肉牛呼吸道疾病综合征的病原学检测
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摘要
牛呼吸道疾病综合征(Bovine respiratory disease complex,BRDC)是严重危害养牛业的一类疾病,试验目的是对四川11个引种肉牛场爆发的BRDC进行病原检测.采集108份BRDC临床样本,分别对IBRV、BVDV、BCo V、BPIV3、BRSV、BAV3、M. Bovis、P. multocida、M. haemolytica和H. somni等10种常见的病原进行PCR检测,并鉴定M. haemolytica的荚膜血清型.IBRV、BVDV、BCo V、BPIV3、BAV3、M. Bovis、P. multocida、M. haemolytica、H. somni的阳性率分别为37%、16. 7%、4. 6%、5. 6%、6. 5%、24. 1%、6. 5%、46. 3%、5. 6%,BRSV未检出;不同牛场检出的病原种类有所差别,但IBRV、M. Bovis和M. haemolytica的个体阳性率和场阳性率较高,且混合感染较严重; M. haemolytica以荚膜血清6型和1型为主(38/50).试验结果表明四川省爆发BRDC的引种肉牛群中,检测出9种病原,以IBRV、M. Bovis和M. haemolytica混合感染较为普遍,为国内BRDC的防控提供了参考.
Bovine respiratory disease complex( BRDC) is a common health problem for the cattle industry,the purpose of this study was to detect the pathogens from clinical samples of BRDC of beef cattle in 11 farms which just introduced cattle from other provinces into Sichuan. Ten commonpathogens,named IBRV,BVDV,BCo V,BPIV3,BRSV,BAV3,M. Bovis,P. multocida,M. haemolyticaand H. somniwere detected from 108 samples collected from the 11 cattle farms by PCR assays,and the capsular serotype of M. haemolyticawas further identified by serotype specific PCR assay. The results of this study showed that the positive rates of IBRV,BVDV,BCo V,BPIV3,BAV3,M. Bovis,P. multocida,M. haemolytica and H. somniwere 37%,16. 7%,4. 6%,5. 6%,6. 5%,24. 1%,6. 5%,46. 3% and 5. 6% respectively,and BRSV was not detected. Differential detecting rate of individual pathogen was foundamongthe farms. However,theindividual positive rates and farm positive rates of IBRV,M. Bovis and M. haemolytica were higher than those of other pathogens,and co-infection of the three pathogens was common. The main capsular serotypes of M. haemolytica were identified as type 6 and 1( 38/50). In this study,nine pathogens were detected from clinical samples of BRDC,and co-infection of IBRV,M. Bovis and M. haemolytica were common in BRDC of beef cattle in Sichuan. The results of this study contribute to the prevention and control of BRDC in China.
Bovine respiratory disease complex( BRDC) is a common health problem for the cattle industry,the purpose of this study was to detect the pathogens from clinical samples of BRDC of beef cattle in 11 farms which just introduced cattle from other provinces into Sichuan. Ten commonpathogens,named IBRV,BVDV,BCo V,BPIV3,BRSV,BAV3,M. Bovis,P. multocida,M. haemolyticaand H. somniwere detected from 108 samples collected from the 11 cattle farms by PCR assays,and the capsular serotype of M. haemolyticawas further identified by serotype specific PCR assay. The results of this study showed that the positive rates of IBRV,BVDV,BCo V,BPIV3,BAV3,M. Bovis,P. multocida,M. haemolytica and H. somniwere 37%,16. 7%,4. 6%,5. 6%,6. 5%,24. 1%,6. 5%,46. 3% and 5. 6% respectively,and BRSV was not detected. Differential detecting rate of individual pathogen was foundamongthe farms. However,theindividual positive rates and farm positive rates of IBRV,M. Bovis and M. haemolytica were higher than those of other pathogens,and co-infection of the three pathogens was common. The main capsular serotypes of M. haemolytica were identified as type 6 and 1( 38/50). In this study,nine pathogens were detected from clinical samples of BRDC,and co-infection of IBRV,M. Bovis and M. haemolytica were common in BRDC of beef cattle in Sichuan. The results of this study contribute to the prevention and control of BRDC in China.
