Abstract:In this study,the toxic mechanism of Patinopecten yessoensis responding to copper exposure was investigated through analyzing the effect of copper on bioaccumulation,activity of antioxidant enzymes and lipid metabolism in the gill or hepatopancreas.The results showed that the maximum accumulation of copper(0.025,0.05 and 0.10 mg/L)in gill occurred at 14 d(347.55 μg/g),6 d(340.72 μg/g)and 6 d(302.30 μg/g)respectively.With the time extension,accumulation of copper has a curvilinear relationship except 0.10 mg/L group.However,concentration of copper in 0.10 mg/L group of hepatopancreas presented liner changes with exposure time and the peak occurred at 14 d(60.88 μg/g).The corresponding changes in 0.025 and 0.05 mg/L were curvilinear and even decreased.Also,SOD,CAT and GPx activities were significantly suppressed in gill following copper exposure.However,in hepatopancreas,0.025 mg/L and 0.05 mg/L groups could induce the expression of antioxidant enzymes in 24 h except GPx in 0.05 mg/L group.The activities of all the three enzymes in 0.10 mg/L group were suppressed during all the experiment.Furthermore,the key regulation factors related to lipid metabolism including PPARs-γ2,6PGD,G6PD,FAS,LDH were found significantly up-regulated in the hepatopancreas,except ACC,CPT1 by using next generation sequencing technology.These results suggest that gill was the main accumulation tissue of copper in P.yessoensis,while the hepatopancreas was more sensitive to higher concentration.Also,the toxicity of copper was more serious than cadmium for P.yessoensis by damaging the activity of antioxidant enzymes in both tissues and inducing the accumulation of lipid in hepatopancreas.

Abstract:Using the monoclonal antibodies(Mab)3F4 against granulocyte of swimming crab(Portunus trituberculatus)to research the cross reaction of other seven species of crustaceans(Penaeus vannamei,Fenneropenaeus chinensis,Metapenaeus ensis,Oratosquilla oratoria,Eriocheir sinensis,Petalomera japonica,Cancer magister).In this paper,laser scanning confocal microscopy(LSCM),Western blot and Flow Cytometry(FCM)were employed to detect the relationship between these seven species.Balb/C mice were immunized by heamocytes of Portunus trituberculatus.Spleens were dissected from the immunized Balb/c mice and the cells were fused with SP2/0 myeloma cell line using polyethylene glycol as fusogen.Hybridomaculture mediums of hybridoma cells were selected using Indirect Immunofluorescence Assay(IFA)screening methods.One positive hybridoma was found which secreted high titer antibody,followed by single-cell cloning and subcloning by limiting dilution method and tested again for their specificity by IFA at the appropriate stage of growth.Single-cell cloning was repeated at least twice so as to prove the monoclonality of the hybridoma culture supernatant.We got the MAb 3F4 specific against granulocyte of swimming crab(Portunus trituberculatus).MAb 3F4 showed strong immunofluorescent reaction with granulocytes,but no reaction with hyalinocytes.In cross-reactivity,LSCM showed the MAb 3F4 could also react with haemocytes of Eriocheir sinensis and Petalomera japonica. Sorted by FCM,MAb 3F4 was positive to the two types of heamocytes of Eriocheir sinensis(hyalinocyte and granulocyte),and the percentage of positive rate were 57.72% and 77.05% respectively.Reacting with haemocyte of Petalomera japonica was mainly positive to granulocyte,whose positive rate was 82.59%,but that to the hyalinocyte was only 9.57%.Analyzed by Western blot,MAb 3F4 was recognized as one heamocyte protein of Eriocheir sinensis,whose molecule weight was 30 ku,but there was no protein band on nitrocellulose membrane of Petalomera japonica.

