Effects of Pseudoalteromonas sp.BC228 on Digestive Enzyme Activity and Immune Response of Juvenile Sea Cucumber （Apostichopus japonicus）

A marine bacterium,Pseudoalteromonas sp.BC228 was supplemented to feed in a feeding experiment aiming to determine its ability of enhancing the digestive enzyme activity and immune response of juvenile Apostichopusjaponicus.Sea cucumber individuals were fed with the diets containing 0 （control）,105,107 and 109 CFU g-1 diet of BC228 for 45 days.Results showed that intestinal trypsin and lipase activities were significantly enhanced by 107 and 109 CFU g-1 diet of BC228 in comparison with control （P〈0.01）.The phagocytic activity in the coelomocytes of sea cucumber fed the diet supplemented with 107 CFUg-1 diet of BC228 was significantly higher than that of those fed control diet （P〈0.05）.In addition,105 and 107 CFUg-1 diet of BC228 significantly enhanced lysozyme and phenoloxidase activities in the coelomic fluid of sea cucumber,respectively,in comparison with other diets （P 〈 0.01）.Sea cucumbers,10 each diet,were challenged with Vibrio splendidus NB 13 after 45 days of feeding.It was found that the cumulative incidence and mortality of sea cucumber fed with BC228 containing diets were lower than those of animals fed control diet.Our findings evidenced that BC228 supplemented in diets improved the digestive enzyme activity of juvenile sea cucumber,stimulated its immune response and enhanced its resistance to the infection of V.splendidus.     

Effect of Bacillus baekryungensis YD13 Supplemented in Diets on Growth Performance and Immune Response of Sea Cucumber（Apostichopus japonicus）

The effect of a potential probiotic on the growth performance and immune response of sea cucumber（Apostichopus japonicus） was investigated. Bacillus baekryungensis YD13 isolated from sea cucumber culturing ponds was added to sea cucumber basal feed as a probiotic in different doses（0, the control; 1×10^4（YD134）, 1×10^6（YD136） and 1×10^8（YD138） CFU g-1 of diet）, and administered orally to A. japonicus（initial mean wet weight 5.44 g ± 0.17 g）. The sea cucumbers were fed in 20 aquaria, 5 each treatment, for 60 d. At the end of growth trial, 20 sea cucumbers from each treatment were challenged with Vibrio splendidus. A. japonicus in YD134 and YD136 exhibited significantly better growth performance than control（P 〈0.05）. Five non-specific immune parameters including lysozyme, acid phosphatase, alkaline phosphatase, superoxide dismutase and catalase in coelomic fluid were measured to evaluate the immune response of A. japonicus to the probiotics. Results showed that all parameters were significantly improved when YD11 was supplemented in the dose of 1×10^6 CFU g^-1（P 〈0.05）. The cumulative incidence and mortality after the Vibrio splendidus challenge decreased significantly in sea cucumbers of YD136. Accordingly, 1×10^6 CFU g^-1 of YD13 in diet was recommended for the growth promotion and immune enhancement of A. japonicus.     

Effect of Bacillus baekryungensis YD13 supplemented in diets on growth performance and immune response of sea cucumber (Apostichopus japonicus)

The effect of a potential probiotic on the growth performance and immune response of sea cucumber(Apostichopus japonicus) was investigated. Bacillus baekryungensis YD13 isolated from sea cucumber culturing ponds was added to sea cucumber basal feed as a probiotic in different doses(0, the control; 1×104(YD134), 1×106(YD136) and 1×108(YD138) CFU g-1 of diet), and administered orally to A. japonicus(initial mean wet weight 5.44 g ± 0.17 g). The sea cucumbers were fed in 20 aquaria, 5 each treatment, for 60 d. At the end of growth trial, 20 sea cucumbers from each treatment were challenged with Vibrio splendidus. A. japonicus in YD134 and YD136 exhibited significantly better growth performance than control(P 0.05). Five non-specific immune parameters including lysozyme, acid phosphatase, alkaline phosphatase, superoxide dismutase and catalase in coelomic fluid were measured to evaluate the immune response of A. japonicus to the probiotics. Results showed that all parameters were significantly improved when YD11 was supplemented in the dose of 1×106 CFU g-1(P 0.05). The cumulative incidence and mortality after the Vibrio splendidus challenge decreased significantly in sea cucumbers of YD136. Accordingly, 1×106 CFU g-1 of YD13 in diet was recommended for the growth promotion and immune enhancement of A. japonicus.     

