Immunomodulatory Effects of Dietary β‐glucan and Vitamin C in Nile Tilapia,Oreochromis niloticus L., Subjected to Cold‐induced Stress or Bacterial Challenge

To determine the dietary β‐glucan and vitamin C levels that would enable the fish to cope with different stress, 252 tilapia (16.86 ± 0.24 g) were distributed in thirty six 250‐L aquaria and fed experimental diets for 60 d. Diets were supplemented with β‐glucan (0.1, 0.2, 0.4, and 0.8%) and vitamin C (400 and 600 mg/kg diet) and distributed in a factorial design that included a control diet (0 β‐glucan/125 mg vitamin C/kg). After growth performance analyses, a group of 108 fish were subjected to cold‐induced stress and another to Aeromonas hydrophila infection. Hematological and immunological parameters were determined before and after the stress. Growth performance was not affected. Red blood cells, hematocrit, mean corpuscular volume, total plasma protein, albumin : globulin ratio and leukocytes, lymphocytes, neutrophils, and monocytes were affected (P < 0.05) by stress and/or diets. The control diet resulted in higher post‐infection cortisol (P < 0.05). Liver vitamin C was directly proportional to dietary vitamin C (P < 0.05). The production of hydrogen peroxide and nitric oxide was also affected by diets and/or bacterial challenge (P < 0.05). In total, 0.1–0.2% β‐glucan and 600 mg/kg vitamin C increased fish resistance to stress and 0.8% β‐glucan resulted in reduced immune responses regardless of the vitamin C supplementation level.     

Effects of different dietary levels of L-ascorbyl-2-polyphosphate on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel)

This experiment was conducted to study the effects of different dietary levels of vitamin C, L‐ascorbyl‐2‐polyphosphate (ASPP), on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel). Fish were fed one of six semi‐purified diets containing an equivalent of 0, 25, 50, 75, 150, or 1500 mg ascorbic acid (AA) kg^?1 diet (C₀, C₂₅, C₅₀, C₇₅, C₁₅₀ or C₁₅₀₀) in the form of ASPP for 12 weeks. Weight gain (WG) and protein efficiency ratio (PER) of fish fed the C₀ diet were significantly lower than those of fish fed the other diets (P < 0.05), and WG and PER of fish fed the C₂₅, C₅₀ and C₇₅ diets were significantly lower than those of fish fed the C₁₅₀₀ diet (P < 0.05). Fish fed the C₀ diet exhibited vitamin C deficiency symptoms such as anorexia, scoliosis, cataract, exophthalmia and fin hemorrhage at the end of the 12‐week test. After 12 weeks of the feeding trial, AA concentrations from gill, kidney, and liver of fish fed the C₀, C₂₅, C₅₀ and C₇₅ diets were significantly lower than those of fish fed the C₁₅₀ and C₁₅₀₀ diets (P < 0.05). Based on broken line analyses for WG and PER, the optimum dietary levels of vitamin C were 91 and 93 mg AA kg^?1 diet respectively. These findings suggest that the dietary vitamin C requirement could be 93 mg AA kg^?1 diet to support reasonable growth, and greater than 150 mg AA kg^?1 diet may be required for AA saturation of major tissues for juvenile olive flounder under experimental conditions.     

The role of dietary L‐ascorbyl‐2‐polyphosphate on the growth and physiological functions of beluga,Huso huso (Linnaeus, 1758)

