Antioxidant effect of dietary tocopherol and ascorbic acid on growth and survival of Litopenaeus vannamei postlarvae

The nutritional effect of vitamin E in dietsfor Litopenaeus vannamei postlarve (PL19)was investigated. Four formulated diets withdifferent combinations of -tocopherylacetate (-TA), ascorbic acid (AA) andhighly unsaturated fatty acids (HUFA) weretested, using four replicates.No significant differences in survival wereobserved among treatments after 34 days offeeding. However, shrimp fed with a dietcontaining 2% fish oil (low n-3 HUFA content),200 mg.kg^–1 -TA and100 mg.kg^–1 AA (diet H/E/C) showedsignificantly better growth than those fed adiet supplemented with 5% fish oil (high n-3HUFA content), 200 mg.kg^–1 -TA and100 mg.kg^–1 AA (diet H+/E/C). Shrimp fedwith a diet containing 5% fish oil,900 mg.kg^–1 -TA and100 mg.kg^–1 AA (diet H+/E+/C) showed a significantly higher tissue level of n-6 PUFAthan postlarvae fed diet H+/E/C. No definiteconclusion could be drawn about a possibleinteraction between -TA and AA, since acomparison of the diet containing 5% fish oil,200 mg.kg^–1 -TA and700 mg.kg^–1 AA (H+/E+/C+) and the dietH+/E/C did not show any significant differencesin any of the measured parameters. Theantioxidative status of the shrimp tissue(measured by means of the thiobarbituric acid(TBA) assay and expressed as nM malonaldehyde(MA) per gramme dry weight) was equal for alltreatments. Nevertheless, there was a slightlylower MA value with the diet H+/E/C+,indicating that AA may be an effectiveantioxidant in the aqueous phase and at thewater/lipid interface of the tissue. The tissuelevels of -T and AA were highlydependent on the amounts in diets and nocorrelation between -T and AAincorporation could be observed.     

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 importance of dietary HUFA for meagre larvae (Argyrosomus regius; Asso, 1801) and its relation with antioxidant vitamins E and C

Despite the interest of meagre (Argyrosomus regius) as a fast‐growing candidate for Mediterranean aquaculture diversification, there is a lack of information on nutrition along larval development. Importance of highly unsaturated fatty acids (HUFA) and the antioxidant vitamins E and vitamin C has not been investigated yet in this species. Six diets with two levels of HUFA (0.4% and 3% dw), two of vitamin E (1500 and 3000 mg kg^?1) and two of vitamin C (1800 and 3600 mg kg^?1) were fed to 15 dah meagre larvae. Larval growth in total length and dry body weight was significantly lowest in larvae fed diet 0.4/150/180 and showed few lipid droplets in enterocytes and hepatocytes and lower HUFA contents than the initial larvae. Increase in dietary HUFA up to 3%, significantly improved larval growth and lipid absorption and deposition. Besides, among fish fed 3% HUFA, increase in vitamin E and vitamin C significantly improved body weight, as well as total lipid, 22:6n‐3 and n‐3 fatty acids contents in the larvae. Thus, the results showed that 0.4% dietary HUFA is not enough to cover the essential fatty acid requirements of larval meagre and a high HUFA requirement in weaning diets is foreseen for this species. Besides, the results also pointed out the importance of dietary vitamin E and C to protect these essential fatty acids from oxidation, increase their contents in the larvae and promote growth, suggesting high vitamin E and C requirements in meagre larvae (higher than 1500 and 1800 mg kg^?1 for vitamin E and vitamin C respectively).     

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.     