Influence of dietary probiotic Bacillus TC22 and Prebiotic fructooligosaccharide on growth, immune responses and disease resistance against Vibrio splendidus infection in sea cucumber Apostichopus japonicus

The effects of probiotic Bacillus TC22 (isolated from intestine of infected sea cucumber) and prebiotic fructooligosaccharide (FOS) on growth, immunity and disease resistance in sea cucumber Apostichopus japonicus were studied. Six experimental diets were formulated with combinations of three levels of TC22 (0, 107 and 109 CFU g-1 diet) and two levels of FOS (0 and 0.5%) in a 3 × 2 factorial experiment. At the end of the 8-week feeding trial,animals were challenged by injecting Vibrio splendidus. The results revealed that the specific growth rates (SGR) of sea cucumbers were not affected by TC22 and FOS, or the interaction between TC22 and FOS (P > 0.05). However,there were significant interactions between TC22 and FOS for immune response and disease resistance in sea cucumbers (P < 0.05). When sea cucumbers were fed with TC22 at 109 CFU g?1 feed and 0.5% FOS alone or in combination, the phagocytosis, respiratory burst and phenoloxidase activity of sea cucumber coelomocytes were significantly enhanced;the disease resistance against V. splendidus infection was also increased significantly (P < 0.05).However, dietary combination of TC22 at 109 CFU g-1 diet and 0.5% FOS did not yield significantly higher efficiency than either TC22 at 109 CFU g-1 diet or 0.5% FOS used alone (P > 0.05). Therefore, further studies should examine the effects of combinations of other levels of FOS(> 0.5% or < 0.5%) and TC22 on the immunity and disease resistance of sea cucumbers.     

Effects of Hanseniaspora opuntiae C21 on the growth and digestive enzyme activity of juvenile sea cucumber Apostichopus japonicas

The effects of a diet containing Hanseniaspora opuntiae C21 on growth and digestive enzyme activity were estimated in juvenile Apostichopus japonicus. Groups of sea cucumbers were fed diets containing H. opuntiae C21 at 0(control), 10 4, 10 5, and 10 6 CFU(colony-forming units)/g feed. Results showed that after 45 d the specific growth rate(SGR) of sea cucumbers fed a C21-supplemented diet at 10 4 CFU/g feed was significantly higher than that of the control( P 0.05). Intestinal trypsin and lipase activities were significantly enhanced by C21 administration at 10 4 and 10 5 CFU/g feed compared with the control( P 0.05). After feeding for 23–42 d, C21 was demonstrated by denaturing gradient gel electrophoresis to be present in the intestine of sea cucumbers. In addition, after feeding the C21-supplemented diets for 15 d, the sea cucumbers were switched to an unsupplemented diet and C21 was confirmed to be capable of colonizing the intestine for at least 31 d after cessation of feeding. In conclusion, C21 was shown to successfully colonize the intestine of juvenile A. japonicus via dietary supplementation, and improve growth and digestive enzyme activity.     

Growth and antioxidant status of oriental river prawn Macrobrachium nipponense fed with diets containing vitamin E

