抗菌肽对凡纳滨对虾生长和机体免疫的影响

基础饲料(对照组)中添加10g/kg抗菌肽制剂喂养体重0.10±0.01g、体长2.17±0.11cm的凡纳滨对虾(Penaeus vannamei)30d,进行10d的水浴攻毒(鳗弧菌10~8/mL),探讨饲料中添加抗菌肽对凡纳滨虾生长、成活率、免疫保护率,以及肌肉中溶菌酶活性、超氧化物歧化酶活性、总抗氧化能力的影响。结果表明:饲料中添加抗菌肽可显著提高凡纳滨对虾相对增重率、成活率和对致病菌的免疫保护率(P<0.05),但对饲料系数无显著影响(P>0.05);未受外界微生物刺激时,饲料中添加抗菌肽对凡纳滨对虾总抗氧化能力有促进作用(P<0.05),受外界微生物刺激后,饲料中添加抗菌肽喂养凡纳滨对虾可提高对虾超氧化物歧化酶活性(P<0.05)和总抗氧化能力(P<0.01)。因此饲料中添加抗菌肽喂养凡纳滨对虾可提高其机体非特异性免疫。     

Impact of Vibrio parahaemolyticus and white spot syndrome virus (WSSV) co-infection on survival of penaeid shrimp Litopenaeus vannamei

White spot syndrome virus(WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect L itopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus(WSSV+Vp), or we fi rst infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection(Vp+WSSV). The effect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of V ibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction(PCR) method. L v My D88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the different mortality rates. Our results show that(1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection;(2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed signifi cant increases in the numbers of Vibrio( P 0.05);(3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10 5 to 10 7 /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.     

Effect of dietary potassium on growth,nitrogen metabolism,osmoregulation and immunity of pacific white shrimp (Litopenaeus vannamei) reared in low salinity seawater

An 8 weeks feeding experiment was conducted to determine the effect of dietary potassium on the growth and physiological 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(P0.05). The shrimps fed the diets containing 0.96-1.48 g K+ per 100 g 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 Cl- were significantly lower than those of the control(P0.05), 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(P0.05). 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.     

蜕壳素对凡纳滨对虾生长及表观消化率的影响

研究了外源性蜕皮激素对凡纳滨对虾生长和表观消化率的影响 ,实验时在基础饲料中分别添加 0 .1%、0 .2 %、0 .4 %的两种市售蜕壳素产品 ,制成 6种外源性脱壳素配合饲料 ,不添加的基础饲料作为对照组 ,试验在 2 6 0L玻璃钢循环水养殖系统中 ,每个剂量组设 3个重复 ,8周生长试验结果表明 :市售蜕壳素产品能有效地促进对虾生长 ,试验结束时 ,试验组虾体增重分别较对照组增加 16 .8%、7.5 2 %、5 .9%、11.4 %、2 9.0 7%、16 .5 3% ,但是凡纳滨对虾对饲料粗蛋白的表观消化率没有明显地提高 ,凡纳滨对虾对脂肪的表观消化率较低。推算凡纳滨对虾饲料中最佳蜕皮酮添加量为 6 0mg/kg。     

Effects of small peptides,probiotics, prebiotics,and synbiotics on growth performance,digestive enzymes,and oxidative stress in orange-spotted grouper,Epinephelus coioides,juveniles reared in artificial seawater

