绵羊肺源性致病性大肠杆菌的分离与鉴定
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摘要
【目的】近年来,羊呼吸系统疾病日益频发,由肠外致病性大肠杆菌感染引起的呼吸道疾病也日渐增多,给养羊业带来了一定经济损失。本文旨在确定新疆石河子地区某羊场表现为呼吸道感染症状的病死羔羊的细菌性病原及其特性。【方法】采用细菌常规分离鉴定结合16S rRNA基因序列分析的方法从发病羔羊的肺脏中分离鉴定细菌,并对分离株进行药敏试验、特异基因PCR检测、小鼠致病性试验及肺脏组织的病理学观察。【结果】从病死羔羊肺脏组织分离得到一株致病性大肠杆菌,该分离株呈多重耐药现象,检测到iutA、fyuA和ireA三种毒力基因;病变肺脏肺泡壁毛细血管充血,界限不清,支气管管腔充血,周围淋巴细胞浸润、增生。【结论】从病死羔羊肺中分离的细菌性病原是肠外致病性大肠杆菌(ExPEC)。
[Objective] In recent years, more and more reported sheep respiratory diseases are partially caused by ExPEC(Extraintestinal pathogenic Escherichia coli), resulting in some economic losses. We determine and characterize the bacterial pathogen from sheep with respiratory tract infection in Shihezi, Xinjiang. [Methods] Bacteria were isolated and identified from the lungs of infected lambs by routine methods combined with 16S rRNA gene sequence analysis. Meanwhile, antimicrobial susceptibility and specific virulence gene of the isolates were tested with K-B and PCR. Pathological examination was carried out from the lung of naturally infected sheep and artificially infected mice. [Results] A pathogenic Escherichia coli was isolated from infected lamb lung, which showed multiple drug resistance and carried 3 virulence genes(iutA, fyuA and ireA). Histology examination showed that the capillary vessel of alveolar wall was congested, bronchial lumen was found hyperemia, infiltration and hyperplasia of lymphocyte could be seen in lung tissue. [Conclusion] The bacterial pathogen isolated from infected lambs was ExPEC.
[Objective] In recent years, more and more reported sheep respiratory diseases are partially caused by ExPEC(Extraintestinal pathogenic Escherichia coli), resulting in some economic losses. We determine and characterize the bacterial pathogen from sheep with respiratory tract infection in Shihezi, Xinjiang. [Methods] Bacteria were isolated and identified from the lungs of infected lambs by routine methods combined with 16S rRNA gene sequence analysis. Meanwhile, antimicrobial susceptibility and specific virulence gene of the isolates were tested with K-B and PCR. Pathological examination was carried out from the lung of naturally infected sheep and artificially infected mice. [Results] A pathogenic Escherichia coli was isolated from infected lamb lung, which showed multiple drug resistance and carried 3 virulence genes(iutA, fyuA and ireA). Histology examination showed that the capillary vessel of alveolar wall was congested, bronchial lumen was found hyperemia, infiltration and hyperplasia of lymphocyte could be seen in lung tissue. [Conclusion] The bacterial pathogen isolated from infected lambs was ExPEC.
引文
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[16]Zhang CC,Liu JH.Isolation and identification of Escherichia coli from calf pneumonia[J].Xinjiang Animal Husbandry,2014(12):22-24(in Chinese)张成成,刘建华.致犊牛肺炎大肠埃希氏菌的分离鉴定[J].新疆畜牧业,2014(12):22-24
[17]Liu ZM,Li JQ,Huang DH,et al.Study on the drug-resistance of Escherichia coli isolated from sheep in Inner Mongolia[J].China Animal Husbandry&Veterinary Medicine,2017,44(3):839-846(in Chinese)刘正明,李金泉,黄德浩,等.内蒙古地区羊源大肠杆菌耐药性研究[J].中国畜牧兽医,2017,44(3):839-846
[18]Xuan HY,Gao C,Lin YJ,et al.Comparison of drug resistance of Escherichia coli from different cattle farms in Kuitun area of Xinjiang[J].China Animal Husbandry&Veterinary Medicine,2016,43(8):2150-2155(in Chinese)轩慧勇,高超,林亚军,等.新疆奎屯地区不同团场牛源大肠杆菌耐药情况比较[J].中国畜牧兽医,2016,43(8):2150-2155
[19]Ma ZJ,Rui P,Lu CX,et al.Isolation and identification of extraintestinal pathogenic Escherichia coli from swine[J].Chinese Journal of Zoonoses,2015,31(2):130-134(in Chinese)马增军,芮萍,逯春香,等.猪源肠外致病性大肠杆菌的分离与鉴定[J].中国人兽共患病学报,2015,31(2):130-134
[20]Clermont O,Olier M,Hoede C,et al.Animal and human pathogenic Escherichia coli strains share common genetic backgrounds[J].Infection,Genetics and Evolution,2011,11(3):654-662
[2]Meng XR,Liu XL,Zhang LY,et al.Virulence characteristics of extraintestinal pathogenic Escherichia coli deletion of gene encoding the outer membrane protein X[J].Journal of Veterinary Medical Science,2016,78(8):1261-1267