A feeding trial was carried out to investigate the dietary vitamin E requirement of the oriental river prawn Macrobrachium nipponense(weight of 0.3–0.4 g) and its effect role on antioxidant activity.Prawns were fed with seven levels of vitamin E(0,25,50,75,100,200,and 400 mg/kg diet) for 60 days.The results show that dietary vitamin E supplementation could significantly increased the prawn weight( P 0.05).The activity of superoxide dismutase(SOD) in the hepatopancreas was significantly higher in prawns fed with diets supplemented with ≤75 mg/kg vitamin E than in those fed with diets supplemented with 100–400 mg/kg vitamin E( P 0.05).The activity of catalase(CAT) in the hepatopancreas decreased significantly as dietary vitamin E supplementation increased( P 0.05),and no significant difference was detected in glutathione peroxidase(GSH-Px) activity between different dietary groups( P 0.05).The contents of vitamin E in the hepatopancreas and in the muscle increased with increasing dietary vitamin E.There was a linear correlation between the vitamin E level in diet and that in muscle,and between the vitamin E level in diet and that in the hepatopancreas.All the above results indicated that dietary vitamin E can be stored in the hepatopancreas and muscle and lower both the activities of SOD and CAT in the hepatopancreas,suggesting that it is a potential antioxidant in M.nipponense.Broken line analysis conducted on the weight gains of prawns in each diet group showed that the dietary vitamin E requirement for maximum growth is 94.10 mg/kg.     

Colonization and Probiotic Effect of Metschnikowia sp. C14 in the Intestine of Juvenile Sea Cucumber,Apostichopus japonicus

Viable cell count was used to determine whether Metschnikowia sp.C14 can colonize the intestine of juvenile sea cucumber Apostichopus japonicus.Sea cucumber individuals were divided into two groups,which were fed the control diet for 38 days or the C14-supplemented diet at 105 cells g−1 diet for 28 days,then the control diet from day 29 to day 38.The number of C14 cells in the intestine of sea cucumber fed the C14-supplemented diet significantly increased from day 7 to day 28,and decreased from day 29 to day 38.Sea cucumber fed with the diet containing C14 showed a significant increase in trypsin activity and lipase activity from day 21 to day 33 compared with the control.Feeding C14 significantly improved the phagocytic activity and respiratory burst in coelomocytes from day 21 to day 35 and from day 14 to day 38,respectively.In addition,there was an obvious enhancement in lysozyme activity(from day 21 to day 38 or day 33),phenoloxidase activity(from day 21 to day 28)and total nitric oxide synthase activity(from day 14 to day 38)in coelomic fluid supernatant and/or coelomocyte cell lysate supernatant compared with the control.There were significant positive correlations between the number of C14 cells colonizing the intestine and trypsin activity of the intestine,lysozyme activity of the coelomic fluid supernatant and coelomocyte lysate supernatant from sea cucumber.These data suggested that the number of C14 cells should be maintained at 105 cfu(colony-forming units)g−1 intestine material for the maximum benefit.     

Proline with or without Hydroxyproline Influences Collagen Concentration and Regulates Prolyl 4-Hydroxylase α(I) Gene Expression in Juvenile Turbot(Scophthalmus maximus L.)

This study was conducted to investigate the effect of dietary proline(Pro), and Pro and hydroxyproline(Hyp) in combination on the growth performance, total Hyp and collagen concentrations of tissues, and prolyl 4-hydroxylase α(I)(P4H α(I)) gene expression in juvenile turbot feeding high plant protein diets. A diet containing 50% crude protein and 12% crude lipid was formulated as the basal and control, on which other two protein and lipid contents identical experimental diets were formulated by supplementing the basal with either 0.75% Pro(Pro-0.75) or 0.75% Pro and 0.75% Hyp(Pro+Hyp). Four groups of fish in indoor seawater recirculating systems, 35 individuals each, were fed twice a day to apparent satiation for 10 weeks. The results showed that dietary Pro and Hyp supplementation had no significant effect on growth performance and feed utilization of juvenile turbot(P 0.05). Total Hyp and collagen concentrations in muscle were significantly increased when dietary Pro and Hyp increased(P 0.05), and fish fed diet Pro+Hyp showed significantly higher free Hyp content in plasma than those fed other diets(P 0.05). The expression of P4 H α(I) gene in liver and muscle was significantly up regulated in fish fed diet Pro-0.75 in comparison with control(P 0.05); however the gene was significantly down regulated in fish fed diet Pro+Hyp in muscle in comparison with fish fed diet Pro-0.75(P 0.05). It can be concluded that supplement of crystal L-Pro and L-Hyp to high plant protein diets did not show positive effects on growth performance of juvenile turbot, but enhanced total collagen concentrations in muscle.     

