1株牛源大肠埃希菌的分离及鉴定
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摘要
目的分离及鉴定内蒙古自治区通辽市某牛场犊牛腹泻致病菌。方法无菌采集犊牛腹泻粪便,通过分离培养、染色镜检、致病性检测、药敏检测、生化鉴定、16S r RNA PCR鉴定、血清型鉴定及系统进化树的构建进行病原研究。结果致病菌的16S rRNA基因序列与大肠埃希菌的相似性在99%以上;生化鉴定该致病菌为大肠埃希菌的置信度为99%,在20种常用药品中仅对阿米卡星、庆大霉素、亚胺培南、美洛培南敏感;血清鉴定该大肠埃希菌菌株的血清型为O17。结论该致病菌鉴定为大肠埃希菌,该菌具有多重耐药性,有成为人畜共患病病原的潜在可能。该研究为通辽地区牛场流行的大肠埃希菌防控提供了参考。
Objective To isolate and identify the pathogen of calf diarrhea in a cattle farm in Tongliao City,Inner Mongolia Autonomous Region,China. Methods The feces of calf with diarrhea were collected aseptically,from which the pathogen was isolated,cultured,stained and examined microscopically,tested for pathogenicity and drug sensitivity,and subjected to biochemical identification,16 S rRNA PCR identification,serotyping and phylogenetic analysis. Results The homology of16 S rRNA gene sequence of pathogen was more than 99% to that of Escherichia coli. The pathogen was identified as E. coli by biochemical identification,with a confidence of 99%. The pathogenic strain was only sensitive to amikacin,gentamycin,imipenem and meropenem among 20 common drugs,which was identified as serotype O17. Conclusion The pathogenic strain was identified as E. coli,which was multiple drug-resistant and might be a potential pathogen of zoonosis.The study provided a reference for prevention and control of E. coli epidemic in cattle farms in Tongliao City.
Objective To isolate and identify the pathogen of calf diarrhea in a cattle farm in Tongliao City,Inner Mongolia Autonomous Region,China. Methods The feces of calf with diarrhea were collected aseptically,from which the pathogen was isolated,cultured,stained and examined microscopically,tested for pathogenicity and drug sensitivity,and subjected to biochemical identification,16 S rRNA PCR identification,serotyping and phylogenetic analysis. Results The homology of16 S rRNA gene sequence of pathogen was more than 99% to that of Escherichia coli. The pathogen was identified as E. coli by biochemical identification,with a confidence of 99%. The pathogenic strain was only sensitive to amikacin,gentamycin,imipenem and meropenem among 20 common drugs,which was identified as serotype O17. Conclusion The pathogenic strain was identified as E. coli,which was multiple drug-resistant and might be a potential pathogen of zoonosis.The study provided a reference for prevention and control of E. coli epidemic in cattle farms in Tongliao City.
引文
[1] MA Z J, RUI P, LU C X, et al. Isolation and identification of extraintestinal pathogenic Escherichia coli from swine[J]. Chin J 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.
[2] MA C B, CHEN W W, WANG Z S, et al. Isolation, identification and drug susceptibility test of E. coli strains isolated from calves with diarrhea[J]. Chin Animal Health Inspect, 2015,32(3):78-82.(in Chinese)马长宾,陈文武,王忠山,等.致犊牛腹泻大肠杆菌的分离鉴定及药敏试验[J].中国动物检疫,2015, 32(3):78-82.
[3] GUO Q Q, KONG J Y, CHAI Y J,et al. Isolation and identification of pathogenic Escherichia coli from sheep lung[J].Microbiol Chin, 2017, 44(12):2805-2811.(in Chinese)郭强强,孔静雅,柴迎锦,等.绵羊肺源性致病性大肠杆菌的分离与鉴定[J].微生物学通报,2017, 44(12):2805-2811.
[4] CHEN F, ZHOU P, WANG Z H, et al. Diagnosis of mixed infection caused by bovine immunosuppressive diseases[J]Heilongjiang Animal Sci Veter Mod,2017(11下):98-101.(in Chinese)陈斐,周澎,王智慧,等.牛免疫抑制性疾病引起混合感染的诊断[J].黑龙江畜牧兽医,2017(11下):98-101.
[5] DANZEISEN J L, WANNEMUEHLER Y, NOLANL K, et al.Comparison of multilocus sequence analysis and virulence genotyping of Escherichia coli from live birds, retail poultry meat,and human extraintestinal infection[J]. Avian Dis,2013,57:104-108.
