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 vitamin A inclusion in practical diets on survival and growth of juvenile crayfish (Pacifastacus leniusculus Dana,Astacidae) from the onset of exogenous feeding

The aim of this study was to evaluate the effects of practical diets with different levels of vitamin A on survival and growth of juvenile crayfish (Pacifastacus leniusculus). An 80‐day trial was conducted with stage 2 juveniles from the onset of exogenous feeding. Four diets differing in the level of retinyl acetate were tested: 0, 0.0025, 0.0050 or 0.0100 g kg^?1 diet, equivalent to 0, 2500, 5000 or 10 000 IU vitamin A kg^?1 diet, respectively. Each diet was tested on grouped or individually isolated crayfish. Crayfish fed the diet with 5000 or 10 000 IU vitamin A kg^?1 diet achieved the highest survival (average of grouped and isolated: 88.9%) and growth (grouped and isolated: 13.45 mm carapace length, 533.4 mg weight). Final growth of isolated crayfish was significantly higher than that of grouped crayfish for all diets. This study provides the first data on vitamin A requirements of freshwater crayfish. A level of 5000 IU vitamin A kg^?1 can be recommended for juvenile P. leniusculus during the first period of intensive rearing.     

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.     

Effects of dietary propolis and vitamin E on growth performance and antioxidant status in juvenile rainbow trout (Oncorhynchus mykiss) under different flow rate

The present study investigated the effects of propolis and vitamin E supplementation in diets of juvenile rainbow trout subjected to two different flow rates (0.9 and 2.1 L min^?1) on growth performance, and vitamin A, C and E concentrations in tissues as well as malondialdehyde (MDA) levels. Juvenile rainbow trout were fed with diets containing 10 and 30 g propolis kg^?1, 60 mg kg^?1 vitamin E (Rovimix E‐50 adsorbate; min.%50 dl‐α‐tokopherly acetate) and without supplemented basal diet for 12 weeks. Weight gain (WG) in the C group was significantly lower (P < 0.05) than P10, P30 and E60 groups at both flow rate treatments. At 2.1 L min^?1, specific growth rate (SGR) in the C group was significantly lower (P < 0.05) than other groups, but at 0.9 L min^?1, SGR of fish did not differ among the diets groups (P > 0.05). Survival rate (SUR) was higher in propolis and vitamin E supplemented diet groups (P < 0.05) than control diet group at 0.9 L min^?1. Fish fed on diet E60 had higher (P < 0.05) tissue vitamin E concentration than fishes fed on other diets groups. Vitamin C concentration in rainbow trout tissues was significantly affected by the 30 g propolis supplemented diet group (P < 0.05), followed by the 10 g propolis supplemented diet group (P < 0.05). MDA level of E60 group was found significantly decreased instead of different than other groups (P < 0.05). The results of Student's t‐test revealed that WG, SGR, SUR values, vitamin (A, C, E) concentrations and MDA levels of tissues were negatively affected by 0.9 L min^?1flow rate treatment in juvenile rainbow trout.     

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).     

Growth and survival of Atlantic salmon, Salmo salar L. fed different dietary levels of astaxanthin. Juveniles

Atlantic salmon, Salmo salar L., juveniles, with a mean initial weight of 1.75 g, were fed casein-based purified diets which had been supplemented with different levels of astaxanthin for a 10-week period. The astaxanthin content of the diets ranged from 0 to 190 mg kg^?1 dry diet. The growth and survival of the juveniles were recorded throughout the experiment. The proximate composition, astaxanthin and vitamin A content were determined from whole-body samples at the start and termination of the experiment. The dietary treatment was found to affect growth significantly (P < 0.05). A reduction in the mean weight of the juveniles was observed in the groups fed the diets without astaxanthin supplementation. There was no difference in growth rate between the fish in the groups fed the diets containing 36 or 190 mg astaxanthin kg^?1 dry diet, whereas the fish in the group fed the diet containing 5.3 mg astaxanthin kg^?1 dry diet had a lower growth rate. There was a tendency to higher survival in the groups fed the diets containing astaxanthin when compared with the groups fed the non-supplemented diets. The moisture and ash contents were significantly lower and the lipid content was higher in the groups fed the astaxanthin-supplemented diets. The astaxanthin and the vitamin A concentrations in the fish were found to be dependent upon the dietary astaxanthin dose; the highest values were found in the fish fed the diet with the highest astaxanthin content. These results strongly indicate that astaxanthin functions as a provitamin A for juvenile Atlantic salmon. The body storage of vitamin A increased in the fish fed the diets containing astaxanthin. However, the increase was low in the fish fed the diet containing 5.3 mg astaxanthin kg^?1 dry diet.     