Abstract:Olfaction is essential for fish to detect odorant elements in environment.The sense of smell plays an important role in locating food,navigating,detecting predators,and spawning migration.The olfactory function is produced by the olfactory transduction pathway and activated by the olfactory receptors through binding of odorant elements.In vertebrates,the olfactory receptor gene families are recently classified into five families,the main olfactory receptor(MOR),vomeronasal type-1 receptor(V1R/ORA),vomeronasal type-2 receptor(V2R/OlfC),trace amine-associated receptor(TAAR)and formyl peptide receptor(FPR).More than 50% of vertebrates are fishes,which own important biological status.However,there are few articles published in this field in China,although more and more attention has been focused and many findings have been reported.This review describes the recent advance in studying the five olfactory receptor gene families in fishes,and makes a discussion and a summary of the problems encountered during the research,as well as the corresponding measures to resolve them.Finally we open an outlook to the further study in olfactory receptor genes.We aim to provide references for the research on olfactory receptors of fish in China.

Abstract:In order to explore the differences between Portunus trituberculatus “Keyong 1”(referred to as “Keyong 1”)and the 1st generation of wild crab(referred to as“common crab”)in terms of growth rate,external characterization and Vibrio alginolyticus resistance,“Keyong 1” and common crab living in the same growth conditions were measured for each morphological trait on the 30th,60th,90th,120th,150th days and according to the injection target body weight,injected into the body cavity with 1.14×10⁷ cfu/mL bacteria of V.alginolyticus at 10 μL/g dose on the 60th,90th,120th,150th days.The results show that “Keyong 1” group was significantly greater than common crab group in body weight on the 60th,90th,120th,150th days;The difference between “Keyong 1” group and common crab group is not significant in external characterization;“Keyong 1” group was significantly stronger than common crab group in resistance to V.alginolyticus on the 60th,90th,120th,150th days.To the same breed P.trituberculatus,the same concentration of V.alginolyticus according to the injection target body weight was injected into the body cavity with 1.14×10⁷ cfu/mL bacteria of V.alginolyticus at 10 μL/g dose had a relatively stable mortality from the 60th to 150th days.

Abstract:To study migratory routes of the largehead hairtail,Trichiurus lepturus,in the East China Sea(ECS),the Yellow Sea(YS)and the Bohai Sea(BS),the fishing records were analyzed with the data collected in the years 1971,1972,1981 and 1982.Firstly,moving images of largehead hairtail were plotced.Based on the images,migratory routes of the fish were further studied in the area.The results showed that wintering ground,spawning ground and feeding ground of the fish were wide and overlapped in space with each other.Furthermore,these migrations of the fish were overlapped in month,then showed its characteristics of batch migration.Overlapping in ground spaces and migrating of batch were two important and ecological characteristics for largehead hairtail.The wintering grounds of north population of the fish was located at 32°00'-36°00'N,124°00'-126°00'E,in the waters of yellow sea warm current.In March-July of each year,the largehead hairtail groups entered Haizhou Bay,Rushan Bay,Sea Island and bays in Bohai,the places were spawning ground and feeding ground for the fish.In Septamber-November,the fish groups moved out of Bohai,passed the Chengshan Cape,collected the fish groups from Haizhou Bay,Rushan Bay,Sea Island along waters of the forbidden fishing zone line,on the way to go to its wintering ground.When the groups arrived at 34°00'N,the batch groups turned to the southeast,to the wintering ground lies in the south-east of the Yellow Sea.The wintering grounds of the south population were mainly located in the waters near the forbidden fishing zoneline in the mid-southern East China Sea,followed by the waters in west of Jeju Island.From March-April,the fish groups moved north-east towards west waters of the Zhoushan Fishing Ground and the Changjiang Estuary Fishing Ground roughly at the speed of one degree of latitude for every two months until May-July.The fish finished spawning migration by reaching the Zhoushan Fishing Ground and the Changjiang Estuary Fishing Ground which were their spawning grounds.During July-August,after spawning,feeding groups migrated north to Dasha Fishing Ground at the south of YS for feeding and growing.In September,the groups arrived near the forbidden fishing zoneline at 35°00'N,the northern feeding ground of south population.Under influence of cold northwest monsoon after October,the population returned to the Changjiang Estuary.From there,the fish split into two groups,the major one migrated southward back to wintering ground in nearshore waters in the mid-southern East China Sea,and the small group moved back to the wintering ground in offshore waters.The main water masses affecting wintering migratory route of the fish are the Taiwan Warm Current and the Yellow Sea Warm Current.