Effect of Different Dietary Protein and Lipid Levels on the Growth,Body Composition,and Intestinal Digestive Enzyme Activities of Juvenile Yellow Drum Nibea albiflora(Richardson)

An 8-week feeding trial was conducted to determine the optimal dietary protein-to-lipid ratio for juvenile Nibea albiflora with an initial weight of(11.76 ± 0.20) g.Nine experimental diets containing different concentrations of protein(40%,47%,or 54%) and lipids(5%,9%,or 13%) in a 3 × 3 factorial experimental design were tested in triplicate groups of fish,while the protein-to-energy(P/E) ratios of the diets varied in the range of 19.74–28.32 mg k J~(-1).Results showed that fish fed diets containing 9% or 13% lipids with 54% protein exhibited significantly higher weight gains and specific growth rates than those fed other diets.The feed conversion rate of fish fed the diet with 40% protein and 5% lipids was significantly poorer than that of fish fed other diets.The protein efficiency rate of fish fed diets with 5% lipids was significantly lower than that of fish fed 9% or 13% lipid diets.Carcass lipid and energy contents were positively correlated with dietary lipid level regardless of protein level.Fish fed a 54% protein diet showed the highest trypsin activity.The intestinal lipase activity of fish fed the diet containing 13% lipids was significantly higher than that of fish fed 5% or 9% lipid diets.These results demonstrate the high protein dietary requirements of N.albiflora.A diet containing 54% protein and 9%–13% lipids with a P/E ratio of 26.2–27.81 mg protein k J~(-1) can be considered optimal for juvenile N.albiflora.     

Effect of Dietary Lipid on the Growth,Fatty Acid Composition and Δ5 Fads Expression of Abalone(Haliotis discus hannai Ino) Hepatopancreas

This study investigated the effect of dietary lipid on the growth, fatty acid composition and Δ5 fatty acyl desaturase genes(Fads) expression of juvenile abalone(Haliotis discus hannai Ino) hepatopancreas. Six purified diets were formulated to contain tripalmitin(TP), olive oil(OO, 72.87% 18:1n-9), grape seed oil(GO, 68.67% 18:2n-6), linseed oil(LO, 70.48% 18:3n-3), ARA oil(AO, 41.81% ARA) or EPA oil(EO, 44.09% EPA and 23.67% DAH). No significant difference in survival rate was observed among abalone fed with different diets. Weight gain rate(WGR) and daily growth rate of shell length(DGRSL) were significantly increased in abalone fed with diets containing OO, AO and EO, but decreased in abalone fed with LO diet(P 0.05) in comparison with those fed with TP. High level of dietary 18:2n-6 resulted in higher content of n-6 polyunsaturated fatty acids(PUFAs) in abalone fed with GO than those fed with TP, OO, LO and EO(P 0.05). n-3 PUFAs in abalone fed with LO was significantly higher than those in abalone fed with TP, OO, GO and AO(P 0.05). The highest contents of 20:1n-9 and 22:1n-9 were observed in abalone fed with OO. The expression of Δ5 Fads in hepatopancreas of abalone was enhanced by high concentration of 18:3n-3 and suppressed by dietary LC-PUFAs; however it was not affected by dietary high concentration of 18:1n-9 or 18:2n-6. These results provided valuable information for understanding the synthesis of LC-PUFAs and nutritional regulation of Δ5 Fads expression in abalone.     

