Growth Characteristics of Bred Marsupenaeusjaponicus in Polyculture Pond Mixed with Bostrychus sinensis

对与中华乌塘鳢(bOSTryCHuS SInEnSIS)混养模式下的日本囊对虾(MArSuPEnAEuS JAPOnICuS)选育新品系的体长、体质量等生长特性进行了100d的跟踪测量分析,结果表明:经过100d的养殖,虾鱼混养模式下日本囊对虾选育新品系平均体长和体质量分别由8MM和0.006g增加至112.73MM和18.35g,其生长分为快速生长期(30~50日龄)、稳定生长期(50~70日龄)和缓慢生长期(70日龄以后),在30~50日龄间,体长的生长速度较快,随着日龄的增加,体长生长速度变缓,体质量的生长拐点出现在55d左右;体质量和体长呈幂函数关系,关系式为W=1.0x10-5 l3.053,l的指数接近3,呈等速生长;混养模式下日本囊对虾选育新品系体长和体质量的VOn bErTAllAnffy生长方程为:l=127.45x(1-E-0.024T+0.212),W=26.77x(1-E-0.024T+0.212)3.053.研究结果为探索日本囊对虾适宜的养殖模式和选育新品系的选育效果提供了基础资料.The growth characteristics of bred Marsupenaeus japonicus in M.japonicus-Bostrychus sinensis polyculture ponds were studied according to the measurement of body length and body weight.Results showed that the average body length and body weight of bred M.japonicus were 112.73 mm and 18.35 g for 100-day cultivation respectively.The growth period of shrimps was devided into three stages:fast growth stage(30-50days),steady growth stage(50-70days),and aging growth stage(after 70days).The body length increased fast in 30-50 days,then slowed down gradually with age.The inflection of body weight growth was found in about 55days;the relationship between body length and body weight was described by apower function:W =1.0×10-5 L3.053,where the index of Lwas close to 3,indicating an isometric growth.The von Bertallanffy equations were expressed as follows:L=127.45×(1-e-0.024t+0.212),W =26.77×(1-e-0.024t+0.212)3.053.In sum,this study provides fundamental information to help explore appropriate aquaculture mode and bred effect of M.japonicus.国家高技术研究发展计划(863计划)(2011AA10A409); 国家“虾产业体系”项目(CARS-47); 厦门市南方海洋中心项目(14CZY033HJ07

Cloning and Sequence Analysis of the apo-14 Gene in Marsupenaeus japonicus

以日本囊对虾(MArSuPEnAEuS JAPOnICuS)肝胰腺组织为实验材料,提取总rnA,通过3'以及5'-rACE基因克隆技术,首次在日本囊对虾中克隆到载脂蛋白APOlIPOPrOTEIn-14(APO-14)基因的全长序列,包含完整的Orf以及3'-uTr和5'-uTr。日本囊对虾APO-14CdnA序列全长为784bP,其中Orf长度为420bP,编码139个氨基酸。对虾APO-14氨基酸序列与其他鱼类APO-14氨基酸序列有较高的相似性。Full length of apolipoprotein-14 (apo-14) gene was cloned from the total RNA of Marsupenaeus japonicus.Sequence analysis showed that the full cDNA sequence was 784 bp in length including the coding region of 420 bp and encoded 139 amino acids.Homology analysis of the apolipoprotein-14 amino acid sequence indicated high sequence similarity in apo-14 amino acid sequences between M.japonicus and other fish.国家虾产业技术体系岗位专家项目:“日本囊对虾育种”(CARS-47

Genetic structure analysis between two generations of cultured Marsupenaeus japonicas by microsatellite markers

