Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38222
Cai Qiufeng, Wang Xichang, G. Cheng, Shen Haiwang, Liu Guangming, Cao Min-jie
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.
{"title":"Effects of processing methods on the immunoreactivity of silver carp parvalbumin","authors":"Cai Qiufeng, Wang Xichang, G. Cheng, Shen Haiwang, Liu Guangming, Cao Min-jie","doi":"10.3724/SP.J.1231.2013.38222","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38222","url":null,"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.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"297-302"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69963860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38290
Weihua Lv, Y. Kuang, Xianhu Zheng, D. Cao, Chao Li, Cui-yun Lu, Xiaowen Sun
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.
{"title":"Quantitative trait locus analysis of four economic traits in one kind of common carp","authors":"Weihua Lv, Y. Kuang, Xianhu Zheng, D. Cao, Chao Li, Cui-yun Lu, Xiaowen Sun","doi":"10.3724/SP.J.1231.2013.38290","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38290","url":null,"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.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"161"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69964656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38304
J. Long, Xinping Zhu, Yan Shi, Jian Zhao, X. Hong
Macrophage inflammatory protein-3α is a kind of inducible secreted protein which plays an important role in the immune response and inflammation damage in many diseases through the corresponding receptors on a variety of immune cells to produce chemokines.We obtained the ORF of MaCCL20 by RT-PCR,inserted into the expression vector pET-32a,and then transformed into E.coli BL21.We expressed the fusion protein pET-32a-CCL20 after IPTG induction,prepared the polyclonal antibodies and did research on Western blot and antimicrobial activity of it.The experiments show that the fusion protein is highly expressed in the conditions of 37 ℃,0.8 mmol/L IPTG for 4 hours.The SDS-PAGE gel electrophoresis figure shows that the relative molecular weight of the fusion protein is 30 ku which is the same as predicted before.We got a single band after His Bind nickel column purification indicating the high purity of MaCCL20 recombinant protein.The antibacterial experiments show that MaCCL20 recombinant protein has strong antibacterial effects on Staphylococcus aureus,Serratia marcescens,and Aeromonas hydrophila.
{"title":"Recombinant expression and antimicrobial activity analysis of macrophage inflammatory protein-3 α in Mauremys mutica","authors":"J. Long, Xinping Zhu, Yan Shi, Jian Zhao, X. Hong","doi":"10.3724/SP.J.1231.2013.38304","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38304","url":null,"abstract":"Macrophage inflammatory protein-3α is a kind of inducible secreted protein which plays an important role in the immune response and inflammation damage in many diseases through the corresponding receptors on a variety of immune cells to produce chemokines.We obtained the ORF of MaCCL20 by RT-PCR,inserted into the expression vector pET-32a,and then transformed into E.coli BL21.We expressed the fusion protein pET-32a-CCL20 after IPTG induction,prepared the polyclonal antibodies and did research on Western blot and antimicrobial activity of it.The experiments show that the fusion protein is highly expressed in the conditions of 37 ℃,0.8 mmol/L IPTG for 4 hours.The SDS-PAGE gel electrophoresis figure shows that the relative molecular weight of the fusion protein is 30 ku which is the same as predicted before.We got a single band after His Bind nickel column purification indicating the high purity of MaCCL20 recombinant protein.The antibacterial experiments show that MaCCL20 recombinant protein has strong antibacterial effects on Staphylococcus aureus,Serratia marcescens,and Aeromonas hydrophila.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"593"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69964853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38314
Xia Li, Y. Guo, Yan-jie Qin, Zhiqiang Jiang
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 2n=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.
