Abstract:13 half-sib families and 39 full-sib families of Mytilus edulis were established with the balance nest design in which each male mated with three females and the artificial fertilization technique.The shell height and length of each family were measured in 2,3,5,6 months.The heritability of the growth was estimated using the full- sib families theories.The results show that the maternal genetic variance component estimates are all significantly greater than those of paternal ones of each growth trait(P<0.05).The female genetic variance components have significant maternal effects(P<0.05),suggesting that the large non- additive genetic effects could not be differentiated from the available data,so the heritability of paternal half-sib families estimates in the narrow sense were precise unbiased estimator.The shell height heritability of paternal half-sib families estimates are 0.29,0.85,0.84,0.87 respectively.The shell length heritability are 0.27,0.87,0.86,0.85 respectively,during 2,3,5,6 months.There were significant additive genetic variance of the growth and different heritability of shell height and shell length,indicating that there is a great potential genetic improvement for the growth of M.edulis using the selective breeding.

Abstract:This experiment was conducted to study the correlation between texture and sensory evaluation of salted fish by sensory evaluation and the instrumental texture analyses.Three kinds of salted fish,11 samples were used as the study object.Several sensory texture attributes of samples were evaluated by trained experts and TPA(Texture Profile Analysis)instrumental texture analyses,respectively.One-way analysis of variance(ANOVA)was used to analyse the correlation of sensory texture attributes and the instrumental texture test data.The results suggested that there were significant difference(P<0.05)between the sensory texture attributes of the 11 samples,and the difference between the TPA parameters were also notable.The result of correlation analysis showed that there was a close correlation between the sensory texture attributes and instrumental texture test parameters.The instrumental test parameters of hardness was significantly(P<0.05)or very significantly(P<0.01)correlated with all the sensory texture attributes with the coefficient between 0.398-0.859(r=0.398-0.859),and chewiness was just presented correlation with the sensory hardness with the coefficient of 0.650(r=0.650),there were widespread correlation among all the other parameters and sensory attributes(r=-0.355-0.829).The results of the correlation analysis between the sensory evaluation total score and the TPA showed that the hardness and chewiness were notably positively correlated with sensory texture evaluation total score(P<0.05,R=0.974-0.994),while the cohesiveness,springiness,adhesiveness showed significantly negatively correlated with sensory texture evaluation total score(P<0.05,R=0.937-0.998).The study showed that there was high correlation between the values of the sensory evaluation and the instrumental texture analyses of the three salted fish,which provided an important theory for the application of texture analyzer in the quality evaluation of salted fish.

Abstract:The Pacific abalone Haliotis discus hannai is a commercially important marine mollusk.To develop a rapid,economically efficient and robust approach for genetic studies in this species,twelve microsatellites were used to optimize four multiplex PCRs for the H.discus hannai,and the results obtained indicate the usefulness of the four multiplexed microsatellite systems in parentage assignment.We evaluated and validated these multiplex PCRs in 12 full-sib families.The average polymorphism information content(PIC)was 0.82.Inheritance analyses in the 144 tests showed that 4.9% microsatellite loci deviated from Mendelian ratios.The frequency of null alleles was estimated at 10.6% of all the alleles segregating based on a within-family analysis of Mendelian segregation patterns.Simulations and parentage analysis showed that with the most informative multiplex set 4,the assignment success could be 86% and 90% respectively,and 100% of the offspring were correctly allocated to their parents when two or more multiplex PCRs were used.Our results show that four multiplex PCR panels of microsatellites are possible and can be used for rapid and highly efficient parentage assignment in the Pacific abalone.

Abstract:The survey experiment of two different type of gillnets was carried out at rocky reef area to find the suitable survey nets for rocky habitat during July and August in 2009.Indices such as Sorensen similarity,Margalef’s species richness index,Shannon-Wiener diversity,Peilou’s species evenness index were calculated,and the constitution and difference of catch were analyzed.Results showed that (1)Catch and species of European gillnet were lower than multi-mesh trammelnet.20 species was caught by European monolayer gillnet,and total weight was 8 572.6 g while 29speceies and 20 793 g catch by multi-mesh trammelnet.(2)Dominant species of European gillnets were:Sebastiscus marmoratus,Hexagrammos agrammus,Collichthys lucidus and multi-mesh trammelnets’ dominant species were:Sebastiscus marmoratus,Hexagrammos agrammus,Collichthys lucidus,Scomber japonicas.The selectivity of European gillnets was much higher than multi mesh trammelnets.(3)European gillnet and multi mesh trammelnets had no significant difference in community structure index,but index of Sorensen similarity was only 0.41,which showed that the species of two gillnets is different.(4)It is suggested that the selectivity of two gillnets was different,the average length of catch was 106.1 mm for European gillnet and 119.1 mm for trammelnets.(5)Different species are distributed in different mesh gillnets,the dominant species such as Sebasticus marmoratus,Hexagrammos agrammus distributed mainly in five mesh gillnets.Our research indicates that the indices of fish community have no significant difference,however,multi mesh trammelnet has a high catch efficiency,and is more durable than European monolayer gillnets for survey in rock reef area.

