Dietary probiotic supplementation in juvenile rainbow trout (Oncorhynchus mykiss) reared under cage culture production: effects on growth,fish welfare,flesh quality and intestinal microbiota

This study was conducted to evaluate the effects of dietary probiotic supplementation on growth performance, fish welfare, sensory evaluation, freshness during ice storage and changes in intestinal microbiota of rainbow trout (Oncorhynchus mykiss). The 9‐week feeding trial was conducted in cages under commercial conditions during the winter season, when water temperature decreased from 14°C to 7°C. Fish were fed either a diet without probiotic supplementation (control) or with 0.2% (2 × 10⁹ CFU kg^?1 diet) of multi‐strain probiotic bacteria (Bacillus sp., Pediococcus sp., Enterococcus sp., Lactobacillus sp.). Fish weight was recorded at 0, 2, 4, 6 and 9 weeks. Growth performance was not affected by dietary probiotic supplementation. Alternative complement activity (ACH50) was significantly higher in fish fed the diet with probiotic (58.4 ± 3.7 units mL^?1) than in the control group (46.9 ± 3.1 units mL^?1) (P = 0.03). Several biochemical markers were altered by the dietary treatment. 7‐ethoxyresorufin‐O‐deethylase (EROD) (P < 0.01), glutathione S‐transferase (GST) (P < 0.001), glutathione peroxidase (GPx) (P < 0.05) increased and lipid peroxidation (LP) (P < 0.01) decreased with dietary probiotic supplementation. Flesh quality and shelf‐life of fish stored on ice was not affected by dietary probiotic supplementation. Probiotic supplementation had a limited effect on the intestinal microbiota. Signal strength of the observed operational taxonomic units (OTUs) differed between the control and treatment samples and could indicate variations in the abundance of certain OTUs. Microbiota adhering to the intestinal mucus varied between sampling points, indicating that changes were associated with water temperature.     

Estimation of dietary vitamin A requirement of juvenile soft‐shelled turtle,Pelodiscus sinensis

A feeding trial was conducted to evaluate the dietary vitamin A requirement of the soft‐shelled turtle Pelodiscus sinensis. Turtles with an approximate body weight of 6.8 g were fed diets containing 0–4.58 mg kg^?1 (15 251 IU) vitamin A (retinyl acetate) for 8 weeks. Turtles fed diets without vitamin A showed the lowest growth performance. The animals gradually gained weight when dietary vitamin A increased from 0 to 3.6 mg kg^?1. Liver vitamin A gradually increased with supplementation of dietary vitamin A of up to 2.7 mg kg^?1. Beyond this concentration, the vitamin A in the liver significantly increased. Data analysis using a polynomial regression or exponential model showed that the estimated dietary vitamin A requirement for juvenile soft‐shelled turtles based on weight gain, red blood cell count and liver vitamin A measurements was 3.48, 2.84 and 2.58 mg kg^?1, respectively.     

Efficacy and tolerance of lutein as colourant in diet of juvenile soft‐shelled turtle Pelodiscus sinensis

The effects of dietary supplementation of lutein on the growth, haematological profile and pigmentation in target tissues of the soft‐shelled turtles were investigated. Five experimental diets were prepared with lutein levels at 1.16 (D1, control), 70.3 (D2), 132 (D3) and 239 (D4) mg kg^?1, and one more high inclusion level of 3410 mg kg^?1 (D5) was included for tolerance test. Each diet was fed to 24 juvenile turtles for 8w. Specific growth rate (SGR) in D2, D3 and D4 was significantly higher than that of group D1 and D5. Skin yellowness (b*) and chroma (C*) of both carapace and plastron skin were all significantly increased with higher dietary lutein levels. Dietary lutein supplementation turned the H° values of skins into yellowness. The accumulation of lutein in skin and muscle also augmented significantly with increasing lutein levels and fit a logarithmic correlation with dietary lutein levels. There were almost no differences in haematological parameters of all turtles. Lutein is an efficient and safe colourant in diet of soft‐shelled turtle. The optimal inclusion levels for the highest SGR and desired skin colour in carapace and plastron of soft‐shelled turtle were 162, 142 and 143 mg kg^?1, respectively.     

