某牛场牛病毒性腹泻病毒、牛传染性鼻气管炎病毒和大肠杆菌混合感染的诊断及分析
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摘要
为了确定甘肃省张掖市某奶牛场犊牛严重腹泻的病因,试验无菌采集发病犊牛血液、鼻黏液及粪便样品,通过病毒特异性引物的PCR检测、细菌分离鉴定及16S r DNA PCR检测及测序分析等方法对发病犊牛的病料进行检测,并对分离菌进行药敏试验。结果表明:全血及粪便等样品中牛病毒性腹泻病毒(BVDV)和牛传染性鼻气管炎病毒(IBRV)呈阳性;从粪便中分离出大肠杆菌,且分离株对氨苄西林、头孢氨苄、环丙沙星高敏,对其他抗菌药物分别呈现不同程度的耐药。说明该奶牛场犊牛腹泻是由BVDV、IBRV和大肠杆菌混合感染引起的,应结合药敏试验结果用药,同时配合中药辅助治疗,以提高临床治疗效果。
In order to determine the cause of serious diarrhea of calves in a dairy farm in Zhangye City,Gansu Province,blood,nasal mucous and feces samples were collected aseptically in this experiment,PCR assay using virus specific primers,bacterial isolation and identification,and PCR detection based on 16 S r DNA and sequencing analysis were used for for the determination of the clinical samples from the diseased calves.The results showed that Bovine viral diarrhea virus(BVDV) and Infectious bovine rhinotracheitis virus(IBRV) were positive in the whole blood and faces samples;a dominant strain of Escherichia coil was isolated from feces sample,which was highly sensitive to ampicillin,cephalexin and ciprofloxacin and showed different degrees of resistance to other antibiotics. The results suggested that the calf diarrhea in the dairy farm was caused by mixed infection of BVDV,IBRV and Escherichia coli. It should be combined with the results of drug sensitivity test for medication and the adjuvant treatment of traditional Chinese medicine to improve the clinical treatment effect.
In order to determine the cause of serious diarrhea of calves in a dairy farm in Zhangye City,Gansu Province,blood,nasal mucous and feces samples were collected aseptically in this experiment,PCR assay using virus specific primers,bacterial isolation and identification,and PCR detection based on 16 S r DNA and sequencing analysis were used for for the determination of the clinical samples from the diseased calves.The results showed that Bovine viral diarrhea virus(BVDV) and Infectious bovine rhinotracheitis virus(IBRV) were positive in the whole blood and faces samples;a dominant strain of Escherichia coil was isolated from feces sample,which was highly sensitive to ampicillin,cephalexin and ciprofloxacin and showed different degrees of resistance to other antibiotics. The results suggested that the calf diarrhea in the dairy farm was caused by mixed infection of BVDV,IBRV and Escherichia coli. It should be combined with the results of drug sensitivity test for medication and the adjuvant treatment of traditional Chinese medicine to improve the clinical treatment effect.
引文
[1]荔霞,王胜义,刘永明,等.犊牛腹泻病因及其药物防治研究进展[J].中国畜牧兽医,2010,37(10):161-165.
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[5]柳强,侯喜林,车车,等.牛冠状病毒和轮状病毒的双重RT-PCR检测方法的建立及应用[J].中国预防兽医学报,2009,31(9):701-704.
[6] BIUK-RUDAN N,CVELNI S,MADI J,et al. Prevalence of antibodies to IBR and BVD viruses in dairy cows with reproductive disorders[J]. Theriogenology,1999,51(5):875-881.
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[8] CHASE C C. The impact of BVDV infection on adaptive immunity[J].Biologicals,2013,41(1):52-60.
[9] PEDRERA M,G MEZ-VILAMANDOS J C,RISALDE M A,et al.Characterization of apoptosis pathways(intrinsic and extrinsic)in lymphoid tissues of calves inoculated with non-cytopathic bovine viral diarrhoea virus genotype-1[J]. J Comp Pathol,2012,146(1):30-39.
[10] SHAHRIAR F M,CLARK E G,EUGENE J,et al.Coinfection with bovine viral diarrhea virus and mycoplasma bovis in feedlot cattle with chronic pneumonia[J]. Can Vet J,2002,43(11):863-868.
[11] POTGIETER L N,MCCRACKEN M D,HOPKINS F M,et al.Comparison of the pneumopathogenicity of two strains of bovine viral diarrhea virus[J].Am J Vet Res,1985,46(1):151-153.
[12] BAKER J C. The clinical manifestations of bovine viral diarrhea infection[J]. Vet Clin North Am Food Anim Pract,1995,11(3):425-445.