Effects of dietary ascorbic acid (AA) supplementation were carried out on growth performance and physiological indices in juvenile beluga Huso huso fed different levels of dietary L‐ascorbyl‐2‐polyphosphate as AA source consisting of 0 (control), 100, 200, 400, 800 and 1600 mg AA kg^?1 diet for 16 weeks. No significant differences were found in growth performances between treated groups except for condition factor. Using AA caused significant differences in haemoglobin, number of red blood cells, white blood cells, aspartate aminotranspherase and lactate dehydrogenase, while no significant difference was found in haematocrit, mean corpuscular haemoglobin concentration, differential white blood cells count and alkaline phosphatase activity at the end of the experiment. Lysozyme activity after 8 weeks was significantly affected by dietary treatments with the lowest value in fish fed control diet, but not at completion of 16‐week feeding trial. Liver AA concentration showed a significant increase in correlation with dietary AA increase. Air exposure stress test showed significant differences among the treatments in case of cortisol and glucose concentrations. Also between pre‐ and post stress, significant changes were observed in some of treatments. The lowest and highest levels of cortisol were 9.2 ± 3 (in 200 mg kg^?1) and 16.8 ± 5.1 ng mL^?1 (in control), 6 h post stress (P < 0.05) respectively. Also, the lowest and highest levels of glucose were measured in fish fed 200 and 800 mg AA kg^?1 respectively. The obtained results showed that most growth parameters were not affected by dietary AA, but some physiological and immunological parameters of this species were influenced by this vitamin. We suggest that under rearing conditions, AA should be added to the diet especially during early life stages for improving health status and resistance to stressors.     

Effect of Dietary Vitamin C on Weight Gain,Tissue Ascorbate Concentration,Stress Response,and Disease Resistance of Channel Catfish Ictalurus punctatus1

Juvenile channel catfish Ictalurus punctatus (average initial weight, 6.5 g/fish) were fed twice daily to apparent satiation with practical-type diets containing 0, 50, 150, or 250 mg supplemental vitamin C/kg from L-ascorbyl-2-polyphosphate for 10 wk under laboratory conditions. At the end of the feeding period, one half of the fish were stressed for 2 h by confinement and both stressed and nonstressed fish were exposed to a virulent strain of Edwardsiella ictaluri. Weight gain and feed conversion efficiency were lower for fish fed the basal diet than those fed diets containing supplemental vitamin C. No differences were observed in weight gain and feed conversion among fish fed diets containing supplemental vitamin C. There were no differences in feed consumption and survival (prior to experimental infection) among treatments. No vitamin C deficiency signs except reduced weight gain were observed in fish fed the basal diet. Serum cortisol concentrations were higher in stressed fish than in non-stressed fish. Dietary vitamin C level had no effect on serum cortisol concentration. As dietary vitamin C increased, ascorbate concentration in serum and liver increased. Confinement stress had no effect on serum and liver ascorbate concentrations. Cumulative mortality of channel catfish 21 d subsequent to experimental infection with E. ictaluri was higher for stressed fish than for nonstressed fish. Regardless of stress or nonstress, overall mortality for fish fed the basal diet was lower than the fish fed diets containing supplemental vitamin C. There were no differences in post-infection antibody levels among treatments or between stressed and nonstressed fish. Results from this study indicate that channel catfish require no more than 50 mg/kg dietary vitamin C for normal growth, stress response, and disease resistance.     

Effects of the supplementation of vitamin D<Subscript>3</Subscript> on the growth and vitamin D metabolites in juvenile Siberian sturgeon (<Emphasis Type="Italic">Acipenser baerii</Emphasis>)

The objective of this study was to evaluate the effects of the supplementation of vitamin D₃ on the growth, vitamin D metabolites, and osteocalcin secretion in juvenile Siberian sturgeon (Acipenser baerii). A 90-day growth trial was conducted with juvenile Siberian sturgeon (initial body weight 3.47 ± 0.14 g) fed seven isonitrogenous and isoenergetic practical diets (45% CP and 13% lipid) containing 60 (basal diet), 240, 450, 880, 1670, 3300, or 1.0 × 10⁵ IU/kg feed (D60~D 1.0 × 10⁵) vitamin D₃. The results showed that weight gain and specific growth rate increased as the dietary vitamin D₃ levels increased from 450 to 3300 IU/kg (P < 0.05). The fish fed with D1670 and D3300 diets had higher crude lipid and ash levels than the fish fed the D60 diet (P < 0.05). The fish fed D880, D1670, or D3300 diets had higher 25-OH-D₃ and 1,25-(OH)₂-D₃ levels than the fish fed the D60 diet (P < 0.05). The fish fed D880, D1670, D3300, or D1.0 × 10⁵ diets had higher osteocalcin levels than the fish fed the D60 diet (P < 0.05). Based on the broken line method analysis of weight gain and osteocalcin, the dietary vitamin D₃ requirement of juvenile Siberian sturgeon was estimated to be 1683.30 and 1403.27 IU/kg per diet, respectively.     