利用8对微卫星引物对日本对虾养殖群体两个世代的遗传结构进行了分析,共检测到108个等位基因,等位基因长度为196-528 bP,各位点等位基因数为6-23个。亲本和子代群体每个位点平均等位基因数目分别为8.5个和11.5个,平均观测杂合度分别为0.721和0.632,平均期望杂合度分别为0.775和0.764,平均多态信息含量分别为0.732和0.729。根据标记信息,利用最大似然法对两个世代的个体分别进行系谱结构推断,亲本群体划分为6个群体,群体间遗传距离为0.277-2.356,均值为1.510;子代群体划分为17个群体,群体间遗传距离为0-2.593,均值为1.113。亲本和子代分群体间的遗传距离变化范围为0-3.089,均值为1.238。根据各分群体间的nEI'S遗传距离,用非加权组平均法(uPgMA)构建了日本对虾两个世代的遗传进化树。其中18个分群体(4个亲本群体和14个子群体)经过聚类后形成3个分支,其余5个分群体(两个亲本群体和3个子群体)明显偏离于主支。研究发现,子代的观测杂合度明显低于亲本,说明子代在继承亲本遗传信息的过程中已经丢失了一定的杂合性。The genetic structure for two generations of cultured Marsupenaeus japonicas was investigated using eight pairs of microsatellite primers.In this study,a total of 108 alleles were tested,and the length of the alleles ranged from 196 bp to 528 bp; the number of alleles per locus ranged from 6 to 23 with an average of 13.5.For the parents and offspring populations,the average number of alleles was 8.5 and 11.5,respectively; the observed heterozygosity was 0.721 and 0.632,respectively; the average expected heterozygosity was 0.775 and 0.764,respectively; and the average polymorphism information content was 0.732 and 0.729,respectively.The pedigree structure for the parents and progeny populations was inferred by maximum likelihood method according to the microsatellite marker information.The parent population was divided into six groups,and the genetic distance among them ranged from 0.277 to 2.356 with an average of 1.510.The offspring population was divided into 17 groups,and the genetic distance among them ranged from 0 to 2.593 with an average of 1.113.The genetic distance between parents and offspring sub-groups ranged from 0 to 3.089 with an average of 1.238.An evolutionary tree for the 23 sub-group populations was drawn by unweighted pair-group method with arithmetic means( UPGMA) according to Nei's genetic distance.Eighteen sub-groups aggregated into the main stretch after forming three sub-stretches.However,five sub-groups( O15, O12,O13,P21 and P18) were obviously away from the main stretch.In this study,the heterozygosity and polymorphism of the offspring population were lower than their parents.The result showed that the heterozygosity of the parents have partially lost during the genetic inbreeding.科技部863项目“主要养殖甲壳类良种培育”(2012AA10A409)资

Genetic Parameters Estimation for Growth Traits of the G1 Marsupenaeus japonicus

采用自然交尾的方式,于2009年同期构建了30个日本囊对虾(MArSuPEnAEuS JAPOnICuS)g1群体(第1世代选育群体)的全同胞家系,利用荧光体内标记技术进行家系标记后,在室外高位池混养至体质量(21.14±5.77)g时进行生长测定,每个家系随机挑选30尾个体测量其体长、体质量等生长性状.通过分析生长数据遗传变异的方差组分,计算遗传力、表型相关和遗传相关.结果显示:日本囊对虾g1群体体长的遗传力为0.43±0.11,体质量的遗传力为0.44±0.12.体长和体质量的表型相关为0.95±0.03,遗传相关为0.96±0.02,显著性统计检验的P值均小于0.001.结果表明:日本囊对虾体长、体质量生长性状的遗传力均属高遗传力,可采用家系选育结合家系内个体选择的方法来提高育种效果;体长、体质量高度遗传相关,可选择将体质量作为生长选育的重点性状,纳入综合选择指数中,结合抗逆性、存活率等性状,对生长性状进行选育改良,提高其生产性能.Thirty full-sib families of kuruma shrimp Marsupenaeus japonicus were constructed by group mating in door scaffolding shrimp pond.Subsequently,thirty individuals were randomly selected from each of these visible implant elastomer(VIE)tagged fullsib families after being cultured in higher-place ponds for 103 days.Body length and body weight were measured to estimate the genetic parameters for growth traits of the G1 population of M.japonicus.And then the heritability of body length and body weight,and the genetic and phenotypic correlation between body length and body weight were evaluated using an animal model and the restricted maximum likelihood method.Data showed that heritability for body length and body weight was 0.43±0.11 and 0.44±0.12,respectively.Genetic and phenotypic correlation between body length and body weight were 0.96±0.02 and 0.95±0.03,respectively.The statistical tests were significant(p<0.001).To conclude,high heritability estimations for growth of M.japonicus demonstrated a potential for family based selection of growth traits in M.japonicus,and the high correlation between body length and body weight indicated each of them could be used as an index to integrate into the effectiveness traits to achieve genetic progress for agrowth breeding scheme in M.japonicus.国家高技术研究发展计划(863计划)(2012AA10A409); 国家虾产业技术体系项目(CARS-47); 厦门市南方海洋中心项目(14CZY033HJ07