Abstract:This study aims to analyze and compare the community structure of planktonic ostracods assemblages between Nansha reef area and open water area,and the distribution pattern influenced by ocean current movement was discussed.During April and September in 2013,27 sampling stations were set up in Nansha Islands Waters,and planktonic ostracods were collected at depths from 200 m to the surface using medium plankton net.Samples were treated and counted according to the standard methods.The results are the following findings:(1)A total,of 45 planktonic ostracods species were identified,belonging to 19 genera,4 subfamilies,and 2 families.Twenty-one species only occurred in spring and 38 species only in autumn,and 14 species occurred in both seasons.(2)The dominant species were different between two seasons;Euconchoecia maimai,Porroecia porrecta,and Conchoecetta acuminata dominated in spring,while Cypridina nami,Euconchoecia bifurata,Porroecia spinirostris,Microconchoecia curta,Metaconchoecia macromma dominated in autumn.All dominant species are either eurythemal-euryhaline or warm-hyperhaline.Studies show that,(1)There are remarkable differences in the density of planktonic ostracods between two seasons.In spring,density ranged from 0.03 to 8.25 ind/m³,averaging 1.4 ind/m³;in autumn,density ranged from 0.05 to 18.92 ind/m³ with an average of 4.11 ind/m³.High density occurred in certain limited areas due to ocean circulation.In spring,three high density areas were located in the west,northwest and southeast corners of Nansha Islands waters,respectively.In autumn high density area occupied the vast east part.(2)Species richness,diversity and evenness were 0.59,1.37 and 0.55,respectively,with much higher values in spring than in autumn.(3)Planktonic ostracods can be divided into two communities in both seasons characterizing,reefs and open waters,and seasonal variation in community distribution was impacted strongly by ocean current change.(4)The tropical cyclonic and anticyclonic circulation driven by monsoon can be regarded as the dominant factor to influence the structure of planktonic ostracods community.

Abstract:The distribution of zooplankton in the area of radiate sand ridges in north Jiangsu shoal was investigated during May and September in 2012.The relationship between the distribution of zooplankton,topography and some other environmental factors were analyzed by using indicators and methods of gathering strength,variance contribution,dominance index,stepwise regression,etc..The results showed that both of the biomass and abundance of zooplankton have obvious seasonal variation.Spatially,the quantity of zooplankton was higher in sand ridges than that in tidal creeks.In spring,the average biomass of different areas were 606.63 mg/m³(sand ridges),428.00 mg/m³(shallow tidal creeks)and 178.55 mg/m³(deep tidal creeks).The average abundance were 560.76 ind/m³(sand ridges),389.39 ind/m³(shallow tidal creeks)and 161.28 ind/m³(deep tidal creeks).However,in autumn,the average biomass of different areas were 208.01 mg/m³(sand ridges),181.30 mg/m³(shallow tidal creeks)and 60.47 mg/m³(deep tidal creeks).And the average abundance were 184.15 ind/m³(sand ridges),56.42 ind/m³(shallow tidal creeks)and 112.14 ind/m³(deep tidal creeks).The study showed that the Copepods was predominant species in two seasons.The research indicated that the distribution of zooplankton was associated with topography and tidal waves in the north Jiangsu shoal.Some reasons can explain it.Firstly,the water velocity was influenced by the topography,where the water velocity was higher in sand ridges than tidal creeks.Secondly,the East China Sea tidal wave and the South Yellow Sea tidal wave converged in north Jiangsu shoal.Sediment can be deposited easily in low flow velocity water,and upwelling waters brought a wealth of nutrients.It would be beneficial to the aggregation and feed of zooplankton,which lead to high biomass and abundance of zooplankton in sand ridges.The dominant species tend to be highly aggregated,which contributed to the main component of economic fishes' bait.The food availability and quality of north Jiangsu shoal were higher than other water areas,which played a crucial role in the formation of Lüsi Fishing Ground.

