鸡传染性支气管炎病毒的分离鉴定及其致病性的研究
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摘要
本研究从江苏省某鸡场疑似鸡传染性支气管炎(IB)的病料中,利用SPF鸡胚尿囊腔接种的方法分离出1株传染性支气管炎病毒(IBV),命名为CK/CH/JS/TZHY48。经RT-PCR检测,该株IBV的S1基因全长1 638 bp。遗传进化分析显示,分离株属于基因Ⅵ型,与常规疫苗株H120和H52的同源性较远。分离株对鸡胚有明显的致矮小化作用;对新城疫病毒有显著的干扰作用。动物回归试验结果显示,该分离株对7日龄SPF鸡的发病率为100%,致死率为40%,发病鸡出现气管啰音,甩头流鼻涕等呼吸道症状;病死鸡的剖检病理变化主要表现为肾苍白肿大,花斑肾,有大量明显的白色尿酸盐沉积,气管环出血。本研究可为IBV的遗传进化、致病机制以及免疫防控提供参考。
In this study, an avian infectious bronchitis virus designated as CK/CH/JS/TZHY48 was isolated and identified from a commercial chickens farm in Jiangsu Province through being inoculated into SPF eggs via the allantoic cavity.The length of S1 gene of the isolate was 1 638 bp by RT-PCR.Based upon data arising from S1 sequence and phylogenetic analyses,the IBV isolate was classified into Gene type Ⅵ,which was genetically different from vaccine isolates H120 和 H52.The pathogenicity of CK/CH/JS/TZHY48 strain in 7-day-old SPF chickens was characterized.The morbidity of challenged chickens was 100% and the mortality was 40%.The infected chickens had tracheal rales,head-twitch,snoring etc.of respiratory symptoms.Furthermore, the typical nephhropothhogenic lesions,oedema with some necrotic foci and obvious urate aggradation observed in kidneys and tracheal ring bleeding occurred in those infected dead chickens.The above-mentioned results can provide references for the genetic evolution,pathogenesis and immune control of IBV.
In this study, an avian infectious bronchitis virus designated as CK/CH/JS/TZHY48 was isolated and identified from a commercial chickens farm in Jiangsu Province through being inoculated into SPF eggs via the allantoic cavity.The length of S1 gene of the isolate was 1 638 bp by RT-PCR.Based upon data arising from S1 sequence and phylogenetic analyses,the IBV isolate was classified into Gene type Ⅵ,which was genetically different from vaccine isolates H120 和 H52.The pathogenicity of CK/CH/JS/TZHY48 strain in 7-day-old SPF chickens was characterized.The morbidity of challenged chickens was 100% and the mortality was 40%.The infected chickens had tracheal rales,head-twitch,snoring etc.of respiratory symptoms.Furthermore, the typical nephhropothhogenic lesions,oedema with some necrotic foci and obvious urate aggradation observed in kidneys and tracheal ring bleeding occurred in those infected dead chickens.The above-mentioned results can provide references for the genetic evolution,pathogenesis and immune control of IBV.
引文
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[17]LIU S W,KONG X G.A new genptype of nephropathogenic infectious bronchitis virus circulating in vaccinated and nonvaccinated flocks in China[J].Avian Pathology,2004,33(3):321-327.
[18]MARK P A,MIEKE G R,DAPHNE V H,et al.The role of phagocytic cells in enhanced susceptibility of broilers to colibacillosis after infectious bronchitis virus infectious[J].Veterinary Immunology and Immunopathology,2008,123(3-4):240-250.
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[20]TERREGINO C,TOFFAN A,BEATO M S,et al.Pathogenicity of a QX strain of infectious bronchitis virus in specitic pathogen free and commercial broiler chickens,and evaluation o f protein induced by a vaccinaton programme based on the Ma5 and 4/91serotypes[J].Avian Pathology,2008,37(5):487-493.
[21]THOR S W,HILT D A,KISSINGER J C,et al.Recombination in avian gamma-coronavirus infectious bronchitis virus[J].Viruses,2011,3(9):1777-1799.
[22]JI J,XIE J,CHEN F,et al.Phylogenetic distribution and predominant genotype of the avian infectious bronchitis virus in China during 2008-2009[J].Virology Journal,2011,8(1):184.
[23]ZAAGSTRA K A,ZEIJST B A M,KUSTWRERS J G.Rapid detection and identification of avian infectious bronchitis virus[J].Journal of Clinical Microbiology,1992,30:79-84.
[24]SOE H S,WANG L,SMITH R.The carboxy-terminal 120-residue polypeptide of infectious bronchitis virus from nucleocapsid induces cytotoxic T lymphocytes and protects chickens acute infectious[J].Virology Journal,1997,71(10):7889-7894.
[25]WEI Y Q,GUO H C,DONG H,et al.Development and characterization of a recombinant infectious bronchitis virus expressing the ectodomain region of S1 gene of H120 strain[J].Applied Microbiology and Biotechnology,2014,98(4):1727-1735.
[2]殷震,刘景华.动物病毒学[M].2版.北京:科学技术出版社,1997:675-680.YIN Zhen,LIU Jing-hua.Animal Virtual[M].2th ed.Beijing:Science and Technology Press,1997:675-680.(in Chinese)
[3]JACKWOOD M W,HILT D A,WILLIAMS S M,et al.Molecular and serologic characterization,pathogenicity,and protection studies with infectious bronchitis virus field isolates from Caklfornia[J].Avian Diseases,2007,51(2):527-533.
[4]CAVANAGH D.Coronavirus IBV:Structural characterization of the spike protein[J].Journal of General Virology,1983,64(12):2577-2583.