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.     

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.     

Effects of different dietary vitamin C supplementations on growth performance,mucus immune responses and antioxidant status of loach (Misgurnus anguillicaudatus Cantor) juveniles

A 60‐day feeding trial was conducted to determine the effects of different dietary vitamin C levels on growth performance, immune response and antioxidant capacity of loach juveniles. Six isonitrogenous (58.6% of crude protein), isoenergetic (17.5 kJ g^?1) practical diets supplemented with 0 (VC0), 100 (VC100), 200 (VC200), 500 (VC500), 1000 (VC1000) and 5000 mg kg^?1 (VC5000) of VC (35% ascorbic acid equivalent) were fed to fish (mean initial weight 0.11 ± 0.02 g) in triplicate. Results showed that fish fed VC0 diet had significantly lower body weight gain (BWG) and survival rate (SR). However, BWG and SR improved significantly in fish fed VC100 and VC200 diet respectively. Whole body ascorbic acid concentration increased with incremental dietary VC levels from 0 to 100 mg kg^?1. The activity of mucus alkaline phophatase was significantly increased by the dietary VC level. Incremental levels of VC significantly reduced activities of glutathione peroxidase (GPx) and catalase. Moreover, fish fed diets containing more than 100 mg kg^?1 VC significantly down‐regulated the superoxide dismutase and GPx mRNA expression in liver. Meanwhile, the expressions of liver heat shock protein (HSP70) and nuclear factor‐erythroid 2‐related‐2 (Nrf2) were affected by fish fed diets containing VC from 100 to 5000 mg kg^?1. In conclusion, VC requirement of loach juveniles for optimum growth and functionally preventing lipid peroxidation was more than 200 mg kg^?1 of diet. Moreover, high dose of VC supplementation did not show any detrimental effects on loach growth performance.     

Bioavailability and interactions with other micronutrients of three dietary iron sources in Atlantic salmon, Salmo salar, smolts

Atlantic salmon, Salmo salar L., with mean initial weight of 60 g were fed a diet based on cod muscle meal supplemented with elemental iron, iron sulphate or haem-bound iron in concentrations of 0, 25, 50, 100, 500 and 1500 mg iron kg^?1 for 8 weeks. No significant differences in growth or mortality were found, except in fish fed 1500 mg haem iron kg^?1, which showed reduced growth. In fish fed diets supplemented with elemental iron below 1500 mg iron kg^?1, blood haemoglobin and hepatic iron concentration decreased compared with fish fed the unsupplemented diet. Fish fed diets supplemented with iron sulphate showed increased blood haemoglobin and hepatic iron concentrations between 25 and 100 mg iron kg^?1. Fish fed diets supplemented with haem-bound iron showed increased hepatic iron at all dietary iron levels, while blood haemoglobin concentration decreased in the group fed 1500 mg haem iron kg^?1. The bioavailability of haem iron relative to sulphate iron was calculated by the slope ratio method to be 239% and 148% using blood haemoglobin and hepatic iron, respectively. Relative bioavailability of elemental iron was zero when dietary supplementation levels were between 25 and 500 mg iron kg^?1, while a small part was utilized when 1500 mg elemental iron kg^?1 was supplemented. Additions of 500 and 1500 mg haem-bound iron kg^?1 resulted in a complete loss of ascorbic acid in these diets. When these groups were discounted, no significant relationship between hepatic iron and hepatic ascorbic acid was found. There was no significant effect of dietary iron on whole-body manganese concentration and only a weak effect on whole-body zinc concentration. No significant correlations between dietary iron and hepatic copper concentration were found in any of the dietary treatments. This study also showed that the level of inorganic iron supplementation may be reduced by inclusion of 20 g blood meal kg^?1 in the diet.     