Path Analysis of the Effects of Morphometric Traits on Body Weight for 2-month-old Marsupenaeus japonicus

选取人工养殖两月龄日本囊对虾442尾,共测量了11个形态性状.采用多元分析的方法,分别计算得出了各个形态性状对体质量(y)的相关系数、通径系数以及决定系数,并且对于形态性状对体质量的直接作用和间接作用进行了进一步的分析.结果表明,在相关分析中,所有形态性状与体质量的相关系数均达到极显著水平(P0.85).Indicating that the morphometric traits included in this study were major factors influencing body weight.We established the regression equation of the four correlated traits to body weight Y=-1.862+0.352X1+1.087X3+0.983X5+0.528X11.This research result would support the breeding program of M.japonicus.国家虾产业技术体系岗位专家项目(CARS-47

Microsatellites in the growth-related genes of female andmale Tongue sole Cynoglossus semilaevis andtheir application in the population genetic analysis

利用在生长激素(gH)、生长激素释放激素(gHrH)和垂体腺苷酸环化酶激活多肽(PACAP)基因中发现的7个微卫星位点,分析了半滑舌鳎两个野生群体(渤海群体和黄海群体)和1个养殖群体间以及各群体内雌雄个体间的遗传多态性差异。结果表明,7个位点中有4个位点表现出多态性,在3个群体中的等位基因数的分布范围为2~37,平均为9.5;有效等位基因数分布范围为2~28.9,平均为8.4。各位点的平均观测杂合度、平均期望杂合度和平均多态信息含量分布范围分别为0.5145~0.7738、0.5690~0.8671和0.4829~0.8314。群体间的成对fST值及个体分配分析的结果表明,半滑舌鳎野生群体和养殖群体之间存在显著性遗传差异,而在两个野生群体之间差异不显著。此外,等位基因分布和双倍体基因型分布的差异性检测结果表明,这4个多态性位点在3个群体的雌、雄性别间均不存在显著性差异。Seven microsatellite loci were identified in the GH,GHRH and PACAP genes of Cynoglossus semilaevis,and were used as polymorphic markers to analyze the genetic diversity and genetic structure of two wild populations(BS and YS) and a cultured population(HS).The results revealed that no genetic polymorphism was detected in three loci out of seven.The numbers of alleles in the other four loci ranged from 2 to 37,with an average of 9.5,and the numbers of effective alleles ranged from 2 to 28.9,with an average of 8.4.The observed and expected heterozygosities ranged from 0.514 5 to 0.773 8 and from 0.569 0 to 0.867 1,respectively.The average polymorphism information content per locus ranged from 0.482 9 to 0.831 4.The results of pairwise FST and individual distribution analysis demonstrated that there was no significant difference between the wild populations,while significant differences were observed between the wild and the cultured populations.Moreover,based on the distribution of alleles and diploid genotypes,no significant difference was observed between female and male in any of the three populations.国家自然科学基金(30871913);山东省泰山学者工程专项(2009.03-2014.02);山东省自主创新成果转化重大专项(2009ZHZX1A1201)共同资

Chromosome karyotype of sevenband grouper Epinephelus septemfasciatus (Thunberg,1793)