Abstract:Grass carp(Ctenopharyngodon idella)is one of the most important cultured fishes in China.During transportation in summer,massive mortality often occurs.As molecular chaperones,Hsps assist cells in their recovery from stress and promote cytoprotection.Limited reports described the expression of Hsps in grass carp at high temperatures,and the relationship between oxygen consumption,expression of heat shock protein and mortality needs to be investigated based on the oxygen- and capability- limited thermal tolerance theory(OCLTT)to further elucidate the thermal adaptation of this species to high temperature in view of integrated biology.In the present study,the expressions of hsp70 and hsp90 after rapid temperature increase were studied.Grass carps were acclimated at 20 ℃ and then exposed to designated temperatures(20,22,24,26,28,30,32,34 ℃)for 3 h heat shock.After 2 h- recovery at 20 ℃,liver,muscle and gill of tested fish were sampled and hsp70 and hsp90 were determined using semi-quantitive real time PCR.Expression of hsps increased with temperature increase until 32 ℃ in muscle and gill,and then decreased at 34 ℃.In liver,expression of hsps kept increasing in all temperature treatments.These results indicated that hsp70 and hsp90 were sensitive to temperature increase in grass carp.Analyzed heat shock proteins data with previous results of oxygen consumption and lethal temperatures of grass carp,the physiological adaptations of grass carp could be explained by oxygen and capability- limited thermal tolerance theory(OCLTT),which pointed out that the imbalance between oxygen demand and tissue oxygen supply ability was the primary factor which limited organisms’ thermal tolerance and the ability of aerobic metabolism greatly limited the survival of organism beyond critical temperatures.With temperature increase,the maximum value of oxygen consumption in grass carp occurred at 28 ℃,and then the oxygen consumption decreased when temperature was beyond 28 ℃,indicating a transformation from aerobic metabolism to anaerobic metabolism at this critical temperature.The expressions of heat shock proteins kept increasing until 32 ℃ in muscle and gill in grass carp,and provided defense against high temperature.However,this protection of heat shock proteins was limited and then death occurred when temperature was beyond 32 ℃.The change of metabolism,increase of oxygen free radicals and the consumption of energy due to stress proteins synthesis of grass carp were closely related to the decrease of capability of immune response when water temperature was beyond 28 ℃.In general,high water temperature could alter the metabolic pathway,induce heat shock protein expression,reduce immune response,and then cause large scale mortality.Therefore,appropriate ways should be applied to maintain water temperature under 28 ℃ to avoid large-scale mortality during transportation.

Abstract:In this study,the samples of Argentine shortfin squid Illex argentinus were collected by the Chinese squid jigger fleet during January to March in 2010,and the dorsal mantle length(DML)of the samples ranged from 166 mm to 266 mm.The total of 264 pairs of beaks(143 pairs for female and 121 pairs for male)were extracted,and the morphological indices in lower beak were measured,including Lower Hood Length(LHL),Lower Crest Length(LCL),Lower Rostral Length(LRL),Lower Lateral Wall Length(LLWL)and Lower Wing Length(LWL).The stages of beak pigmentation of those samples were checked based on the standard of Stage 0 to Stage 7.Combined with the ontogenesis,maturity stage of I.argentinus and growth of beak,the variation of beak pigmentation stages were preliminarily analyzed.The results showed that Stage 3 was the dominant proportion,accounting for 35.98% of the total samples during January to March.The monthly average level of beak pigmentation increased with the time passage from January to March.The beak pigmentation degrees of the samples increased with the growth of DML and body weight,and there were the significant linear relationships between beak pigmentation degrees and DML,beak pigmentation degrees and body weight(P<0.01).The beak pigmentation degrees had a significant relationship with the sexual maturity stage for female squid(P<0.01),but had not significant relationship for male squid(P>0.05).The differences existed in the relationships between different morphological parameters of beaks(LHL,LCL,LRL,LLWL and LWL)and pigmentation stages,in which the significant relationships with LHL,LLWL and LWL are provided