Dietary vitamin E could improve growth performance,lipid peroxidation and non‐specific immune responses for juvenile cobia (Rachycentron canadum)

An 8‐week feeding trial was conducted to establish the dietary vitamin E requirement of juvenile cobia. The basal diet was supplemented with 10, 20, 30, 40, 60, 120 mg vitamin E kg^?1 as all‐rac‐α‐tocopheryl acetate. The results indicated that fish fed the diets supplemented vitamin E had significantly higher specific growth rate, protein efficiency ratio, feed efficiency and survival rate than those fed the basal diet. It was further observed that vitamin E concentrations in liver increased significantly when the dietary vitamin E level increased from 13.2 to 124 mg kg^?1. Fish fed the basal diet had significantly higher thiobarbituric acid‐reactive substances concentrations in liver than those fed the diets supplemented vitamin E. Fish fed the diets supplemented with 45.7 and 61.2 mg kg^?1 vitamin E had significantly higher red blood cell and haemoglobin than those fed the basal diet, while fish fed the diets supplemented with 61.2 and 124 mg kg^?1 vitamin E had higher immunoglobulin concentration than those fish fed the basal diet. Lysozyme and superoxide dismutase were significantly influenced by the dietary vitamin E level. The dietary vitamin E requirement of juvenile cobia was established based on second‐order polynomial regression of weight gain and lysozyme to be 78 or 111 mg all‐rac‐α‐tocopheryl acetate kg^?1 diet, respectively.     

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.     

The pigmenting effect of different carotenoids on fancy carp (Cyprinus carpio)

The optimal concentration of a panel of individual and combined carotenoid sources on skin pigmentation in fancy carp was investigated by nine experimental diets that were formulated and supplemented with astaxanthin at 25 mg kg^?1, lutein at 25 and 50 mg kg^?1, β‐carotene at 25, 50 and 75 mg kg^?1, and lutein combined with β‐carotene at 25 : 25 and 50 : 50 mg kg^?1, while a diet without supplemented carotenoid served as a control. The results showed that serum TC of fish fed diets containing supplemented with lutein plus β‐carotene at 25 : 25; 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet were higher than the other treatments (P ≤ 0.05). Serum TC of the respective treatments was 6.2 ± 2.0, 7.8 ± 3.3 and 7.3 ± 1.9 μg mL^?1 serum, respectively. Fish fed diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet had serum astaxanthin concentrations similar to fish fed the diet with astaxanthin alone at 25 mg kg^?1. Serum astaxanthin concentrations was 0.7 ± 0.01, 0.9 ± 0.01, 0.4 ± 0.02 and 1.7 ± 0.18 μg mL^?1 serum, respectively. The chromaticity of fish body skin of red and white position was assessed by colourimetry using the CIE L*a*b (CIELAB) system. Pigmentation response of skin redness of fancy carp fed with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were higher than other treatments (P ≤ 0.05) but they were similar to fish fed with 25 mg kg^?1 astaxanthin diet. The redness (a* values) of fish fed diets with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were 28.3 ± 0.53, 29.9 ± 1.38, 28.8 ± 3.95 and 28.5 ± 2.49, respectively. After 3 weeks of feeding the experimental diets, the fish fed on a diet without carotenoid supplement for one week demonstrated that the same three groups still retained their redness and had an overall tendency to improve skin colouring. Finally, concentrations 50 mg kg^?1 of lutein, or the combination of lutein and β‐carotene at 25 : 25 mg kg^?1 showed the highest efficiency for improving skin pigmentation and redness of skin.     