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

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

Effects of dietary β‐carotene levels on growth and liver vitamin A concentrations of the soft‐shelled turtle,Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to evaluate the effect of dietary β‐carotene level on the growth and liver vitamin A concentrations in soft‐shelled turtles, Pelodiscus sinensis, fed a vitamin A‐free diet. Soft‐shelled turtles were fed diets containing 0, 14.5, 26.5, 47.5, 87.3, 112.8 and 163.8 mg β‐carotene kg^?1 for 10 weeks. Although it was not statistically significant due to high deviation within each group, mean weight gain of soft‐shelled turtles fed the diet without β‐carotene supplementation was the lowest among all test groups. Vitamin A concentrations in liver of turtles significantly (P<0.05) increased when dietary β‐carotene level reached 47.5 mg kg^?1 indicating that soft‐shelled turtles were capable of converting β‐carotene to vitamin A. Analysed by regression modelling, dietary β‐carotene levels for optimal growth and maximal liver vitamin A contents of juvenile soft‐shelled turtles fed the vitamin A‐free diets were 49.1 and 88.7 mg kg^?1 respectively.     

Effects of dietary live or heat‐inactivated autochthonous Bacillus pumilus SE5 on growth performance,immune responses and immune gene expression in grouper Epinephelus coioides

To evaluate the possible dietary application of live and heat‐inactivated probiotic Bacillus pumilus SE5 in grouper Epinephelus coioides, juveniles (14.6 ± 0.2 g) were fed either a basal control diet (without probiotic) or the basal diet supplemented with 1.0 × 10⁸ CFU g^?1 live (T1) and heat‐inactivated B. pumilus SE5 (T2). The heat‐inactivated probiotic significantly improved the final weight, weight gain (WG) and specific growth rate (SGR) at day 60 and significantly decreased the feed conversion ratio (FCR) at day 30 and 60, while the viable probiotic significantly decreased the FCR at day 60 (P < 0.05). Phagocytic activity, serum complement C3 and IgM levels as well as SOD activity elevated significantly in fish fed the heat‐inactivated probiotic for 60 days (P < 0.05). Furthermore, the heat‐inactivated probiotic remarkably up‐regulated expression of TLR2 and pro‐inflammatory cytokines (IL‐8 and IL‐1β) in head kidney (P < 0.05), but the viable probiotic failed to do so. These results indicated that heat‐inactivated B. pumilus SE5 can effectively improve the growth performance and immune responses of E. coioides.     

Effects of dietary magnesium on the growth,carapace strength and tissue magnesium concentrations of soft‐shelled turtle,Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to evaluate the effects of dietary magnesium on the growth, carapace strength, tissue and serum Mg concentration of soft‐shelled turtles, Pelodiscus sinensis (Wiegmann). Juvenile soft‐shelled turtles of approximate 5.4 g body weight were fed diets with seven levels of Mg (48, 206, 369, 670, 955, 1195 and 1500 mg Mg kg^?1) for eight weeks. No significant difference (P ≥ 0.05) was found in weight gain (WG), feed conversion ratio or protein efficiency ratio among treatments. However, the WG of turtles continued to increase with increasing dietary Mg levels up to 670 mg kg^?1, beyond which the WG levelled off. The plasma alkaline phosphatase activity and the muscle, bone Mg concentrations of the turtles increased with the increasing dietary Mg levels between 48 and 955 mg kg^?1, beyond which the tissue Mg concentrations remained relatively constant. Furthermore, the carapace strengths of turtles fed with the control diet of 48 mg Mg kg^?1 were significantly weaker (P < 0.05) than that of turtles fed with diets containing higher Mg levels. Based on a broken‐line modelling analysis, the required dietary Mg level for the optimal WG of juvenile soft‐shelled turtles was estimated to be approximately 650 mg kg^?1. By contrast, the required dietary Mg levels for turtles to reach the optimal muscle and bone Mg concentrations were 1050 and 1000 mg kg^?1 respectively. The required dietary Mg level for maximal alkaline phosphatase activity was approximately 980 mg kg^?1.     

