DISEASE RESISTANCE AND HUMORAL IMMUNOMODULATORY EFFECTS OF VITAMIN C ON GROUPER, EPINEPHELUS AWOARA

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Growth and antioxidant status of oriental river prawn <Emphasis Type="Italic">Macrobrachium nipponense</Emphasis> 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.     

Effects of Dietary Protein Level on Growth and Utilization of Protein and Energy by Juvenile Mangrove Red Snapper （ Lutjanus argentimaculatus ）

A feeding trial was conducted in a recirculating water system to investigate the effects of dietary protein levels on growth, feed utilization, hepatosomatic index and liver lipid deposition of juvenile red snapper, Lutjanus argentimaculatus (average initial wet weight 8.0 ± 0.39 g and total length 3.14 ± 0.3 cm). In the experiment, six fishmeal-based diets were formulated to contain various protein levels (20% to 45% in 5% increments), with dietary energy ranging from 2210.7kJ lOOg to 2250.2kJlOOg dry matter. The protein to energy ratios of diets ranged from 8.58 mg protein kJ⁻¹ to 20.03 mg protein kJ⁻¹. Diets were fed for 90d to triplicate groups of fish stocked in 0.128m³ seawater tanks, 25 individuals each. The daily ration of 2% wet body weight was offered to the fish thrice a day. The fish at the end of the study had more than ten-fold (77.0g) increase in weight compared to the initial (8.0g). Fish fed diets of 40% and 45% protein produced significantly (P<0.05) higher weight gain of 77.2g and 76.5g, and specific growth rate (SGR) of 2.65% and 2.62% than those of 67.0 g and 68.3g, and 2.49% and 2.51% of the other diets. The broken-line regression of SGR against dietary protein level yielded an optimum dietary protein requirement of 42.6% (Y=−1.6295 + 0.1114 X ²,P<0.05). Survival remained 100% among groups. Feed conversion ratio decreased from 0.45 for fish fed 20% dietary protein to 0.35 for fish fed 45% dietary protein. Nitrogen intake increased with an increase in dietary protein, which in turn resulted in an increase in nitrogen gain of fish whole body. Fish fed 40% and 45% protein diets showed higher (P<0.05) nitrogen gain (0.27g and 0.26g) than those (0.23g and 025g) fed all other diets. Gross energy intake (GEI) in fish fed 45% protein was lower (600.67kJ) than that (607.97 kJ) of 40% protein diet, though the differences were not statistically significant (P>0.05); GEI ranging from 677.31 kJ to 663.20 kJ at remaining four diets (20% to 35% protein) did not appear to differ significantly (P>0.05). The highest energy gain of 518.33 kJ was obtained with fish fed 40% protein, resulting in the highest energy retention efficiency of 85.26%. The hepatosomatic index of fish fed diets of 20%, 25%, 30% and 35% protein were significantly (P<0.05) higher (2.09% to 2.57%) than those (1.44% and 1.41%) of fish fed diets containing 40% and 45% protein. Liver lipid contents decreased from 8.72% to 7.0% in fish fed dietary protein of 20% to 45% in 5% increments. Results suggest that the diet containing 40% to 42.6% protein with a P/E ratio of 17.6 mg protein kJ⁻¹ is required for good growth of L. argentimaculatus weighing between 8.0 g and 85.2 g under the culture conditions of the present study.     

Effect of dietary phosphorus on the growth and body components of juvenile Synechogobius hasta