Aquaculture production efficiency may increase by using feed additives. This study investigated the effects of different dietary additives [w/w: 2% small peptides, 0.01% probiotics( Bacillus licheniformis) and 0.2% prebiotics(inulin)] on growth performance, digestive enzyme activities, and oxidative stress in juvenile Epinephelus coioides reared in artificial seawater of two salt concentrations(13.5 vs. 28.5). Weight gain rate was significantly higher in fish fed the diet supplemented with small peptides, B. licheniformis, inulin, or synbiotics than that in fish fed the basal diet; the greatest weight gain rate was found in fish fed the small peptide treatment [56.0% higher than basal diet]. Higher feed efficiency was detected in fish fed the diet supplemented with small peptides than that of fish in the other dietary treatments. Total protease activity in the stomach and intestines was highest in fish fed the small peptide-treated diet, whereas lipase activity was highest in those fed synbiotics(combination of Bacillus licheniformis and inulin) than that in fish fed the other treatments. Antioxidant enzyme(total superoxide dismutase and catalase) activities and hepatic malondialdehyde content were higher in fish receiving the dietary supplements and maintained in artificial seawater containing 13.5 salinity compared with those in the control(28.5). Hepatic catalase activity in grouper fed the diets with small peptides or synbiotics decreased significantly compared with that in control fish. Overall, the three types of additives improved growth rate of juvenile grouper and digestive enzymes activities to varying degrees but did not effectively improve antioxidant capacity under low-salinity stress conditions.     

大黄对凡纳滨对虾生长和非特异性免疫指标的影响

在凡纳滨对虾饲料中分别添加大黄0、0.5、1.0、5.0、10.0和20.0g/kg,研究大黄对凡纳滨对虾(初始体重为0.34±0.004g)生长及非特异性免疫指标的影响。结果表明,大黄对凡纳滨对虾成活率、增重率、特定生长率、饲料系数、蛋白质效率和蛋白质累积率的影响不显著(P>0.05),对全虾和尾肌肉的灰分、脂肪和粗蛋白含量影响显著,全虾的粗蛋白和粗脂肪含量以1.0g/kg组最高(P<0.05),对凡纳滨对虾血清碱性磷酸酶、酸性磷酸酶、酚氧化酶、超氧化物歧化酶、溶菌酶以及血清总蛋白量等的影响显著,溶菌酶活性以1.0g/kg组最高(P<0.05);细菌感染实验中以1.0g/kg组存活率最高(P<0.05)。以非特异性免疫反应指标及感染实验存活率为指标,凡纳滨对虾饲料中大黄的适宜添加量为1.0g/kg。     

REQUIREMENTS OF SHRIMP, PENAEUS CHINENSIS O''''SBECK FOR POTASSIUM, SODIUM, MAGNESIUM AND IODINE

Potassium, sodium, magnesium and iodine requirements of shrimp, Penaeus chinensis were studied.Orthogonal design was employed in this experiment. The composition of the basal diet consisted of fishmeal, peanut cake, corn meal, soybean cake, wheat bran, vitamin mix and mineral mix, and supple-mentations of potassium, sodium, magnesium and iodine in the basal diet were made according to theL_9(3~4) orthogonal table. The results indicatal that iodine supplementation improved growth of the shrimpsignificantly and raised survival very significanly, iodine requirement of the shrimp was 0.003%, sodiumrequirement was 0.87% or less; and that 1.1-1.3% potassium and 0.18-0.38% magnesium in the dietwere proper nutrition supplements for the shrimp.     

Effects of small peptides,probiotics, prebiotics,and synbiotics on growth performance,digestive enzymes,and oxidative stress in orange-spotted grouper, <Emphasis Type="Italic">Epinephelus coioides</Emphasis>, juveniles reared in artificial seawater

Aquaculture production efficiency may increase by using feed additives. This study investigated the effects of different dietary additives [w/w: 2% small peptides, 0.01% probiotics (Bacillus licheniformis) and 0.2% prebiotics (inulin)] on growth performance, digestive enzyme activities, and oxidative stress in juvenile Epinephelus coioides reared in artificial seawater of two salt concentrations (13.5 vs. 28.5). Weight gain rate was significantly higher in fish fed the diet supplemented with small peptides, B. licheniformis, inulin, or synbiotics than that in fish fed the basal diet; the greatest weight gain rate was found in fish fed the small peptide treatment [56.0% higher than basal diet]. Higher feed efficiency was detected in fish fed the diet supplemented with small peptides than that of fish in the other dietary treatments. Total protease activity in the stomach and intestines was highest in fish fed the small peptide-treated diet, whereas lipase activity was highest in those fed synbiotics (combination of Bacillus licheniformis and inulin) than that in fish fed the other treatments. Antioxidant enzyme (total superoxide dismutase and catalase) activities and hepatic malondialdehyde content were higher in fish receiving the dietary supplements and maintained in artificial seawater containing 13.5 salinity compared with those in the control (28.5). Hepatic catalase activity in grouper fed the diets with small peptides or synbiotics decreased significantly compared with that in control fish. Overall, the three types of additives improved growth rate of juvenile grouper and digestive enzymes activities to varying degrees but did not effectively improve antioxidant capacity under low-salinity stress conditions.     