{"title":"Limited subculture and biological characteristic analysis of three cell lines from fat greenling(Hexagrammos otakii)","authors":"Xia Li, Y. Guo, Yan-jie Qin, Zhiqiang Jiang","doi":"10.3724/SP.J.1231.2013.38314","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38314","url":null,"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 2n=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.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"168"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69965237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38366
Lijing Chen, Z. Mei, Youjia Kong, Qigen Liu
As a dominant group of zooplankton in fresh water worldwide,Cladocera plays an important role in the material circulation and energy flow of water ecosystem,and it is not only a consumer for small animals and algae,but also food for zooplanktivorous fish.Many current studies have showed that non-classical bio-manipulation,a way to prevent water bloom and improve water environment by controlling fierce fish and stocking planktonivorous filter-feeding fish(such as Hypophthalmichthys molitrix and Aristichys mobilis),can be effective.However,to some extent,phytoplankton and community structure of cladocera may be varied with different stocking patterns,fish density and lake type.Lake Gehu,a total area of 164 km2 crossing Wujin and Yixing,is located in the south of Jiangsu province.In recent years,more attention was paid to the researches about the influence of H.ypophthalmichthys molitrix and Aristichys mobilis on lake ecosystems,of which the past domestic studies were mainly focused on aquatic plants,physical and chemical factors,the control of eutrophication and phytoplankton,however,no reports have been made in the bio-manipulation fish pen of Lake Gehu about the influences of abiotic environmental factors and filter-feeding fish on cladocera,and that is our work to fill in the blanks.Thus,from December 2009 to November 2010,a large bio-manipulation fish pen with the stock of silver carp(H.molitrix)and bighead carp(A.mobilis)inside was built to control the cyanobacterial bloom in Lake Gehu,an investigation and laboratory analysis was carried out to compare the variation of the cladoceran community structure between region A outside of the fish pen and regions B,D,C with different ratios of fish(1∶ 4,1∶ 2,1∶ 1.5),which was combined to discuss the influences of bio-manipulation fish pen on cladocera and the effects on improving eutrophic water.Through the identification,32 species of cladocera,belonging to 7 families,14 genera were collected inside and outside of the fish pen where there were no differences between species,and the annual average density and biomass of region A were both higher than any region inside of the fish pen,having a significant difference(P0.05)with region B,respectively.The main dominant species were Bosmina longirostris,Ceriodaphnia cornuta and Moina sp..In August,the density of Moina sp.in region A had significant differences from region B and D,while 3 regions inside of fish pen had no apparent differences,et al.,the results of which indicated that Moina sp.with a larger volume was easier to be preyed by fish than the other two small species.Moreover,some characteristic parameters of water were investigated to depict the correlation between cladoceran and environmental variables.CCA(Canonical Correspondence Analysis)showed that water temperature,permanganate index(CODMn),nitrate nitrogen(NO3-N)and water transparency(SD)were the main abiotic factors affecting the community structure of cladocera.
{"title":"The influences of silver carp and bighead carp in bio-manipulation pen on the community structure of cladocera in Lake Gehu","authors":"Lijing Chen, Z. Mei, Youjia Kong, Qigen Liu","doi":"10.3724/SP.J.1231.2013.38366","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38366","url":null,"abstract":"As a dominant group of zooplankton in fresh water worldwide,Cladocera plays an important role in the material circulation and energy flow of water ecosystem,and it is not only a consumer for small animals and algae,but also food for zooplanktivorous fish.Many current studies have showed that non-classical bio-manipulation,a way to prevent water bloom and improve water environment by controlling fierce fish and stocking planktonivorous filter-feeding fish(such as Hypophthalmichthys molitrix and Aristichys mobilis),can be effective.However,to some extent,phytoplankton and community structure of cladocera may be varied with different stocking patterns,fish density and lake type.Lake Gehu,a total area of 164 km2 crossing Wujin and Yixing,is located in the south of Jiangsu province.In recent years,more attention was paid to the researches about the influence of H.ypophthalmichthys molitrix and Aristichys mobilis on lake ecosystems,of which the past domestic studies were mainly focused on aquatic plants,physical and chemical factors,the control of eutrophication and phytoplankton,however,no reports have been made in the bio-manipulation fish pen of Lake Gehu about the influences of abiotic environmental factors and filter-feeding fish on cladocera,and that is our work to fill in the blanks.Thus,from December 2009 to November 2010,a large bio-manipulation fish pen with the stock of silver carp(H.molitrix)and bighead carp(A.mobilis)inside was built to control the cyanobacterial bloom in Lake Gehu,an investigation and laboratory analysis was carried out to compare the variation of the cladoceran