Abstract:The samples of stomach content of Pelteobagrus nitidus were collected at 6 stations in Dianshan Lake from August 2009 to July 2010,and the food composition was investigated based on the analysis of stomach contents.Results showed that P.nitidus preyed around the year and its feeding intensity had significant seasonal variations with the highest index of fullness in autumn and the lowest in summer and with the highest rate of empty stomach in spring and the relatively lower in the other seasons.Seven broad diet categories(11 small diet categories)were identified.Cluster analysis showed that the occurrance rate(F%),numerical percentage(N%)and percent index of relative importance(IRI%)were consistent in measuring contribution of each food category to P.nitidus.There were significant differences between weight percentage(W%)and each of the above three indices in measuring the contribution.The polychaete,benthic crustacea,aquatic insect and zooplankton were the main food sources for P.nitidus in terms of the above-mentioned four indices.The numerical and gravimetric compositions both significantly differed between seasons,the former also conspicuously differed among stations,but the latter did not display marked spatial variation with benthic crustacea as the predominant food source for P.nitidus caught at each station.There were obvious spatial changes in diversity of food compositon with the lowest in winter(0.15),and the largest H' at Station 5(1.52)and the smallest at Station 6(0.72).P.nitidus displays high plasticity in food composition by comparison with the feeding spectrum of this species from the other lakes.

Abstract:Behavioral preferences of juvenile Scapharca broughtonii with 10 mmS.broughtonii to five kinds of substrates(mud group added)was investigated.The results showed that 10 mm

Abstract:The traditional morphological methods implemented in mathematical statistic software,including clustering analysis,principal component analysis and discriminant analysis,have been widely used in dealing with morphological data in fishes.However,a series of molecular genetics software have also been developed coupling with development of modern molecular biology techniques.In this paper,in order to study the applicability of the molecular population genetics methods in analyzing fish morphological data,the morphological traits of nine mating combinations from a complete diallel cross of Xingguo red common carp(C.carpio var.singuonensis),Glass red common carp(C.carpio var.wananensis)and Purse red common carp(C.carpio var.wuyuanensis),were used to conduct comparative analysis of traditional morphological methods and molecular population genetics methods.The results showed that the same profiles of clustering analysis were found from the two kinds of methods.The principal component analysis indicated that the traditional morphological methods can better reflect the morphological variations among individuals,while the molecular population genetics methods can better reflect the morphological differences among populations.The discriminant analysis showed the similar discriminant accuracy of the two kinds of methods.Furthermore,the genetic clustering analysis results using software STRUCTURE illustrated the consistency of the two kinds of methods.Consequently,the study produced the same results from the traditional morphological methods and molecular population genetics methods in dealing with morphological data of red common carp.It is feasible to analyze morphological data of common carp using molecular population genetics methods.

Abstract:A feeding experiment was conducted to investigate the effects of lysine,arginine and their interaction on the growth performance,whole body composition and muscle amino acid levels of juvenile turbot(Scophthalmus maximus L.)[initial weight(18.48±0.16)g].Nine isonitrogenous and isoenergetic diets according to a 3×3 factorial were designed:the basal diet as the control diet supplemented with three levels of arginine(0%,0.9% and 2.0%)crossed with lysine(0%,1.19% and 2.39%).Each diet was randomly allocated to triplicate groups of juvenile turbots for 10 weeks.The results showed that the weight gain rate(WGR)and specific growth rate(SGR)were significantly increased at the level of lysine(1.19%)supplement diets(P<0.05),while the addition of arginine had no additional effect(P>0.05).Dietary lysine and arginine had significant interactions on feed efficiency rate(FER),protein efficiency rate(PER),protein retention(PR)and crude protein content(P<0.05),the maximal value of which was obtained at the levels of lysine(1.19%)and arginine(0.9%)supplement diet.However,weight gain(WGR),the specific growth rate(SGR),crude lipid content,ash and body indexes were not affected by the interaction of arginine and lysine(P>0.05).Crude lipid content and moisture were decreased with the increased levels of lysine(P<0.05),while the gradient arginine showed no significant effect(P>0.05).Hepatosomatic indexes(HSI)and visceral indexes(VSI)were greatly reduced with the increased levels of both lysine and arginine(P<0.05).Dietary lysine and arginine had significant interactions on majority muscle individual amino acid levels,the minimum value of which was obtained at the levels of lysine(2.39%)and arginine(0.9%)supplement diet(P<0.05).It was concluded that lysine and arginine had interaction for the aspects of feed efficiency,body protein retention and muscle individual amino acid levels,while the interaction effect of the growth and body index does not exist;juvenile turbot maintained maximal growth and feed efficiency at the supplement levels of lysine(1.19%)and arginine(0.9%);compared to arginine,lysine seems to be the major factor,proper supplementation promotes growth,while excessive level inhibits turbot growth,feed efficiency and muscle amino acids retention through the antagonism with arginine.