Effects of exogenous xylanase supplementation in plant protein‐enriched diets on growth performance,intestinal enzyme activities and microflora of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 900 juvenile Jian carp (Cyprinus carpio var. Jian) (7.99 ± 0.02 g) were fed diets containing graded levels of xylanase at 220 (unsupplemented control), 650, 1070, 1480, 1810 and 2470 U kg^?1 diet for 10 weeks to investigate the effects of xylanase levels on growth performance, intestinal enzyme activities and microflora. The per cent weight gain, feed efficiency, protein efficiency ratio, protein production value, lipid production value, ash production value, calcium production value and phosphorus retention ratio were significantly improved with increasing levels of xylanase up to a point, and thereafter declined (P < 0.05). The activities of trypsin, chymotrypsin, lipase and amylase in the hepatopancreas and intestine, activities of alkaline phosphatase, Na⁺, K⁺‐ATPase, creatine kinase and γ‐glutamyl transpeptidase in three intestinal segments were improved by dietary xylanase (P < 0.05). The amounts of Lactobacillus, Escherichia coli and Aeromonas were significantly affected by dietary xylanase levels (P < 0.05). In conclusion, xylanase supplementation improved growth performance, enhanced intestinal enzyme activities and influenced the balance of intestinal microflora of juvenile Jian carp. The optimal level of xylanase in juvenile Jian carp (7.99–99.16 g) based on PWG was 1259 U kg^?1 diet by the quadratic regression analysis.     

Effects of dietary choline supplementation on growth performance,lipid deposition and intestinal enzyme activities of blunt snout bream Megalobrama amblycephal fed high‐lipid diet

This study aimed to investigate the effects of dietary choline supplementation on growth, lipid deposition and intestinal enzyme activities of Megalobrama amblycephala. Fish were fed four diets with two lipid levels (50 and 150 g kg^?1) and two choline supplementations (600 and 1600 mg kg^?1) for 8 weeks. Feed conversion ratio (FCR), viscerosomatic index (VSI), hepatosomatic index (HSI), intraperitoneal fat (IPF) ratio, whole‐body and muscle lipid contents, intestinal lipase activities and lipoprotein lipase (LPL) activities all increased significantly (P < 0.05) as lipid levels increased, whereas the opposite was true for whole‐body and muscle moisture contents and intestinal amylase activities. VSI, IPF ratio and whole‐body lipid contents all decreased significantly (P < 0.05) with increasing dietary choline supplementations. Weight gain, muscle moisture content all increased significantly (P < 0.05) with increasing dietary choline supplementations when dietary lipid levels reached 150 g kg^?1, whereas the opposite was true for FCR, IPF ratio, IPF and liver LPL activities. In addition, abnormal hepatocytes were found in the liver of fish fed 150 g kg^?1 lipid with 600 mg kg^?1 choline supplementation. The result of this study indicated that extra choline supplementation can improve growth performance, intestinal enzymes activities and reduce excessive lipid deposition of M. amblycephala fed high lipid.     

Effect of replacing fish meal with soybean meal and of DL‐methionine or lysine supplementation in pelleted diets on growth and nutrient utilization of juvenile golden pompano (Trachinotus ovatus)