Vitamin E requirement of sea cucumber (Apostichopus japonicus) and its' effects on nonspecific immune responses

This feeding trial was conducted to determine the vitamin E requirement of sea cucumber (Apostichopus japonicus) and to study the effects of vitamin E on nonspecific immune parameters. Commercial sea cucumber diets supplemented with 0, 50, 100, 200, 500 or 1000 mg kg^?1 vitamin E were fed to juvenile sea cucumber (7.96 ± 0.01 g) for 60 days. Specific growth rate and total coelomocyte counts were maximized in sea cucumber fed diets supplemented with 100 mg kg^?1 vitamin E. Reactive oxygen species production was maximized at 50 mg kg^?1 vitamin E, whereas superoxide dismutase activity was the highest in sea cucumber fed diets supplemented with 100 mg kg^?1 vitamin E. The vitamin E concentration in the body wall of the sea cucumber increased as the vitamin E level in the diet increased. Analysis of specific growth rate and total coelomocyte counts of the sea cucumber indicated that 88–92 mg kg^?1 vitamin E is required for maximal growth and nonspecific immune responses of A. japonicus and that 114.7 mg kg^?1 vitamin E is required to minimize lipid peroxidation levels.     

Effect of vitamin C and astaxanthin on stress and disease resistance of postlarval tiger shrimp, Penaeus monodon (Fabricius)

Postlarvae of tiger shrimp, Penaeus monodon (Fabricius), were fed semipurified diets supplemented with various levels of astaxanthin (AX) and ascorbic acid-polyphosphate (ApP): three groups were fed 230 mg AX kg^?1 diet combined with 100, 1700 and 3400 mg ascorbic acid (AA) kg^?1 diet, respectively; two diets contained 810 mg AX kg^?1 mixed with 200 and 1700 mg AA kg^?1, respectively. Each treatment was run in four replicates. Incorporated levels of AA and AX, production output, and physiological condition were recorded after 4 weeks of feeding. Whole-body AA (21-47 μg g^?1) and AX, concentrations (19-35 μgg^?1) were linked to dietary ApP and AX supply, respectively, although not significantly for the latter. The biomass of the group receiving the lower dietary ApP-AX combination was significantly lower than for all other treatments, i.e. 3.1 versus 3.9 g, respectively. In the groups fed 230 mg AX kg^?1 diet, significant differences in stress resistance were observed according to the dietary ApP level, i.e. raising the vitamin C content in the feed from 100 to 3400 mg AA kg^?1 resulted in a concomitant drop in mortality after an osmotic shock. For the treatments receiving 810 mg AX kg^?1 diet, the beneficial effect of extra dietary vitamin C was not significant. An increase in the dietary AX for shrimp fed comparable ApP levels resulted in a significant drop of the stress index from 56 to 33 (cumulative mortality index). An increased resistance to salinity shock was demonstrated in association with supplementation of high dietary AA or AX levels. No conclusive results regarding possible improved disease resistance could be made since no mortality was observed after a disease challenge with Vibrio harveyi.     

The effect of vitamin E on growth, survival and hepatopancreas structure of the Argentine red shrimp Pleoticus muelleri Bate (Crustacea, Penaeidea)

Two trials were conducted to evaluate the growth, survival and hepatopancreas histology of the Argentine red shrimp Pleoticus muelleri (Bate, 1888) fed different levels of vitamin E and butylated hydroxytoluene (BHT) in a semipurified diet. The diets contained 0, 100, 600 or 1500 mg vitamin E kg^?1 and 16 mg BHT kg^?1 diet (trial 1) and 0, 1250, 1500, 1750 or 2000 mg vitamin E kg^?1 diet, squid mantle and vitamin‐free diet as a control (trial 2). After 30 days (trial 1), survival ranged between 43% and 64%, and the percentage weight gain of the shrimp varied from 22% to 31% with no significant differences among treatments (P<0.05). After 40 days (trial 2), survival of shrimp fed the diet with no vitamin E and squid mantle was significantly lower (62%) than the other treatment (86–90%). Shrimp fed diets containing vitamin E from 1250 to 1750 mg kg^?1 exhibited increased weight gain (34–65%); however, a significant difference was observed for shrimp fed the diet containing 2000 mg kg^?1. Histological results yielded differences among treatments. In shrimp fed 1750 mg kg^?1 of vitamin E, the functional morphology of the organ was normal, with abundant secretion in the tubules. Signs of malnourishment such as cellular and nuclear retraction, desquamation of cells and hipertrofia, were evident in the hepatopancreas of shrimp fed the other diets. The results indicate that optimal vitamin E requirement for P. muelleri under the present experimental conditions appears to be approximately 1750 mg vitamin E kg^?1 diet.     

