一株鸡大肠杆菌的分离鉴定与耐药性研究
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摘要
无菌采集病死鸡的肝脏、肾脏、脾脏、肺脏等组织,接种于麦康凯平板和普通琼脂,进行细菌分离与培养,并对分离菌进行生化试验和16s rDNA基因序列分析,同时用14种抗菌药物对分离菌株进行药敏试验。结果显示,分离菌在麦康凯平板上呈红色圆形菌落;镜检结果为革兰氏阴性杆菌;分离菌株能够发酵葡萄糖、棉籽糖、乳糖、木糖等糖类并产酸产气,鸟氨酸及赖氨酸试验阳性,苯丙氨酸脱氨酶及硫化氢阴性,符合大肠杆菌的生化特性;16s rDNA基因序列测定结果显示分离菌株为大肠杆菌;分离菌株对菌必治、美洛西林、恩诺沙星、诺氟沙星的敏感率较高,而对萘啶酸、青霉素等耐药。本研究为鸡大肠杆菌的诊断和防治提供了依据。
Livers, kidneys, spleens, lungs and other tissues of dead chickens were inoculated onto MacConkey plates and common agar plates for bacterial isolation. The biochemistry reactions, PCR ampli?cation for 16s rDNA were used to identifying the obtained isolates. As a result, one isolate was obtained, which had red round colonies on MacConkey plates and was Gram-negative in microscopic examination. Results showed that the isolate fermented glucose, raffinose, lactose and xylose but did not produce hydrogen sulfide.Meanwhile, 16s rDNA sequencing indicated that this isolate was E. coli. Additionally, the drug susceptibility test showed that this isolate was highly sensitive to ceftriaxone, mezlocillin, nor?oxacin and enro?oxacin but resistant to nalidixan, penicillin and trimethoprim.
Livers, kidneys, spleens, lungs and other tissues of dead chickens were inoculated onto MacConkey plates and common agar plates for bacterial isolation. The biochemistry reactions, PCR ampli?cation for 16s rDNA were used to identifying the obtained isolates. As a result, one isolate was obtained, which had red round colonies on MacConkey plates and was Gram-negative in microscopic examination. Results showed that the isolate fermented glucose, raffinose, lactose and xylose but did not produce hydrogen sulfide.Meanwhile, 16s rDNA sequencing indicated that this isolate was E. coli. Additionally, the drug susceptibility test showed that this isolate was highly sensitive to ceftriaxone, mezlocillin, nor?oxacin and enro?oxacin but resistant to nalidixan, penicillin and trimethoprim.
引文
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[11]陈晨,彭珂,王海印,等.基于16S rDNA数据库的细菌在线分类鉴定平台的构建[J].疾病监测,2013,28(3):236-240.
[12]Li W,Lu S,Cui Z,et al.PulseNet China,a model for future laboratory-based bacterial infectious disease surveillance in China[J].Front Med,2012,6(4):366-375.
[13]刘蔚雯,汪洋.细菌对氟苯尼考的耐药机制研究进展[J].中国动物传染病学报,2018,26(1):1-6.
[14]中华人民共和国卫生部医政司.全国临床检验操作规程[M].3版.南京:东南大学出版社,2006:890-923.
[15]牛建宁.鸡源大肠杆菌的分离鉴定及四环素类耐药基因检测研究[D].杨凌:西北农林科技大学,2014.
[2]郝海红,王玉莲,黄玲利,等.国外禽用抗菌药的耐药性监测与控制[J].中国兽药杂志,2013,47(1):60-63,67.
[3]王利勤.鸡源致病性大肠埃希菌耐药基因及毒力基因检测研究[D].杨凌:西北农林科技大学,2012.
[4]郑蓓,常维山.2011年我国部分地区鸡源大肠杆菌耐药性分析[J].山东畜牧兽医,2012,33(5):80-82.
[5]Liu C,Chen Z,Tan C,et al.Immunogenic characterization of outer membrane porins OmpC and OmpF of porcine extraintestinal pathogenic Escherichia coli[J].FEMSMicrobiol Lett,2012,337(2):104-111.
[6]宿志民.泰安地区鸡源大肠杆菌的分离鉴定与耐药性分析[D].泰安:山东农业大学,2015.
[7]徐睿,季芳,李梦龙.鸡致病性大肠杆菌16S rRNA PCR鉴定方法的建立[J].现代畜牧兽医,2016(7):1-5.
[8]李淑芳,曹慧,黄燕霞,等.禽源大肠杆菌和沙门氏菌的分离鉴定与耐药性检测[J].中国动物检疫,2013,30(7):63-67.
[9]王宾宾,闫大为,倪欣涛,等.鸭坦布苏病毒分离鉴定及全基因组序列分析[J].中国动物传染病学报,2017,25(6):8-14.
[10]于可响,侯艳霞,胡峰,等.Ⅰ群4型禽腺病毒卵黄抗体在鸡体内的消长规律[J].中国动物传染病学报,2017,25(6):28-31.
[11]陈晨,彭珂,王海印,等.基于16S rDNA数据库的细菌在线分类鉴定平台的构建[J].疾病监测,2013,28(3):236-240.
[12]Li W,Lu S,Cui Z,et al.PulseNet China,a model for future laboratory-based bacterial infectious disease surveillance in China[J].Front Med,2012,6(4):366-375.
[13]刘蔚雯,汪洋.细菌对氟苯尼考的耐药机制研究进展[J].中国动物传染病学报,2018,26(1):1-6.
[14]中华人民共和国卫生部医政司.全国临床检验操作规程[M].3版.南京:东南大学出版社,2006:890-923.
[15]牛建宁.鸡源大肠杆菌的分离鉴定及四环素类耐药基因检测研究[D].杨凌:西北农林科技大学,2014.