp,Ctenopharyngodon idella

A 63‐day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant‐based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day^?1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP‐ and 28CP‐diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P < 0.05), and the interaction between dietary protein level and amino acid supplementation was noted between WG and SGR (P < 0.05). Feed intake (FI) was significantly increased with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05), but feed conversion ratio (FCR) showed a significant decreasing trend (P < 0.05). Two days after total ammonia nitrogen (TAN) concentration test, the values of TAN discharged by the fish 8 h after feeding were 207.1, 187.5, 170.6, 157.3, 141.3 and 128.9 mg kg^?1 body weight for fish fed 32CP, 32AA, 30CP, 30AA, 28CP and 28AA diets, respectively. TAN excretion by grass carp was reduced in plant‐based practical diets with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05). The results indicated that lysine and methionine supplementation to the plant protein sources‐based practical diets can improve growth performance and feed utilization of grass carp, and the dietary crude protein can be reduced from 320 g kg^?1 to 300 g kg^?1 through balancing amino acids profile. The positive effect was not observed at 280 g kg^?1 crude protein level.     

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.     

Effects of dietary vitamin E level on growth and tissue lipid peroxidation of soft-shelled turtle, Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to determine the effect of dietary vitamin E supplementation on growth, liver lipid peroxidation and liver and muscle vitamin E level of soft‐shelled turtle, Pelodiscus sinensis. Eight experimental diets analysed to contain 0–457 IU vitamin E kg^?1 were fed to juvenile soft‐shelled turtle of 4.8 g initial body weight for 12 weeks. Weight gain (WG) of the turtles fed the diet containing no vitamin E was significantly lower than those fed diets containing 83–457 IU vitamin E kg^?1 (P<0.05). Feed conversion ratio and protein efficiency ratio showed similar trends to that of WG. No significant difference (P>0.05) was found in whole‐body composition among turtles fed the different diets. Dietary vitamin E requirement using WG as the response and estimated using the broken‐line regression model is approximately 88 IU kg^?1. Liver and muscle vitamin E content increased when dietary vitamin E level increased. Ascorbate‐induced lipid peroxidation in liver tissue of turtles fed diets containing 0 and 17 IU vitamin E kg^?1 was significantly (P<0.05) greater than those fed diets containing high vitamin E (≥35 IU kg^?1).     

Effects of extruded and pelleted diets with differing lipid levels on growth,nutrient retention and serum biochemical indices of tilapia (Oreochromis aureus × Tilapia nilotica)

This study was conducted to evaluate the effects of extruded diets and pelleted diets with varying dietary lipid levels on growth performance and nutrient utilization of tilapia. Six diets, containing three levels of lipid at 40, 60 or 80 g kg^?1 (with the supplemental lipid of 0, 20 or 40 g kg^?1, respectively), were prepared by extruding or pelleting and then fed to tilapia juveniles (8.0 ± 0.1 g) in cages (in indoor pools) for 8 weeks. The results indicated that the fish that were fed the diet with 60 g kg^?1 of lipid had a higher weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER), lipid retention (LRE), energy retention (ERE), apparent protein digestibility, apparent dry matter digestibility and a lower feed conversion ratio (FCR) than those fed the diet with 40 g kg^?1 lipid in both the extruded diet and pelleted diet (P < 0.05). As the dietary lipid level increased from 60 to 80 g kg^?1, these parameters were not further improved, even digestibilities of the crude protein and dry matter decreased (P < 0.05). With the dietary lipid level increased, whole‐body lipid content significantly increased (P < 0.05), serum aspartate aminotransferase, alkaline phosphatase, total cholesterol and low‐density lipoprotein cholesterol (LDL‐C) tended to increase (P > 0.05), whereas whole‐body protein content, serum triglyceride (TG), high‐density lipoprotein cholesterol (HDL‐C) and HDL‐C/LDL‐C tended to decrease (P > 0.05). Fish fed with the extruded diets had a higher WG, SGR, hepatosomatic index (HSI), PER, protein retention (PRE), LRE, ERE, TG, apparent digestibility of protein and dry matter, as well as a lower FCR, than those fed with the pelleted diets at the same dietary lipid level (P < 0.05). These results suggested that tilapia fed with the extruded diets had a better growth and higher nutrient utilization than fish fed with the pelleted diets, when dietary lipid level ranged from 40 to 80 g kg^?1 and at dietary crude protein level was 280 g kg^?1. The optimum dietary lipid level was 60 g kg^?1 in both the pelleted and extruded diets, and extrusion did not affect dietary lipid requirement of the tilapia.     