The effect of dietary phosphorus on the growth and body components of juvenile Synechogobius hasta was determined. Different percentages of dietary phosphorus (0.63, 0.77, 0.93, 1.06, 1.22 and 1.36) were tested by feeding the fish (body weight, 15.81 g ± 0.32 g; 20 individuals each group; 3 groups each percentage) at a surplus of 5%–10% above satiation for 35 d. Dietary phosphorus did not significantly affect the specific growth rate, feed intake, feed conversion ratio and protein efficiency rate. Nitrogen retention was found to be the highest in fish fed the diet containing 1.06% of phosphorus; however, this was not significantly different from other diets. Fish fed the diet containing 0.93% of phosphorus showed the highest phosphorus retention; similar phosphorus retention rates were found in fish fed the diets containing 0.77% and 1.06% of phosphorus. Fish fed the diet containing the highest percentage of dietary phosphorus were found to contain the least whole body lipid, lower than fish fed other diets (P<0.05). The protein content increased from 18.59% to 19.55% (although not significant) with the decrease of body lipid content (P>0.05). The contents of the whole body ash, whole body phosphorus and vertebrae phosphorus increased with dietary phosphorus percentage up to 1.06 (P<0.05), reaching a plateau after that. Dietary phosphorus did not significantly influence the muscle components (protein, lipid and moisture). Condition factor and hepatosomatic index were the highest in fish fed the diet containing 0.63% of dietary phosphorus; however, this was not significantly different from those of other diets. The second-order polynomial regression of phosphorus retention against dietary phosphorus identified a breakpoint at 0.88% of dietary phosphorus. However, the dietary requirement of phosphorus for maintaining maximum phosphorus storage determined by broken-line analysis of the contents of whole body phosphorus, and ash and vertebrae phosphorus was 1.06% of the diet.     

Effects of dietary vitamin C on the immune function of shrimps, Penaeus chinensis

Prepared in this experiment were six groups of diets, i.e. VC0, VC1, VC2, VC3, VC4 and VC5 with the contents of vitamin C (VCmg(100 g)⁻¹ diet) of 0, 100, 200, 400, 800, and 1200 respectively. It was found that vitamin C increased the concentrations of immunoglobulin-like (IgG-like, IgA-like and IgM-like) substances in the serum of Penaeus chinensis after a feeding period of 3 weeks. The differences among groups were significant (P<0.01), but there was no difference in the contents of complement3-like and complement4-like substances in the serum (P>0.05). Phenoloxidase (PO) activity in the serum of VC3 group shrimps was higher than that of VC0 and other groups, but no significant difference was observed between VC0 group and other groups. Furthermore, bactericidal activity of the serum to Vibrio parahaemolyticus in shrimps fed with the VC1 diet was higher than that in the other groups (P<0.01), while no difference was demonstrated among all groups for the bactericidal activity to Vibrio alginolyticus (P>0.05). It is, therefore, suggested that vitamin C (100–400 mg(100 g)⁻¹ diets) could be used as an immunostimulant of P. chinensis.     

Effects of diet and body size on phosphorus utilization of Liza haematocheila T. ＆ S.

A 21-d laboratory experiment was conducted to study, the phosphorus （P） utilization of two different diets by redlip mullet Liza haematocheila T. ＆ S. Sand-filtered water in salinity 30 and temperatare 25℃ was used. Twenty-nine fish individuals were divided into three groups： 11 to group 1 （G1） fed on diet 1, 11 to group 2 （G2） fed on diet 2, and 7 to contrast group. Diet 1 was a commercial feed, more valuable in nutrition than diet 2 that similar to natural detritus. The results show the intake phosphorus （IP） of G1 was significantly higher than that of G2, and both increased linearly with body size at a certain amount of diet. The retention phosphorus （RP） in fish of G1 was lower than G2. The relationship between retention phosphorus and body size was positive and stronger in G2. Significant difference in faecal phosphorus （FP） was found between G1 and G2. Body size significantly impacted the excretion phosphorus （EP） in G1 but G2. The loss of intake phosphorus in G1 was 10.83-20.27 mg per g fish weight gain, higher than that in G2 for 6.63-9.56. Of the phosphorus, about 10% was allocated into growth, 50% in faeces, and the rest lost in excretion. The main part of phosphorus was lost in faeces but excretion. The phosphorus budget of the fish could be described as 100IP = 7.40RP ＋ 47.39FP ＋ 36.63EP （Diet 1） or 100IP = 11.93RP ＋ 56.64FP ＋ 21.76EP （Diet 2）.     

