广东部分地区鸭疫里默氏杆菌的分离鉴定和耐药性分析  

Isolation,Identification and Drug Resistance Analysis of Riemerella anatipestifer from Some Areas in Guangdong

作　　者：陈亦杏 张美琳 李仕楷 杨智灿 李宛珈 陈济铛[1] 朱婉君 张济培[1,2] CHEN Yixing;ZHANG Meilin;LI Shikai;YANG Zhican;LI Wanjia;CHEN Jidang;ZHU Wanjun;ZHANG Jipei(Foshan University,Foshan 528225,China;Foshan Tianmu Biotechnology Co.,Ltd.,Foshan 528225,China)

机构地区：[1]佛山科学技术学院,佛山528225 [2]佛山万牧洲生物科技有限公司,佛山528225

出　　处：《中国畜牧兽医》2024年第7期3185-3194,共10页China Animal Husbandry & Veterinary Medicine

基　　金：广东省普通高校科研项目重点领域专项(2020ZDZX1018);横向课题(BKH209059)。

摘　　要：【目的】了解广东部分地区鸭疫里默氏杆菌(Riemerella anatipestifer,RA)的流行、耐药及耐药基因携带情况。【方法】本试验于2020-2022年从广东地区采集101份疑似RA感染病死鸭、鹅的肝脏、心脏、脑等组织,对采集的组织进行细菌分离,通过革兰染色、生化试验及PCR鉴定分离株。对分离鉴定为RA的菌株进行血清型鉴定,并使用K-B纸片扩散法和PCR对分离株进行药敏试验和相应的耐药基因检测,采用统计学软件SPSS 22.0对分离株的耐药表型及耐药基因型进行统计,分析其相关性。【结果】分离菌在含5%新生牛血清的TSA平板培养后可见圆形、稍突、表面有光泽、乳脂状小菌落,革兰染色镜检为阴性短小杆菌。生化鉴定结果显示,分离株不发酵碳水化合物,氧化酶阳性,硝酸盐还原、西蒙氏枸椽酸盐、蛋白胨水、MR试验、VP试验、H_(2)S阴性,部分菌株能液化明胶和产生尿素酶。经PCR鉴定ompA基因阳性,本试验共分离得到51株RA。对分离株进行血清型鉴定,共检测到5种血清型,分别为血清1、2、4、11、18型,分析不同宿主来源的分离株血清型发现,血清18型仅在鹅上检测到。药敏试验结果显示,选择的25种药物中,RA对氟苯尼考、氧氟沙星、多西环素、大观霉素、头孢噻呋和头孢拉定6种药物敏感;对β-内酰胺类、氨基糖苷类耐药性最为严重,其中,对β-内酰胺类大部分药物耐药率在50.0%以上,对氨基糖苷类药物耐药率在80.0%以上。多重耐药统计结果显示,三重及以上耐药性菌株高达96.08%。分析不同宿主来源耐药性发现,鹅源RA分离株对青霉素、阿莫西林、氨苄西林、头孢唑啉4种药物的耐药率高于鸭源,其余药物鸭源和鹅源差异不明显。对15种耐药基因进行检测,其中blaCTX-M、aph(3′)Ⅱa、qnrB、qnrS、floR、tetM、ermF基因检出率较高,检出率分别为70.59%、68.63%、76.47%、96.08%、88.24%、82.35%和96.08%。耐药�【Objective】The experiment was aimed to understand the prevalence of Riemerella anatipestifer(RA),drug resistance and drug resistance gene carrier in some areas of Guangdong province.【Method】In this study,101 liver,heart,brain and other tissues suspected to be infected with RA were collected from Guangdong during 2020-2022.Bacteria were isolated from the collected tissues,and the isolates were identified by Gram staining,biochemical experiments and PCR.The serotype of isolates identified as RA was identified,and K-B disk diffusion method and PCR were used to conduct drug sensitivity tests and corresponding drug resistance gene detection for isolates.Statistical software SPSS 22.0 was used to analyze the drug-resistant phenotypes and drug-resistant genotypes of these strains.【Result】The small round,slightly protruding,shiny and creamy colonies of the isolated bacteria were found in the plate culture containing 5%newborn bovine serum TSA.Gram-stained microscopic examination showed that they were negative short bacilli.The results of biochemical identification showed that the isolates did not ferment carbohydrates,and were positive for oxidase,negative for nitrate reduction,Simon’s citrate,peptone water,MR,VP,H_(2)S,and some strains could liquify gelatin and produce urease.The ompA gene was identified by PCR,and 51 strains of RA were isolated in this experiment.Serotype identification of the isolates showed that 5 serotypes were detected,which were serotypes 1,2,4,11 and 18,respectively.Analysis of serotypes of isolates from different host sources showed that serotype 18 was detected only in geese.The results of drug sensitivity test showed that RA was sensitive to flufenicol,ofloxacin,doxycycline,daguamycin,ceftiofurme and cephradine among the 25 drugs selected.The resistance of RA toβ-lactam and aminoglycoside was the most serious,among which the resistance rate ofβ-lactam and aminoglycoside were above 50.0%and 80.0%,respectively.The results of multi-drug resistance showed that the strains with thre

关 键 词：鸭疫里默氏杆菌 分离鉴定 耐药性 耐药基因 

分 类 号：S852.61[农业科学—基础兽医学]