Dietary methionine requirement of juvenile Pseudobagrus ussuriensis

An 8‐week feeding trial was conducted to determine the optimum dietary methionine (Met) requirement of juvenile Pseudobagrus ussuriensis with an initial average weight of 0.60 g reared in indoor flow‐through and aerated aquaria. Six isonitrogenous (430 g kg^?1 protein) and isolipidic (50 g kg^?1 lipid) test diets were formulated to contain graded levels of crystalline L‐methionine (4.9, 9.0, 11.8, 14.2, 18.1 and 20.8 g kg^?1 dry diets, respectively) at a constant dietary cystine level of 2.5 g kg^?1 dry diets. Equal amino acid nitrogen was maintained by replacing methionine with non‐essential amino acid mixture. Fish were randomly allotted to 18 aquaria (1.0 × 0.5 × 0.8 m) with 50 fish to each glass aquarium. Fish were fed twice daily (08:00 and 16:00) to apparent satiation. No significant difference was observed in survival of fish (84.67–91.33%). Specific growth rate (SGR), weight gain (WG), feed conversion ratio (FCR), protein productive value (PPV) and protein efficiency ratio (PER) were significantly affected by different dietary methionine levels (P < 0.05). WG, SGR PPV and PER increased, while FCR decreased with increasing dietary methionine level from 4.9 to 11.8 g kg^?1 (P < 0.05). However, with further increase from 11.8 to 20.8 g kg^?1, WG, SGR PPV and PER significantly decreased, FCR increased (P < 0.05). The whole body and muscle composition were affected by different dietary methionine levels (P < 0.05). Condition factor (CF) increased with increasing dietary methionine levels up to 11.8 g kg^?1 (P < 0.05) and after 11.8 g kg^?1 methionine diet, but not significant, declines were observed (P > 0.05). Hepatosomatic index (HSI) of the 4.9, 9.0, 11.8 and 14.2 g kg^?1 Met diets was significantly higher than that of fish fed diets 18.1 and 20.8 g kg^?1 Met diets (P < 0.05). Viscerosomatic index (VSI) of the 4.9, 9.0 and 11.8 g kg^?1 Met diets was significantly higher than that of fish fed diets 14.2, 18.1 and 20.8 g kg^?1 Met diets (P < 0.05). Quadratic regression analysis of WG and PER against dietary methionine levels indicated that the optimal dietary methionine requirement for maximum growth and feed utilization of juvenile Pseudobagrus ussuriensis was 14.3 and 14.1 g kg^?1 dry diet (35.3 and 34.8 g kg^?1 dietary protein), respectively, in the presence of 2.5 g kg^?1 dry diets cystine.     

Effects of dietary inorganic copper on growth performance and immune responses of juvenile beluga,Huso huso

A 12‐week feeding trial was conducted to evaluate the optimum dietary inorganic copper (copper sulphate) in juvenile beluga, Huso huso. Eight semi‐purified diets containing 1.1 (Cu_1.0), 3.5 (Cu_4.0), 7.1 (Cu_7.0), 9.7 (Cu₁₀), 13.1 (Cu₁₃), 25.1 (Cu₂₅), 49.9 (Cu₅₀) and 195 (Cu₁₉₅) mg Cu kg^?1 diet in the form of CuSO₄.5H₂O were fed to fish of initial body weight 8.49 ± 0.32 g and length 11.85 ± 0.66 cm (mean ± SD) in triplicate groups in a flow‐through system. Weight gain (WG) of fish fed Cu₁₀ and Cu₁₃ diets was significantly higher than that of fish fed Cu_1.0, Cu_4.0, Cu₂₅, Cu₅₀ and Cu₁₉₅ diets (P < 0.05). Whole‐body and muscle crude protein increased with dietary Cu up to the supplementation level of 13.1 mg kg^?1 diet and then decreased. Whole‐body lipid content was negatively correlated, while whole‐body ash was positively correlated with dietary copper concentration. Hepatic copper–zinc superoxide dismutase activity of fish fed Cu₁₀ and Cu₁₃ diets was significantly higher than that of fish fed Cu_1.0, Cu_4.0 and Cu₁₉₅ diets. Hepatic thiobarbituric acid‐reactive substances of fish fed Cu₁₃ diet was significantly lower than those of fish fed the other diets except for that of fish fed Cu₁₀ diet. Aspartate aminotransferase, alanine aminotransferase and copper accumulation in tissues increased with dietary copper. Broken‐line analysis of WG suggested that the optimum dietary Cu level was 10.3 mg Cu kg^?1 diet. Therefore, these results may indicate that the optimum dietary Cu levels could be greater than 10.3 mg Cu kg^?1 diet but less than 13.1 mg Cu kg^?1 diet in juvenile beluga, when copper sulphate is used as the dietary source of inorganic copper.     