community structure between region A outside of the fish pen and regions B,D,C with different ratios of fish(1∶ 4,1∶ 2,1∶ 1.5),which was combined to discuss the influences of bio-manipulation fish pen on cladocera and the effects on improving eutrophic water.Through the identification,32 species of cladocera,belonging to 7 families,14 genera were collected inside and outside of the fish pen where there were no differences between species,and the annual average density and biomass of region A were both higher than any region inside of the fish pen,having a significant difference(P0.05)with region B,respectively.The main dominant species were Bosmina longirostris,Ceriodaphnia cornuta and Moina sp..In August,the density of Moina sp.in region A had significant differences from region B and D,while 3 regions inside of fish pen had no apparent differences,et al.,the results of which indicated that Moina sp.with a larger volume was easier to be preyed by fish than the other two small species.Moreover,some characteristic parameters of water were investigated to depict the correlation between cladoceran and environmental variables.CCA(Canonical Correspondence Analysis)showed that water temperature,permanganate index(CODMn),nitrate nitrogen(NO3-N)and water transparency(SD)were the main abiotic factors affecting the community structure of cladocera.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"545"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69965623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38401
Ji-hong Zhang, Tao Wu, Ya-Jie Gao, W. Tang
5 species bivalves' feeding behavior on different food,including flounder(Paralichthys olivaceus)faeces,residul feed and deposition was studied so as to discuss the potential IMTA based on filter-feeding bivalves.Results showed the 5 species bivalves could feed on fish faeces,residul feed and deposition.However,different species showed different feeding behaviors.Ingestion rates of oyster,clam and scallop were significantly higher than the other 2 species(two-way ANOVA,P0.01),and absorption efficiency(AE)and absorption rate(AR)of the 5 species bivalves were not significantly different.Diet had significant influence on the feeding behavior.With particle organic matter(POM)increasing,AE increased.There were significantly linear relationship between AR and POM.TPM threshold for pseudo-feces production of oyster and scallop and was 26.24,21.64,27.00 mg/L,respectively,and the 5 species bivalves could regulate food intake by pseudo-feces production.The shellfish+fish IMTA mode,clam Ruditapes philippinarum,oyster Crassostrea gigas and scallop Chlamys farreri are relatively good candidate species.
{"title":"Feeding behavior of 5 species filter-feeding bivalves on Paralichthys olivaceus feed,fecal and sediment particulates in cage farming area","authors":"Ji-hong Zhang, Tao Wu, Ya-Jie Gao, W. Tang","doi":"10.3724/SP.J.1231.2013.38401","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38401","url":null,"abstract":"5 species bivalves' feeding behavior on different food,including flounder(Paralichthys olivaceus)faeces,residul feed and deposition was studied so as to discuss the potential IMTA based on filter-feeding bivalves.Results showed the 5 species bivalves could feed on fish faeces,residul feed and deposition.However,different species showed different feeding behaviors.Ingestion rates of oyster,clam and scallop were significantly higher than the other 2 species(two-way ANOVA,P0.01),and absorption efficiency(AE)and absorption rate(AR)of the 5 species bivalves were not significantly different.Diet had significant influence on the feeding behavior.With particle organic matter(POM)increasing,AE increased.There were significantly linear relationship between AR and POM.TPM threshold for pseudo-feces production of oyster and scallop and was 26.24,21.64,27.00 mg/L,respectively,and the 5 species bivalves could regulate food intake by pseudo-feces production.The shellfish+fish IMTA mode,clam Ruditapes philippinarum,oyster Crassostrea gigas and scallop Chlamys farreri are relatively good candidate species.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"727"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69966079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38420
K. Lian, Li-li Zhao, Linlin Zhang, P. Jia, Lijie Tang, Junwei Ge, L. Yijing, Min Liu
The recombinant protein IPNV VP2 was used as immunogen after purification by Ni-NTA.The 8-week-old BALB/c mice were intraperitoneally immunized with the VP2 protein for three times,then myeloma cells SP2/0 were fused with the spleen cells of the immunized BALB/c mice.Two hybridoma cell lines against the VP2 protein were obtained by screening with the indirect ELISA and limiting dilution assay,which were identified to be IgG1 subtype and named 5G10,5F3.The numbers of chromosomes of the two hybridomas were in the range of 75 to 120.Their antibody titers of cells culture supernatant were 1∶ 105,1∶ 102 respectively.Their titers of ascites were 1∶ 108,1∶ 104 respectively.Western-blot analysis and indirect immunofluorescence showed that the two McAbs could react with IPNV specificity.The 2 McAbs had no reactive capability with IHNV,VHSV,SVCV and HRV by the indirect ELISA.Antibodies additivity assay demonstrated that 5G10 and 5F3 recognized the different epitopes of IPNV VP2 nucleoprotein.We detected the clinical suffering from IPN rainbow trout liver tissue material,and the results confirmed that the 2 McAbs can be used for follow-up testing.