Abstract:Inimicus japonicus is one of the most commercially important marine fishes in China and Japan.The embryonic and morphological development of larvae of this species has been studied,but it is incompletely known about its osteological development.In order to obtain the knowledge of the normal onset of skeletal structures throughout development,the developmental sequences of vertebral column and the appendicular skeleton were examined in laboratory-reared larvae of Inimicus japonicus from hatching through 35 days after hatching(d.a.h)by the the clearing and staining technique of cartilage and bone.According to the observation,the vertebral column started to develop from haemal arches and neural arches at 10 d.a.h and 11 d.a.h respectively.Then,centrum,medullary spines,and pulses spines were formed at 20 d.a.h.and they were completely ossified by 35 d.a.h with trunk vertebrae flexion.The development sequence and ossification of centrum,medulllary spines,and pulses spines both started from anterior to posterior.The appendicular skeleton developed in following sequences:the pectoral fin is the first to develop,followed by caudal fins,anal fins,dorsal fins,and then ventral fins.The pectoral fin started to develop from the cleithrum and the fin plate at 3 d.a.h.Cartilages of pterygiophore of pectoral fin were formed at 11 d.a.h and some fin supports of pectoral fins separated at 35 d.a.h.Pelvic girdle and ventral fin appeared right at 18 d.a.h,later than pectoral fin,but they are rapidly developmented.Dorsal and anal fin appeared at 20 d.a.h,and dorsal and anal fin-ray complements completed at 23 d.a.h.However,anal fin developed posterior from the middle section.The development of the caudal fin began with the appearance of the hypural at 8 d.a.h,and then caudal fin rays and fin-supports begin to develop.Caudal fin was basically formed at 15 d.a.h with two leaves a fin bone formation,which divided the caudal fin rays into two parts.The caudal fins bone formed and started preliminary ossification at 35 d.a.h.Previously recognized critical periods for Inimicus japonicas,corresponded to transformation phases from the primitive,basic modes to stable,more functional modes in both swimming functional development and changing in living habits.The study of the development of the vertebral column and the appendicular skeleton of Inimicus japonicas will provide the basis for the functional adaptation during early development.

Abstract:An electron probe microanalyzer was used to analyze the element Sr and Ca accumulation patterns in otoliths of two individuals of Coilia nasus from the Xingzi area in the Poyang Lake, China. Results of both the line transect and mapping analysis confirmed objectively, for the first time, the coexistence of resident and anadromous forms of Coilia nasus in the Poyang Lake. This fact will provide important information not only to more effectively evaluate and protect the resources of Coilia nasus, but also to rationally plan and guide future ecological economic development in the Poyang Lake.

Abstract:This study was to investigate the effect of dietary carbohydrate levels on juvenile tilapia(Oreochromis niloticus)growth performance and liver metabolism functions.Eight groups of 1 200 tilapias,in triplicate,with initial body weight of(0.36±0.01)g were fed eight experimental diets(similar protein and lipid contents but carbohydrate levels were 10%,15%,20%,25%,30%,35%,40%,45%,respectively).After 7 weeks feeding,the results showed that:(1)dietary carbohydrate level had significant effects on weight gain rate,survival rate,feed conversion ratio and hepato-somatic index(HSI)in tilapia juvenile(P<0.05).The growth rate of the fish fed the diet of 35% carbohydrate level was the highest among the treatments,and the feed conversion ratio of the fish fed the diet of 40% carbohydrate level was the lowest.(2)With increasing of dietary carbohydrate,the muscle glycogen and crude fat content gradually increased.However,the glycogen content rose initially and then declined.(3)The activities of serum alanine aminotransferase,aspartate aminotransferase,phosphofructokinase,and the concentrations of serum total protein and total bile acid were all significantly increased with the elevated levels of carbohydrate(P<0.05).However,the activities of liver esterase and lipoprotein lipase rose initially and then declined.(4)Liver tissues displayed a few lipid vacuolations in hepatocyte cells at 35%.At the carbohydrate level over 40%,liver histological sections showed a large number of hepatocyte lipid droplets vacuolar degeneration,nuclear migrations and cytoplasm disappearances.The lipid vacuolation areas in liver histological sections basically tallied with the lipid content of the liver,which were consistent with the severity of fatty liver.In conclusion,according to the observation of growth and prevention of late fatty liver disease,the most appropriate proportion of carbohydrate in the fish diet is around 29.10%-35.00%.