Influences of dietary fatty acid profile on growth, body composition and blood chemistry in juvenile fat cod (Hexagrammos otakii Jordan et Starks)

This study was conducted to investigate the influence of dietary lipid source and n‐3 highly unsaturated fatty acids (n‐3 HUFA) level on growth, body composition and blood chemistry of juvenile fat cod. Triplicate groups of fish (13.2 ± 0.54 g) were fed the diets containing different n‐3 HUFA levels (0–30 g kg^?1) adjusted by either lauric acid or different proportions of corn oil, linseed oil and squid liver oil at 100 g kg^?1 of total lipid level. Survival was not affected by dietary fatty acids composition. Weight gain, feed efficiency and protein efficiency ratio (PER) of fish fed the diets containing squid liver oil were significantly (P < 0.05) higher than those fed the diets containing lauric acid, corn oil or linseed oil as the sole lipid source. Weight gain, feed efficiency and PER of fish increased with increasing dietary n‐3 HUFA level up to 12–16 g kg^?1, but the values decreased in fish fed the diet containing 30 g kg^?1 n‐3 HUFA. The result of second‐order polynomial regression showed that the maximum weight gain and feed efficiency could be attained at 17 g kg^?1 n‐3 HUFA. Plasma protein, glucose and cholesterol contents were not affected by dietary fatty acids composition. However, plasma triglyceride content in fish fed the diet containing lauric acid as the sole lipid source was significantly (P < 0.05) lower than that of fish fed the other diets. Lipid content of fish fed the diets containing each of lauric acid or corn oil was lower than that of fish fed the diets containing linseed oil or squid liver oil only. Fatty acid composition of polar and neutral lipid fractions in the whole body of fat cod fed the diets containing various levels of n‐3 HUFA were reflected by dietary fatty acids compositions. The contents of n‐3 HUFA in polar and neutral lipids of fish increased with an increase in dietary n‐3 HUFA level. These results indicate that dietary n‐3 HUFA are essential and the diet containing 12–17 g kg^?1 n‐3 HUFA is optimal for growth and efficient feed utilization of juvenile fat cod, however, excessive n‐3 HUFA supplement may impair the growth of fish.     

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.     

Effect of dietary n‐3 HUFA on growth performance and tissue fatty acid composition of gibel carp Carassius auratus gibelio

A 12‐week growth trial was conducted with gibel carp Carassius auratus gibelio (initial weight: 2.69 g) to evaluate the effects of dietary n‐3 highly unsaturated fatty acids (n‐3 HUFA) on growth performance and tissue fatty acid composition. Five diets of different n‐3 HUFA levels from 0 to 17 g kg^?1 diet were supplemented at 80 g kg^?1 dietary lipid by including fish oil (FO) at 0, 25, 50, 75 and 100% of supplemental lipid. The remainder was coconut oil. The results showed that fish fed FO₂₅ and FO₅₀ obtained highest specific growth rate and lowest with FO₀. Feed efficiency was highest at FO₁₀₀ and lowest at FO₀. Apparent digestibility coefficient of lipid increased with increasing dietary n‐3 HUFA. The fish fed FO₀ diet had the lowest thiobarbituric acid reactive substance in serum and muscle and highest moisture and lowest lipid content in viscera. Fatty acid compositions of muscle and liver were correlated with dietary fatty acids. Fish muscle concentration of 20:5n‐3 increased with increasing dietary n‐3 HUFA while the concentration of 22:6n‐3 was distinctly reduced in FO₀ group. It suggested that 4 g kg^?1 n‐3 HUFA in diet could permit gibel carp normal growth performance and provide considerable n‐3 HUFA in fish muscle. Excessive n‐3 HUFA showed impact on growth performance of gibel carp.     