Abstract:For preliminary research on the biology function of MKK4 in Fenneropenaeus chinensis in this study,full-length cDNA sequence of MKK4 gene in F.chinensis was cloned using RACE method and was named FcMKK4.The full-length of cDNA sequence of MKK4 is 2 064 bp,which contains a 214 bp 5'-UTR,629 bp 3'-UTR and 1 221 bp open reading frame(ORF)that encodes 406 amino acid residues.Its isoelectric point(PI)was 8.50 and molecular mass was 45.94 ku.Homology analysis revealed that the amino acid sequences of FcMKK4 were highly homogenous with MKK4 of other species,80% with Ixodes Scapularis,and 78% with Harpegnathos saltator.The phylogenetic analysis showed that F.chinensis FcMKK4 was in the same class with other arthropods' MKK4.The expression level of FcMKK4 gene in different tissues was analyzed by quantitative real-time PCR.The results indicated that the highest expression level of FcMKK4 gene was in muscle,and that in hepatopancreas was the second.Real-time PCR analysis showed that the expression level of FcMKK4 was up-regulated significantly in muscle,hepatopancreas,hemocytes,gill,heart,intestine and stomach after stimulation with ammonia-N stress,and FcMKK4 showed different expression profiles in different tissues.The results implied that FcMKK4 might play an important role in abiotic stress response in F.chinensis.

Abstract:In order to investigate the effects of grass carp diets,which were added with stickwater meal and fish oil to supplement fish required part of growth factor,on grass carp growth,liver fat content and serum indicators.In practical formulation mode,six experimental diets added 6% pork powder meal(6P),3% pork powder meal +3% stickwater meal(3P3SW),6% stickwater meal(6SW),3% pork powder meal +3% stickwater meal +2% fish oil(3P3SW2O),6% stickwater +2% fish oil(6SW2O),6% fish meal(6F control)were prepared.All the experimental diets were isonitrogenous and isoenergetic.The diets were fed to grass carps[mean initial body weight of(80.0±0.2)g]in pond cages for 72 d.The results showed that,there were no significant differences in SGR,FCR,PRR,ERR and HSI between 6F and 6P.After adding stickwater meal,SGR of 3P3SW and 6SW improved by 9.1%-9.6% compared with 6F,when it comes to FCR,it decreased by 8.5%-11.2%,the differences were significant(P<0.05),but there were no significant differences between 3P3SW and 6SW(P>0.05),while in PRR,FRR and ERR terms,3P3SW,6SW have some improvements.After adding fish oil,compared 3P3SW2O,6SW2O with 3P3SW,6SW,the SGR decreased by 9.7%,15.4%,the FCR increased by 6.9%,31.1%,the differences were significant(P<0.05),except the FCR of 3P3SW2O and 3P3SW.In terms of 3P3SW2O and 6SW2O,PRR and ERR declined;at the same time,FRR and liver fat content increased,and HSI,TBA,CHOL,HDL and LDL with liver fat showed the same trend,and the highest group of these indicators was 6SW2O which had significant differences(P<0.05),compared with all other groups.In the correlation analysis of free amino acids,biogenic amines with FCR and SGR,Pearsion correlation coefficients of free lysine,taurine and putrescine were all greater than 0.8,and presented a quadratic function.So,in the effect of promoting growth,stickwater meal is better than fish meal.Adding small amount of stickwater meal to diets which have no fish meal can not only meet the needs of grass carp but also save the amount of fish meal,and this growth-promoting effect is closely related with free Lys,Tau and Put.Fish oil promotes fat deposition in the body and liver,burdens the lipid metabolism.Together with its easily oxidized characteristics,its use in aquaculture feed is limited.