The experiment was conducted to determine the effect of dietary DL‐methionine addition in fish meal reduction diet for juvenile golden pompano (Trachinotus ovatus). The trial comprises the following of 11 diet treatments. (i) D1 diet as the standard was formulated with fish meal as the main protein source. (ii) D2–D5 diets were formulated to replace fish meal with soybean meal at 60, 120, 180 and 300 g kg^?1, respectively, and the amino acid profiles (including methionine and lysine) of D2–D5 were referred to the amino acid profile of D1. (iii) D6‐D9 diets were the same as D2–D5, respectively, but without methionine supplementation. (iv) D10 and D11 diets were the same as D3 and D5 diets, respectively, but without lysine supplementation. Each diet was randomly fed to groups of 20 fish (initial average weight about 18 g) per net cage (1.0 × 1.0 × 1.5 m) in triplicate and the feeding experiment lasted for 56 days. Weight gain (WG, %) and final body weight (FBW, g) of fish fed D3 diet were significantly higher than those of fish fed D8 and D9 diets (P < 0.05), but without significant differences with fish fed other diets (P > 0.05). Feed conversion ratio (FCR) of D3, D4 and D5 diet treatments were significantly lower than that of D11 diet treatment (P < 0.05), but without significant difference with other diet treatments (P > 0.05). Whole‐body protein contents of fish fed D3–D5 were significantly higher than those of fish fed D7–D9 (P < 0.05), but without significant differences with fish fed other diets (P > 0.05). In conclusion, the replacement of fish meal by soybean meal could reach 300 g kg^?1 without detrimental but profitable effect on growth and survival when enough methionine and lysine were supplemented in the diet; moreover, lysine is not the first‐limiting amino acid relative to methionine for juvenile T. ovatus from the present growth and proximate composition results.     

Effect of dietary calcium and phosphorus supplementation in fish meal-based diets on the growth of soft-shelled turtle Pelodiscus sinensis (Wiegmann)

A growth experiment was conducted to determine the effect of supplementing dietary calcium in fish meal‐based diets on the growth of cultured soft‐shelled turtle Pelodiscus sinensis. Juvenile soft‐shelled turtles of 4.1 g mean body weight were fed nine diets containing two levels of phosphorus (2.7% or 3.0%) and analysed calcium levels ranging from 4.7% to 6.6% for 10 weeks. The growth of the turtles was enhanced when inorganic calcium was added to the diets. The weight gain of the turtles fed the control diet containing calcium solely from fish meal was the lowest among the test groups, and was significantly lower than those fed the diet containing 5.7% calcium at the 3.0% phosphorus level (P<0.05). Feed conversion and protein efficiency ratios were not affected by different dietary treatments. Whole‐body moisture and crude protein contents of turtles were not affected by different dietary treatments. The body ash of turtles fed 3.0% phosphorus diets tended to be higher than turtles fed 2.7% phosphorus diets. The body calcium to phosphorus ratio of turtles fed 3.0% phosphorus diets was greater than that of turtles fed diets containing 2.7% phosphorus. Supplementation of Ca in a fish meal‐based practical diet is required for the optimum growth of soft‐shelled turtles.     

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

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

Effects of dietary mannan oligosaccharide on growth,survival, physiological,immunological and gut morphological conditions of black tiger prawn (Penaeus monodon Fabricius 1798)

The aim of this experiment was to determine the effects of dietary inclusion with mannan oligosaccharide (Bio‐Mos, Alltech, Nicholasville, KY, USA) on growth, survival, physiological and immunological conditions and gut morphology of the black tiger prawn (Penaeus monodon). Five diets supplemented with MOS at 0 g kg^?1 (control diet), 1, 2, 4 and 8 g kg^?1 were fed to the prawn juveniles (0.4 ± 0.06 g, total weight) for the duration of 63 days. Growth was the highest (P < 0.05) when the prawns were fed the 1 g kg^?1 MOS included diet. Wet tail muscle index (Tw/B), dry tail muscle index (Td/B) and tail muscle protein (Tmp) were higher (P < 0.05) in the prawns fed MOS included diets MOS compared with the prawns fed the control diet. Total haemocyte counts (THCs) of the prawns fed MOS included diets were higher (P < 0.05) than THCs of the prawns fed the control diet. Epithelium layer and epidermal cell density of the gut of the prawns fed 1 g kg^?1 and 2 g kg^?1 MOS diets were better than the prawns fed the control and other MOS diets. The results imply a positive effect of dietary supplementation of 1–2 g kg^?1 MOS in the culture of black tiger prawns.     