[3]Bélanger L,Garenaux A,Harel J,et al.Escherichia coli from animal reservoirs as a potential source of human extraintestinal pathogenic E.coli[J].FEMS Immunology&Medical Microbiology,2011,62(1):1-10
[4]Xu DD,Yang MW,Guli ZP,et al.Isolation,identification and histopathological observation of E.coli causing calf encephalitis[J].Chinese Journal of Veterinary Science,2016,36(2):252-255(in Chinese)许丹丹,杨铭伟,古丽孜帕,等.犊牛脑炎大肠杆菌的分离鉴定及病理组织学观察[J].中国兽医学报,2016,36(2):252-255
[5]Liu CY,Zheng HJ,Yang MJ,et al.Genome analysis and in vivo virulence of porcine extraintestinal pathogenic Escherichia coli strain PCN033[J].BMC Genomics,2015,16:717
[6]Guo X,Kong LC,Liu SM,et al.Advances in studies on virulence factors of intestinal pathogenic Escherichia coli from animal[J].Chinese Journal of Preventive Veterinary Medicine,2015,37(5):405-408(in Chinese)郭霞,孔令聪,刘树明,等.动物源肠外致病性大肠杆菌毒力因子研究进展[J].中国预防兽医学报,2015,37(5):405-408
[7]Tao L,Lan XL,Li J,et al.Pathogen analysis of goat’s respiratory diseases[J].China Animal Husbandry&Veterinary Medicine,2016,43(4):1072-1078(in Chinese)陶立,蓝显利,李军,等.山羊呼吸系统疾病病原分析[J].中国畜牧兽医,2016,43(4):1072-1078
[8]Jakobsen L,Spangholm DJ,Pedersen K,et al.Broiler chickens,broiler chicken meat,pigs and pork as sources of Ex PEC related virulence genes and resistance in Escherichia coli isolates from community-dwelling humans and UTI patients[J].International Journal of Food Microbiology,2010,142(1/2):264-272
[9]Obeng AS,Rickard H,Ndi O,et al.Antibiotic resistance,phylogenetic grouping and virulence potential of Escherichia coli isolated from the faeces of intensively farmed and free range poultry[J].Veterinary Microbiology,2012,154(3/4):305-315
[10]Johnson JR,Stell AL.Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise[J].The Journal of Infectious Diseases,2000,181(1):261-272
[11]Hodgins DC,Shewen PE.Pneumnic Pasteurellosis of Cattle[A]//Coetzer JAW,Tustin RC.Infectious Disease of Livestock,3[M].2nd Edition.Cape Town,South Africa:Oxford University Press,2004
[12]Bi BF,Tao L,Li J,et al.Identification of Mycoplasma pneumonia pathogen in Mashan black goat[J].China Animal Husbandry&Veterinary Medicine,2014,41(9):249-253(in Chinese)闭炳芬,陶立,李军,等.马山黑山羊支原体肺炎病原的鉴定[J].中国畜牧兽医,2014,41(9):249-253
[13]Ban GX,Luo DB,Zhao FQ.Comprehensive prevention and control measures of streptococcus goat[J].Chinese Abstracts of Animal Husbandry and Veterinary Medicine,2013,29(3):89(in Chinese)班国祥,罗德彪,赵福琼.山羊链球菌病的综合防治措施[J].中国畜牧兽医文摘,2013,29(3):89
[14]Chen XZ.Isolation and identification of a pathogen of pasteurella goat disease[J].Fujian Journal of Animal Husbandry and Veterinary Medicine,2013,35(4):58-59(in Chinese)陈新梓.一例山羊巴氏杆菌病的病原分离及鉴定[J].福建畜牧兽医,2013,35(4):58-59
[15]Zhou YL,Li GJ,Li Y,et al.Isolation and identification of infection pathogens between Mirenella dauricus and Escherichia coli[J].Heilongjiang Animal Science and Veterinary Medicine,2007(11):80-82(in Chinese)周玉龙,李国军,李阳,等.奶牛曼氏杆菌与大肠杆菌混合感染病原分离鉴定[J].黑龙江畜牧兽医,2007(11):80-82
[16]Zhang CC,Liu JH.Isolation and identification of Escherichia coli from calf pneumonia[J].Xinjiang Animal Husbandry,2014(12):22-24(in Chinese)张成成,刘建华.致犊牛肺炎大肠埃希氏菌的分离鉴定[J].新疆畜牧业,2014(12):22-24
[17]Liu ZM,Li JQ,Huang DH,et al.Study on the drug-resistance of Escherichia coli isolated from sheep in Inner Mongolia[J].China Animal Husbandry&Veterinary Medicine,2017,44(3):839-846(in Chinese)刘正明,李金泉,黄德浩,等.内蒙古地区羊源大肠杆菌耐药性研究[J].中国畜牧兽医,2017,44(3):839-846
[18]Xuan HY,Gao C,Lin YJ,et al.Comparison of drug resistance of Escherichia coli from different cattle farms in Kuitun area of Xinjiang[J].China Animal Husbandry&Veterinary Medicine,2016,43(8):2150-2155(in Chinese)轩慧勇,高超,林亚军,等.新疆奎屯地区不同团场牛源大肠杆菌耐药情况比较[J].中国畜牧兽医,2016,43(8):2150-2155
[19]Ma ZJ,Rui P,Lu CX,et al.Isolation and identification of extraintestinal pathogenic Escherichia coli from swine[J].Chinese Journal of Zoonoses,2015,31(2):130-134(in Chinese)马增军,芮萍,逯春香,等.猪源肠外致病性大肠杆菌的分离与鉴定[J].中国人兽共患病学报,2015,31(2):130-134
[20]Clermont O,Olier M,Hoede C,et al.Animal and human pathogenic Escherichia coli strains share common genetic backgrounds[J].Infection,Genetics and Evolution,2011,11(3):654-662