Alteration of liver and muscle fatty acid composition in gilthead seabream (Sparus aurata) juveniles held at high stocking density and fed an essential fatty acid deficient diet

The aim of the present study was to determine the combined effect of both stress and EFA deficiency on several biological and biochemical parameters. Fish were fed during 15 weeks two isocaloric and isoproteic diets: a control diet based on fish oil and formulated to meet the n-3 HUFA requirements for this species (1.5% of n-3 HUFA) and a deficient diet containing beef tallow and formulated to be deficient in n-3 HUFA. Each experimental diet was evaluated both at high and low stocking densities (10 and 3.2 kg m^–3 of initial density, respectively).High stocking density produced a chronic stress situation with elevation of plasma cortisol levels. It also caused a reduction in hepatosomatic index and liver lipid contents, increasing the oleic acid/n-3 HUFA ratios in the polar lipids. Fish fed the EFA deficient diet at low stocking density showed common deficiency symptoms. High stocking density in fish fed the EFA deficient diet induced a higher degree of EFA deficiency symptoms leading to mortality, liver steatosis, liver lipid deposition, reduced muscle lipid and reduced n-3 HUFA contents, which particularly affected EPA, but not DHA, suggesting a preferential retention of the latter fatty acid, specially in the phosphoglycerides fraction.     

Evaluation of fish-meal free diets for rainbow trout, Oncorhynchus mykiss

Eight experimental diets were formulated for rainbow trout using agricultural byproducts as major ingredients. Each experimental diet contained varying amounts of corn grain, corn gluten meal, corn gluten feed and one of the following: 200 g kg^?1 peanut meal, 200 or 400 g kg^?1 soybean meal (SBM), 390 g kg^?1 low-allergen soy flour, 310 g kg^?1 soy protein concentrate, 300 g kg^?1 low-allergen soy protein concentrate or 200 g kg^?1 SBM + 110 g kg^?1 blood meal. One diet contained 200 g kg^?1 SBM and canola oil as the main lipid source. The remaining diets contained 95 g kg^?1 menhaden oil. Fish fed a commercial trout diet exhibited significantly greater weight gain (322%), and a lower feed conversion ratio (0.89) but significantly lower protein efficiency ratio (2.18) than fish fed the experimental diets. Within the experimental diets, fish fed the 400 g kg^?1 soy flour diet and the 400 g kg^?1 soybean meal diet had significantly higher weight gains (276% and 268%) and protein efficiency ratios (2.58 and 2.52), and lower feed conversion ratios (1.02 and 1.03) than fish fed other experimental diets. Fillet flavour varied between treatments. Most notable was the lower fishy flavour and higher chicken flavour of fish fed the diet that contained canola oil rather than menhaden oil. Microscopic evaluation of the liver and five sections of the gastrointestinal tract failed to demonstrate any differences between treatment groups. The ingredient costs of several experimental diets were lower than the estimated cost of a standard commercial trout diet. However, the superior feed conversion ratios of fish fed the control diet resulted in lower feed costs per unit of fish produced.     

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.     

Effects of diets containing linseed oil on fatty acid desaturation and oxidation in hepatocytes and intestinal enterocytes in Atlantic salmon (Salmo salar)

We hypothesized that replacing fish oil with 18:3n-3-rich linseed oil may enable salmon to maintain the levels of tissue n-3HUFA levels through a combination of increased desaturation activity and increased substrate fatty acid provision. To this end we investigated desaturation/elongation of [1-¹⁴C18:3n-3 in hepatocytes and intestinal enterocytes, and determined the extent to which 18:3n-3 was oxidized and desaturated by measuring both simultaneously in a combined assay. Salmon smolts were stocked randomly into five seawater pens and fed for 40 weeks on diets in which the fish oil was replaced in a graded manner by linseed oil. At the end of the trial, fatty acyl desaturation/elongation and oxidation activities were determined in isolated hepatocytes and intestinal enterocytes using [1-¹⁴C]18:3n-3 as substrate, and samples of liver and intestinal tissue were collected for analysis of lipid and fatty acid composition. The results showed that, despite increased desaturation of [1-¹⁴C]18:3n-3 in hepatocytes, provision of dietary 18:3n-3 did not prevent the decrease in tissue n-3HUFA in fish fed linseed oil. Intestinal enterocytes were a site of significant fatty acid desaturation but, in contrast to hepatocytes, the activity was not increased by feeding linseed oil and was generally lower in fish fed linseed oil compared to fish fed only fish oil. In contrast, oxidation of [1-¹⁴C]18:3n-3 in enterocytes was generally increased in fish fed linseed oil compared to fish fed the diet containing only fish oil. However, oxidation of [1-¹⁴C]18:3n-3 in hepatocytes was 4- to 8-fold lower than in enterocytes and was not affected by diet. Furthermore, oxidation of [1-¹⁴C]18:3n-3 in enterocytes exceeded desaturation irrespective of dietary treatment, whereas similar amounts of [1-¹⁴C]18:3n-3 were desaturated and oxidized in hepatocytes from fish fed only fish oil and desaturation exceeded oxidation by 3-fold in fish fed the diet containing 100% linseed oil. The molecular mechanisms underpinning these results were discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Utilization of different dietary lipid sources at high level in herbivorous grass carp (Ctenopharyngodon idella): mechanism related to hepatic fatty acid oxidation