Dietary selenium requirement for on‐growing gibel carp (Carassius auratus gibelio var. CAS III)

A 12‐week growth trial was conducted in a flow‐through system to determine dietary selenium (Se) requirement for on‐growing gibel carp (initial body weight: 76.2 ± 0.05 g, mean ± SEM). Selenomethionine was supplemented to the basal diet to formulate seven semi‐purified diets containing 0.26, 0.58, 0.72, 1.14, 1.34, 1.73 and 2.09 mg Se kg^?1 diet. The results showed that plasma superoxide dismutase (SOD) activity significantly increased when fish were fed with 0.58 mg Se kg^?1 diet (P < 0.05) and then decreased at 2.09 mg Se kg^?1 diet (P < 0.05). Plasma T‐AOC activity was higher in fish fed with 0.72 mg Se kg^?1 diet (P < 0.05) and plasma malondialdehyde (MDA) was higher in fish fed with 0.26 mg Se kg^?1 diet (P < 0.05). When fish were fed 1.14 mg Se kg^?1 diet, hepatic GSH‐Px, T‐AOC, GSH and CAT activities were significantly higher than those fed with 0.26 mg Se kg^?1 diet (P < 0.05). Hepatic superoxide dismutase (SOD) activity was higher at 1.34 mg Se kg^?1 diet (P < 0.05). Fish liver Se concentrations were significantly higher when fed with 0.72 mg Se kg^?1 diet (P < 0.05) and then kept constant when Se ≥ 0.72 mg kg^?1 (P > 0.05). Whole‐body and muscle Se concentrations were higher when fed with 1.34 mg Se kg^?1 diet (P < 0.05) and kept a plateau when Se ≥ 1.34 mg kg^?1 (P > 0.05). In conclusion, based on broken‐line regression of hepatic Se concentrations, hepatic SOD activity and hepatic T‐AOC activity, dietary Se requirements for on‐growing gibel carp was 0.73 mg kg^?1, 1.12 mg kg^?1 and 1.19 mg kg^?1 diet respectively.     

The effects of Spirulina platensis meal on proximate composition,fatty acid profile and lipid peroxidation of rainbow trout (Oncorhynchus mykiss) muscle

This study evaluated the effects of diets containing 0, 25, 50, 75 and 100 g kg^?1 Spirulina platensis on proximate composition, fatty acid profile and lipid peroxidation of rainbow trout (Oncorhynchus mykiss). Supplementation of S. platensis did not change moisture and protein contents, but fish fed 50 and 100 g kg^?1 S. platensis had lower muscle lipid content than those fed control diet (P < 0.05). Fish fed 100 g kg^?1 of S. platensis contained lower percentages of saturated and monounsaturated fatty acid and a higher percentage of polyunsaturated fatty acid than those fed control diet (P < 0.05). The n‐3/n‐6 ratio of the fatty acid increased and muscle atherogenic and thrombogenic indices were significantly decreased as the dietary supplement of S. platensis increased. Furthermore, lipid peroxidation of the fillet significantly decreased with increasing dietary S. platensis at 4 °C and at ?20 °C (P < 0.05). The results of this study show that supplementation of S. platensis to the diet improves muscle quality of the rainbow trout.     

Effects of dietary organic selenium on immune responses,total selenium accumulation and digestive system health of marron,Cherax cainii (Austin, 2002)