Dietary vitamin A,ascorbic acid and α-tocopherol affect the gonad development and reproductive performance of starry flounder Platichthys stellatus broodstock

The present trial was conducted with starry fl ounder Platichthys stellatus broodstock to evaluate the effects of dietary vitamin A,ascorbic acid,and α-tocopherol on the gonadal development and reproductive performance.8 000 IU/kg diet vitamin A(VA group),500 mg/kg diet ascorbic acid(Vcpp group),or 250 mg/kg diet α-tocopherol(α-TA group)was added into basal diet to create 3 vitamin experimental diets,respectively.Each diet was fed to 450 starry fl ounder broodstock for 104 days.Samples were collected weekly.The gonadosomatic index(GSI)of 3 vitamin experimental groups fi rst decreased and then increased.Maximum GSI of Vcpp group was higher than that of α-TA group but lower than that of VA group.The spawning periods of 3 vitamin experimental groups lasted 49,56,and 45 days,respectively.No mature eggs were observed in the control group during the trial.The absolute fecundity(AF)and relative fecundity(RF)of α-TA group was higher than that of Vcpp group but lower than that of VA group.The results suggest that different vitamins play different roles in the fi sh reproductive process.Vitamin A stimulated the maturation of the ovary,ascorbic acid prolonged the spawning period,and α-tocopherol affected the development of the eggs.     

Assessment of nitrogen maintenance requirement in juvenile Black Porgy <Emphasis Type="Italic">Acanthopagrus schlegeli</Emphasis>

Nitrogen balance method and nitrogen-free diet were used in this study to determine nitrogen maintenance requirement (NM) and nitrogen maintenance requirement per unit metabolism body weight (NM′) of black porgy Acanthopagrus schlegeli. Fish with body weight (BW) of 50, 80, 120, 160 and 200 g were fed by the diets containing three graded levels of crude protein (380, 420 and 460 g/kg). The results from nitrogen balance experiment showed that the amount of nitrogen deposition varied from 0.15 to 0.31 mg/g BW per day, accounting for 12.2% to 21.1% of nitrogen intake. The amount of fecal nitrogen excretion varied from 0.21 to 0.32 mg/g BW per day, accounting for 16.3% to 21.6% of nitrogen intake. The endogenous nitrogen excretion, a main part of nitrogen consumption varied from 0.79 to 0.97 mg/g BW per day, accounting for 63.3 % to 68.0% of nitrogen intake. Positive correlation was found between NM and body weight, while a negative correlation was found between NM of unit body weight and the growth duration. No significant differences (p>0.05) were found in NM′ among different growth stages. The average of NM′ was 0.485 7mg/g per day. The results from nitrogen-free diet experiment showed that a negative correlation between NM and feed intake of nitrogen-free diet. NM increased with the decrease of feed intake of fish. The average of NM was 0.482 9 mg/g BW per day that was close to 0.483 8 mg/g BW obtained from fish with 120 g BW in nitrogen balance experiment. The nitrogen balance method is recommended to be a better method for determining NM in consideration of fish stress and result stability. This study also provides a calculated result of the protein content in diets, which is necessary for maintaining fish body protein at different growth stages. The calculation was based on the amount of nitrogen required for maintaining body protein per kg BW. Supported by Scientific Research Project Grant (No.2004C100059) from the city government of Ningbo, China.     

饲料中添加L-肉碱对牙鲆幼鱼生长、生化组成及血液指标的影响

研究0、300、600、900、1 200、1 500 mg/kg 6个L-肉碱添加水平对牙鲆(Paralichthys olivaceus)幼鱼生长、生化组成和血液指标的影响。结果表明:添加量为1 200 mg/kg的处理组牙鲆幼鱼的增重率和特定生长率显著提高,饲料系数显著降低(P<0.05);随着饲料中L-肉碱含量的增加,肝指数呈现下降趋势,且在添加量为900~1 500 mg/kg时,各组较对照组差异显著(P<0.05);饲料中添加L-肉碱显著降低各实验组脏体比(P<0.05),而对各处理组间存活率和肥满度均无影响(P<0.05);肌肉和肝脏中的脂肪含量与L-肉碱添加量呈负相关,当添加量为1 200~1 500 mg/kg时,较对照组差异显著(P<0.05);肌肉中的蛋白含量与L-肉碱添加量呈正相关,但各组差异不显著(P<0.05);饲料中添加L-肉碱降低血清中的谷草转氨酶、胆固醇、甘油三酯和尿素氮含量,且添加量1 200 mg/kg组各指标含量均显著低于对照组(P<0.05);各处理组的血糖含量较对照组均有所增加,其中1 200 mg/kg组血糖含量较对照组提高41.6%(P<0.05);血清中总蛋白和低密度脂蛋白均随着L-肉碱添加量的增加而呈下降趋势,高密度脂蛋白却得以提升,但差异均不显著(P<0.05)。分析认为,在本实验条件下,饲料中添加L-肉碱可促进牙鲆幼鱼生长,且降脂效果显著。建议L-肉碱最适添加量为1 200 mg/kg。     