Replacement of Dietary Fish Oil with Vegetable Oils Improves the Growth and Flesh Quality of Large Yellow Croaker （Larmichthys crocea）

We investigated the effect of the replacement of dietary fish oil with vegetable oils on the growth and flesh quality of large yellow croaker(Larmichthys crocea). The basal diet(FO) was formulated to contain 66.5% fish meal and 6.4% menhaden fish oil; whereas the other 3 experimental diets were formulated by replacing the fish oil with 50% soybean oil(SO50), 100% soybean oil(SO100) and 100% palm oil(PO100), respectively. The 4 diets were randomly assigned to 4 floating sea cages(3.0 m × 3.0 m × 3.0 m), and each was stocked with 250 fish individuals with an initial average weight of 245.29 g ± 7.45 g. The fish were fed to apparent satiation twice a day at 5:00 and 17:00, respectively, for 12 weeks. Experimental analysis showed that the specific growth rate of fish fed SO50 or PO100 were significantly higher than that of fish fed FO or SO100(P0.05), and crude lipid contents of ventral muscle and viscera were significantly lower in fish fed FO than in those fed the other 3 diets(P0.05). No significant differences in condition factor, viscerosomatic index, hepatosomatic index, gutted yield and colorimetric values of fish among the dietary treatments were observed(P0.05). Compared to FO diet, SO50, SO100 and PO100 diets led to substantial decreases in the liquid loss and water loss from fresh fillets(1 d, 4℃)(P0.05). Similarly, thiobarbituric acid reactive substance(TBARS) values of fillets under different storage conditions(1 d, 4℃; 7 d, 4℃; 4 weeks,-20℃; 8 weeks,-20℃) decreased significantly after partial or complete replacement of fish oil with vegetable oils. These findings indicated that the growth performance and selected flesh quality properties(liquid holding capacity and TBARS value) of large yellow croaker were substantially improved by replacing dietary fish oil with vegetable oils.     

Verification of protein sparing by feeding carbohydrate to common carp Cyprinus carpio

A 9-week feeding trial in floating freshwater cages(1.0 m×1.0 m×2.0 m) was conducted to study the effects of different dietary levels of protein and starch on growth,body composition,and gene expression of enzymes in common carp,Cyprinus carpio(mean body weight,36.12±1.18 g) to evaluate the protein-sparing effect of dietary carbohydrate.Four diets were formulated with corn starch as the carbohydrate source to obtain corn starch levels of 6.5%,13%,19.5%,or 26%and protein levels of 30.5%,28.2%,26.4%,and 24.2%.The results showed no differences in growth performance of fish fed the diets with different protein and corn starch levels,but body composition and glucose metabolic enzyme activity of carp were significantly affected by the different diets(P0.05).Weight gain,specific growth rate,and the feed conversion ratio were not different in fish fed the different dietary treatments.Protein efficiency ratio increased significantly as corn starch level increased(P0.05).Whole-body crude lipid composition increased with increasing dietary corn starch level(P0.05).Glucokinase(GK),hexokinase,and pyruvate kinase(PK) activities increased significantly with increasing dietary corn starch level(P0.05),whereas glucose-6-phosphate(G6Pase) activity decreased with increasing dietary corn starch level(P0.05).GK gene expression was significantly higher in fish fed the high-corn starch diet than those fed the low-corn starch diet(P0.05).G6 pase gene expression tended to decrease with increasing starch level(P0.05).In summary,the results indicate a protein-sparing effect by substituting carbohydrate in the diet of common carp.     

Effects of Dietary Protein and Lipid Levels on Growth Performance,Muscle Composition,Immunity Index and Biochemical Index of the Greenfin Horse-Faced Filefish(Thamnaconus septentrionalis) Juvenile