{"title":"Preparation and preliminary application of monoclonal antibodies against VP2 COE protein of infectious pancreatic necrosis virus","authors":"K. Lian, Li-li Zhao, Linlin Zhang, P. Jia, Lijie Tang, Junwei Ge, L. Yijing, Min Liu","doi":"10.3724/SP.J.1231.2013.38420","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38420","url":null,"abstract":"The recombinant protein IPNV VP2 was used as immunogen after purification by Ni-NTA.The 8-week-old BALB/c mice were intraperitoneally immunized with the VP2 protein for three times,then myeloma cells SP2/0 were fused with the spleen cells of the immunized BALB/c mice.Two hybridoma cell lines against the VP2 protein were obtained by screening with the indirect ELISA and limiting dilution assay,which were identified to be IgG1 subtype and named 5G10,5F3.The numbers of chromosomes of the two hybridomas were in the range of 75 to 120.Their antibody titers of cells culture supernatant were 1∶ 105,1∶ 102 respectively.Their titers of ascites were 1∶ 108,1∶ 104 respectively.Western-blot analysis and indirect immunofluorescence showed that the two McAbs could react with IPNV specificity.The 2 McAbs had no reactive capability with IHNV,VHSV,SVCV and HRV by the indirect ELISA.Antibodies additivity assay demonstrated that 5G10 and 5F3 recognized the different epitopes of IPNV VP2 nucleoprotein.We detected the clinical suffering from IPN rainbow trout liver tissue material,and the results confirmed that the 2 McAbs can be used for follow-up testing.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"77 1","pages":"1229"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69966725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Maternal transfer of immune factors from mother to eggs has been reported in many aquatic species,and these factors played important immune roles in developing embryos and larvae.However,the study on maternal immunity of shellfish remains lacking.In this study,maternal transfer and bacteriostasis of a C-type lectin in scallop(Patinopecten yessoensis)were analyzed.We detected the mRNA expression pattern of the C-type lectin gene in ovary and early developing larvae using the real-time quantitative PCR method,and tested the antibacterial activity of maternal C-type lectin in eggs by colony-forming unit(CFU)assay.The results of qRT-PCR showed that the expression of C-type lectin gene in ovary after bacterial challenge was time-dependent,and it was up-regulated gradually from 2 h after injection of bacteria and reached the peak at 8 h which was 6.2 times compared to the control group,and then dropped progressively to the original level.On the other hand,the fact that mRNA was found both in the eggs and larvae indicated that it could be transferred from mother to offspring.The mRNA expression in bacterial challenge group was significantly higher than those in normal group except for the 36 h point.The CFU assay revealed that the cytoplasm of eggs with protein concentration of 200 μg/mL and 400 μg/mL had lethal effect on the bacteria Vibrio anguillarum,and the function weakened significantly once the antibody of C-type lectin was added,which indicated the C-type lectin from mother had antibacterial activity.It is concluded that the maternal C-type lectin can transfer from mother to the offspring and has antibacterial activity.
