泰安地区兔源大肠杆菌的流行情况与耐药性分析
发布时间：2018-07-26 16:49
【摘要】：大肠杆菌广泛存在于人和动物体内,是肠道中的常驻菌也是一种条件性致病菌,一部分特殊血清型的大肠杆菌对人尤其是婴儿和动物具有病原性。兔大肠杆菌病在临床上多混感,是由致病性大肠埃希氏菌及其毒素所引发的具有暴发性的仔兔肠道传染病,并表现出潜伏期短、死亡率高的特点。各个年龄段的兔子对大肠杆菌都比较易感,因此在兔养殖业上给其带来的经济损失明显。本试验主要是为了了解泰安地区兔养殖场大肠杆菌的流行情况与耐药性。在2016年3月份从泰安(宁阳、新泰、东平)三个兔养殖场各采集20份兔子的粪便样品,三个养殖场总共采集样品60份。采集后的样品经过选择培养基并对其进行革兰氏染色和16srRNA分离大肠杆菌,并采用进行生化试验观察大肠杆菌的生化特性。通过对大肠杆菌分离鉴定后,采用纸片琼脂扩散法(K-B法)检测其对常用14种抗生素的敏感性可以表明兔源大肠杆菌分离菌株的耐药性,同时亦可了解大肠杆菌其耐药表型。之后本试验检测了兔源大肠杆菌分离菌株的24种耐药基因的存在情况。然后通过检测细菌Ⅰ类整合子的方法对大肠杆菌分离株进行试验以了解细菌的耐药性。最后通过肠杆菌基因间重复共有序列(ERIC)PCR对兔源大肠杆菌分离菌株进行分子的多样性分析。本试验通过对大肠杆菌的分离鉴定后共分离获得55株非重复大肠杆菌。其药敏试验的结果表明,这55株大肠杆菌对试验所用的14种抗生素表现出不同程度的耐药,敏感性各不相同。对四环素不敏感,耐药率是试验中最高的,为78.2%,但对头孢他啶、头孢曲松、亚胺培南、阿米卡星这四种药物表现为敏感,耐药率低。耐药基因检测结果表明55株兔源大肠杆菌分离菌株中检测出11种耐药基因,其中检出率最高的是β内酰胺酶类的blaTEM和blaCTX-M耐药基因,检出率分别是blaTEM(98.18%)和blaCTX-M(94.5%),其次是磺胺类的耐药基因sul-2(58.18%)。经Ⅰ类整合子检测表明在55株兔源大肠杆菌分离菌株中检测到17株大肠杆菌携带Ⅰ类整合子,检测率为30.91%。本试验在检测到的Ⅰ类整合子中发现其共携带7种耐药基因盒,分别为dfrA17+aadA5,dfrA1+catB3+aac A4,aadA2+LinF,dfrA1+aadA1,aadA22,dfrA12+orfF+addA2,aadA16+dfrA27+arr-3。ERIC-PCR把分离菌株分为23种基因型(A-W),它们的同源性在58%-100%之间。目前大肠杆菌的流行规律、特点及耐药性问题是全球关注的热点,本试验分析兔养殖场中大肠杆菌的耐药特点和整合子特点,对泰安地区兔源大肠杆菌的耐药性方面有了一定的认识,为临床合理选择抗生素提供了可以借鉴的初步依据,对以后在临床工作中能更全面的防治大肠杆菌所引发的感染,从而更有针对性的做出预防对策。
[Abstract]:Escherichia coli exists widely in human and animal body. It is a resident bacteria in intestinal tract and is also a kind of conditional pathogenic bacteria. Some special serotypes of Escherichia coli have pathogenicity to human, especially to infants and animals. Rabbit colibacillosis is a fulminant intestinal infection caused by pathogenic Escherichia coli and its toxin, which has the characteristics of short incubation period and high mortality. Rabbits of all ages are susceptible to Escherichia coli. The purpose of this study was to understand the prevalence and drug resistance of Escherichia coli in rabbit farms in Tai'an area. In March 2016, 20 rabbit feces were collected from three rabbit farms in Taian (Ningyang, Xintai and Dongping), and 60 samples were collected from three farms. The collected samples were separated by Gram staining and 16srRNA. The biochemical characteristics of Escherichia coli were observed by biochemical test. After isolation and identification of Escherichia coli, the susceptibility of Escherichia coli isolates to 14 kinds of antibiotics was detected by disk Agar diffusion method (K-B), which indicated the resistance of Escherichia coli strains isolated from rabbits, and the resistance phenotype of Escherichia coli was also known. After that, the presence of 24 resistant genes in Escherichia coli strains isolated from rabbits was detected. Then we tested the Escherichia coli isolates by detecting the class I integron of bacteria to understand the drug resistance of the bacteria. Finally, the molecular diversity of Escherichia coli isolated from rabbit was analyzed by repeated common sequence (ERIC) PCR of Enterobacter coli. After isolation and identification of Escherichia coli, 55 strains of non-repeated Escherichia coli were isolated. The results of drug sensitivity test showed that the 55 strains of Escherichia coli showed different degrees of resistance to 14 kinds of antibiotics used in the experiment, and the sensitivity was different. Tetracycline was insensitive to tetracycline, and the drug resistance rate was 78.2, but it was sensitive to ceftazidime, ceftriaxone, imipenem and amikacin, and the resistance rate was low. The results of drug resistance gene detection showed that 11 resistant genes were detected in 55 strains of Escherichia coli isolated from rabbits, among which the highest detection rate was detected by 尾 lactamases blaTEM and blaCTX-M genes. The detection rates of blaTEM (98.18%) and blaCTX-M (94.5%), followed by sulfonamides resistant gene sul-2 (58.18%). The detection of class I integron showed that 17 strains of Escherichia coli carrying class I integron were detected in 55 strains of Escherichia coli isolated from rabbits, and the detection rate was 30.91%. In this study, we found that there were 7 drug resistant gene cassettes in the class I integron, which were dfrA17 aadA5, dfrA1 catB3 aac A4A4, LinaadA2 lindfraadA1, aadA22dfrA12 orfF addA2aadA16 dfrA27 arr-3.ERIC-PCR. The isolates were divided into 23 genotypes (A-W), and their homology was between 58% and 100%. At present, the prevalence, characteristics and drug resistance of Escherichia coli are the focus of global attention. The characteristics of drug resistance and integron of Escherichia coli in rabbit farms are analyzed in this experiment. It has a certain understanding of the drug resistance of Escherichia coli from rabbits in Tai'an area, which provides a preliminary basis for clinical rational selection of antibiotics, and can prevent and treat the infection caused by Escherichia coli more comprehensively in clinical work. In order to make more targeted preventive measures.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.291