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 performance of Nile tilapia,Oreochromis niloticus,fed diets containing levels of pyridoxine and haematological response under heat stress

In order to assess the effect of dietary pyridoxine supplementation on the growth performance of Nile tilapia and the haematological response under heat stress, 192 fingerlings (8.41 ± 0.22 g) were randomly distributed into eight tanks and fed practical diets supplemented with increasing levels of pyridoxine (0.0; 5.0; 10.0 and 20.0 mg of pyridoxal HCl kg^?1 diet) for 91 days. The fish were then weighed and the diet was quantified to determine the growth performance [weight gain (WG), feed intake, feed conversion ratio, protein efficiency rate, protein retention (PR) and survival percentage]. Haematological analyses (red blood cell count, haematocrit, haemoglobin, total leucocyte and differentiation, mean corpuscular volume, mean corpuscular haemoglobin concentration, albumin, globulin and albumin/globulin ratio) were carried out and then 32 fish were transferred and subjected to heat stress (32 °C) for 3 days, after which the haematological parameters were analysed. The fish fed the unsupplemented diet showed the lowest WG and PR. For the normal growth and health of the Nile tilapia, the Pyridoxine requirement in a practical diet is 10.0 mg of pyridoxine HCl kg^?1.     

Effects of the different levels of dietary vitamin C on growth and tissue ascorbic acid changes in parrot fish (Oplegnathus fasciatus)

An 11-week feeding trial was conducted to study the effects of the different levels of dietary vitamin C on growth, tissue ascorbic acid concentrations and histopathological changes in parrot fish. Casein and gelatin-based diets were formulated to contain 0, 60, 120, 240, 480 and 2000 mg l-ascorbic acid (AA) per kg diets on AA equivalent basis in the form of l-ascorbyl-2-monophosphate (AMP), 60 and 240 mg AA per kg diet as l-ascorbic acid. However, the analyzed AA levels were 0, 50, 100, 205, 426 and 1869 mg AA per kg diet in AMP supplemented diets; 36 and 149 mg AA per kg diet in l-ascorbic acid supplemented diets. Thus, the diets were designated as AA-free, AMP₅₀, AMP₁₀₀, AMP₂₀₅, AMP₄₂₆, AMP₁₈₆₉, AA₃₆ and AA₁₄₉. Fish averaging 3.9±0.06 g (mean±S.D.) randomly were fed the experimental diets in triplicate groups for 11 weeks. Weight gain (WG) of fish fed AMP₅₀ and AA₃₆ diets were significantly lower than those of the other groups, and fish fed AMP₁₈₆₉ diet showed a higher WG than did fish fed AMP₅₀, AMP₁₀₀, AMP₂₀₅, AA₃₆ and AA₁₄₉ diets (P<0.05). However, there was no significant difference in WG between fish fed diets AMP₄₂₆ and AMP₁₈₆₉, and among fish fed AMP₁₀₀, AMP₂₀₅, AMP₄₂₆ and AA₁₄₉ diets. Fish fed the AA-free diet began to show deficiency signs, such as retarded growth, darkening, anorexia and high mortality, after 3 weeks of feeding trial. After 6 weeks of the feeding trial, liver from fish fed the AA-free diet showed severe atrophy. By the end of the seventh week, all fish fed the AA-free diet were dead. Liver AA concentration from fish fed AMP₅₀ diet was significantly lower than that from fish fed AMP₂₀₅, AMP₄₂₆, AMP₁₈₆₉ and AA₁₄₉ diets, and fish fed AMP₅₀, AMP₁₀₀, AA₃₆ diets showed a significant lower AA concentration in liver than did fish fed AMP₂₀₅, AMP₄₂₆ and AMP₁₈₆₉ diets.Broken line analysis of weight gain indicated that the dietary vitamin C requirement of parrot fish is 118±12 mg AA per kg diet in the form of l-ascorbyl-2-monophosphate for maximum growth.     