饲料中添加虾青素对血鹦鹉皮肤类胡萝卜素含量和体色三刺激值的影响

为探讨投喂增色剂对鱼类增色效果的定量评价方法,分别用添加0、30、100、300、500、700、900 mg/kg虾青素的饲料饲喂血鹦鹉60 d,测定血鹦鹉皮肤中的总类胡萝卜素含量和鱼体体色的三刺激值。结果显示,血鹦鹉皮肤总类胡萝卜素含量随着虾青素添加量的增加而升高,而三刺激值X、Y、Z则逐渐减小。总类胡萝卜素含量300～900 mg/kg各组显著高于对照组(P<0.05),500、700和900 mg/kg组显著高于0、30、100 mg/kg组(P<0.05),而500、700和900 mg/kg各组间差异不显著(P>0.05)。总类胡萝卜素含量与三刺激值间的相关系数分别为-0.966、-0.932、-0.981,均达到极显著水平(P<0.01)。可见,在饲料中添加虾青素可明显提高血鹦鹉皮肤类胡萝卜素含量,改善血鹦鹉体色,血鹦鹉体色的三刺激值与皮肤总类胡萝卜素含量有极高的相关性,可用测色色差计测定血鹦鹉的三刺激值,并以此来推算鱼体皮肤总类胡萝卜素含量,定量评价血鹦鹉的体色。     

Effect of dietary supplementation of vitamin C on growth,reactive oxygen species,and antioxidant enzyme activity of Apostichopus japonicus (Selenka) juveniles exposed to nitrite

Different amounts of vitamin C were added to diets fed to juveniles(2.5 ± 0.15 g) of sea cucumber Apostichopus japonic u s(Selenka) in an attempt to reduce the stress response of specimens exposed to nitrite stress. A commercial feed was used as the control diet and three experimental diets were made by supplementing 1 000, 1 500, or 2 000 mg vitamin C/kg diet to control diet separately in a 45-day experiment. Sea cucumbers were exposed to three different levels(0.5, 1.0, and 1.5 mg/L) of nitrite stress for 4, 8, and 12 h at four time intervals(0, 15, 30, and 45 d). Growth of the animals was recorded during the experiment. Reactive oxygen species(ROS)(i.e. hydroxyl free radical(-OH), malondialdehyde(MDA) and total antioxidant capacity(T-AOC)) and antioxidant enzyme activities(i.e., superoxide dismutase(SOD) and catalase(CAT)) were measured. Response surface methodology(RSM) was used to analyze the effect of multiple factors on ROS indices and enzyme activities. Weight gain(WG) and special growth rate(SGR) of vitamin C supplementation groups were significantly higher than those of control group( P 0.05). The levels of-OH and MDA increased under exposure time extending and nitrite concentration increasing, whereas T-AOC level decreased. SOD and CAT activities increased at 4 h and 8 h and decreased at 12 h. During the days in which the animal consumed experimental diets, the levels of-OH and MDA decreased and that of T-AOC increased. This result suggests that diets containing vitamin C could reduce the nitrite stress response in the animals and increase their antioxidant capacity. The multifactor regression equation of growth performance, ROS indices, and duration of feeding results suggest that vitamin C supplementation of 1 400–2 000 mg/kg diet for 29–35 days could reduce effectively the effects of nitrite exposure.     