The effects of different dietary protein and lipid levels on juvenile greenfin horse-faced filefish(Thamnaconus septentrionalis) was experimentally evaluated in a 56-day study. Fish(initial weight 6.47 ± 0.56 g) were fed with nine different diet combinations containing three varying levels of protein(37%, 43%, 49%) and lipids(4%, 7%, 10%) in triplicates, where the protein-to-energy(P/E) ratios of the diets varied in the range of 19.17–26.99 mg kJ-1. Results showed that fish fed diets containing 49%protein and 10% lipid exhibited significantly lower feed intake than those fed other diets. The weight gain of fish fed diets with 43%protein was significantly higher than that of fish fed 37% or 49% protein diets. While no significant change in weight gain was observed when lipid content was increased from 4% to 7%, weight gain decreased significantly when lipid increased from 7% to 10%.The protein efficiency ratio and feed efficiency were highest when the fish were fed the diet with 49% protein and 7% lipids. Muscle crude lipid content was positively correlated with dietary protein level regardless of lipid level. The diet of 43% protein or 7% lipids significantly reduced the activity of alanine aminotransferase and aspartate aminotransferase in liver. The contents of aspartate aminotransferase, superoxide dismutase and malondialdehyde were significantly increased from 4% to 10% lipids in plasma. The plasma total cholesterol and triglyceride contents were positively correlated with dietary lipid level at the same protein level. Our comprehensive analysis concluded that a diet with 43% protein and 7% lipid, with a P/E ratio of 23.07 mg kJ-1, significantly promote the growth performance, feed utilization and health of juvenile greenfin horse-faced filefish.     

Effects of irradiated Ergosan on the growth performance and mucus biological components of rainbow trout Oncorhynchus mykiss

Effects of irradiated and non-irradiated Ergosan extract(alginic acid) on rainbow trout growth performance and skin mucosal immunity were compared. Ergosan was irradiated at 30 kGy in a cobalt-60 irradiator. A total of 252 fish(128.03±9.4 g) were randomly divided into four equal groups,given the basal diet either unsupplemented with Ergosan(control group) or supplemented with crude Ergosan(5 g/kg),ethanol-extracted Ergosan(0.33 g/kg) or irradiated Ergosan(0.33 g/kg) according to this protocol: basal diet for 15 days,treatment diet for 15 days,basal diet for 10 days and treatment diet for 15 days. Highest growth performance was observed in fish fed irradiated Ergosan(P0.05). Dietary administration of different Ergosan types did not cause any changes in mucus protein level,but improved alkaline phosphatase level and hemagglutination titer compared with the control(basal diet without Ergosan) on day 55 offeeding trial(P0.05). Furthermore,the highest value of lysozyme activity was observed in gamma-irradiated Ergosan on day 55. In conclusion,gamma-irradiated Ergosan at 0.33 g/kg was found to improve growth performance and mucus biological components significantly in comparison with the control group(basal diet without Ergosan).     

Effects of dietary corn gluten meal on growth performance and protein metabolism in relation to IGF-I and TOR gene expression of juvenile cobia (Rachycentron canadum)

A growth experiment was conducted on cobia(Rachycentron canadum,initial weight 108.2 g ± 3.0 g) to investigate the effects of dietary corn gluten meal(CGM) levels on the fish growth,whole body composition and protein metabolism in relation to specific gene expression.Five isonitrogenous(crude protein 45%) and isoenergetic(gross energy 20 kJ g 1) practical diets were formulated by replacing 0%(the control),17.5%,35.0%,52.5%,and 70.0% of fish meal(FM) protein with CGM protein.No significant differences were observed in the survival,feed intake(FI),specific growth rate(SGR),feed efficiency(FE) and protein productive value(PPV) among fish fed diets with 0%,17.5%,35.0%,and 52.5% of CGM protein.However,these indices were significantly lower in fish fed the diet with 70.0% of CGM protein than those in fish fed the control diet(P < 0.05).The whole-body crude protein and lipid contents were significantly lower while the whole-body moisture content was significantly higher in fish fed the diet with 70.0% of CGM protein compared with the control group(P < 0.05).When 70.0% of FM protein was replaced by CGM,plasma total protein and cholesterol contents were significantly lower than those in the control group(P < 0.05).Fish fed the diet with 70.0% of CGM protein had significantly lower hepatic insulin-like growth factor I(IGF-I) expression levels than those in the control group(P < 0.05).However,no significant differences were observed in hepatic target of rapamycin(TOR),dorsal muscle IGF-I and TOR expression levels among dietary treatments.Results of the present study indicated that 52.5% of FM protein could be replaced by CGM in the diets without significant influences on the growth,feed utilization and protein metabolism of juvenile cobia.The present results might be useful for developing cost effective and sustainable cobia dietary formulations.     