Abstract:In order to further study salt tolerance performance of a hybrid by Oreochromis niloticus(♀)×Sarotherodon melanotheron(♂)which had been examined and approved by China National Aquaculture Variety Approval Committee as a new variety for aquaculture and named “GILI tilapia”in 2009,researches have been carried out to study the salt tolerance in the chronic salinity stress experiment to GILI tilapia and its two parents,the differences of salt tolerance performance of three kinds of tilapia are analysed,and the regression model between mortality of salt tolerance in the chronic salinity stress and death of time,death of salinity are established.The results of the study show that:(1)There are significant differences in capacity of salt tolerance among three kinds of tilapia,the performance of salt tolerance of GILI tilapia is close to S.melanotheron,it is much higher than O.niloticus,the average death salinity of three kinds of tilapia when they are domesticated in salt are 57.9,66.7 and 18.5 respectively.(2)There are the greatest differences of death time domesticated among individuals in O.niloticus,it is the least in S.melanotheron,it is medium in GILI tilapia.There are the most significant normal negative deviation in GILI tilapia and S.melanotheron,there are the more values of outlier and extremum,O.niloticus has a significant normal positive deviation,it has outlier values,no extremum values.(3)The regression relationships between the mortality of chronic salinity stress to salt and the death time,death salinity of three kinds of tilapia are fit for single-element regression model,the single element regression model between the salt tolerance mortality(Y)and the domestication time of death(t)and salinity of death(s)of GILI tilapia are growth model Y=e^{(-7.694+0.031t)}(R²=0.979)and quadratic model Y=0.542－0.037s+0.001s²(R^2=0.950)respectively.

Abstract:In this study,the reference panel contained 92 individuals,which were a F₂ family obtained by mating of F1 hybrids derived from crossing Barbless carp(Cyprinus pellegrini)with Hebao cold tolerance red carp(Cyprinus carpio wuyuanensis),and the genetic linkage map was constructed with 300 microsatellites,which was used to detect the QTLs for standard length(SL),body thickness(BT),body height(H)and body weight(W)in the panel by GridQTL software.The results showed that 17 related QTLs distributed in the 6 linkage groups were obtained.Four QTLs related to standard length,of which the linkage group of LG5,LG7,LG19 and LG32 were at 5% significant level,but LG32 was at 1% significant level,accounting 5.14%,10.2%,7.02%,9.36% of phenotypic variation,respectively.Four QTLs were identified for body thickness in LG11,LG19,which were at 5% significant level,QTL based on LG5 1% was at significant level,accounting for 6.54%,6.42% and 8.43% of phenotypic variation,respectively.Six QTLs were associated with body height in LG5,LG7,LG10,LG11,LG19 and LG32.LG7 was at 1% significant level,whereas the others were at 5% significant level,accounting 8.95%,6.34%,5.92%,6.29%,6.78%,7.79% of phenotypic variation.Three QTLs were responsible for body weight and LG5,LG7 were at 5% significant level and LG32 was at 1% significant level,which accounted for 6.44%,5.55%,8.36%.Hybrid progenies crossed by two carp strains have been used in this study,which enriched the diversity of QTL population.It is a foundation for further QTL comparative analysis with whole genome level,and common QTL identification in different strains or breeds of carp.It will also be of great assistance in marker assisted selection(MAS)in common carp.