本文编号：2146650
[Abstract]:Escherichia coli exists widely in human and animal body. It is a resident bacteria in intestinal tract and is also a kind of conditional pathogenic bacteria. Some special serotypes of Escherichia coli have pathogenicity to human, especially to infants and animals. Rabbit colibacillosis is a fulminant intestinal infection caused by pathogenic Escherichia coli and its toxin, which has the characteristics of short incubation period and high mortality. Rabbits of all ages are susceptible to Escherichia coli. The purpose of this study was to understand the prevalence and drug resistance of Escherichia coli in rabbit farms in Tai'an area. In March 2016, 20 rabbit feces were collected from three rabbit farms in Taian (Ningyang, Xintai and Dongping), and 60 samples were collected from three farms. The collected samples were separated by Gram staining and 16srRNA. The biochemical characteristics of Escherichia coli were observed by biochemical test. After isolation and identification of Escherichia coli, the susceptibility of Escherichia coli isolates to 14 kinds of antibiotics was detected by disk Agar diffusion method (K-B), which indicated the resistance of Escherichia coli strains isolated from rabbits, and the resistance phenotype of Escherichia coli was also known. After that, the presence of 24 resistant genes in Escherichia coli strains isolated from rabbits was detected. Then we tested the Escherichia coli isolates by detecting the class I integron of bacteria to understand the drug resistance of the bacteria. Finally, the molecular diversity of Escherichia coli isolated from rabbit was analyzed by repeated common sequence (ERIC) PCR of Enterobacter coli. After isolation and identification of Escherichia coli, 55 strains of non-repeated Escherichia coli were isolated. The results of drug sensitivity test showed that the 55 strains of Escherichia coli showed different degrees of resistance to 14 kinds of antibiotics used in the experiment, and the sensitivity was different. Tetracycline was insensitive to tetracycline, and the drug resistance rate was 78.2, but it was sensitive to ceftazidime, ceftriaxone, imipenem and amikacin, and the resistance rate was low. The results of drug resistance gene detection showed that 11 resistant genes were detected in 55 strains of Escherichia coli isolated from rabbits, among which the highest detection rate was detected by 尾 lactamases blaTEM and blaCTX-M genes. The detection rates of blaTEM (98.18%) and blaCTX-M (94.5%), followed by sulfonamides resistant gene sul-2 (58.18%). The detection of class I integron showed that 17 strains of Escherichia coli carrying class I integron were detected in 55 strains of Escherichia coli isolated from rabbits, and the detection rate was 30.91%. In this study, we found that there were 7 drug resistant gene cassettes in the class I integron, which were dfrA17 aadA5, dfrA1 catB3 aac A4A4, LinaadA2 lindfraadA1, aadA22dfrA12 orfF addA2aadA16 dfrA27 arr-3.ERIC-PCR. The isolates were divided into 23 genotypes (A-W), and their homology was between 58% and 100%. At present, the prevalence, characteristics and drug resistance of Escherichia coli are the focus of global attention. The characteristics of drug resistance and integron of Escherichia coli in rabbit farms are analyzed in this experiment. It has a certain understanding of the drug resistance of Escherichia coli from rabbits in Tai'an area, which provides a preliminary basis for clinical rational selection of antibiotics, and can prevent and treat the infection caused by Escherichia coli more comprehensively in clinical work. In order to make more targeted preventive measures.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.291
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