Effectiveness of l‐ascorbyl‐2‐monophosphate Na/Ca as a vitamin C source for yellowtail Seriola quinqueradiata juveniles

A feeding experiment was conducted to examine the efficacy of a vitamin C (L‐ascorbic acid, AsA) derivative, l ‐ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) by comparing to l ‐ascorbyl‐2‐monophosphate‐Mg (AMP‐Mg) in terms of growth, AsA content in tissues, and hematology in yellowtail Seriola quinqueradiata juvenile (1.10 ± 0.01 g). Furthermore, a stress resistance test for the species fed AMP‐Na/Ca was also conducted. In experiment 1, 11 test pellet diets with different levels of AMP‐Na/Ca (0, 12, 43, 88, 440 and 881 mg AsA kg^?1diet) and AMP‐Mg (0, 16, 52, 106, 595 and 1164 mg AsA kg^?1 diet) were formulated and fed to the yellowtail three times per day for 50 days. In both the vitamin C sources, the survival rates of the fish fed diets without supplemental AsA were lower than 50% at day 20, and more than 50% mortality occurred in fish that fed the diets containing 12 or 16 mg AsA kg^?1 after day 30. However, no significant differences were detected in survival and growth among fish that fed the diets containing more than 43 mg AsA kg^?1. Liver and brain AsA concentrations generally increased with increasing dietary AsA level in both sources. In experiment 2, test diets were formulated to contain 43, 88, 440 and 881 mg AsA kg^?1 using AMP‐Na/Ca, and after 60 days of feeding, yellowtail juveniles were subjected to low salinity and air exposure stress. The fish that received diets with 440 mg AsA kg^?1 showed significantly higher tolerance to low salinity stress and higher survival rate in air exposure stress than those of other groups. The present study demonstrated that yellowtail juveniles could utilize AMP‐Na/Ca as an AsA source like AMP‐Mg, and that supplementation of 43–52 mg AsA kg^?1 diet was optimum for normal growth. However, this study showed that dietary inclusion of 440 mg AsA kg^?1 would be necessary to improve stress resistance of this species.     

Dietary calcium requirements of juvenile tilapia, Oreochromis niloticus × O. aureus, reared in fresh water

A feeding experiment was conducted to determine the dietary calcium (Ca) requirement for juvenile hybrid tilapia, Oreochromis niloticus × O. aureus reared in nature water. Purified diet supplemented with 0, 1, 2, 3, 4, 5, 7 and 10 g Ca kg⁻¹ diet providing of 0.6, 1.6, 2.6, 3.7, 4.7, 5.5, 7.5 and 10.7 g Ca kg⁻¹ diet, respectively, were fed to tilapia (mean initial weight: 0.52 ± 0.01 g, n = 3) for 8 weeks. Each diet was fed to three replicate groups of fish in a closed, recirculating fresh water rearing system. The rearing water contained 27.1–33.3 mg L⁻¹ Ca. The tilapia fed the diets supplemented with ≥3.7 g Ca kg⁻¹ had significantly (P < 0.05) higher weight gain, when compared with fish fed the diet with ≤1.6 g Ca kg⁻¹. Fish fed the unsupplemented control showed significantly lower weight gain when compared with the other groups (P < 0.05). Bone Ca concentration was highest in fish fed the diets with ≥4.7 g Ca kg⁻¹, intermediate in fish fed the diet with 2.6 g Ca kg⁻¹ and lowest in fish fed the control diet. Scale Ca concentration was higher in fish fed the diets with ≥3.7 g Ca kg⁻¹ than in fish fed the diets with ≤2.6 g Ca kg⁻¹. Serum alkaline phosphatase activity was 36% increased in fish fed the diets with ≥2.6 g Ca kg⁻¹ than fish fed the diets with <1.6 g Ca kg⁻¹. Analysis by broken‐line regression of weight gain, bone and scale Ca concentrations indicated that the adequate dietary Ca concentration for tilapia in water containing 27.1–33.3 mg Ca L⁻¹ was 3.5, 4.3 and 4.2 g Ca kg⁻¹ diet, respectively, supplied as Ca‐lactate.     