Abstract:White spot syndrome virus(WSSV)is one kind of destructive virus affecting shrimp industry in the world.Output of Fenneropenaeus chinensis which once was a staple species has been greatly affected by outbreak of WSSV since 1993 in China.To produce WSSV-resistance shrimp strains is an important approach to revive the shrimp industry.To select survived F.chinensis as broodstock via WSSV challenge test can increase the efficiency of breeding.However,mortality of shrimps could reach nearly 100% once they were infected with WSSV,which increases the difficulty for shrimp breeding.In this study for the purpose to reveal effect of different WSSV amount on F.chinensis,juvenile F.chinensis were infected with a certain amount of WSSV individually and the survival time and survival rate were observed.The results showed that survival time of shrimps infected with 5.2×10^8 copies,1.0×10⁹ copies and 2.1×10⁹ copies of WSSV respectively was 389.3,323.3 and 187.3 h and significant different(P<0.01).The cumulative mortalities of all the shrimps finally reached 100%.The survival time increased while shrimps were fed with less WSSV bait.For the purpose to reveal the effect of diets on the ability for disease resistance,shrimps infected with WSSV were fed with different diets of live brine shrimps,commercial bait and fresh clam.It was showed that the survival time of infected shrimps fed with live brine shrimps,fresh clam and commercial bait respectively was 281.7,173.9 and 164.9 h.The survival time of infected shrimp fed with live brine shrimps was significantly higher than that fed by commercial bait and fresh clam respectively(P<0.01).There was no significant difference of survival time between infected shrimps fed with commercial bait and fresh clam(P>0.05).Cumulative mortalities of all the groups of infected shrimps that fed with different diets were 100%.Live brine shrimp is better than commercial bait and fresh clam for the purpose of enhancing shrimp WSSV resistance ability.

Abstract:Fat greenling(Hexagrammos otakii)is one of the important commercial fishes along the northern coast of China,and its artificial breeding and culture have been developed recently.Primary culture of fin,lip and kidney from H.otakii was studied using tissue explant method.Until now cells of three tissues have been subcultured at passage 30 for fin,passage 31 for lip,passage 35 for kidney respectively.The results showed that the fin and lip tissues could be dispersed quickly after being digested with 0.5% hyaluronidase and 0.2% collagenase Ⅱ.The optimal growth conditions for fin,lip and kidney tissues were 20% fetal bovine serum(FBS),5 ng/mL basic fibroblast growth factor(bFGF),20 μg/mL chondroitin sulfate and 40 ng/mL insulin like growth factor-I to medium DMEM/F12(pH 7.2)at 25 ℃.The doubling time of fin,lip and kindey cell numbers was about 58.7 h,50.4 h and 32.9 h at the 20th passage.Karyotype analysis of 100 metaphase plates revealed that the feature diploid chromosome number was 2n=48 in fin,lip and kidney cell at 25th passage.The cell livability of these three kinds of cell were(84.59±1.07)%,(85.75±1.03)% and(87.39±1.05)%,respectively,when recovered after being stored in liquid nitrogen for 60 d at the 20th.Now,the three kinds of cell have been preserved in China Center for Type Culture Collection(CCTCC).The methods of cell culture in vitro could be used to prevent and cure fish diseases and the pathomechanism study.

Abstract:Sea cucumber Apostichopus japonicus(Selenka)is a typical deposit-feeder,that is,food came from some organic matter in sediment such as bacteria,prozotoa,benthic microalgae and detritus of macroalgae.Artificial feeds are applied in some areas for sea cucumber farming to increase production of the pond cultured sea cucumbers,however,to what extent the artificial feed contributes to their growth is still unclear.Study on the food sources by a traditional way of direct gut content analysis has limitations.Stable isotope is a predictable method to evaluate food sources of A.japonicus.The present experiments were conducted to estimate contribution of artificial feed to growth of sea cucumber at five different stocking densities(5,10,15,25 and 35 ind/m^2)using a stable carbon isotope ratio(δ^13C)tracer.Sea cucumbers with an average weight of(4.78±0.58)g were randomly assigned to the enclosures.The animals were fed with artificial feed at 8:00 every day,at a feeding rate of about 5% wet weight.There were 5 treatments and 3 replicates in the experiment that lasted for 6 months.The control treatments without feed were set simultaneously.A lab experiment with four plastic aquaria(100 cm×60 cm×60 cm)was conducted and feeding time and ration were the same as the pond experiment.The results showed that the final weight and δ^13C value of sea cucumbers were affected markedly by both artificial feed and stocking density(P<0.05).With stocking density increasing,final weight of the animals decreased from(35.62±3.50)g to(24.59±4.38)g,and with stocking density increasing from 5 ind/m² to 35 ind/m²,the δ^13C values of sea cucumbers decreased from -13.262‰ to -15.102‰(P<0.05).Contributions of artificial feed to the growth of the sea cucumbers increased with increasing of stocking density,only 3.78%±2.98% at 5 ind/m^2 to 29.48%±3.31% at 35 ind/m².Previous studies showed that artificial feed contributed 23.0%-61.6% to the growth of shrimp and fish,which is higher than that to A.japonicus.This was caused by quality of artificial feed and the physiological characteristics of feeding behavior of sea cucumbers.