采用PHA体内直接注射法制备了七带石斑鱼(EPInEPHEluS SEPTEMfASCIATuS)头肾组织染色体标本并分析其核型。结果表明,七带石斑鱼二倍体染色体数为48,24对染色体均为端部着丝粒染色体,其染色体臂数(nf)为48,核型公式为:2n=48T,nf=48。未发现与性别相关的异型染色体,第24对染色体长度明显小于其他染色体。通过与其他22种石斑鱼染色体核型进行比较,发现七带石斑鱼具有石斑鱼属鱼类的原始核型特征,属于石斑鱼属鱼类的原始类群。The metaphase chromosomes of Epinephelus septemfasciatus were obtained from head kidney tissue by the method of PHA and colchicine injection and the karyotype was analyzed.The results showed that there were 48 telocentric chromosomes in the diploid and their karyotypic formula was 2n=48t,NF=48.The length of the 24th chromosome was distinctively shorter than that of others,while there was no obvious difference in length among the other 23 chromosomes.By comparison with the chromosome karyotypes of other 22 Epinephelus species,it can be concluded E.septemfasciatus has the representative characteristics of ancestral karyotypic pattern and belongs to the primitive grouper.青岛市科技发展计划项目(08-2-3-6-hz);农业部948项目(2008-Z8

Thermal tolerance of Marsupenaeus japonicus compared with that of Litopenaeus vannamei and Exopalaemon carinicauda

为构建日本囊对虾(MArSuPEnAEuS JAPOnICuS)耐高温性状的评价体系,全面了解日本囊对虾的耐热性状,本研究首先采用耐热性(uPPEr THErMAl TOlErAnCE,uTT)作为评定指标,开展不同规格日本囊对虾耐热性状的分析;进一步结合临界温度法(CrITICAl THErMAl METHOdOlOgy,CTM)将日本囊对虾与脊尾白虾(EXOPAlAEMOn CArInICAudA)、凡纳滨对虾(lITOPEnAEuS VAnnAMEI)进行了耐热性状的种间比较。结果显示:(1)不同大小的日本囊对虾个体之间耐热性(uTT)存在差异,其体重与耐热性之间呈负相关,相关系数为-0.142(P<0.05),特别是日本囊对虾仔虾(0.01 g)耐热性显著高于较大规格(1~5 g)的日本囊对虾(P<0.05);(2)受高温刺激,3种虾表现出不同的应激行为,凡纳滨对虾的开始死亡温度与死亡高峰温度最高,日本囊对虾次之,脊尾白虾最低;(3)3种虾的耐热性状存在明显差异,凡纳滨对虾的平均uTT值最高,日本囊对虾次之,脊尾白虾的最低(P<0.01);(4)凡纳滨对虾的临界高温(CrITICAl THErMAl MAXIMuM,CTMAX)最高,日本囊对虾次之,脊尾白虾最低(P<0.05),即与uTT的比较结果一致。本研究为日本囊对虾耐高温品系选育提供了基础科学依据。Relatively high mortality has become a problem during Marsupenaeus japonicus culture in the summer in southern China.Thus, it is necessary to evaluate the thermal tolerance of M.japonicus.The upper thermal tolerance trait(UTT) was determined to evaluate thermal tolerance of different sizes of M.japonicus.We combined the UTT with critical thermal methodology to compare thermal tolerance among three species, such as M.japonicus, Exopalaemon carinicauda, and Litopenaeus vannamei.We observed their behavior under gradually changing temperature and compared the lethal index.A correlation analysis revealed a correlation coefficient of-0.142 between body weight and the UTT of M.japonicus.The UTT of juveniles(<0.01 g) was significantly higher than that of the group weighing 1–5 g(P<0.05).The three species responded differently to high temperature: temperatures for initial mortality and climax mortality of L.vannamei were highest, whereas those for E.carinicauda were the lowest.The mean UTT and critical thermal maximum of L.vannamei were highest, followed by those of M.japonicus and E.carinicauda(P<0.05).In conclusion, the thermal tolerance of M.japonicus was lower than that of L.vannamei but higher than that of E.carinicauda.These results are important data for a comprehensive understanding of M.japonicus thermal tolerance and have practical applications for selecting high-temperature resistant strains.国家虾产业技术体系岗位专家项目(CARS-47); 厦门市南方海洋中心项目(14CZY033HJ07); 福建省海洋高新产业发展专项项目(闽海洋高新[2014]12号