The effects of dietary vitamin C on growth performance,serum enzymes activities and resistance to Vibrio alginolyticus challenge of yellow drum Nibea albiflora

A 9‐week feeding trial was conducted to determine the optimal dietary vitamin C requirement and its effects on serum enzymes activities and bacterial resistance in the juvenile yellow drum Nibea albiflora (initial weight 33.2 ± 0.10 g). Six practical diets were formulated containing vitamin C 2.1, 45.3, 89.6, 132.4, 178.6 and 547.1 mg kg^?1 diet supplied as l ‐ascorbyl‐2‐monophosphate. The fish fed 547.1 mg kg^?1 diet showed a significantly higher survival than that fed 2.1 mg kg^?1 diet. The weight gains and specific growth rate of the fish fed 2.1 mg kg^?1 diet were significantly lower than those of the fish fed 89.6–547.1 mg kg^?1 diets. The liver vitamin C concentration firstly increased with increasing dietary vitamin C supply from 2.1 to 178.6 mg kg^?1 diet and then stabilized. The serum superoxide dismutase activities of the fish fed 547.1 mg kg^?1 diet were significantly lower than those of the fish fed 2.1–89.6 mg kg^?1 diet. The fish fed 2.1 mg kg^?1 diet had a significantly higher alkaline phosphatase activity than those in the other groups except the 45.3 mg kg^?1 group. Fish that received diets containing vitamin C at 547.1 mg kg^?1 had significantly higher nitro blue tetrazolium and lysozyme activity, and fish that received diets containing vitamin C at 45.3–547.1 mg kg^?1 exhibited resistance against Vibrio alginolyticus infection. The dietary vitamin C requirement of the juvenile yellow drum was established based on broken‐line model of weight gain to be 142.2 mg l ‐ascorbyl‐2‐monophosphate kg^?1 diet.     

Dietary digestible lysine requirement and essential amino acid to lysine ratio for pacu Piaractus mesopotamicus

To determine the digestible lysine requirement for pacu juveniles, a dose–response feeding trial was carried out. The fish (8.66 ± 1.13 g) were fed six diets containing the digestible lysine levels: 6.8, 9.1, 11.4, 13.2, 16.1 and 19.6 g kg^?1 dry diet. The gradual increase of dietary digestible lysine levels from 6.8 to 13.2 g kg^?1 did not influence the average values of the parameters evaluated (P > 0.05). The increase of dietary digestible lysine level to 16.1 g kg^?1 significantly improved weight gain (WG), specific growth rate (SGR), protein productive value (PPV), protein efficiency rate (PER), and apparent feed conversion rate (FCR), but was not different from fish fed diets containing 19.6 g kg^?1 lysine. Fish fed diets containing 16.1 and 19.6 g kg^?1 digestible lysine showed lower body lipid contents than fish in the other treatments. The digestible lysine requirement as determined by the broken‐line model, based on average WG values, was 16.4 g kg^?1. The other essential amino acid requirements were estimated based on the ideal protein concept and the value determined for lysine.     

Histologic changes in channel catfish,Ictalurus punctatus Rafinesque,fed diets containing graded levels of gossypol–acetic acid

A study was performed to evaluate the histologic changes among fingerling channel catfish, Ictalurus punctatus Rafinesque, fed purified diets containing gossypol from gossypol–acetic acid. The catfish were maintained on diets with 0, 300, 600, 900, 1200 or 1500 mg gossypol kg^?1 diet for 12 weeks, and histologic samples from the stomach, proximal and distal intestines, pancreas, liver, and spleen were obtained from fish in all groups. Stomach sections exhibited significant gastric gland necrosis in fish fed 600 mg gossypol kg^?1 diet or higher. Non‐existent‐to‐mild enterocyte vacuolization loss, inflammatory cell infiltration, and hyperplastic lamina propria were noted in the intestinal sections from gossypol‐fed fish, but no significant differences in severity scores were noted. The pancreas from fish fed doses of gossypol above 600 mg gossypol kg^?1 diet exhibited significant mild‐to‐severe necrosis, and livers from fish fed 900 and 1500 mg gossypol kg^?1 diet exhibited significantly higher pigment deposition. No other significant histologic differences were observed in the fish fed diets containing gossypol–acetic acid. The data in this study indicates that at least 600–900 mg gossypol kg^?1 purified diet can cause statistically significant histologic changes in fingerling channel catfish, suggesting that gossypol should remain at concentrations below 600 mg gossypol kg^?1 diet.     

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).     