Abstract:FcβGBP-HDL was a pattern recognition receptor(PRR)which was up regulated post WSSV(white spot syndrome virus)challenge.To investigate the mechanism of transcription regulation,primers were designed based on the FcβGBP-HDL cDNA obtained,the gene and 5′-flanking sequences were isolated from the Chinese Shrimp Fenneropenaeus chinensis by genome walking,and the activity of promoter was analyzed by voter construction.The results showed that this gene was composed of only one exon spanning 6 713 bp.The 5′-flanking sequence(1 507 bp)of FcβGBP-HDL gene contained several typical transcription regulation elements,such as one promoter core sequence,two SRF,two TBP,one CTF and one CRE,and several transcription regulation elements of immune-gene in other arthropods,such as two GATA 1,three AP 1 and seven Sp1.The upstream sequence of FcβGBP HDL gene had highest level similarity with FcCTL(similarity 41.4%).The transcription levels of luciferase reporter gene were up regulated in all construction voters,highest in pGL3-FcβGBP2 with the deletion of two AP-1,and lowest in pGL3-FcβGBP4 with the deletion of promoter core sequence.The results indicated that promoter of FcβGBP-HDL might be regulated in immune reaction.

Abstract:Charybdis japonicais an important economic crab,very popular in Chinese coastal cities.But in recent years,the germplasm resources have been declining,and the work of resources proliferation already received great concern.With the development of artificial breeding and farming,the study of disease development and prevention and control measures is inevitable.Currently,few studies have related to the immune aspects of C.japonica.In order to explore the immune status,the non-specific immune function was studied in four populations of C.japonica from Dalian,Laizou Bay,Haizhou Bay and Xiangshan.Four types of tissues(muscle,gill,hepatopancreas and serum)and seven immunity related enzymes(ACP,AKP,LSZ,MDA,SOD,CAT and GSH-px)were detected.The results showed that the seven kinds of enzymes all exist in the four detected tissues,and the activities were different in different groups.The serum ACP,AKP and LSZ activities of Dalian was significantly higher than that of Haizhou Bay population(P<0.05),and there was no significant difference among other populations(P>0.05).The MDA content varied from different tissues.Dalian population was significantly higher than Haizhou Bay population in hepatopancreas and serum(P<0.05),but there were no significant differences in other tissues(P>0.05).For the SOD,CAT and GSH px activity of hepatopancreas and gill,Dalian population and Haizhou Bay population have shown significant difference(P<0.05).The results of the immunity enzyme activities are quite correlated with those of nonspecific immunity abilities of preventing disease and disease resistance evaluation.

Abstract:The scallopChlamys farreri is one of the major species cultured in North China,and its culture in commercial scale has been performed for more than 20 years.However,the great expansion and intensification have induced the occurrence of disease called “acute viral necrosis disease”(AVND)since 1990’s,and the cumulative mortality could be higher than 90%.This disease is caused by a virus called “acute viral necrosis virus”(AVNV),which is a spherical enveloped virus(130 to 170 nm in diameter)with spike-like surface protrusions and has been becoming the major limiting factor in the development of the scallop industry.In order to establish a rapid diagnosis method of AVNV parasitizing on scallop(C.farreri),a pair of primers of nested-PCR were developed by Accelrys gene 2.5 based on the conserved region of the AVNV genome(GenBank accession number:GQ153938)in this study.The reactive conditions such as concentration of Mg^2+,dNTPs and annealing temperature were optimized for the PCR system,and the expected products of the external and internal primers were 979 and 548 bp respectively.The results showed that the primers were specific for AVNV and did not amplify marine aquaculture animals’s and bacteria’s genome DNA,and the method can be stably amplified 5×10 copies virus particles in 5 pg total nucleic acid of scallop tissue.Also this method was successfully applied to the Sanggou Bay scallop samples detected.Therefore,it is confirmed that the method will be very useful for sensitive and specific detection of AVNV in the laboratory,and has high specificity,good reproducibility,which will definitely facilitate the monitoring of the epidemic disease in the future.