凡纳滨对虾不同生长阶段食物组成结构的研究

利用稳定同位素碳比率δ13C作为天然示踪物,对高位养殖池生态系统中人工饲料和天然饵料提供的能源物质对养殖凡纳滨对虾生长的贡献比率进行研究。结果表明:虾体的δ13C值受食物δ13C值的影响,由于其摄食饵料的δ13C值逐渐减小,虾体的δ13C值也逐渐降低,逐渐接近于最后人工饲料的δ13C值,试验虾体的δ13C值由养殖开始时的-19.52‰±0.09‰变为实验结束时的-21.74‰±0.17‰;对虾的食性在10d左右时发生转变,34d左右时人工饲料和浮游动物对虾体的能量贡献基本相同,养殖结束时对虾生长能量的97.29%±0.98%来源于人工饲料。     

植酸酶对凡纳滨对虾生长性能和体成分的影响

在凡纳滨对虾饲料中添加不同量微生物植酸酶(0,500,1000,2000 U/kg),观察虾的存活率、增重率、饲料系数及虾体和虾壳成分,研究饲料中添加植酸酶对凡纳滨对虾生长性能的影响。结果表明:饲养8周后,饲料中添加500~2000 U/kg植酸酶对凡纳滨对虾增重率、存活率和饲料利用无显著影响(P>0.05);植酸酶添加各组与对照组虾体的水分、粗蛋白、脂肪、灰分、总钙和总磷含量差异不显著;虾壳中粗灰分和钙含量在500和1000 U/kg组显著高于对照组和2000 U/kg组(P<0.05),虾壳磷含量在各饲料组间差异不显著。血清磷浓度在对照组和2000 U/kg组显著高于500 U/kg组(P<0.05),血清钙浓度在组间差异不显著,血清碱性磷酸酶活性以对照组最高,显著高于500和1000 U/kg组。结果说明,在特定试验条件下,饲料中添加植酸酶对凡纳滨对虾幼虾的虾壳和血清成分有显著影响(P<0.05),对幼虾生长性能影响不显著(P>0.05)。     

PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp.on intestinal microbial diversity in kuruma shrimp(Marsupenaeus japonicus)

In this study,the intestinal microbiota of kuruma shrimp(Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects of Bacillus spp.on intestinal microbial diversity.Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp.amended feed.PCR and denaturing gradient gel electrophoresis(DGGE) analyses were then performed on DNA extracted directly from the guts.Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S rDNA amplicons,and distinct bands in the gels were sequenced.The results suggested that the gut of kuruma shrimp was dominated by Vibrio sp.and uncultured gamma proteobacterium.Overall,the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity of shrimp.     

Growth and feed efficiency of juvenile shrimp Litopenaeus vannamei fed formulated diets containing different levels of poultry by-product meal

This feeding trial was conducted to evaluate the potential of poultry by-product meal (PBM) as a protein source in the culture ofLitopenaeus vannamei. Seven isonitrogenous and isoenergetic diets were formulated to near to commercial diet with about 40% protein and 7.5% lipid. Fish meal was replaced by 0, 30%, 40%, 50%, 60%, 70% and 100% of PBM (diets 1-7). The diet with 100% fish meal was used as a control (diet 1). Post-larvae were reared in an indoor semi-closed re-circulating system. Each dietary treatment was tested in 4 replicate tanks (260 L) of 40 shrimp, arranged in a completely randomized design. The shrimps were hand-fed for three times a day to near-satiation (0700, 1200 and 1800) for 60d. Percentage weight gain, survival, feed conversion ratio (FCR), protein efficiency ratio (PER) and body composition of shrimps were measured. There were no significant differences (P>0.05) in growth performance among shrimps fed diets 1-5 (0-60% fish meal replacement). However, shrimps fed diet 7 (100% fish meal replacement) had significantly lower (P<0.05) growth than those fed diets 1-5 (0-60% fish meal replacement). Shrimp fed diets 2-4 (30%-50% fish meal replacement) showed significantly higher growth than those fed diets 6 and 7 (70% and 100% fish meal replacement, respectively). Survival ranged from 94.7% to 100.0% and did not differ significantly (P>0.05) among different experimental diets. No differences in body composition were found among shrimps fed different diets. These results showed that up to 70% of fish meal protein can be replaced by PBM without adversely affecting the growth, survival, FCR, PER and body composition of Litopenaeus vannamei.     