Effects of acute thermal stress on acid–base regulation,haematology, ion‐osmoregulation and aerobic metabolism in Channel Catfish (Ictalurus punctatus)

The Channel Catfish Ictalurus punctatus is an important ecological, recreational and aquaculture species. In both natural and pond culture systems, Channel Catfish are periodically exposed to acute increase in temperature, especially in the summer, which may induce a stress response. With the effects of global climate change, the impact of rising temperatures on aquatic systems is expected to be exacerbated in the future. Hence, there is the need to understand the physiological mechanisms controlling the thermal stress response in fish. Acid–base, haematological, osmotic and metabolic regulations were investigated following acute temperature increase from 28°C to either 32°C or 36°C at a rate of 2°C/hr to assess the physiological response of Channel Catfish to temperature increase. Blood was sampled 1 hr after treatment temperatures were reached. When temperature increased from 28 to 32°C, blood bicarbonate () decreased while cortisol, partial pressure of carbon dioxide (pCO₂) and glucose increased. When temperature increased from 28 to 36°C, blood pH, , chloride, total proteins and mean corpuscular haemoglobin concentration decreased while cortisol, pCO₂, haematocrit, haemoglobin, glucose, lactate, mean corpuscular volume and mean corpuscular haemoglobin all increased. The changes observed are characteristic of acute stress responses in fish and suggest that acute temperature increases to both 32 and 36°C are physiologically stressful to Channel Catfish.     

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.     

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.     

Dietary Vitamin C Requirement of Fingerling,Cirrhinus mrigala (Hamilton), Based on Growth,Feed Conversion,Protein Retention,Hematological Indices,and Liver Vitamin C Concentration

This study was conducted to quantify dietary vitamin C requirement of fingerling, Cirrhinus mrigala, (0.79 ± 0.07 g; 3.51 ± 0.15 cm) by feeding casein‐gelatin based purified diets (400 g/kg crude protein; 3.45 kcal/g digestible energy) containing nine levels of vitamin C as l‐ascorbyl‐2‐polyphosphate (0.0, 5, 15, 25, 35, 45, 55, 75, and 95 mg vitamin C equivalent/kg diet) to triplicate groups of fish to apparent satiation for 16 wk. Absolute weight gain (AWG, g/fish), feed conversion ratio (FCR), protein retention efficiency (PRE%), RNA/DNA ratio, hemoglobin (Hb, g/dL), and hematocrit value (Hct%) were taken as the response criteria to determine vitamin C requirement of mrigal. Fish fed diet with 35 mg/kg vitamin C had significantly higher AWG (9.94 g/fish), FCR (1.39), PRE (27.72%), RNA/DNA ratio (4.18), Hb (11.15 g/dL), and Hct (34.44%) values. However, liver vitamin C concentration was found to be higher (64.92 µg/g wet tissue) in diet containing 45 mg vitamin C/kg. Broken‐line regression analysis of AWG data estimated the requirement of 35.65 mg/kg, whereas that of the liver vitamin C concentration data projected the requirement to 41.99 mg/kg.     

Effect of Dietary Vitamin C on Tissue Vitamin C Concentration in Channel Catfish,Ictalurus punctatus,and Clearance Rate at Two Temperatures-A Preliminary Investigation