Dietary leucine requirement for juvenile large yellow croaker <Emphasis Type="Italic">Pseudosciaena crocea</Emphasis> (Richardson, 1846)

Dietary leucine requirement for juvenile large yellow croaker, Pseudosciaena crocea Richardson 1846 (initial body weight 6.0 g ± 0.1 g) was determined using dose-response method. Six isonitogenous (crude protein 43%) and isoenergetic (19 kJ g⁻¹) practical diets containing six levels of leucine (Diets 1–6) ranging from 1.23% to 4.80% (dry matter) were made at about 0.7% increment of leucine. Equal amino acid nitrogen was maintained by replacing leucine with glutamic acid. Triplicate groups of 60 individuals were fed to apparent satiation by hand twice daily (05:00 and 17:30). The water temperature was 26–32°C, salinity 26–30 and dissolved oxygen approximately 7 mg L⁻¹ during the experimental period. Final weight (FW) of large yellow croaker initially increased with increasing level of dietary leucine but then decreased at further higher level of leucine. The highest FW was obtained in fish fed diet with 3.30% Leucine (Diet 4). FW of fish fed the diet with 4.80% Leucine (Diet 6) was significantly lower than those fed Diet 4. However, no significant differences were observed between the other dietary treatments. Feed efficiency (FE) and whole body composition were independent of dietary leucine contents (P > 0.05). The results indicated that leucine was essential for growth of juvenile large yellow croaker. On the basis of FW, the optimum dietary leucine requirement for juvenile large yellow croaker was estimated to be 2.92% of dry matter (6.79% of dietary protein).     

Vaccination in three different ways against vibriosis of <Emphasis Type="Italic">Seriola dumerili</Emphasis> caused by <Emphasis Type="Italic">Vibrio hollisae</Emphasis>

Bacterin was prepared by formalin-inactivating the virulent strain of Vibrio hollisae isolated from diseased Seriola dumerili （amberjack） suffering from vibriosis. Healthy S. dumeriIi were vaccinated by respective procedures of intramuscular injection, immersion, and orally administration. Results of the three different vaccinations were compared. Blood was drawn from the vaccinated fish every 7 days, and the antibody titers and lysozyme activities of the sera were determined. The antibody titer of injected fish was 1：40 at 7 d, and reached its peak of 1：320 at 28 d, while the fish vaccinated by immersion and orally administration exhibited weak antibody responses, the antibody titres of 〈1：10, 1：20, 1：160 were observed at 7 d, 14 d, 35 d respectively. Compared with the control, the vaccinated fish exhibited significantly higher lysozyme activities （P〈0.05）. Upon challenge with virulent strain, the relative percent survival （RPS） of injected, immersed and oral administrated fish were 75%, 45%, and 40% respectively, and the injected fish showed significantly higher RPS than immersed and oral administrated fish. The results suggested that vaccination of S. dumerili by the injection would be the best strategy to prevent the vibriosis in S. dumerili farm.     

Immunosuppressive activities of fucoidan from Laminaria japonica

Effects of fucoidan fromLaminaria japonica on 2,4-dinitrochlorobenzene induced delayed-type hypersensitivity (DTH) reaction and the serum levels of IgG, IgM, complement C3 and C4 were investigated in the present study. Results showed that oral administration of fucoidan at dose of 150 and 300 mg/(kg· d) for 9 days before the hapten challenge significantly inhibited 2,4-dinitrochlorobenzene induced delayed-type hypersensitivity reaction; and also inhibited the humoral immunity. Serum C3 and C4 levels were markedly reduced by fucoidan at dose of 150 and 300 mg/kg; and serum IgG and IgM levels were reduced by fucoidan at dose of 300mg/kg. The inhibitory effects of fucoidan on delayed-type hypersensitivity suggested that it may be potential medication for chronic inflammatory diseases in the future. This study was supported by National Foundation for New Drug Research & Development (NFND) No.144.     