引文
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[15]周玉龙，吴丹丹，周金玲，等．牛呼吸道综合症多重PCR检测方法的建立[J]．黑龙江八一农垦大学学报，2016,28(6):97-101.
[16]张颖慧，王牧川，张斌，等．检测牛传染性鼻气管炎病毒和牛支原体的双重PCR方法的建立及应用[J]．中国兽医科学，2018,48(7):824-829.
[17]吴翠兰，彭昊，李军，等．2016—2017年广西牛呼吸道疾病综合征病原学的调查研究[J]．中国畜牧兽医，2018,45(12):238-247.
[18]陈新诺，张朝辉，徐林，等．青藏高原牦牛感染BVDV和BEV的分子流行病学调查[J]．动物医学进展，2016,37(9):35-38.
[19]何琪富，郭紫晶，李然，等．牛冠状病毒RT-PCR检测方法的建立及应用[J]．畜牧兽医学报，2018,49(10):235-241.
[20]刘晓乐，张敏敏，陈颖钰，等．牛副流感病毒3型RT-PCR检测方法的建立[J]．中国奶牛，2011,22:1-4.
[21]URBAN-CHMIEL R,WERNICKI A,GROOMS D L,et al. Rapid detection of bovine respiratory syncytial virus in poland using a human patient-Side Diagnostic Assay.[J]. Transboundary and Emerging Diseases,2015,62(4):407-410.
[22]冯旭飞，刀筱芳，李定菲，等．溶血性曼氏杆菌和多杀性巴氏杆菌双重PCR检测方法的建立及应用[J]．中国兽医科学，2014(6):624-629.
[23]MADAMPAGE C A,RAWLYK N,CROCKFORD G,et al. Single nucleotide polymorphisms in the bovine Histophilus somni genome; a comparison of new and old isolates[J]. Canadian Journal of Veterinary Research=Revue Canadienne de Recherche Veéteérinaire,2015,79(3):190-200.
[24]KLIMA C L,ZAHEER R,BRIGGS R E,et al. A multiplex PCR assay for molecular capsular serotyping of Mannheimia haemolytica,serotypes 1,2,and 6[J]. Journal of Microbiological Methods,2017,139:155-160.
[25]KLIMA C L,ZAHEER R,COOK S R,et al. Pathogens of bovine respiratory disease in North American feedlots conferring multidrug resistance via integrative conjugative elements.[J]. Journal of Clinical Microbiology,2014,52(2):438-448.
[26]AGNES J T,ZEKARIAS B,SHAO M,et al. Bovine respiratory syncytial virus and Histophilus somni interaction at the alveolar barrier[J]. Infection and Immunity,2013,81(7):2592-2597.
[27]刘畅，王军，杨雨江，等．利用PCR-DGGE技术分析肉牛阴道菌群结构[J]．中国兽医学报，2016,36(1):35-39.
[2]NG T F,KONDOV N O,DENG X,et al. A metagenomics and casecontrol study to identify viruses associated with bovine respiratory disease[J]. Journal of Virology,2015,89(10):5340-5349.
[3]PARKER A M,SHEEHY P A,HAZELTON M S,et al. A review of mycoplasma diagnostics in cattle[J]. Journal of Veterinary Internal Medicine,2018,32(3):1241-1252.
[4]HEADLEY S A,OKANO W,BALBO L C,et al. Molecular survey of infectious agents associated with bovine respiratory disease in a beef cattle feedlot in southern Brazil[J]. Journal of Veterinary Diagnostic Investigation:Official Publication of the American Association of Veterinary Laboratory Diagnosticians,Inc,2018,30(2):249-251.
[5]CUSACK P M,MCMENIMAN N,LEAN I J. The medicine and epidemiology of bovine respiratory disease in feedlots[J]. Australian Veterinary Journal,2003,81(8):480–487.
[6]张越杰，曹兵海．2018年肉牛牦牛产业发展趋势与政策建议[J]．中国畜牧业，2018(8):22-24.
[7]杨春，王明利．当前我国肉牛养殖业发展形势及未来趋势[J]．农业经济与管理，2013(6):68-74.
