鹅致病性大肠杆菌的分离鉴定、毒力基因和耐药基因的检测及药敏试验
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摘要
为了确定发病鹅感染的病原菌及了解病原菌的耐药情况,试验检查发病鹅的临床症状及剖检病变,采集发病鹅的肝脏、脾脏等器官进行病原菌的分离培养、生化鉴定,同时对分离的病原菌进行16S rDNA序列分析、动物致病性试验、毒力基因和耐药基因检测及药敏试验。结果表明:此次分离到一株优势菌株,命名为CL-1,是短杆状、中等大小的革兰氏阴性菌,且菌落形态特征和生化鉴定结果与大肠杆菌相符。经16S rDNA序列分析进一步证实分离菌株CL-1为大肠杆菌。分离菌株CL-1能够引起雏鹅和小白鼠感染发病,且雏鹅的感染症状与临床自然感染病例的症状相似。分离菌株CL-1携带eaeA、irp2、FyuA毒力基因,同时携带blaCTX-M、blaTEM、blaTEM1、blaOXA1、strA-strB、sul1耐药基因。分离菌株CL-1对大观霉素敏感;对环丙沙星、青霉素、阿莫西林、多西环素、磺胺二甲氧嘧啶、恩诺沙星、土霉素、新霉素、林可霉素耐药。说明发病鹅为大肠杆菌感染,且该分离菌株有较强的致病性,毒力基因及耐药基因检测结果可能与分离菌株的强致病性和多重耐药性有关,可以选择敏感药物大观霉素进行防控。
In order to determine the pathogenic bacteria of geese and to understand the drug resistance of pathogens, the clinical symptoms and pathological changes of the sick geese were examined, and the liver and spleen organs of the sick geese were collected for isolation, culture and biochemical identification of pathogenic bacteria. At the same time, 16 S rDNA sequence analysis, animal pathogenicity test, virulence gene and drug resistance gene detection and drug sensitivity test were carried out. The results showed that a dominant strain was isolated and named CL-1. It was a short-rod, medium-sized Gram-negative bacterium, and the colony morphological characteristics and biochemical identification were consistent with that of Escherichia coli. The strain CL-1 was confirmed to be Escherichia coli by further sequence analysis of 16 S rDNA. Animal pathogenicity test results showed that the isolated strains CL-1 could cause the infection of goose and mice, and goose infection symptoms was similar to clinical natural infection symptoms. The virulence gene test of showed that Escherichia coli CL-1 carried eaeA, irp2, and FyuA virulence genes. The detection results of drug resistance genes showed that Escherichia coli CL-1 carried blaCTX-M, blaTEM, blaTEM1, blaOXA1, strA-strB, and sul1 drug resistance genes. The results of drug sensitivity test showed that Escherichia coli CL-1 was sensitive to spectinomycin and resistant to ciprofloxacin, penicillin, amoxicillin, doxycycline, sulfadimethoxine, enrofloxacin, oxytetracycline, neomycin and lncomycin. The results indicates that the goose is a coliform infection, and the isolates have strong pathogenicity. The detection results of virulence gene and drug resistance gene may be related to the strong pathogenicity and multiple drug resistance of the isolates. The sensitive drug spectinomycin can be selected for prevention and control.
In order to determine the pathogenic bacteria of geese and to understand the drug resistance of pathogens, the clinical symptoms and pathological changes of the sick geese were examined, and the liver and spleen organs of the sick geese were collected for isolation, culture and biochemical identification of pathogenic bacteria. At the same time, 16 S rDNA sequence analysis, animal pathogenicity test, virulence gene and drug resistance gene detection and drug sensitivity test were carried out. The results showed that a dominant strain was isolated and named CL-1. It was a short-rod, medium-sized Gram-negative bacterium, and the colony morphological characteristics and biochemical identification were consistent with that of Escherichia coli. The strain CL-1 was confirmed to be Escherichia coli by further sequence analysis of 16 S rDNA. Animal pathogenicity test results showed that the isolated strains CL-1 could cause the infection of goose and mice, and goose infection symptoms was similar to clinical natural infection symptoms. The virulence gene test of showed that Escherichia coli CL-1 carried eaeA, irp2, and FyuA virulence genes. The detection results of drug resistance genes showed that Escherichia coli CL-1 carried blaCTX-M, blaTEM, blaTEM1, blaOXA1, strA-strB, and sul1 drug resistance genes. The results of drug sensitivity test showed that Escherichia coli CL-1 was sensitive to spectinomycin and resistant to ciprofloxacin, penicillin, amoxicillin, doxycycline, sulfadimethoxine, enrofloxacin, oxytetracycline, neomycin and lncomycin. The results indicates that the goose is a coliform infection, and the isolates have strong pathogenicity. The detection results of virulence gene and drug resistance gene may be related to the strong pathogenicity and multiple drug resistance of the isolates. The sensitive drug spectinomycin can be selected for prevention and control.