湛江港区底泥悬浮物对凡纳滨对虾仔虾成活的影响

模拟吹填、疏浚等海上施工过程所产生的底泥悬浮物浓度的变化规律,探讨不同底泥悬浮物浓度下对凡纳滨对虾仔虾成活影响。结果表明:海水中底泥悬浮物浓度超过10mg/L,持续时间超过12h时,将对凡纳滨对虾成活率产生影响,随着悬浮物实验浓度的递加,仔虾的死亡率从13.33%增至96.67%;超过80mg/L,且持续时间超过48h,将对凡纳滨对虾产生严重影响,甚至导致全部仔虾的死亡;悬浮物对凡纳滨对虾的12h、24h、36h和48h半致死量(LC50)分别为54.56、40.29、25.8和19.18mg/L。     

Application of Wet Waste from Shrimp (Litopenaeus vannamei) with or Without Sea Mud to Feeding Sea Cucumber (Stichopus monotuberculatus)

In the present study, the applicability of the wet waste collected from shrimp(Litopenaeus vannamei) to the culture of sea cucumber(Stichopus monotuberculatus) was determined. The effects of dietary wet shrimp waste on the survival, specific growth rate(SGR), fecal production rate(FPR), ammonia- and nitrite-nitrogen productions of sea cucumber were studied. The total organic matter(TOM) level in the feces of sea cucumber was compared with that in corresponding feeds. Diet C(50% wet shrimp waste and 50% sea mud mash) made sea cucumber grow faster than other diets. Sea cucumber fed with either diet D(25% wet shrimp waste and 75% sea mud mash) or sole sea mud exhibited negative growth. The average lowest total FPR of sea cucumber occurred in diet A(wet shrimp waste), and there was no significant difference in total FPR between diet C and diet E(sea mud mash)(P > 0.05). The average ammonia-nitrogen production of sea cucumber in different diet treatments decreased gradually with the decrease of crude protein content in different diets. The average highest nitrite-nitrogen production occurred in diet E treatment, and there was no significant difference in nitrite-nitrogen production among diet A, diet B(75% wet shrimp waste and 25% sea mud mash) and diet C treatments(P > 0.05). In each diet treatment, the total organic matter(TOM) level in feces decreased to different extent compared with that in corresponding feeds.     

Optimal content and ratio of lysine to arginine in the diet of Pacific white shrimp, Litopenaeus vannamei

The optimal quantity of dietary lysine (Lys) and arginine (Arg), and the optimal ratio of dietary Lys to Arg for Pacific white shrimp Litopenaeus vannamei were investigated. Coated Lys and Arg were added to a basal diet (37.99% crude protein and 7.28% crude lipid) to provide graded levels of Lys and Arg. The experimental diets contained three Lys levels (2.51%, 2.11%, and 1.70% of total diet), and three Arg levels (1.41%, 1.80%, and 2.21% of total diet) and all combinations of these levels were tested. Pacific white shrimp, with a mean weight of 3.62±0.1 g, were randomly distributed in 36 fiberglass tanks with 30 shrimp per tank and reared on the experimental diets for 50 days. After the feeding trial, the growth performance, survival, feed conversion rate (FCR), body composition and protease and lipase activities in the hepatopancreases of the experimental shrimps were determined. The results show that weight gain (WG), specific growth rate (SGR), FCR, body protein, body Lys and Arg content were significantly affected by dietary Lys and Arg (P <0.05) and improved when dietary Lys and Arg levels were 2.11% ～ 2.51% and 1.80%～2.21%, respectively. Protease and lipase activities in the hepatopancreases of the shrimps appeared higher when dietary Lys and Arg quantities were 2.11% ～2.51% and 1.80%～2.21%, although the difference was not statistically significant (P >0.05). Therefore, according to our results, the optimal Lys and Arg quantities in the diet of Pacific white shrimp, L. vannamei, were considered to be 2.11%–2.51% and 1.80%–2.21%, respectively, and the optimal ratio to be 1:0.88–1:1.05.     