The effect of dietary methionine concentrations on growth performance of juvenile Nile tilapia (Oreochromis niloticus) fed diets with two different digestible energy levels

A growth trial was conducted to examine the effect of dietary digestible energy (DE) content on methionine (Met) utilization and requirement in juvenile Nile tilapia (Oreochromis niloticus). Ten iso‐nitrogenous (288 g kg^?1 protein) practical diets, with two DE levels (10.9 MJ kg^?1; 12.4 MJ kg^?1) and five methionine supplementation levels (0, 1, 2, 4 and 6 g kg^?1), were hand‐fed twice daily to triplicate groups of Nile tilapia (initial body weight 8.95 ± 0.06 g) for 8 weeks. Weight gain (WG) and specific growth rate (SGR) increased significantly with increasing dietary methionine concentration at the same DE content (P < 0.001). At the same dietary methionine level, WG and SGR of fish fed high‐DE diets were significantly higher than that of fish fed low‐DE diets (P = 0.0001), although no interaction was found between dietary DE and methionine supplementation. Based on quadratic regression analysis between dietary methionine concentration and weight gain, optimal methionine requirement for maximum growth, expressed as g Met required kg^?1 diet (low‐ versus high‐DE diets), increased as diet DE concentration increased (7.34 versus 9.90 g kg^?1 diet, respectively; with cysteine 4.70 g kg^?1 diet). The results indicated that diet DE content affects methionine utilization and requirement in juvenile Nile tilapia, fish fed high‐DE diets required more methionine for maximum growth.     

Quantifying the dietary niacin requirement of the Indian catfish, Heteropneustes fossilis (Bloch), fingerlings

A growth study was conducted to determine the dietary niacin requirement of the Indian catfish, Heteropneustes fossilis (Bloch), fingerlings (Mean weight 9.41 ± 0.18 g). Semi‐purified diets with five levels (0, 5, 10, 20 and 40 mg kg^?1 diet) of supplemental niacin were fed to H. fossilis for 15 weeks. Each diet was fed to three replicate groups of fish. Results indicated that the highest (P < 0.05) weight gain was for the fish fed the diet supplemented with 20 mg niacin kg^?1, followed by fish fed the diets with 40, 10 and 5 mg niacin kg^?1, and the lowest in fish fed the unsupplemented control diet. Patterns of specific growth rate (SGR) and protein efficiency ratio (PER) were similar to those of the weight gain. Survival of fish fed the control diet and niacin‐supplemented diet was 58% and 91–100% respectively. Niacin deficiency signs such as anaemia, anorexia, lethargy and skin haemorrhage were observed in fish fed the control diet. The haematocrit values (Ht) were higher (P < 0.05) in fish fed the diets supplemented with niacin than in fish fed the control diet. The hepatosomatic indexes (HSI) of fish fed with or without niacin‐supplemented diets were not significantly (P > 0.05) different from each other. Both body protein and lipid content were higher (P < 0.05) in fish fed the diet supplemented with 20 and 40 mg niacin kg^?1, respectively, than those fish fed other diets. The niacin content in liver significantly (P < 0.05) reflected the supplementation level in the diet and ranged from 29.11 to 40.31 mg g^?1 tissue. The associated liver niacin content for growth was about 47 μg g^?1 tissue. Quadratic regression analysis showed that the dietary niacin requirement for maximal growth of H. fossilis under these experimental conditions was about 25 mg kg^?1 diet.     

Dietary vitamin E requirement for maximizing the growth,conversion efficiency,biochemical composition and haematological status of fingerling Channa punctatus

The effect of feeding graded levels of vitamin E (E₀, E₂₀, E₄₀, E₆₀, E₁₀₀, E₁₄₀, E₁₈₀, E₂₂₀, E₂₆₀) in nine casein–gelatin‐based isonitrogenous (450 g kg^?1 crude protein) and isoenergetic (17.97 kJ g^?1 gross energy) experimental diets was evaluated in fingerling Channa punctatus for 12 weeks. Growth, nutritional and haematological parameters were studied. Hepatic lipid peroxidation as thiobarbituric acid‐reactive substances (TBARS) was also assayed. The maximum absolute weight gain (AWG g/fish, 55), best feed conversion ratio (FCR, 1.32), protein retention efficiency (PRE, 40%) and energy retention efficiency (ERE, 76%) were achieved in fish fed on a diet supplemented with 140 mg vitamin E kg^?1 diet (E₁₄₀). A consistent decline in the hepatic TBARS concentration and an improvement in haematocrit (Hct) and haemoglobin (Hb) were displayed in fish fed on diets with increasing concentrations of vitamin E up to 140 mg kg^?1 (E₀–E₁₄₀), beyond which (E₁₈₀–E₂₆₀) a reverse trend in these parameters was evident. Based on the broken‐line regression and exponential analyses of AWG, FCR, PRE, ERE, Hb and Hct data, diets for fingerling C. punctatus should contain vitamin E in the range of 140–169 mg kg^?1 to maintain satisfactory fish performance.