Glutamate dehydrogenase and Na+-K+ ATPase expression and growth response of Litopenaeus vannamei to different salinities and dietary protein levels

Improvement in the osmoregulation capacity via nutritional supplies is vitally important in shrimp aquaculture.The effects of dietary protein levels on the osmoregulation capacity of the Pacific white shrimp(L.vannamei) were investigated.This involved an examination of growth performance,glutamate dehydrogenase(GDH) and Na+-K+ ATPase mRNA expression,,and GDH activity in muscles and gills.Three experimental diets were formulated,containing 25%,40%,and 50% dietary protein,and fed to the shrimp at a salinity of 25.After 20 days,no significant difference was observed in weight gain,though GDH and Na+-K+ ATPase gene expression and GDH activity increased with higher dietary protein levels.Subsequently,shrimp fed diets with 25% and 50% dietary protein were transferred into tanks with salinities of 38 and 5,respectively,and sampled at weeks 1 and 2.Shrimp fed with 40% protein at 25 in salinity(optimal conditions) were used as a control.Regardless of the salinities,shrimp fed with 50% dietary protein had significantly higher growth performance than other diets;no significant differences were found in comparison with the control.Shrimp fed with 25% dietary protein and maintained at salinities of 38 and 5 had significantly lower weight gain values after 2 weeks.Ambient salinity change also stimulated the hepatosomatic index,which increased in the first week and then recovered to a relatively normal level,as in the control,after 2 weeks.These findings indicate that in white shrimp,the specific protein nutrient and energy demands related to ambient salinity change are associated with protein metabolism.Increased dietary protein level could improve the osmoregulation capacity of L.vannamei with more energy resources allocated to GDH activity and expression.     

Effect of Dietary Potassium on Growth,Nitrogen Metabolism,Osmoregulation and Immunity of Pacific White Shrimp （Litopenaeus vannamel） Reared in Low Salinity Seawater

An 8 weeks feeding experiment was conducted to determine the effect of dietary potassium on the growth and physio-logical acclimation of Pacific white shrimp （Litopenaeus vannamei） reared in diluted seawater （salinity 4）. Six semi-purified practical diets containing 0.59, 0.96, 1.26, 1.48, 1.74, and 2.17 g potassium K＋ per 100 g diet were formulated, respectively. The survival and feed conversion rate did not show significant difference among groups of shrimps given these diets （P〉0.05）. The shrimps fed the diets containing 0.96-1.48 g K＋ per 100g diet gained the highest weight, specific growth rate, and protein efficiency ratio. Their ammonium-N excretion rate as well as hemolymph concentration of Na＋ and C1- were significantly lower than those of the control （P〈0.05）, but a reverse trend was observed for their gill Na＋/K＋-ATPase. Moreover, the shrimps fed with 1.48 g K＋ per 100 g diet were the highest in hemolymph urea level, and the phenoloxidase and lysozyme activities were significantly higher than those of the control （P〈0.05）. The growth and physiological response of the test shrimps suggested that diet containing 1.48 g K＋ per 100 g diet improved the growth of L. vannamei in low-salinity seawater, and enhanced the physiological acclimation of the organism.     

Effect of Dietary Olaquindox on the Growth of Large Yellow Croaker（Pseudosciaena crocea R.） and the Distribution of Its Residues in Fish Tissues