Four trials were conducted with channel catfish, Ictalurus punctatus, under controlled laboratory conditions or in earthen ponds, to evaluate effects of dietary vitamin C concentration and feeding period on liver and anterior kidney vitamin C concentration. Clearance rate of vitamin C in liver and anterior kidney was also evaluated in unfed fish or fish fed a vitamin C-free diet. Results indicated that tissue vitamin C concentration is generally proportional to deitary vitamin C concentration, and that it may reach a maximum concentration within 2 weeks in fish fed diets containing vitamin C. Tissue vitamin C may be reduced to a minimum concentration within 2 weeks when fish were not fed or fed with a vitamin C-free diet at a high temprerature (31°). As temperature was reduced from 31 to 10°C there was a 44% and 13.6% drop in liver and anterior kidney vitamin C concentrations, respectively. At 10°C tissue vitamin C remained relatively constant (100 to 120 mg/g for liver, and 200 to 250 mg/g for anterior kidney) for at least 12 weeks if tissue vitamin C was saturated prior to reducing water temperature. Liver vitamin C appeared to be affected more dramatically by dietary vitamin C concentration and cleared more rapidly than anterior kidney during times of vitamin C deficiency.     

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.     

Spirulina as a natural carotenoid source on growth, pigmentation and reproductive performance of yellow tail cichlid Pseudotropheus acei

A feeding trial on yellow tail cichlid Pseudotropheus acei (Regan 1922) was undertaken to asses the effect of dietary Spirulina meal as a natural carotenoid source. Four experimental diets were formulated to progressively replace 0% (C), 2.5% (SP2.5), 5% (SP5) and 10% (SP10) of fish meal weight. Ten fish per tank (initial weight 3.75?±?0.02?g) were randomly distributed into twelve 80?l fiberglass tanks connected to a closed recirculation system (temperature 26.7?±?0.06°C). The diets were tested in triplicate for 12?weeks. The specific growth rate of fish fed all Spirulina diets were significantly higher compared to diet C. Feed intake (FI) tended to increase with dietary Spirulina level, and fish fed diet SP10 had significantly higher FI values compared to diet C. No significant differences in feed conversion ratio were observed among these groups. Although the protein efficiency ratio of fish fed diet C was lower than that of all Spirulina diets, no significant differences were observed among these groups (P?>?0.05). The total egg production and hatching rate (%) of eggs derived from all fish fed with Spirulina diets was significantly higher compared those from fish fed diet C. The yellow and blue coloration of yellow tail cichlid fed the diet containing Spirulina meal was enhanced and inclusion of dietary Spirulina meal was elevated carotenoids in skin. Results of the present study indicated that Spirulina meal has the potential to enhance the growth, reproductive performance and coloration on yellow tail cichlid.     

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.     

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.     

Effects of dietary propolis supplementation on growth performance, immune responses, disease resistance and body composition of juvenile eel, Anguilla japonica

This study was conducted to investigate the effects of dietary propolis supplementation on growth performance, immune responses, disease resistance and body composition of juvenile eel, Anguilla japonica, in order to evaluate its bioavailability as a feed additive for this species. A total of 540 fish averaging 7.7?±?0.22?g (mean?±?SD) were randomly distributed into 18 tanks in groups of 30, and each tank was then randomly assigned to one of three replicates of six diets containing 0 (P₀), 0.25 (P_0.25), 0.5 (P_0.5), 1.0 (P₁), 2.0 (P₂) and 4.0 (P₄) % dietary propolis. At the end of 12?weeks of feeding trial, weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and protein efficiency ratio (PER) of fish fed P_0.5 diet were significantly higher than those of fish fed P₀, P₁, P₂ and P₄ diets (P?<?0.05). These parameters were 148.9%, 0.72% day^?1, 94.4% and 2.9, respectively for fish fed P_0.5 diet. Serum lysozyme activity of fish fed P_0.5 (105.7 units mL^?1) and P₁ (106.0 units mL^?1) diets were significantly higher than those of fish fed P₀, P_0.25, P₂ and P₄ diets. Mucus lysozyme activity of fish fed P₁ (8.4 units 10?cm^?2) diet was significantly higher than those of fish fed P₀, P₂ and P₄ diets. Results indicated that the optimum dietary propolis supplementation levels could be 0.25?C0.5% for optimum growth and feed efficiency, and 0.5?C1% for enhanced immune responses and disease resistance in eel, A. japonica. This study may suggest that the dietary propolis level for the optimum immune responses could be higher than the level for the optimum growth of eel.