Abstract:Cyclin B gene was cloned from shrimp Metapenaeus affinis using RACE.The results showed that the full-length cDNA of cyclin B gene is 1 667 bp,including 1 209 bp coding sequence,the open reading frame encodes a putative peptide of 402 amino acid.The encoded protein does not contain signal peptides in this sequence.BLAST search has shown that the deduced peptide shared 90% sequence identity with that of Metapenaeus ensis.Gene expression profiling by semi quantitative reverse transcriptase polymerase chain reaction(RT- PCR)demonstrated that cyclin B expression levels are the highest in ovary and muscle,then in thoracic ganglion and heart,but almost no expression in gill,hepatopancreas and eyestalk.The results suggest that cyclin B expression levels may relate to its function as a regulator of cell division during cell cycle.Result of prokaryotic expression showed that,the recombinant protein was expressed with expression plasmid pET32a.At last we got a high quantity of recombinant protein(67 ku)in the optimum induction conditions,when the concentration of IPTG was 0.2 mmol/L for 4 h at 25 ℃ for 32a-MaCB.

Abstract:In order to use two-dimensional electrophoresis(2-DE)analysis in the muscle proteins of Chinese shrimp(Fenneropenaeus chinensis),its muscle proteins were separated using immobilized pH gradient 2-DE after dissolving.By optimizing different extraction methods,pH of IPG gel strips and its length,isoelectric focusing programs,and loading amount,etc,the proteins were successfully extracted from Chinese shrimp muscle and were separated by 2-DE.After silver staining or Coomassie brilliant blue staining,PDQuest image analysis software was applied to analyze the 2-DE images.The results showed that most of shrimp muscle protein isoelectric points were between 4 and 7.The 2 DE related techniques was constructed and optimized in muscle proteome of Chinese shrimp by comparative tests on different extraction methods,IPG gel strips,isoelectric focusing programs,salt bridge,and sample volume,etc.The results showed that the resolution and reproducibility of 2- DE profiles were significantly improved by adding thiourea,CHAPS and DTT in lysis buffer,active rehydrating of 17 cm(pH 4-7)IPG gel strips,loading the sample 120 μg,prolonging the time of desalting,increasing the voltage and power of isoelectric focusing,employing the salt bridge,preparing 12.5% SDS- PAGE gel,and dying the gels by the silver staining.It shows that the repeatability and stability are good enough.

Abstract:The swimming crab is widely distributed in the coastal waters of China,Japan and Korea.This species is one of the most common edible marine crab in the East Asia.To assess the fatty acid composition and variation of swimming crab populations and to develop a genetic resources identification technique based on fatty acids biomarkers,a multivariate analysis method was used to study fatty acid composition difference of swimming crab P.trituberculatus along China coast,the six geographical populations from China Sea(Dalian,Dongying,Lianyungang,Zhangzhou,Zhoushan and Zhanjiang)in autum and winter.A set of fatty acids as fingerprint technique was developed for identifying different populations.The crabs from the six population all contained 27 kinds of fatty acids among which the total volume of five kinds of fatty acids C18∶1n9t,C16∶0,C22∶1n9,DHA and EPA was over 82%,and multivariate ANOVA revealed that there were significant differences among the six populations(P<0.01).As fatty acid fingerprint markers,the five kinds of fatty acids C18∶1n9c,C20∶1,C22∶1n9,ARA and EPA,were effective to identify the six-populations of crabs for the total discriminant accuracy amounted to 88.46%.The results of cluster analysis indicated that there were no significant correlations between geographical distance and Euclidean distance based on 27 kinds of fatty acids.The reason of population differences of fatty acids was probably related with feed composition in habitat of the swimming crabs.

Abstract:Parvalbumin(PV)is the major allergen in fish products,which could induce serious IgE-mediated food anaphylaxis when sensitive people encountered with it.A variety of clinical symptoms including angioedema,respiratory symptoms,gastrointestinal symptoms,and even death could be caused after ingestion or contact with fish or fish products or even just inhalation of fish cooking vapors.Besides monitoring the allergens in foods,reducing allergenicity of fish products is another important way to decrease the incidence of fish allergy.Therefore,the purpose of this study was to analyze the change of immunoreactivity and digestion stability of parvalbumin after different food processings.Four processing methods including surimi preparation,roast,fried,high pressured methods were evaluated in this study.Tricine-SDS-PAGE and Western-blotting were used to determine the variation of contents,immunoreactivity and digested stability of parvalbumin in the processed products.The results showed that,though surimi prepartion,roast,and fried processing could increase the gastric stability,high pressure processing could effectively reduce the content and immunoreactivity of parvalbumin in fish products.And the significance of these findings remains to be established.