[6] HAO Y L, YAN G Q, WANG J M, et al. Isolation and identification of E. coli inducing diarrhea of calf and is biological characteristics in Northern Xinjiang[J]. Chin J Veter Sci, 2012, 32(6):848-851.(in Chinese)郝彦龙,剡根强,王静梅,等.新疆北疆地区致犊牛腹泻大肠杆菌的分离鉴定及部分生物学特性[J].中国兽医学报,2012,32(6):848-851.
[7] ZHANG T, WANG C G, WANG Q, et al. Serological identification and surveillance of susceptibility to antibiotics and isolate of E.scherichia coli from Swine[J]. Chin Agricultural Sci Bull, 2005, 12(21):23-25.(in Chinese)张铁,王春光,王谦,等.猪源大肠杆菌的分离、鉴定及耐药性检测[J].中国农学通报,2005, 12(21):23-25.
[8] ZHAO X S,SUN Y,JI X,et al. Antibiotic resistance of Escherichia coli isolated from dogs[J]. Chin J Zoonoses,2014,30(3):268-272.(in Chinese)赵相胜,孙洋,纪雪,等.犬源大肠杆菌分离鉴定及耐药性分析[J].中国人兽共患病学报,2014, 30(3):268-272.
[9] LI S X, XI M L, WANG P et al. Identification and antimicrobial resistance of Escherichia coli isolates from chicken in different provinces[J]. Chin J Zoonoses,2018,34(2):158-164.(in Chinese)李穗霞,席美丽,王盼盼,等.8省市鸡肉源性大肠杆菌分离鉴定及其耐药性检测[J].中国人兽共患病学报,2018. 34(2):158-164.
[10] NI K, SHEN L P, WANG X, et al. Isolation, identification and antimicrobial susceptibility test to Escherichia coli of animal origin in Shanghai City[J]. Chin Animal Health Inspect,2016, 33(9):23-26.(in Chinese)宁昆,沈莉萍,王晓旭,等.上海市动物源性大肠杆菌的分离鉴定和药敏试验[J].中国动物检疫,2016, 33(9):23-26.
[11] ZHANG L, JIANG Y, SHENG P F, et al. Serotypes, genotypes,antimicrobial resistance and resistant genes of pathogenic Esche-richia coli from diarrheic calves[J]. Chin J Veter Med, 2013,49(9):15-17.(in Chinese)张陆,蒋月,盛鹏飞,等.腹泻犊牛大肠杆菌血清型、抗菌药物、耐药性耐药基因鉴定及脉冲场凝胶电泳分型[J].中国兽医杂志,2013, 49(9):15-17.
[12] FANG G Y, ZHANG L, LU Y C, et al. Isolation, identification and resistance testing of E. coli isolated from diarrheal calves[J]. Chin Animal Husbandry&Veter Med, 2014, 41(9):273-277.(in Chinese)方光远,张莉,陆宇超,等.犊牛腹泻大肠杆菌的分离鉴定和耐药性检测[J].中国畜牧兽医,2014, 41(9):273-277.
[13] YANG S Q, AO R G L, WANG C J, et al. Prevalence of serotypes virulence genes and antimicrobial susceptibility of bovine pathogenic Escherichia coli isolates in inner Mongolia[J]. Chin J Prevent Veter Med, 2015, 37(10):761-764.(in Chinese)杨斯琴,敖日格乐,王纯洁,等.内蒙古呼和浩特地区牛源致病性大肠杆菌的血清型、毒力基因检测及耐药性分析[J].中国预防兽医学报,2015, 37(10):761-764.
[14] ZHANG S X, ZHANG X Z, LI Y Y, et al. Analysis on serotypes, virulence genes and pathogenicity of local strains of pathogenic Echerichia coli from chicken[J]. Chin J Veter Sci,2015, 37(12):2260-2265.(in Chinese)张召兴,张香斋,李蕴玉,等.鸡致病性E.coli地方株血清型的鉴定、毒力基因及致病性检测[J].中国兽医学报,2015,37(12):2260-2265.
[2] MA C B, CHEN W W, WANG Z S, et al. Isolation, identification and drug susceptibility test of E. coli strains isolated from calves with diarrhea[J]. Chin Animal Health Inspect, 2015,32(3):78-82.(in Chinese)马长宾,陈文武,王忠山,等.致犊牛腹泻大肠杆菌的分离鉴定及药敏试验[J].中国动物检疫,2015, 32(3):78-82.
[3] GUO Q Q, KONG J Y, CHAI Y J,et al. Isolation and identification of pathogenic Escherichia coli from sheep lung[J].Microbiol Chin, 2017, 44(12):2805-2811.(in Chinese)郭强强,孔静雅,柴迎锦,等.绵羊肺源性致病性大肠杆菌的分离与鉴定[J].微生物学通报,2017, 44(12):2805-2811.