[13]童光志.牛传染性鼻气管炎病毒的持续感染及其防治[J].中国畜禽传染病,1992(1):60-62.
[14]李艳婷,孙光野,张露,等.犊牛轮状病毒和大肠杆菌混合感染检测[J].中国畜牧兽医,2017,44(1):248-253.
[15]王丹阳,王旭荣,张康,等.牛病毒性腹泻病毒、大肠杆菌和奇异变形杆菌混合感染致犊牛腹泻的研究[J].中国畜牧兽医,2018,45(1):189-195.
[16]夏利宁,向发,郭庆勇,等.新疆不同地区牛源大肠杆菌耐药性分析[J].中国畜牧兽医,2014,41(2):203-206.
[17]张伟,崔冰冰,孙武文,等.北方不同地区犊牛腹泻大肠杆菌耐药性和耐药基因携带率的监测分析[J].黑龙江畜牧兽医,2016(11上):162-165.
[18]安慧慧,李晓娜,余婷,等.银川地区鸡源与牛源大肠杆菌耐药性比较[J].黑龙江畜牧兽医,2014(09上):125-127.
[2] SMITH D R. Field disease diagnostic investigation of neonatal calf diarrhea[J]. Vet Clin North Am Food Aim Pract,2012,28(3):465-481.
[3]季新成.牛传染性鼻气管炎病毒和牛病毒性腹泻病毒分子生物学检测技术研究[D].杨凌:西北农林科技大学,2010.
[4]王嵩,林红丽,王宇鹏,等.牛传染性鼻气管炎病毒PCR检测方法的建立及初步应用[J].黑龙江八一农垦大学学报,2014,26(3):26-29.
[5]柳强,侯喜林,车车,等.牛冠状病毒和轮状病毒的双重RT-PCR检测方法的建立及应用[J].中国预防兽医学报,2009,31(9):701-704.
[6] BIUK-RUDAN N,CVELNI S,MADI J,et al. Prevalence of antibodies to IBR and BVD viruses in dairy cows with reproductive disorders[J]. Theriogenology,1999,51(5):875-881.
[7] YESILBAG K,FORSTER C,OZYIGIT M O,et al. Characterisation of bovine viral diarrhoea virus(BVDV)isolates from an outbreak with haemorrhagic enteritis and severe pneumonia[J]. Vet Microbiol,2014,169:42-49.
[8] CHASE C C. The impact of BVDV infection on adaptive immunity[J].Biologicals,2013,41(1):52-60.
[9] PEDRERA M,G MEZ-VILAMANDOS J C,RISALDE M A,et al.Characterization of apoptosis pathways(intrinsic and extrinsic)in lymphoid tissues of calves inoculated with non-cytopathic bovine viral diarrhoea virus genotype-1[J]. J Comp Pathol,2012,146(1):30-39.
[10] SHAHRIAR F M,CLARK E G,EUGENE J,et al.Coinfection with bovine viral diarrhea virus and mycoplasma bovis in feedlot cattle with chronic pneumonia[J]. Can Vet J,2002,43(11):863-868.
[11] POTGIETER L N,MCCRACKEN M D,HOPKINS F M,et al.Comparison of the pneumopathogenicity of two strains of bovine viral diarrhea virus[J].Am J Vet Res,1985,46(1):151-153.
[12] BAKER J C. The clinical manifestations of bovine viral diarrhea infection[J]. Vet Clin North Am Food Anim Pract,1995,11(3):425-445.
[13]童光志.牛传染性鼻气管炎病毒的持续感染及其防治[J].中国畜禽传染病,1992(1):60-62.
[14]李艳婷,孙光野,张露,等.犊牛轮状病毒和大肠杆菌混合感染检测[J].中国畜牧兽医,2017,44(1):248-253.
[15]王丹阳,王旭荣,张康,等.牛病毒性腹泻病毒、大肠杆菌和奇异变形杆菌混合感染致犊牛腹泻的研究[J].中国畜牧兽医,2018,45(1):189-195.
[16]夏利宁,向发,郭庆勇,等.新疆不同地区牛源大肠杆菌耐药性分析[J].中国畜牧兽医,2014,41(2):203-206.
[17]张伟,崔冰冰,孙武文,等.北方不同地区犊牛腹泻大肠杆菌耐药性和耐药基因携带率的监测分析[J].黑龙江畜牧兽医,2016(11上):162-165.
[18]安慧慧,李晓娜,余婷,等.银川地区鸡源与牛源大肠杆菌耐药性比较[J].黑龙江畜牧兽医,2014(09上):125-127.