血清5型和13型副猪嗜血杆菌二价灭活苗的制备和免疫效果评价
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摘要
【目的】制备一种用于预防副猪嗜血杆菌病的二价灭活苗,并对其免疫效力进行评价。【方法】选择四川地区流行的副猪嗜血杆菌(Haemophilus parasuis,HPS)血清5型MC-203株和血清13型XJ-103株,用甲醛灭活后,经PBS重悬,将2种菌液与佐剂(铝胶)按1:1:2的比例混合制出二价灭活苗。随后用制备的灭活苗免疫仔猪进行相关安全性检验,抗体滴度检测。同时为了对比制备疫苗与国产商品疫苗和进口商品疫苗的免疫效果,进行了仔猪免疫保护效果评价实验。【结果】(1)用0.2%的甲醛在37℃作用20 h可将HPS完全灭活。(2)灭活苗免疫仔猪后未造成不良反应,安全性好。(3)免疫仔猪后14 d即可在体内检测到HPS特异性抗体,196 d的血清中仍可检测到HPS抗体。(4)仔猪免疫保护效力试验表明,所制备的灭活疫苗、进口商品疫苗和国产商品疫苗的免疫保护率分别为80%、80%、60%。【结论】制备的HPS灭活苗免疫仔猪,可以产生较好的免疫应答,并且能在一定程度上抵御HPS的感染。
【Objective】The goal of this study is to prepare a bivalent inactivated vaccine for controlling Haemophilus parasuis infection and evaluate its immune efficacy.【Method】HPS serovar 5 strain MC-203 and serovar 13 strain XJ-103 prevalent in Sichuan province were inactivated by 0.2% formaldehyde and then resuspended in PBS,after which,the MC-203,XJ-103 and alhydrogel adjuvant were mixed by 1:1:2 to inoculate piglets for security and antibody titer inspection. This newly prepared vaccine was also compared with imported and domestic inactivated vaccines in their immune protection.【Result】(1)By the use of 0.2% formaldehyde at 37 ℃ for 20 h,HPS can be completely inactivated.(2)After the inactivated vaccine was immunized to piglets,no adverse reaction was caused,thus its safety was good.(3)The specific antibody against HPS could be measured at 14 days after inoculation and last at least for 196 days.(4)The newly prepared inactivated vaccine,imported inactivated vaccine and domestic inactivated vaccine have a protection of 80%,80% and 60%,respectively.【Conclusion】We can conclude that the bivalent inactivated vaccine prepared in this study presents a desired immune efficacy and a promissing role against HPS infection.
【Objective】The goal of this study is to prepare a bivalent inactivated vaccine for controlling Haemophilus parasuis infection and evaluate its immune efficacy.【Method】HPS serovar 5 strain MC-203 and serovar 13 strain XJ-103 prevalent in Sichuan province were inactivated by 0.2% formaldehyde and then resuspended in PBS,after which,the MC-203,XJ-103 and alhydrogel adjuvant were mixed by 1:1:2 to inoculate piglets for security and antibody titer inspection. This newly prepared vaccine was also compared with imported and domestic inactivated vaccines in their immune protection.【Result】(1)By the use of 0.2% formaldehyde at 37 ℃ for 20 h,HPS can be completely inactivated.(2)After the inactivated vaccine was immunized to piglets,no adverse reaction was caused,thus its safety was good.(3)The specific antibody against HPS could be measured at 14 days after inoculation and last at least for 196 days.(4)The newly prepared inactivated vaccine,imported inactivated vaccine and domestic inactivated vaccine have a protection of 80%,80% and 60%,respectively.【Conclusion】We can conclude that the bivalent inactivated vaccine prepared in this study presents a desired immune efficacy and a promissing role against HPS infection.
引文
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[2]蔡旭旺,陈焕春,吴斌,等.副猪嗜血杆菌病的危害与防治[J].养殖与饲料,2003(5):37-39.