Herbivorous grass carp (Ctenopharyngodon idella) has been reported to exhibit low capacity to utilize high dietary lipid, but different lipid sources might affect this limited capacity. In order to compare the effects of different lipid sources with different lipid levels, juvenile grass carp were fed one of nine diets containing three oils [lard, plant oil mixed by maize and linseed oil, and n‐3 high unsaturated fatty acid‐enriched (HUFA‐enriched) fish oil] at three lipid levels (20, 60 and 100 g kg^?1 dry diet) for 8 weeks. Decreased feed intake, poor growth performance, hepatic pathology and higher blood lipid peroxidation were found in 60 and 100 g kg^?1 fish oil groups. Conversely, in lard and plant oil groups, even at 100 g kg^?1 dietary lipid level, feed intake and growth performance did not decrease, despite histological observation revealed hepatic pathology in these groups. Plasma triglyceride and cholesterol contents increased significantly in all 100 g kg^?1 dietary lipid groups. In the comparison of hepatic FA β‐oxidation among three oil groups at 60 g kg^?1 dietary lipid level, impaired mitochondrial and peroxisomal FA oxidation capacity was observed in fish oil group. The results confirmed the relatively low capacity of grass carp to utilize high dietary lipid, and furthermore excess HUFA intake will result in more serious adverse effects than other FA.     

Partial substitution of fish oil with rapeseed, linseed and olive oils in diets for European sea bass (Dicentrarchus labrax L.): effects on flesh fatty acid composition, plasma prostaglandins E2 and F2α, immune function and effectiveness of a fish oil finishing diet

Triplicate groups of European sea bass (Dicentrarchus labrax L.), of initial weight 90 g, were fed four practical‐type diets in which the added oil was 1000 g kg^?1 fish oil (FO) (control diet), 600 g kg^?1 rapeseed oil (RO) and 400 g kg^?1 FO, 600 g kg^?1 linseed oil (LO) and 400 g kg^?1 FO, and 600 g kg^?1 olive oil (OO) and 400 g kg^?1 FO for 34 weeks. After sampling, the remaining fish were switched to the 1000 g kg^?1 FO diet for a further 14 weeks. Fatty acid composition of flesh total lipid was influenced by dietary fatty acid input but specific fatty acids were selectively retained or utilized. There was selective deposition and retention of docosahexaenoic acid (DHA; 22:6n‐3). Eicosapentaenoic acid (EPA; 20:5n‐3) and DHA were significantly reduced and linolenic (LNA; 18:3n‐3), linoleic (LA; 18:2n‐6) and oleic (OA; 18:1n‐9) acids significantly increased in flesh lipids following the inclusion of 600 g kg^?1 RO, LO and OO in the diets. No significant differences were found among different treatments on plasma concentrations of prostaglandin E₂ and prostaglandin F_2α. Evaluation of non‐specific immune function, showed that the number of circulating leucocytes was significantly affected (P < 0.001), as was macrophage respiratory burst activity (P < 0.006) in fish fed vegetable oil diets. Accumulation of large amounts of lipid droplets were observed within the hepatocytes in relation to decreased levels of dietary n‐3 HUFA, although no signs of cellular necrosis was evident. After feeding a FO finishing diet for 14 weeks, DHA and total n‐3 HUFA levels were restored to values in control fish although EPA remained 18% higher in control than in the other treatments. This study suggests that vegetable oils such as RO, LO and OO can potentially be used as partial substitutes for dietary FO in European sea bass culture, during the grow out phase, without compromising growth rates but may alter some immune parameters.     

