3株鸭疫里氏杆菌的分离鉴定与药敏试验

doi:10.16431/j.cnki.1671-7236.2023.03.039

摘要/Abstract

摘要： 【目的】了解湖北地区鸭疫里氏杆菌病的分布流行与耐药情况,为该地区肉鸭养殖场鸭疫里氏杆菌病的临床用药提供参考。【方法】对武汉周边区县(江夏区、黄陂区、黄梅县)肉鸭养殖场的鸭疫里氏杆菌疑似病例进行采样并分离病原菌,对分离菌的形态学、染色特性、生化特性、16S rRNA进行鉴定,并对鉴定为鸭疫里氏杆菌的分离株进行血清型和耐药性分析。利用标志基因整合酶的特异性引物PCR鉴定分离菌基因组,判断是否整合有50K基因岛。【结果】分离鉴定了3株鸭疫里氏杆菌,3株分离菌均能在TSA平皿培养基中生长为光滑的半透明状菌落,革兰氏染色为阴性、瑞氏染色可观察到两极浓染;生化试验结果显示,3株鸭疫里氏杆菌均不发酵多种糖类,过氧化氢酶阳性,不产硫化氢,不能液化明胶。16S rRNA鉴定结果显示,获得大小为1480 bp的目的条带,血清型均为1型;50K基因岛标志基因PCR扩增结果显示,仅黄梅分离株(HM株)含有该基因岛。最小抑菌浓度(minimum inhibitory concentration,MIC)试验与纸片扩散(Kirby-Bauer,K-B)试验测定耐药性结果显示,3株分离菌株均对青霉素、阿莫西林、头孢曲松、头孢噻肟、氟苯尼考及庆大霉素敏感,对萘啶酮酸、壮观霉素、恩诺沙星、多黏菌素B表现出不同程度的耐药性。【结论】分离鉴定了3株血清1型鸭疫里氏杆菌,药敏试验结果对本地区鸭疫里氏杆菌病的防治及流行病学研究提供一定的理论指导。

关键词: 鸭疫里氏杆菌; 分离; 鉴定; 血清型; 50K基因岛; 药物敏感性

Abstract: 【Objective】 The purpose of this study was to understand the distribution and prevalence and drug resistance of Riemerella anatipestifer (R.anatipestifer) in some areas of Hubei,and further to provide a reference for the clinical medication of this disease on duck farms in these regions.【Method】 The suspected cases of R.anatipestifer from meat duck farms in districts and counties around Wuhan (Jiangxia district,Huangpi district,Huangmei county) were sampled and the pathogenic bacteria were isolated.The morphology,dyeing characteristics,biochemical characteristics and 16S rRNA of the isolated bacteria were identified.The serotype and drug resistance of the isolated strains identified as R.anatipestifer were analyzed.The genomes of the isolated strains were identified by specific primer PCR of marker gene integrase to determine whether 50K gene island was integrated.【Result】 A total of three R.anatipestifer strains were identified.All the three strains could grow as the morphology of smooth translucent colonies on TSA plate medium.The results of Gram staining for the three isolated strains were negative,and the results of Wright staining showed bipolar dyeing.Biochemical test results showed that the three strains of R.anatipestifer did not ferment glucose,were catalase positive,did not produce hydrogen sulfide,and could not liquefy gelatin.16S rRNA identification results showed that the target band with a size of 1 480 bp was obtained,and the serotype was type 1.PCR amplification results showed that only the isolated HM strain contained the 50K genomic island.Minimum inhibitory concentration and Kirby-Bauer tests showed that these strains were sensitive to penicillin,amoxicillin,ceftriaxone,cefotaxime,florfenicol and gentamicin,and vary degrees of resistance to nalidixic acid,spectinomycin,enrofloxacin and polymyxin B.【Conclusion】 In this study,three strains of serotype 1 R.anatipestifer strains were isolated and identified,the results of drug sensitivity could provide some theoretical guidance for the prevention,treatment and epidemiological study of R.anatipestifer in these regions.

Key words: Riemerella anatipestifer; isolation; identification; serotype; 50K genomic island; drug sensitivity

中图分类号:

S852.61

陈栋, 张言浩, 张阳, 李森, 周祖涛, 李自力. 3株鸭疫里氏杆菌的分离鉴定与药敏试验[J]. 中国畜牧兽医, 2023, 50(3): 1250-1258.

CHEN Dong, ZHANG Yanhao, ZHANG Yang, LI Sen, ZHOU Zutao, LI Zili. Isolation,Identification and Drug Sensitivity of 3 Strains of Riemerella anatipestifer[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1250-1258.

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