The toxic effects of ozone on the embryonic development and postembryonic larvae of Epinephelus coioide

为了评价臭氧消毒技术及其对石斑鱼(EPInEPHEluS)潜在的毒性,首先评价了两种质量浓度(0.05 Mg/l和0.30Mg/l)臭氧对亚硝酸盐和弧菌的水处理效果,并以斜带石斑鱼(E.COIOIdE)受精卵为实验材料,探讨臭氧对胚胎发育和初孵仔鱼的毒性作用。结果显示,臭氧的水处理效果良好,0.30 Mg/l的臭氧在3 H内对水中亚硝酸盐的去除率可达72.29%;0.05 Mg/l的臭氧处理20 MIn后也能完全杀灭弧菌。但高浓度臭氧(0.30 Mg/l)对胚胎发育具有一定的毒性,会造成胚胎油球数目增多、萎缩、解体和死亡,而低浓度组(0.05 Mg/l)臭氧处理后22 H胚胎孵化率高达(87.14±7.20)%,与对照组无显著差异。与胚胎相比,初孵仔鱼对臭氧更加敏感,0.05 Mg/l的臭氧处理24 H也会导致初孵仔鱼的全部死亡。本研究结果旨在为石斑鱼育苗及养殖过程中正确使用臭氧消毒技术提供重要的数据参考和理论依据。Epinephelus coioide is one of the most important seawater fish that cultured in China.In recent years, with the breakthrough of grouper artificial seedling rearing, the farming scale of grouper has expanded rapidly, and the gross of cultured grouper reached 59 534 tons in 2011.The effective prevention and control of virus, vibrio, and the effective treatment for harmful water quality factors is the key link in grouper artificial seedling rearing.In traditional pond farming mode, strong oxidizers, such as chloride, iodine and potassium permanganate, have played a significant role in disease prevention and water treatment.In recent years, flatfish industrial farming system, as a mature mode, has been driving the technology intergration of grouper recirculating aquiculture system.The solid oxidants mentioned above were greatly limited in the advanced farming mode, while ozone, a kind of gas disinfectant, not only can remove toxic substance such as nitrite, hydrogen sulfide, ammonia nitrogen and killed pathogenic bacteria effectively, but also will not cause secondary pollution, has been widely applied in water treatment of recirculating aquiculture system in the European Union, the United States, Japan and other developed countries.Although the efficiency and stability of ozone disinfection was widely recognized, its security, especially its toxic effects on the cultured livestock, cannot be ignored.In order to evaluate the ozone disinfection and the potential toxicity of ozone to E.coioide, the efficiency of ozone at two kind of concentration(0.05 mg/L and 0.30 mg/L) on water treatment, such as removing vibrio and nitrite, was assessed.Furthermore, using the zygote of E.coioide as material, the toxic effects of ozone on the embryonic development and post-embryonic larvae were discussed.Our results demonstrated that the efficiency of ozone on water treatment was favorable, when the concentration of ozone reached 0.30 mg/L, the removal rate of nitrite can be up to 72.29% within 3 h; at the applied ozone dose of 0.05 mg/L and a treatment time of 20min, the vibrio all can be killed, while the amount of vibrio in the hatching water that was not disinfected by ozone showed a trend of increase, which was fatal to the embryonic development and post-embryonic larvae.However, the higher concentration of ozone had toxic effects on the embryonic development to some extent, which can caused the number of oil globules increased and led to analosis, collapse and death of the embryo, while the hatching rate can be up to(87.14±7.20)% when the embryo treated with the lower concentration of ozone(0.05 mg/L), suggesting the lower concentration of ozone(0.05 mg/L) had no significant effects on the embryo.The post-embryonic larvae were more sensitive to ozone compared with the embryo, all the post-embryonic larvae died when they exposed to the lower concentration of ozone(0.05 mg/L) for 24 h.The results of this study provided some important reference data and theoretical basis for using the ozone disinfection technology properly in the process of seedling rearing and cultivating of E.coioide, and was significant to the sustainable and healthy development of grouper rearing.国家质检公益项目资助(201210055); 国家支撑计划项目资助(2011BAD13-B08