[5]孙丽娜,江勇,胡建民,等.鸡传染性支气管炎病毒分子变异机制的研究[J].中国畜牧兽医,2005,32(3):24-26.SUN Li-na,JIANG Yong,HU Jian-min,et al.The study on molecular variation mechanism of avian infectious bronchitis virus[J].China Animal Husbandry&Veterinary Medicine,2005,32(3):24-26.(in Chinese)
[6]余娟,刘兴友,王玲丽,等.鸡传染性支气管炎病毒致病机理的研究进展[J].中国畜牧兽医,2011,38(12):160-162.YU Juan,LIU Xing-you,WANG Ling-li,et al.Pathogenesis progression of avian infectious bronchitis virus[J].China Animal Husbandry&Veterinary Medicine,2011,38(12):160-162.(in Chinese)
[7]WANG H N,WU Q Z,HUAN G Y,et al.Isolation and identification of infectious bronchitis virus from chickens in Sichuan,China[J].Avian Diseases,1997,41(2):279-282.
[8]任涛,廖明,罗开健,等.鸡传染性支气管炎病毒中国地方流行株的分离与鉴定[J].畜牧兽医学报,2002,33(4):362-365.REN Tao,LIAO Ming,LUO Kai-jian,et al.The isolation and characterization of avian infectious bronchitis virus isolates in China[J].Chinese Journal of Animal and Veterinary Sciences,2002,33(4):362-365.(in Chinese)
[9]HAN Z,SUN C,YAN B,et al.A 15-year analysis of molecular epidemiology of avian infectious bronchitis coronavirus in China[J].Infection Genetics and Evolution,2011,11(1):190-200.
[10]刘胜旺.我国鸡传染性支气管炎流行现状及原因分析[J].中国家禽,2010,16:5-9.LIU Sheng-wang.Epidemiology and analysis of avian infectious bronchitis in China[J].China Poultry,2010,16:5-9.(in Chinese)
[11]曹永长.鸡传染性支气管炎的控制[J].中国家禽,2013,18:37-38.CAO Yong-chang.Control of infectious bronchitis in chicken[J].China Poultry,2013,18:37-38.(in Chinese)
[12]于冬玲,谭春萍,张艳雯.广西2013年-2015年鸡传染性支气管炎病毒S1基因分子流行病学分析[J].动物医学进展,2016,37(8):19-23.YU Dong-ling,TAN Chun-ping,ZHANG Yan-wen.Molecular epidemiological analysis of S1 gene of avian infectious bronchitis virus isolated in guangxi province during 2013-2015[J].Progress in Veterinary Medicine,2016,37(8):19-23.(in Chinese)
[13]FENG K,XUE Y,WANG J,et al.Development and efficacy of a novel live-attenuated QX-like nephropathogenic Infectious bronchitis virus vaccine in China[J].Vaccine,2015,33(9):1113-1120.
[14]陈锋,吴孔兴,张齐,等.5株传染性支气管炎病毒分离株的鉴定和血清分型[J].华南农业大学学报,2005,26(4):92-95.CHEN Feng,WU Kong-xing,ZHANG Qi,et al.Identification and serotyping of five infectious bronchitis virus isolates[J].Journal of South China Agricultural University,2005,26(4):92-95.(in Chinese)
[15]LUO H,QIN J,CHEN F,et al.Phylogenetic analysis of the S1 glycoprotein gene of infectious bronchitis viruses isolated in China during 2009-2010[J].Virus Genes,2012,44(1):19-23.
[16]DE WIT J J,JANE K A,COOK J K,et al.Infectious bronchitis virus variants:A review of the history,current situation and control measure[J].Avian Pathology,2011,40(3):223-235.
[17]LIU S W,KONG X G.A new genptype of nephropathogenic infectious bronchitis virus circulating in vaccinated and nonvaccinated flocks in China[J].Avian Pathology,2004,33(3):321-327.
[18]MARK P A,MIEKE G R,DAPHNE V H,et al.The role of phagocytic cells in enhanced susceptibility of broilers to colibacillosis after infectious bronchitis virus infectious[J].Veterinary Immunology and Immunopathology,2008,123(3-4):240-250.
[19]LI M,WANG X Y,WEI P,et al.Serotype and genotype diversity of infectious bronchitis viruses isolated during 1985-2008 in Guangxi,China[J].Archives of Virology,2012,157(3):467-474.
[20]TERREGINO C,TOFFAN A,BEATO M S,et al.Pathogenicity of a QX strain of infectious bronchitis virus in specitic pathogen free and commercial broiler chickens,and evaluation o f protein induced by a vaccinaton programme based on the Ma5 and 4/91serotypes[J].Avian Pathology,2008,37(5):487-493.
[21]THOR S W,HILT D A,KISSINGER J C,et al.Recombination in avian gamma-coronavirus infectious bronchitis virus[J].Viruses,2011,3(9):1777-1799.
[22]JI J,XIE J,CHEN F,et al.Phylogenetic distribution and predominant genotype of the avian infectious bronchitis virus in China during 2008-2009[J].Virology Journal,2011,8(1):184.
[23]ZAAGSTRA K A,ZEIJST B A M,KUSTWRERS J G.Rapid detection and identification of avian infectious bronchitis virus[J].Journal of Clinical Microbiology,1992,30:79-84.
[24]SOE H S,WANG L,SMITH R.The carboxy-terminal 120-residue polypeptide of infectious bronchitis virus from nucleocapsid induces cytotoxic T lymphocytes and protects chickens acute infectious[J].Virology Journal,1997,71(10):7889-7894.
[25]WEI Y Q,GUO H C,DONG H,et al.Development and characterization of a recombinant infectious bronchitis virus expressing the ectodomain region of S1 gene of H120 strain[J].Applied Microbiology and Biotechnology,2014,98(4):1727-1735.