Effects of Different Diets on the Dietary Attractability and Selectivity of Chinese Shrimp, Fenneropenaeus chinensis

Attractabilities of different diets and dietary selectivity of Chinese shrimp, Fenneropenaeus chinensis were studied through behavior observation and feeding experiment, respectively. The five diets used in the experiment are: Fish Flesh (FF), Shrimp Flesh (SF), Clam Foot (CF), Polychaete Worm (PW), and Formulated Diet (FD). No significant differences of attractability exist between any two different diets when every two natural diets or all five diets are provided simultaneously. On the other hand, significant differences of attractability exist between FD and every single natural diet when they are provided simultaneously. Results of behavioral observation indicate that natural diets are more attractive than FD. In feeding experiment, Chinese shrimp has distinct selectivity on different diets. It positively selects CF and PW, negatively selects FF and SF, and excludes FD absolutely. The results of the present studies indicate that the dietary selectivity of shrimp was based not only on the attractabilities of the diets, but also on the responses such as growth and food conversion.     

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.     

Requirements of shrimp,Penaeus chinensis O'sbeck for potassium,sodium, magnesium and iodine

Potassium, sodium, magnesium and iodine requirements of shrimp,Penaeus chinensis were studied. Orthogonal design was employed in this experiment. The composition of the basal diet consisted of fish meal, peanut cake, corn meal, soybean cake, wheat bran, vitamin mix and mineral mix, and supplementations of potassium, sodium, magnesium and iodine in the basal diet were made according to the L₉(3⁴) orthogonal table. The results indicated that iodine supplementation improved growth of the shrimp significantly and raised survival very significantly, iodine requirement of the shrimp was 0.003%; sodium requirement was 0.87% or less; and that 1.1–1.3% potassium and 0.18–0.38% magnesium in the diet were proper nutrition supplements for the shrimp. Contribution No. 2342 from the Institute of Oceanology, Chinese Academy of Sciences.     

贮藏时间和加热温度对凡纳滨对虾虾仁ATP关联化合物及品质的影响

【目的】研究贮藏时间和加热温度对凡纳滨对虾(Litopenaeus vannamei)虾仁三磷酸腺苷(ATP)关联化合物及虾仁品质的影响。【方法】利用高效液相色谱法分析凡纳滨对虾中6种ATP关联化合物,通过比较不同贮藏时间的凡纳滨对虾虾仁在加热过程中的ATP关联化合物,分析加热前后虾仁的感官品质,以及鱼虾类鲜度指标K值、G值、P值的变化。【结果与结论】随贮藏时间的增加,ATP、二磷酸腺苷(ADP)、腺苷酸(AMP)含量呈下降趋势,肌苷酸(IMP)先上升后下降,次黄嘌呤(HX)和肌苷(HXR)上升;随着加热温度的增加,ATP和ADP下降,AMP增长,IMP、HX、HXR先增加后降低。贮藏2 d后的生虾和贮藏3 d后再加热的虾仁均将达到最大感官可接受上限。随着贮藏时间的增加,加热前后K值、G值和P值线性增加,但生虾的G值与鲜度的相关性较差,不适宜作为鲜度指标,K值和P值可作为生虾的鲜度指标。加热后的K值、G值、P值与贮藏时间、感官质量评定值(QI值)的线性回归方程的决定系数较好,表明K值、G值、P值可以用来表征熟虾的品质。