引文
[1] 黄宇翔, 张军, 李洪彬, 等. 雏鹅大肠杆菌的分离鉴定及药敏试验[J]. 中国兽药杂志, 2017,51(8):22-25.
[2] 严玉霖,雷华,杨尚斌,等. 鹅大肠杆菌的分离鉴定及药敏试验[J]. 中国畜牧兽医, 2011,38(10):160-162.
[3] 吴海滨, 杨娟, 周杰珑, 等. 鸡源大肠杆菌的分离鉴定及耐药特性分析[J]. 黑龙江畜牧兽医, 2017(05下):97-99.
[4] 谢永平, 杨威. 雏鹅大肠杆菌病的诊治[J]. 广西畜牧兽医, 2000,16(5):21-22.
[5] 毛福超, 郁川, 韩璐, 等. 豫西地区禽源大肠杆菌的分离鉴定与耐药性分析[J]. 河南农业科学, 2016,45(1):127-130.
[6] 陆承平. 兽医微生物学[M]. 4版.北京:中国农业出版社, 2007.
[7] 童秋廷. 一株雏鹅大肠杆菌的分离鉴定及其药敏试验[J]. 中国动物保健, 2017,19(10):91-92.
[8] 武军元, 姚礼文, 康强, 等. 新疆阿克苏地区奶牛乳房炎源大肠杆菌毒力基因检测及耐药性分析[J]. 黑龙江畜牧兽医, 2017(05下):110-112.
[9] 杨小平, 徐建国. 携带小肠结肠炎耶尔森菌HPI毒力岛大肠杆菌的毒力基因分析[J]. 中国热带医学, 2007,7(9):1508-1510.
[10] 洪婷, 缪萍, 林云, 等. 尿感方对大肠杆菌生物膜形成及与毒力基因fim、usp、hlyA表达的影响[J]. 辽宁中医杂志, 2012,39(11):2319-2323.
[11] 蒋月, 盛鹏飞. 鸡源大肠杆菌中HPI和LEE毒力岛相关基因的检测[J]. 中国家禽, 2014,36(21):54-56.
[12] 王春莲, 陈志军, 魏晓巍, 等. 承德滦平地区鹅致病性大肠杆菌的分离鉴定与药敏试验分析[J]. 黑龙江畜牧兽医, 2017(05下):126-128.
[2] 严玉霖,雷华,杨尚斌,等. 鹅大肠杆菌的分离鉴定及药敏试验[J]. 中国畜牧兽医, 2011,38(10):160-162.
[3] 吴海滨, 杨娟, 周杰珑, 等. 鸡源大肠杆菌的分离鉴定及耐药特性分析[J]. 黑龙江畜牧兽医, 2017(05下):97-99.
[4] 谢永平, 杨威. 雏鹅大肠杆菌病的诊治[J]. 广西畜牧兽医, 2000,16(5):21-22.
[5] 毛福超, 郁川, 韩璐, 等. 豫西地区禽源大肠杆菌的分离鉴定与耐药性分析[J]. 河南农业科学, 2016,45(1):127-130.
[6] 陆承平. 兽医微生物学[M]. 4版.北京:中国农业出版社, 2007.
[7] 童秋廷. 一株雏鹅大肠杆菌的分离鉴定及其药敏试验[J]. 中国动物保健, 2017,19(10):91-92.
[8] 武军元, 姚礼文, 康强, 等. 新疆阿克苏地区奶牛乳房炎源大肠杆菌毒力基因检测及耐药性分析[J]. 黑龙江畜牧兽医, 2017(05下):110-112.
[9] 杨小平, 徐建国. 携带小肠结肠炎耶尔森菌HPI毒力岛大肠杆菌的毒力基因分析[J]. 中国热带医学, 2007,7(9):1508-1510.
[10] 洪婷, 缪萍, 林云, 等. 尿感方对大肠杆菌生物膜形成及与毒力基因fim、usp、hlyA表达的影响[J]. 辽宁中医杂志, 2012,39(11):2319-2323.
[11] 蒋月, 盛鹏飞. 鸡源大肠杆菌中HPI和LEE毒力岛相关基因的检测[J]. 中国家禽, 2014,36(21):54-56.
[12] 王春莲, 陈志军, 魏晓巍, 等. 承德滦平地区鹅致病性大肠杆菌的分离鉴定与药敏试验分析[J]. 黑龙江畜牧兽医, 2017(05下):126-128.