A 90‐day feeding trial was conducted to determine the effects of dietary organic selenium (OS) supplementation on immune responses, total selenium accumulation in various tissues and the digestive system health of marron Cherax cainii. A group of marron (initial weight 3.65 ± 0.05 g) were fed 0.2 g kg^?1 of Sel‐Plex^® as a source of OS and were compared to a control group of marron that were not fed OS. After 90 days of feeding OS, total haemocyte counts (THC) and the percentages of granular and semigranular cells in marron were significantly higher than in marron fed a control diet. However, dietary OS did not affect the proportion of hyaline cells. The marron fed OS had significantly lower bacteraemia and longer neutral red retention times than the control group. Total selenium levels in the haemolymph, hepatopancreas and muscle tissues of the OS‐fed marron group were significantly higher (P < 0.05) than in control diet group. The highest total soluble selenium levels were found in the muscle tissues of OS‐fed marron on the final day of the feeding trial. Amylase and protease activity, soluble protein, and numbers of microvilli were significantly higher (P < 0.05) in marron fed OS. These findings demonstrate the benefits of including 0.2 g kg^?1 OS in marron diet to enhance immune parameters, increase total selenium accumulation in muscle and improve marron digestive system health.     

Dietary choline requirement for juvenile blunt snout bream,Megalobrama amblycephala

A 12‐week feeding trial was conducted to determine dietary choline requirement for juvenile Megalobrama amblycephala. The basal diet was formulated to contain 310 g kg^?1 diet from vitamin‐free casein and gelatine. Choline chloride was supplemented to the basal diet to formulate six purified diets containing 0, 250, 500, 1000, 2000 and 4000 mg kg^?1, respectively. Each diet was randomly fed to quadrupled groups of Megalobrama amblycephala with initial average weight 1.84 ± 0.04 g in a flow‐through system. Results showed weight gain was increased significantly with increasing dietary choline levels (P < 0.01). Lipid content of liver decreased significantly as dietary choline concentration increased (P < 0.01), whereas lipid content of dressed carcass showed opposite trend (P < 0.01), and lipid content of whole‐body was unaffected by dietary choline supplementation. Broken‐ line regression of weight gain, liver and muscle choline concentration showed choline requirements of Megalobrama amblycephala of 1198, 1525 and 1365 mg kg^?1, respectively. In addition, dietary choline supplementation significantly improved lipid content of dressed carcass but not the content of whole body of blunt snout bream.     

The effect of vitamin C on stress withstanding capability in the juvenile soft-shelled turtle (Pelodiscus sinensis)

The study was conducted to determine the effect of dietary supplementation of vitamin C on anti‐acid stress ability in juvenile soft‐shelled turtle. The soft‐shelled turtles were fed vitamin C supplemented diets at levels of 0, 250, 500, 2500, 5000 and 10 000 mg kg^?1 for 4 weeks. The results showed that the phagocytic rate of blood cell in the group fed vitamin C deficient diets, the serum bacteriolytic activity in the two groups fed vitamin C supplemented diet at 0 and 250 mg kg^?1 and the serum bactericidal activity in all groups after acid stress significantly decreased compared with those of before stress (P < 0.1). The phagocytic rate of blood cells in the two groups fed vitamin C supplemented diet at 2500 and 5000 mg kg^?1 were significantly higher than those of the four groups fed at 0, 250, 500 and 10 000 mg kg^?1 (P < 0.01). The serum bacteriolytic activity in the groups fed vitamin C supplemented diet at 500, 2500, 5000 and 10 000 mg kg^?1 were significantly higher than that of the group fed vitamin C deficient diets (P < 0.05), and the two groups fed vitamin C at 2500 and 5000 mg kg^?1 were significantly higher than those of the groups fed at 250 and 10 000 mg kg^?1 (P < 0.05). The serum bactericidal activities in the four groups fed vitamin C supplemented diet at 500, 2500, 5000 and 10 000 mg kg^?1 were significantly higher than those of the two groups fed at 0 and 250 mg kg^?1 diet (P < 0.01), and the two groups fed vitamin C at 2500 and 5000 mg kg^?1 were significantly higher than those of the two groups fed at 500 and 10 000 mg kg^?1 (P < 0.1). These results suggest that supplementation of vitamin C higher than 250 mg kg^?1 is necessary to reduce the adverse effects of acid stress.     