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 ascorbic acid/iron ratio on some production traits, lipid peroxide state and amount/activity of the glutathione redox system in African catfish Clarias gariepinus (Burchell) fingerlings

African catfish Clarias gariepinus (Burchell) fingerlings (3.16–3.92 g initial body weight) were investigated for 30 days in four different groups using different amounts of l ‐ascorbic acid (AA) and iron (supplied as FeC₆H₅O₇) in their feedings. Diet 1 (control): no addition of AA or iron; diet 2 (H‐AA/FE): high (600 mg kg^?1) AA and low (218 mg kg^?1) iron; diet 3 (H‐HE/AA): high (364 mg kg^?1) iron and low (200 mg kg^?1) AA; and an unfed group, which was investigated only for 15 days due to high mortality. The live weight gain, feed intake, specific growth rate (SGR; % body weight day^?1) and feed conversion rate (FCR) were measured or calculated. At the end of the experimental period, the whole body content of AA, iron, reduced glutathione (GSH), glutathione disulphide (GSSG) and malondialdehyde (MDA), as well as the glutathione peroxidase (GSHPx) activity, were measured. The production traits did not differ significantly as a result of the different AA and iron contents of the feed. AA content increased significantly in all the groups as compared with the initial value, except in the unfed group. The difference between the treated groups as compared with the control, with regard to the two AA/iron treated groups, was also significant. The iron content in the fish body increased significantly compared with the initial value, except in the unfed group. The difference compared with the control was significant only in the H‐FE/AA group. The difference between the groups that consumed low and high iron content diets was also significant. The GSH and GSSG content, as well as the GSH/GSSG ratio and GSHPx activity of the fish body, did not differ significantly as compared with the initial value or with the control. The lipid peroxide status, as measured by the MDA content, did not differ significantly either as an effect of the AA and iron supplementation, but decreased as an effect of ageing and starvation. It may be concluded that, under the present experimental conditions, the C. gariepinus fingerling tissue stores of AA and/or iron increased as a result of feed supplementation, but without altering the actual lipid peroxide status and the amount/activity of the glutathione redox system.     

Effect of dietary lipid oxidation with vitamin C and E supplementation on fillet quality of red sea bream,Pagrus major (Temminck & Schlegel) during storage

An experiment was conducted to determine the effects of different levels of dietary vitamin C (VC) and E (VE) supplementation on fillet quality of red sea bream fed oxidized fish oil (OFO). Fish with an average body weight of 205.0 g were fed four test diets for 9 weeks. Control diet contained fresh fish oil (FFO) with 100 mg kg^?1 of VE and 500 mg kg^?1 of VC (FFO100E/500C). The other three diets contained OFO with varying levels of VE (mg kg^?1) and VC (mg kg^?1) (OFO100E/500C, OFO200E/500C and OFO200E/1000C). After feeding trial, two fillets from each fish by hand filleting were stored in a refrigerator at 4°C for 96 h during analyses. Results showed that fish fed OFO increased fillet thiobarbituric acid reactive substances (TBARS) and K‐value, and decreased fillet VC and VE concentrations during storage time. Supplementation of VC did not have any detectable effect on fillet quality. Increasing dietary VE supplementation increased fillet VE concentrations, reduced fillet TBARS and K‐value values of red sea bream. Therefore, we suggest that dietary supplementation of 200 mg kg^?1 of vitamin E could improve fillet oxidative stability of red sea bream fed OFO.     

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.     

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.     

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.     

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.