Effect of supplemental dietary zinc sources on the growth and carbohydrate utilization of tilapia Smith 1840, Oreochromis niloticus × Oreochromis aureus

An 8‐week feeding trial was conducted to investigate the effect of supplemental dietary zinc sources on the growth performance and carbohydrate utilization of juvenile tilapia Smith 1840, Oreochromis niloticus × O. aureus. The goal was to compare the bioavailability of two zinc sources, zinc sulphate (ZnSO₄) or zinc methionine (ZnMet), by using two practical basal diets with 350 g kg^?1 (C350) or 400 g kg^?1 (C400) carbohydrates based on wheat as the carbohydrate source. The results showed that fish fed with a diet supplemented with 60 mg kg^?1 Zn from either ZnSO₄ or ZnMet had a significantly (P < 0.05) greater specific growth rate and protein efficiency ratio than those fed with the diets of ≤30 mg kg^?1 Zn. The composition of tilapia carcass was also found to be influenced by various levels of dietary zinc from the two zinc sources. The G6P‐DH in fish fed with the 20 mg kg^?1 ZnMet diet and the PK levels in fish fed with 20 mg kg^?1 ZnSO₄ and 30 mg kg^?1 ZnMet diet were significantly (P < 0.05) higher than those in fish fed with the C400 diet. The data suggest that supplemental dietary zinc from either ZnMet or ZnSO₄ significantly affects the growth performance and carbohydrate utilization of tilapia.     

Histology and growth performance in rainbow trout (Oncorhynchus mykiss) in response to increasing dietary concentration of sparteine,a common alkaloid in lupins

This study evaluated the effects of sparteine, a lupin alkaloid, on growth and organ histology in rainbow trout. Eight diets with increasing concentrations of sparteine (0, 50, 100, 250, 500, 1000, 2500 and 5000 mg kg^?1) were fed to triplicate groups of 61‐g fish for 62 days. Survival was not affected by the dietary treatments. Weight gain, feed conversion efficiency and feed intake decreased in a quadratic manner for sparteine levels above 100 mg kg^?1. No effects were observed at inclusion levels ≤100 mg kg^?1. Whole‐body lipid and dry matter was depleted in a quadratic manner, while protein and ash were not affected. No tissue alteration attributable to sparteine alkaloid intake was found in liver, kidney, spleen or mid and distal intestines. At sparteine levels above 1000 mg kg^?1 of diet, the weight of the intestines and the liver relative to body weight decreased, whereas the weight of the spleen increased. These effects were likely associated with low feed intake and starvation. These results indicate that dietary sparteine alkaloid primarily reduced palatability, but did not impair the health of the fish. The level of sparteine should be kept below 100 mg kg^?1 in trout diets to ensure rapid growth.     

Effect of dietary iron supplement on growth, haematology and microelements of juvenile grouper, Epinephelus coioides

An experiment was conducted to investigate the effect of dietary iron supplement on growth, haematology and microelements of juvenile grouper, Epinephelus coioides. Casein–gelatine‐based diets supplemented with 0, 50, 100, 150, 200 and 250 mg kg⁻¹ iron from ferrous sulphate were fed to grouper (mean initial weight: 21.0 ± 0.2 g) for 8 weeks. Weight gain was highest in fish fed the diet supplemented with 100 mg kg⁻¹ iron, intermediate in fish fed diets with 50, 150, 200 and 250 mg kg⁻¹ iron and lowest in fish fed the basal diet. Feed efficiency followed a similar trend except that the lowest value was in fish fed the basal diet and the diet supplemented with 250 mg kg⁻¹ iron. Hepatic iron was highest in fish fed diets supplemented with iron ≥100 mg kg⁻¹, followed by fish fed diet with 50 mg kg⁻¹ iron and lowest in fish fed the basal diet. The whole‐body iron was lowest in fish fed the basal diet but not significantly different from other groups, as judged by anova . Iron supplement to the basal diet had no significant effect on haematological parameters (red blood cell count, haematocrit and haemoglobin), hepatic copper concentration or manganese, zinc concentration in liver and whole body. Broken‐line analysis of hepatic iron indicated that iron supplementation of 100 mg kg⁻¹ satisfied the hepatic iron storage and that further supplementation did not expand the iron status.