据报道，在许多水生物种中，母体将免疫因子转移到卵中，这些因子在胚胎和幼虫的发育中起着重要的免疫作用。然而，对贝类母体免疫的研究仍然缺乏。本研究分析了一种c型凝集素在扇贝(Patinopecten yessoensis)中的母体转移和抑菌作用。采用实时定量PCR方法检测c型凝集素基因在卵巢和早期发育幼虫中的mRNA表达谱，并采用菌落形成单位(colony forming unit, CFU)法检测母体c型凝集素在卵中的抑菌活性。qRT-PCR结果显示，细菌攻毒后卵巢c型凝集素基因的表达具有时间依赖性，从注射细菌后2 h开始逐渐上调，8 h达到峰值，为对照组的6.2倍，之后逐渐回落至初始水平。另一方面，在卵和幼虫中都发现了mRNA，这表明它可以从母亲转移到后代。除36 h点外，细菌攻毒组mRNA表达量均显著高于正常组。CFU实验结果表明，蛋白浓度为200 μg/mL和400 μg/mL的卵细胞质对鳗弧菌均有致死作用，且加入c型凝集素抗体后作用明显减弱，说明母体c型凝集素具有抑菌活性。由此可见，母体c型凝集素可从母体传给子代，并具有抗菌活性。
{"title":"A preliminary study on the maternal transfer and bacteriostasis of a C-type lectin in scallop(Patinopecten yessoensis)","authors":"Biao Wu, Chang Chi, A. Yang, Shicui Zhang, Zhihong Liu, Liqing Zhou","doi":"10.3724/SP.J.1231.2013.38464","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38464","url":null,"abstract":"Maternal transfer of immune factors from mother to eggs has been reported in many aquatic species,and these factors played important immune roles in developing embryos and larvae.However,the study on maternal immunity of shellfish remains lacking.In this study,maternal transfer and bacteriostasis of a C-type lectin in scallop(Patinopecten yessoensis)were analyzed.We detected the mRNA expression pattern of the C-type lectin gene in ovary and early developing larvae using the real-time quantitative PCR method,and tested the antibacterial activity of maternal C-type lectin in eggs by colony-forming unit(CFU)assay.The results of qRT-PCR showed that the expression of C-type lectin gene in ovary after bacterial challenge was time-dependent,and it was up-regulated gradually from 2 h after injection of bacteria and reached the peak at 8 h which was 6.2 times compared to the control group,and then dropped progressively to the original level.On the other hand,the fact that mRNA was found both in the eggs and larvae indicated that it could be transferred from mother to offspring.The mRNA expression in bacterial challenge group was significantly higher than those in normal group except for the 36 h point.The CFU assay revealed that the cytoplasm of eggs with protein concentration of 200 μg/mL and 400 μg/mL had lethal effect on the bacteria Vibrio anguillarum,and the function weakened significantly once the antibody of C-type lectin was added,which indicated the C-type lectin from mother had antibacterial activity.It is concluded that the maternal C-type lectin can transfer from mother to the offspring and has antibacterial activity.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"777"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69967774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38654
房逢立, 吴洪, 周志刚
Acyl-CoA:diacylglycerol acyltransferase(DGAT;EC 2.3.1.20)is regarded as a key enzyme in triacylglycerol(TAG)biosynthesis of microalgae and other plants.In order to understand the biosynthesis of TAG in Myrmecia incisa,we found a putative full-length DGAT2 cDNA sequence in the homology search of a pyrosequencing transcriptome of this microalga.The full-length cDNA sequence was composed of 1 997 bp.It comprised a 44-bp 5′-untranslated region(UTR),a 897-bp 3′-UTR with a typical poly A tail,and a 1 056 bp open reading frame(ORF)encoding a 351-amino-acid protein with a putative molecular weight of 39.43 ku and pI at 9.46.Neighbor-Joining(NJ)phylogenetic tree inferred from the putative proteins of DGAT genes indicated that this gene belongs to DGAT2 gene family,significantly different from DGAT1 and DGAT3 families.Multiple sequence alignment of amino acids indicated that a conserved and characteristic sequence HPHG of the DGAT2's was present in this gene.Therefore,this gene was designated as MiDGAT2.Compared with the DNA sequence of MiDGAT2,it was found that its coding region was interrupted by 6 introns with all splicing sites well matching the GT-AG rule.In order to investigate the function of MiDGAT2,its open reading frame was amplified by RT-PCR and sub-cloned into the shuttle vector pYES2 to generate the recombinant vector pY-MiDGAT2.This recombinant plasmid was transformed into a TAG-defective mutant H1246 of Saccharomyces cerevisiae for expression by electroporation.The target gene integrated in the yeast genome was confirmed by sequencing and a transformant with pY-MiDGAT2 was screened out.This yeast transformant was cultured in SC medium with galactose as an inducer.Thin layer chromatogram(TLC)analysis of yeast lipids showed that the TAG-defective mutant H1246 transformed with MiDGAT2 could restore the ability to synthesize TAG,indicating that MiDGAT2 encodes a DGAT enzyme involved in the biosynthesis of TAG.When the yeast cells were stained with fluorescent dye Bodipy,it was found that lipid droplets were present in the TAG-defective mutant H1246 transformed with MiDGAT2,although the diameter of lipid droplets was smaller than that of wild type.