[3]MARTINEZ-MARTINEZ S,FRANDOLOSO R,RODRIGUEZ FE,et al.Immunoproteomic analysis of the protective response obtained with subunit and commercial vaccines against Glasser's disease in pigs[J].Vet Immunol Immunopathol,2013,151(3/4):235-247.
[4]BROCKMEIER S L,LOVING C L,MULLINS M A,et al.Virulence,transmission,and heterologous protection of four isolates of Haemophilus parasuis[J].Clin Vaccine Immunol,2013,20(9):1466-1472.
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[6]ROSNER H,KIELSTEIN P,MULLER W,et al.[Relationship between serotype,virulence and SDS-PAGE protein patterns of Haemophilus parasuis[J].Dtsch Tierarztl Wochenschr,1991,98(9):327-330.
[7]LIU H,XUE Q,ZENG Q,et al.Haemophilus parasuis vaccines[J].Vet Immunol Immunopathol,2016,180:53-58.
[8]MINIATS O P,SMART N L,ROSENDAL S.Cross protection among Haemophilus parasuis strains in immunized gnotobiotic pigs[J].Can J Vet Res,1991,55(1):37-41.
[9]蔡旭旺,刘正飞,陈焕春,等.副猪嗜血杆菌的分离培养和血清型鉴定[J].华中农业大学学报,2005,24(1):55-58.
[10]ZHANG J,XU C,GUO L,et al.Prevalence and characterization of genotypic diversity of Haemophilus parasuis isolates from southern China[J].Can J Vet Res,2012,76(3):224-229.
[11]代军,刘君,黄倩,等.副猪嗜血杆菌分型血清制备及其四川地区流行病学[J].中国兽医学报,2016(1):24-29.
[12]OLIVEIRA S,PIJOAN C.Computer-based analysis of Haemophilus parasuis protein fingerprints[J].Can J Vet Res,2004,68(1):71-75.
[13]王金合,石冬梅,郭素琴,等.副猪嗜血杆菌的分离鉴定及自家灭活苗的研制[J].中国畜牧兽医,2009,36(8):144-147.
[14]BAK H,RIISING H J.Protection of vaccinated pigs against experimental infections with homologous and heterologous Haemophilus parasui[sJ].Vet Rec,2002,151(17):502-505.
[15]MOROZUMI T,NICOLET J.Some antigenic properties of Haemophilus parasuis and a proposal for serological classification[J].J Clin Microbiol,1986,23(6):1022-1025.
[16]王乐,赵战勤,薛云,等.副猪嗜血杆菌国内流行血清型菌株的致病性比较研究[J].中国兽医学报,2014(11):1748-1752.
[17]徐蓉,白娟,刘捷,等.猪圆环病毒2型和副猪嗜血杆菌5型二联灭活疫苗研制与免疫效力研究[J].畜牧与兽医,2015,47(3):1-6.
[18]宋凤香,陆桂英,肖丽辉,等.副猪嗜血杆菌灭活疫苗对仔猪免疫保护效力的研究[J].山东畜牧兽医,2010,31(6):8-9.
[19]谭实勇.副猪嗜血杆菌广东流行株的分离鉴定、优势血清型定型及灭活苗的研制[D].广州:华南农业大学兽医学院,2010.
[20]孙守勤.副猪嗜血杆菌分离鉴定及其灭活苗的研制[D].成都:四川农业大学,2009.
[21]YUAN F,FU S,HU J,et al.Evaluation of recombinant proteins of Haemophilus parasuis strain SH0165 as vaccine candidates in a mouse mode[lJ].Res Vet Sci,2012,93(1):51-56.
[22]FUENTE A J M D L,MART魱N C B G,MART魱NEZ C P,et al.Effect of different vaccine formulations on the development of Gl覿sser's disease induced in pigs by experimental Haemophilus parasuis infection[J].Journal of Comparative Pathology,2009,140(2/3):169.
[2]蔡旭旺,陈焕春,吴斌,等.副猪嗜血杆菌病的危害与防治[J].养殖与饲料,2003(5):37-39.