[4] CHEN F, ZHOU P, WANG Z H, et al. Diagnosis of mixed infection caused by bovine immunosuppressive diseases[J]Heilongjiang Animal Sci Veter Mod,2017(11下):98-101.(in Chinese)陈斐,周澎,王智慧,等.牛免疫抑制性疾病引起混合感染的诊断[J].黑龙江畜牧兽医,2017(11下):98-101.
[5] DANZEISEN J L, WANNEMUEHLER Y, NOLANL K, et al.Comparison of multilocus sequence analysis and virulence genotyping of Escherichia coli from live birds, retail poultry meat,and human extraintestinal infection[J]. Avian Dis,2013,57:104-108.
[6] HAO Y L, YAN G Q, WANG J M, et al. Isolation and identification of E. coli inducing diarrhea of calf and is biological characteristics in Northern Xinjiang[J]. Chin J Veter Sci, 2012, 32(6):848-851.(in Chinese)郝彦龙,剡根强,王静梅,等.新疆北疆地区致犊牛腹泻大肠杆菌的分离鉴定及部分生物学特性[J].中国兽医学报,2012,32(6):848-851.
[7] ZHANG T, WANG C G, WANG Q, et al. Serological identification and surveillance of susceptibility to antibiotics and isolate of E.scherichia coli from Swine[J]. Chin Agricultural Sci Bull, 2005, 12(21):23-25.(in Chinese)张铁,王春光,王谦,等.猪源大肠杆菌的分离、鉴定及耐药性检测[J].中国农学通报,2005, 12(21):23-25.
[8] ZHAO X S,SUN Y,JI X,et al. Antibiotic resistance of Escherichia coli isolated from dogs[J]. Chin J Zoonoses,2014,30(3):268-272.(in Chinese)赵相胜,孙洋,纪雪,等.犬源大肠杆菌分离鉴定及耐药性分析[J].中国人兽共患病学报,2014, 30(3):268-272.
[9] LI S X, XI M L, WANG P et al. Identification and antimicrobial resistance of Escherichia coli isolates from chicken in different provinces[J]. Chin J Zoonoses,2018,34(2):158-164.(in Chinese)李穗霞,席美丽,王盼盼,等.8省市鸡肉源性大肠杆菌分离鉴定及其耐药性检测[J].中国人兽共患病学报,2018. 34(2):158-164.
[10] NI K, SHEN L P, WANG X, et al. Isolation, identification and antimicrobial susceptibility test to Escherichia coli of animal origin in Shanghai City[J]. Chin Animal Health Inspect,2016, 33(9):23-26.(in Chinese)宁昆,沈莉萍,王晓旭,等.上海市动物源性大肠杆菌的分离鉴定和药敏试验[J].中国动物检疫,2016, 33(9):23-26.
[11] ZHANG L, JIANG Y, SHENG P F, et al. Serotypes, genotypes,antimicrobial resistance and resistant genes of pathogenic Esche-richia coli from diarrheic calves[J]. Chin J Veter Med, 2013,49(9):15-17.(in Chinese)张陆,蒋月,盛鹏飞,等.腹泻犊牛大肠杆菌血清型、抗菌药物、耐药性耐药基因鉴定及脉冲场凝胶电泳分型[J].中国兽医杂志,2013, 49(9):15-17.
[12] FANG G Y, ZHANG L, LU Y C, et al. Isolation, identification and resistance testing of E. coli isolated from diarrheal calves[J]. Chin Animal Husbandry&Veter Med, 2014, 41(9):273-277.(in Chinese)方光远,张莉,陆宇超,等.犊牛腹泻大肠杆菌的分离鉴定和耐药性检测[J].中国畜牧兽医,2014, 41(9):273-277.
[13] YANG S Q, AO R G L, WANG C J, et al. Prevalence of serotypes virulence genes and antimicrobial susceptibility of bovine pathogenic Escherichia coli isolates in inner Mongolia[J]. Chin J Prevent Veter Med, 2015, 37(10):761-764.(in Chinese)杨斯琴,敖日格乐,王纯洁,等.内蒙古呼和浩特地区牛源致病性大肠杆菌的血清型、毒力基因检测及耐药性分析[J].中国预防兽医学报,2015, 37(10):761-764.
[14] ZHANG S X, ZHANG X Z, LI Y Y, et al. Analysis on serotypes, virulence genes and pathogenicity of local strains of pathogenic Echerichia coli from chicken[J]. Chin J Veter Sci,2015, 37(12):2260-2265.(in Chinese)张召兴,张香斋,李蕴玉,等.鸡致病性E.coli地方株血清型的鉴定、毒力基因及致病性检测[J].中国兽医学报,2015,37(12):2260-2265.