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 dietary carbohydrates sources on lipids compositions in abalone, Haliotis discus hannai Ino

A study was conducted to evaluate the effects of dietary carbohydrates on triglyceride, cholesterol and fatty acid concentrations in abalone, Haliotis discus hannai Ino. Six semi-purified diets with different carbohydrates (dextrin, heat-treated wheat starch, wheat starch, corn starch, tapioca starch and potato starch, respectively), all containing a carbohydrate level of 33.5%, were fed to abalone (initial shell length: 29.98 mm ± 0.09 mm; initial weight: 3.42 g ± 0.02 g) for 24 weeks in a recirculation system. The results indicate that serum triglyceride concentrations were significantly (P < 0.05) higher in the abalone fed with dextrin, heat-treated wheat starch and wheat starch than those fed with corn starch, and serum cholesterol concentrations were significantly (P < 0.05) higher in the abalone fed with dextrin, heat-treated wheat starch than those fed with corn starch. Fatty acid C20:4n-6 in the foot muscles were significantly (P < 0.05) lower in the abalone fed with dextrin than those fed with wheat starch, corn starch, tapioca starch and potato starch. Fatty acid C20:4n-6 in hepatopancreas was significantly (P < 0.05) lower in abalone fed with heat-treated wheat starch than those fed with corn starch, tapioca starch and potato starch. Fatty acid C22:6n-3 in the foot muscles were significantly (P < 0.05) lower in the abalone fed with dextrin and heat-treated wheat starch than those fed with wheat starch and potato starch.     

A study on the meat and bone meal and poultry by-product meal as protein substitutes of fish meal in practical diets for Litopenaeus vannamei juveniles

A study was conducted to evaluate the effects of meat and bone meal (MBM) and poultry by-product meal (PBM) as the replacement of fish meal in the diets on the growth performance, survival and apparent digestibility coefficients (ADC) of Litopenaeus vannamei. The basal diets were formulated with 22% fish meal and other ingredients which provided about 40% protein and 9% lipid in the diet. The experimental diets included MBM or PBM to replace 0, 20%, 40%, 60% and 80% of total fish meal respectively. All diets were iso-nitrogenous and isocaloric in gross terms. The results showed that there were no significant differences (Pτ;0.05) in growth performance and ADC among the treatments fed with the diets in which 0–60% fish meal had been replaced with MBM, while the percent weight gain (WG, %), body length gain (BLG, %) and ADC significantly decreased when the MBM was up to 80% of the fish meal. There were no significant differences (Pτ;0.05) in growth performance and ADC among all the treatments fed with the diets in which 0–80% fish meal had been replaced with PBM.     

Effects of irradiated Ergosan on the growth performance and mucus biological components of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

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 (P <0.05). 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 of feeding trial (P <0.05). 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 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.     

Steroid sex hormone dynamics during estradiol-17β induced gonadal differentiation in Paralichthys olivaceus (Teleostei)

Steroid sex hormones, such as estradiol-17β (E2) and testosterone (T), are important regulators of sex change in fish. In this study, we examined the effects of E2 treatment on the dynamics of E2 and T during gonadal differentiation in the olive flounder Paralichthys olivaceus using histology and radioimmunoassay (RIA). Flounder larvae were divided into five groups (G0-G4), and fed with 0 (control), 0.2, 2, 20 and 100 mg E2/kg feed from 35 to 110 day post hatching (dph). Fish growth in the G1 and G2 groups was not significantly different from that of the control group (P>0.05), while fish in the G3 and G4 groups were less active and showed growth depression and high mortality. The gonads of fish in the G3 and G4 groups were smaller and surrounded by hyperplastic connective tissue. The frequency of females in the G0-G4 groups was 54.5%, 75.0%, 100%, 100% and 93.3%, respectively. The RIA analyses of E2 and T showed that T levels decreased during gonadal differentiation, and increased slightly at the onset of ovarian differentiation, while E2 levels increased gradually and peaked at the onset of ovarian differentiation in the control group. In the E2-treated groups, T levels decreased before the onset of ovarian differentiation. E2 levels were high on the 48 dph, but declined to a lower level on the 54 dph, and then increased gradually during gonadal differentiation. And a sharp increase of E2 levels were observed in all E2-treated groups at the onset of ovarian differentiation. The data suggest that T and E2 play important roles during gonadal differentiation, and an E2 dose of 2 mg/kg feed could induce sex reversal in P. olivaceus.