[3]MARTINEZ-MARTINEZ S,FRANDOLOSO R,RODRIGUEZ FE,et al.Immunoproteomic analysis of the protective response obtained with subunit and commercial vaccines against Glasser's disease in pigs[J].Vet Immunol Immunopathol,2013,151(3/4):235-247.
[4]BROCKMEIER S L,LOVING C L,MULLINS M A,et al.Virulence,transmission,and heterologous protection of four isolates of Haemophilus parasuis[J].Clin Vaccine Immunol,2013,20(9):1466-1472.
[5]KIELSTEIN P,RAPP-GABRIELSON V J.Designation of 15serovars of Haemophilus parasuis on the basis of immunodiffusion using heat-stable antigen extracts[J].J Clin Microbiol,1992,30(4):862-865.
[6]ROSNER H,KIELSTEIN P,MULLER W,et al.[Relationship between serotype,virulence and SDS-PAGE protein patterns of Haemophilus parasuis[J].Dtsch Tierarztl Wochenschr,1991,98(9):327-330.
[7]LIU H,XUE Q,ZENG Q,et al.Haemophilus parasuis vaccines[J].Vet Immunol Immunopathol,2016,180:53-58.
[8]MINIATS O P,SMART N L,ROSENDAL S.Cross protection among Haemophilus parasuis strains in immunized gnotobiotic pigs[J].Can J Vet Res,1991,55(1):37-41.
[9]蔡旭旺,刘正飞,陈焕春,等.副猪嗜血杆菌的分离培养和血清型鉴定[J].华中农业大学学报,2005,24(1):55-58.
[10]ZHANG J,XU C,GUO L,et al.Prevalence and characterization of genotypic diversity of Haemophilus parasuis isolates from southern China[J].Can J Vet Res,2012,76(3):224-229.
[11]代军,刘君,黄倩,等.副猪嗜血杆菌分型血清制备及其四川地区流行病学[J].中国兽医学报,2016(1):24-29.
[12]OLIVEIRA S,PIJOAN C.Computer-based analysis of Haemophilus parasuis protein fingerprints[J].Can J Vet Res,2004,68(1):71-75.
[13]王金合,石冬梅,郭素琴,等.副猪嗜血杆菌的分离鉴定及自家灭活苗的研制[J].中国畜牧兽医,2009,36(8):144-147.
[14]BAK H,RIISING H J.Protection of vaccinated pigs against experimental infections with homologous and heterologous Haemophilus parasui[sJ].Vet Rec,2002,151(17):502-505.
[15]MOROZUMI T,NICOLET J.Some antigenic properties of Haemophilus parasuis and a proposal for serological classification[J].J Clin Microbiol,1986,23(6):1022-1025.
[16]王乐,赵战勤,薛云,等.副猪嗜血杆菌国内流行血清型菌株的致病性比较研究[J].中国兽医学报,2014(11):1748-1752.
[17]徐蓉,白娟,刘捷,等.猪圆环病毒2型和副猪嗜血杆菌5型二联灭活疫苗研制与免疫效力研究[J].畜牧与兽医,2015,47(3):1-6.
[18]宋凤香,陆桂英,肖丽辉,等.副猪嗜血杆菌灭活疫苗对仔猪免疫保护效力的研究[J].山东畜牧兽医,2010,31(6):8-9.
[19]谭实勇.副猪嗜血杆菌广东流行株的分离鉴定、优势血清型定型及灭活苗的研制[D].广州:华南农业大学兽医学院,2010.
[20]孙守勤.副猪嗜血杆菌分离鉴定及其灭活苗的研制[D].成都:四川农业大学,2009.
[21]YUAN F,FU S,HU J,et al.Evaluation of recombinant proteins of Haemophilus parasuis strain SH0165 as vaccine candidates in a mouse mode[lJ].Res Vet Sci,2012,93(1):51-56.
[22]FUENTE A J M D L,MART魱N C B G,MART魱NEZ C P,et al.Effect of different vaccine formulations on the development of Gl覿sser's disease induced in pigs by experimental Haemophilus parasuis infection[J].Journal of Comparative Pathology,2009,140(2/3):169.