Effects of dietary choline on growth performance,lipid deposition and hepatic lipid transport of grouper (Epinephelus coioides)

The number of 360 individuals with an average initial weight of 87.8 ± 0.04 g was fed six diets containing graded levels of choline at 8.1 (control group), 602.5, 1119.0, 1511.5, 1970.0 and 4029.0 mg choline kg^?1 diet, respectively, to investigate the effects of dietary choline on growth performance, lipid deposition and hepatic lipid transport for grouper, Epinephelus coioides. Dietary methionine was estimated to be 10.02 g kg^?1, less than the requirement (13.10 g kg^?1). The results of 10‐week study period indicated that the best values of specific growth rate (SGR), feed conversion rate (FCR) and protein efficiency rate (PER) all occurred in 1119.0 mg choline kg^?1 diet (P < 0.05). The survival range increased from 8.1 to 1511.5 mg choline kg^?1 diet and then plateaued. Dietary choline supplementation significantly decreased the liver lipid content of grouper (P < 0.05), but the lipid content of the muscle tended to be increased firstly and then decreased (P < 0.05). Liver choline concentration reached a plateau in 1511.5 mg choline kg^?1 diet and then levelled off (P < 0.05). Serum high density lipoprotein‐cholesterol (HDL‐C) and total cholesterol (TCHO) levels were firstly decreased and then increased with dietary choline supplementation. A reversed tendency, however, was found in triglyceride. Broken‐line regression analysis of SGR and liver choline content indicated that choline requirement of grouper was 1093.7 and 1579.7 mg kg^?1 diet, respectively.     

Effects of dietary manganese source and supplemental levels on growth performance,antioxidant enzymes activities,tissue Mn concentrations and cytosolic manganese superoxide dismutase (cMnSOD) mRNA expression level of juvenile Litopenaeus vannamei

This experiment was conducted to study the effects of different forms and levels of manganese (Mn) on the growth performance, antioxidant activities, tissue Mn content and cytosolic manganese superoxide dismutase (cMnSOD) gene expression of Litopenaeus vannamei. Treatments consisted of 0, 10, 20, 30, 40 and 60 mg Mn kg^?1 from manganese sulphate (Mn‐S) and manganese methionine (Mn‐Met), providing the actual dietary value of 5.17, 15.62, 25.55, 34.22, 44.48 and 67.90 mg Mn kg^?1 Mn‐S, and 5.17, 15.71, 25.36, 35.86, 45.16 and 65.06 mg Mn kg^?1 Mn‐Met, respectively. Each diet was fed to triplicate groups of L. vannamei (initial body weight: 1.925 ± 0.002 g) in a recirculated fresh water rearing system for 8 weeks. Weight gain rate (WGR) increased in prawns provided with from 25.55 to 44.48 mg Mn kg^?1 Mn‐S and 15.71 to 45.16 mg Mn kg^?1 Mn‐Met and then declined above these levels. The lowest protein efficiency ratio (PER) and the highest feed conversion rate (FCR) were observed in prawns fed the control diet (P < 0.05) and showed no significant differences among other treatments (P > 0.05). Survival rate (SR) was not affected by the dietary treatments (P > 0.05). Total SOD and Mn‐SOD activities were higher in the hepatopancreas of prawns fed with Mn‐supplemented diets from 15.71 to 44.48 mg Mn kg^?1 (P < 0.05). On the contrary, malondialdehyde (MDA) content was lower in the hepatopancreas of prawns fed the basal diet (P < 0.05). Mn concentrations in the hepatopancreas and muscles increased with increasing levels of dietary Mn supplementation. Moreover, Mn accumulation was lower in the muscle than in the hepatopancreas of the prawns. The mRNA expression of cMnSOD gene in the hepatopancreas of prawns was upregulated with increasing dietary Mn levels of Mn‐S from 25.55 to 44.48 mg Mn kg^?1, Mn‐Met from 15.71 to 45.16 mg Mn kg^?1 and then plateaued above these levels. Broken‐line regression analysis of WGR indicated that the optimal dietary Mn requirements for juvenile L. vannamei were 32.26 mg Mn kg^?1 Mn‐S and 23.90 mg Mn kg^?1 Mn‐Met, respectively.