[8]BEHURA S K,TIZIOTO P C,KIM J,et al. Tissue tropism in host transcriptional response to members of the bovine respiratory disease Complex[J]. Scientific Reports,2017,7(1):17938.
[9]陈颖钰，郭爱珍．疾病防控能力决定肉牛产业的核心国际竞争力[J]．中国奶牛，2015(14):7-13.
[10]王洪梅，赵贵民，侯佩莉，等．牛呼吸道疾病综合征流行现状及防控技术研究进展[J]．中国畜牧杂志，2015,51(16):33-39.
[11]范颖，张继瑜，周绪正，等．牛呼吸道疾病的病原学与防制研究进展[J]．中国畜牧兽医，2013,40(1):157-163.
[12]冉艾，张斌，岳华，等．肉牛溶血性曼氏杆菌的分离鉴定及耐药性分析[J]．中国预防兽医学报，2018,40(4):301-305.
[13]于作，朱远茂，蔡红，等．一株3型牛腺病毒的分离与鉴定[J]．中国预防兽医学报，2011,33(3):169-172.
[14]沈付娆．牛冠状病毒实时荧光定量PCR检测方法的建立及应用[D]．济南:山东师范大学，2015.
[15]周玉龙，吴丹丹，周金玲，等．牛呼吸道综合症多重PCR检测方法的建立[J]．黑龙江八一农垦大学学报，2016,28(6):97-101.
[16]张颖慧，王牧川，张斌，等．检测牛传染性鼻气管炎病毒和牛支原体的双重PCR方法的建立及应用[J]．中国兽医科学，2018,48(7):824-829.
[17]吴翠兰，彭昊，李军，等．2016—2017年广西牛呼吸道疾病综合征病原学的调查研究[J]．中国畜牧兽医，2018,45(12):238-247.
[18]陈新诺，张朝辉，徐林，等．青藏高原牦牛感染BVDV和BEV的分子流行病学调查[J]．动物医学进展，2016,37(9):35-38.
[19]何琪富，郭紫晶，李然，等．牛冠状病毒RT-PCR检测方法的建立及应用[J]．畜牧兽医学报，2018,49(10):235-241.
[20]刘晓乐，张敏敏，陈颖钰，等．牛副流感病毒3型RT-PCR检测方法的建立[J]．中国奶牛，2011,22:1-4.
[21]URBAN-CHMIEL R,WERNICKI A,GROOMS D L,et al. Rapid detection of bovine respiratory syncytial virus in poland using a human patient-Side Diagnostic Assay.[J]. Transboundary and Emerging Diseases,2015,62(4):407-410.
[22]冯旭飞，刀筱芳，李定菲，等．溶血性曼氏杆菌和多杀性巴氏杆菌双重PCR检测方法的建立及应用[J]．中国兽医科学，2014(6):624-629.
[23]MADAMPAGE C A,RAWLYK N,CROCKFORD G,et al. Single nucleotide polymorphisms in the bovine Histophilus somni genome; a comparison of new and old isolates[J]. Canadian Journal of Veterinary Research=Revue Canadienne de Recherche Veéteérinaire,2015,79(3):190-200.
[24]KLIMA C L,ZAHEER R,BRIGGS R E,et al. A multiplex PCR assay for molecular capsular serotyping of Mannheimia haemolytica,serotypes 1,2,and 6[J]. Journal of Microbiological Methods,2017,139:155-160.
[25]KLIMA C L,ZAHEER R,COOK S R,et al. Pathogens of bovine respiratory disease in North American feedlots conferring multidrug resistance via integrative conjugative elements.[J]. Journal of Clinical Microbiology,2014,52(2):438-448.
[26]AGNES J T,ZEKARIAS B,SHAO M,et al. Bovine respiratory syncytial virus and Histophilus somni interaction at the alveolar barrier[J]. Infection and Immunity,2013,81(7):2592-2597.
[27]刘畅，王军，杨雨江，等．利用PCR-DGGE技术分析肉牛阴道菌群结构[J]．中国兽医学报，2016,36(1):35-39.
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