Abstract:Acetyl-CoA C-acetyltransferase(AACT)is the starting enzymes in mevalonate pathway of terpenoids biosynthesis.To reveal the function of this enzyme during the ovarian development of the crustaceans,we used reversed transcript PCR(RT-PCR)and rapid amplification of cDNA ends(RACE)techniques to clone the AACT(GeneBank accession number:KM033231).The full length of Pt-AACT is 1 630 bp,including a 5'-untranslated region(UTR)of 101 bp,a 3'-UTR of 395 bp and an opening reading frame(ORF)of 1 134 bp encoding 377 amino acid protein.The structures of AACT proteins have two special conserved sequences of thiolase,belonging to stable hydrophobic proteins without an obvious transmembrane region.The deduced amino acid sequence of Pt-AACT exhibited the highest identity(72%)with AACT of Nasonia vitripennis and Aedes aegypti.The phylogenetic tree analysis showed that Pt-AACT was clustered in insect AACTs.Tissue distribution by quantitative real-time PCR(qRT-PCR)illustrated that the Pt-AACT was most highly expressed in the MOs.During the ovarian development,expression of Pt-AACT in MOs significantly remained the highest level at stage Ⅰ.Therefore,AACT plays an important role in ovarian development regulation of P.Trituberculatus.

Abstract:Insulin like growth factor binding protein(IGFBP)is a member of IGFBP family participating in transportation,localization and biological regulation of IGFs.Here,we conducted RACE-PCR and LONG-PCR for the cloning of full length cDNA and DNA of IGFBP gene in Sinonovacula constricta.Quantitative real-time PCR analysis revealed that IGFBP transcript was expressed in a wide range of tissues and developmental stages.Additional growth-linked SNP were discovered based on association study.The full length of IGFBP cDNA sequence consists of 631 base pairs(bp),comprising a 60 bp 5' untranslated region(UTR),a 136-bp 3' UTR,and an open reading frame(ORF)which is 435 bp in length.The ORF encodes 144 amino acids with a signal peptide(18 amino acids)and 12 cysteine residues in the conserve N-terminal.There is one intron(2 687 bp)and two exons(200 and 245 bp)in the sequence of IGFBP gene(3 122 bp).Based on quantitative real-time PCR analysis,we found the highest level of IGFBP expression in the digestive gland.Also the transcript was highly detected in juvenile stages rather than other stages.Four SNPs were identified in IGFBP genomic DNA and one of them was significantly associated with the shell length and body weight.Our results indicated that IGFBP gene plays important roles in the growth and development in S.constricta.

Abstract:A 3-week feeding experiment and a 2-week Vibrio parahaemolytics infection experiment were conducted to study the effects of adding probiotics to the feed on non-specific immune gene expression and resistance to V.parahaemolytics of L.vannamei[initial body weight(6.95±1.20)g].Litopenaeus vannamei was fed four different diets:Control group(commercial fodder)and experimental group(feed supplemented with Bacillus licheniformis;or with Bacillus subtilis and with mixed diet comprising Bacillus licheniformis/Bacillus subtilis at the ratio of 1:1),the final concentration of probiotics was 10⁷ cfu/g.And real-time PCR method was used to study the non-specific immune gene expression of L.vannamei in Bacillus licheniformis/Bacillus subtilis experimental group,which is the highest protection group after being infected with V.parahaemolytics.The results showed that all probiotics groups can effectively improve the ability of shrimp to resist the V.parahaemolytics's infection,and compound probiotics can better resist the V.parahaemolytics infections,its relative protection ratio is 31.11%.After infection with the V.parahaemolytics,the relative expressions of IMD,penaiedin 3a,proPO,LZM and Crustin were obviously up-regulated in hemolymph of shrimp fed diets supplemented with the compound probiotics(Bacillus licheniformis/Bacillus subtilis)compared to the control during the immunization.In hemolymph,the relative expression of immune deficiency gene,penaiedin 3a,prophenoloxidase,Lysozyme and Crustin were increased to the maximum levels at 18-24 h.The experimental result indicates:adding probiotics to the feed can effectively improve the ability of L.vannamei to resist the V.parahaemolytics infection,and the improvement of this ability may be made through enhancing the disease resistance related gene expression.