{"title":"缺刻缘绿藻二酰甘油酰基转移酶2（DGAT2）的基因特性与功能鉴定","authors":"房逢立, 吴洪, 周志刚","doi":"10.3724/SP.J.1231.2013.38654","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38654","url":null,"abstract":"Acyl-CoA:diacylglycerol acyltransferase(DGAT;EC 2.3.1.20)is regarded as a key enzyme in triacylglycerol(TAG)biosynthesis of microalgae and other plants.In order to understand the biosynthesis of TAG in Myrmecia incisa,we found a putative full-length DGAT2 cDNA sequence in the homology search of a pyrosequencing transcriptome of this microalga.The full-length cDNA sequence was composed of 1 997 bp.It comprised a 44-bp 5′-untranslated region(UTR),a 897-bp 3′-UTR with a typical poly A tail,and a 1 056 bp open reading frame(ORF)encoding a 351-amino-acid protein with a putative molecular weight of 39.43 ku and pI at 9.46.Neighbor-Joining(NJ)phylogenetic tree inferred from the putative proteins of DGAT genes indicated that this gene belongs to DGAT2 gene family,significantly different from DGAT1 and DGAT3 families.Multiple sequence alignment of amino acids indicated that a conserved and characteristic sequence HPHG of the DGAT2's was present in this gene.Therefore,this gene was designated as MiDGAT2.Compared with the DNA sequence of MiDGAT2,it was found that its coding region was interrupted by 6 introns with all splicing sites well matching the GT-AG rule.In order to investigate the function of MiDGAT2,its open reading frame was amplified by RT-PCR and sub-cloned into the shuttle vector pYES2 to generate the recombinant vector pY-MiDGAT2.This recombinant plasmid was transformed into a TAG-defective mutant H1246 of Saccharomyces cerevisiae for expression by electroporation.The target gene integrated in the yeast genome was confirmed by sequencing and a transformant with pY-MiDGAT2 was screened out.This yeast transformant was cultured in SC medium with galactose as an inducer.Thin layer chromatogram(TLC)analysis of yeast lipids showed that the TAG-defective mutant H1246 transformed with MiDGAT2 could restore the ability to synthesize TAG,indicating that MiDGAT2 encodes a DGAT enzyme involved in the biosynthesis of TAG.When the yeast cells were stained with fluorescent dye Bodipy,it was found that lipid droplets were present in the TAG-defective mutant H1246 transformed with MiDGAT2,although the diameter of lipid droplets was smaller than that of wild type.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"1162-1172"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69970172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38740
J. Ni, Yuhe Yu
In order to assess the changes of the intestinal microbial community after the feeding conversion of grass carp(Ctenopharyngodon idellus),we analyzed the intestinal microbial communities of grass carp by PCR-DGGE. The results indicated that there were significant differences among the intestinal microbial communities from the grass carps caught in different months. However,the sample size and fullness were not detected to have significant correlation with the intestinal microbial community,which were generally recognized as the factors influencing the intestinal microbial community. These results implied that environmental difference probably is the most important factor influencing the intestinal microbial community in a short-term. Our results will provide the basic information to elucidate the change rule of intestinal microbial community of grass carp,and the action mechanism of the intestinal probiotics,the relationship between intestinal microbes and the fish diseases.
{"title":"Intestinal microbiota changes of grass carp(Ctenopharyngodon idellus)in different months","authors":"J. Ni, Yuhe Yu","doi":"10.3724/SP.J.1231.2013.38740","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38740","url":null,"abstract":"In order to assess the changes of the intestinal microbial community after the feeding conversion of grass carp(Ctenopharyngodon idellus),we analyzed the intestinal microbial communities of grass carp by PCR-DGGE. The results indicated that there were significant differences among the intestinal microbial communities from the grass carps caught in different months. However,the sample size and fullness were not detected to have significant correlation with the intestinal microbial community,which were generally recognized as the factors influencing the intestinal microbial community. These results implied that environmental difference probably is the most important factor influencing the intestinal microbial community in a short-term. Our results will provide the basic information to elucidate the change rule of intestinal microbial community of grass carp,and the action mechanism of the intestinal probiotics,the relationship between intestinal microbes and the fish diseases.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"1558"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69970649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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