Olaquindox（OLA）, one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial enteritis. In this study, we evaluated the effect of dietary OLA on the growth of large yellow croaker（Pseudosciaena crocea R.） and the histological distribution of OLA and its metabolite 3-methyl-quinoxaline-2-carboxylic acid（MQCA） in fish tissues. Four diets containing 0（control）, 42.5, 89.5 and 277.2 mg kg^-1 OLA, respectively, were formulated and tested, 3 cages（1.0 m × 1.0 m × 1.5 m） each diet and 100 juveniles（9.75 ± 0.35 g） each cage. The fish were fed to satiation twice a day at 05：00 am and 17：00 pm for 8 weeks. The survival rate of fish fed the diet containing 42.5 and 89.5 mg kg^-1 OLA was significantly higher than that of fish fed the diet containing 0 and 277.2 mg kg^-1 OLA（P 0.05）, while the weight gain rate of fish fed the diet containing 42.5 and 89.5 mg kg^-1 OLA was significantly higher than that of fish fed the diet without OLA（control）（P0.05）, but similar to that of fish fed the diet with 277.2 mg kg^-1 OLA. Fish fed the diet with 277.2 mg kg^-1 OLA had the highest content of OLA and MQCA in liver（3.44 and 0.39 mg kg^-1, respectively）, skin（0.46 and 0.09 mg kg^-1, respectively） and muscle（0.24 and 0.06 mg kg^-1, respectively）. In average, fish fed the diet containing OLA had the highest content of OLA and MQCA in liver which was followed by skin and muscle（P〈 0.05）, whereas OLA and MQCA were not detectable in control. Our findings demonstrated that OLA and MQCA accumulated in large yellow croaker when it was fed with the diet containing OLA, thus imposing a potential safety risk to human health.     

Effects of supplemental coated or crystalline methionine in low-fishmeal diet on the growth performance and body composition of juvenile cobia Rachycentron canadum （Linnaeus）

We evaluated the effects of supplemental coated and crystalline methionine （Met） on the growth performance and feed utilization of juvenile cobia （Rachycentron canadum Linnaeus） in a 60-d feeding trial. Fish groups were fed one of six isonitrogenous and isolipidic diets： 1） fishmeal control; 2） un- supplemented experimental （low-fish-meal diet deficient in Met）; or 3） one of four Met diets supplemented with crystalline L-Met, cellulose-acetate-phthalate coated L-Met, acrylic-resin coated L-Met, or tripalmitin- polyvinyl alcohol coated L-Met. The test diets were fed to triplicate groups of cobia （initial body weight 5.40-4-0.07 g） twice a day. The weight gain and specific growth rate of the fish fed the RES diet were highest among the Met-supplemented groups and were 23.64% and 7.99%, respectively, higher than those of the fish fed with the un-supplemented experimental diet （P〈0.05）. The protein efficiency ratio of the fish fed the MET diet was significantly higher than that of the fish fed the un-supplemented experimental diet and the fish in the other methionine supplementation groups （P〈0.05）. Our results suggest that supplementation of crystalline Met in low-fish-meal diets promotes the growth performance of juvenile cobia.     

Effects of Dietary Carbohydrate-to-Lipid Ratio on the Growth Performance and Feed Utilization of Juvenile Turbot(Scophthalmus maximus)

A 9-week feeding trial was conducted to investigate the effects of dietary carbohydrate to lipid ratio(CHO:LIP) on the growth performance and feed utilization of juvenile turbot Scophthalmus maximus(initial body weight 8.75 g ± 0.04 g). Four isonitrogenous and isoenergetic low protein level(39%) diets were formulated with increasing ratios of dietary carbohydrate to lipid(2:18, 6:18, 18:12 and 28:6). A high protein level(50%) diet with the 2:12 ratio of carbohydrate to lipid was used as the control. Results showed that the survival rate, contents of moisture, crude protein and ash in muscle were not significantly affected by dietary treatments. With the dietary CHO:LIP ratio increased from 2:18 to 18:12, weight gain rate significantly increased(P 0.05). Higher dietary CHO:LIP ratio(28:6) resulted in the significantly decreased weight gain rate(P 0.05). Meanwhile, this treatment also resulted in the highest daily feed intake and liver glycogen content, as well as the lowest feed efficiency(P 0.05). Muscle glycogen content in fish fed the diet with 2:12 or 2:18 CHO:LIP ratio was significantly lower than those fed with the other three diets(P 0.05). The present results confirmed that the juvenile turbot can utilize carbohydrate. Furthermore, the appropriate ratio of dietary carbohydrate to lipid was important to the growth and feed utilization of turbot. The proper CHO:LIP ratio based on the growth performance in the present study was determined to be 18:12 when the dietary protein level was 39%.