黑斑蛙普通变形杆菌的分离鉴定及药物敏感性分析  

Isolation,identification and susceptibility of a pathogenic Proteus vulgaris isolated from diseased Pelophylax nigromaculatus

作　　者：张昱泽 张德福 杨秋红[1] 周顺 艾晓辉[1] 董靖[1] ZHANG Yuze;ZHANG Defu;YANG Qiuhong;ZHOU Shun;AI Xiaohui;DONG Jing(Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China;College of Food Science and Engineering,Bohai University,Jinzhou 121013,China)

机构地区：[1]中国水产科学研究院长江水产研究所,武汉430223 [2]渤海大学食品科学与工程学院,锦州121013

出　　处：《华中农业大学学报》2025年第2期251-257,共7页Journal of Huazhong Agricultural University

基　　金：“十三五”国家重点研发计划项目(2019YFD0901702)。

摘　　要：为探明养殖场黑斑蛙大量死亡的原因,从患病黑斑蛙的脑、脊及肝肾中分离纯化一株致病菌B20200824,经过生理生化测定、16S rRNA序列测序及进化树构建对该菌株进行鉴定,利用纸片扩散法和肉汤微量稀释法分别测定该菌株的敏感性和最小抑菌浓度(minimal inhibitory concentrations,MICs)。经鉴定,发现该菌株为革兰氏阴性短杆菌,生化特征与普通变形杆菌一致,16S rRNA序列分析和进化树研究发现该菌株与普通变形杆菌有较高亲缘性,基因相似性达到99%以上,因此将该病原菌鉴定为普通变形杆菌。药敏试验结果显示,该菌株对诺氟沙星、阿莫西林和恩诺沙星等6种药物耐药,对多西环素、红霉素2种药物中介,对阿奇霉素、头孢噻肟和氟苯尼考等5种药物敏感;13种药物的MICs试验结果显示,氟苯尼考最小抑菌质量浓度为0.25μg/mL,亚胺培南、左氧氟沙星和阿奇霉素的最小抑菌质量浓度为0.125μg/mL,头孢噻肟的最小抑菌质量浓度为0.061 25μg/mL,该测定结果与药敏试验结果基本一致。以上结果表明,普通变形杆菌可导致养殖黑斑蛙发病和死亡,在生产中可使用氟苯尼考进行防控。To clarify the cause of mortality in Pelophylax nigromaculatus,a pathogenic bacterial strain designated B20200824 was isolated from the brain,spine,liver,and kidney of diseased P.nigromaculatus.The strain was identified through physiological and biochemical assays,16S rRNA sequencing,and phylogenetic analysis..Subsequently,the susceptibility and minimal inhibitory concentrations(MICs) were determined using the Kirby-Bauer(K-B) and broth micro-dilution methods,respectively.After identification,the strain was characterized as a Gram-negative short bacillus with biochemical properties consistent with those of Proteus vulgaris.Moreover,16S rRNA sequencing and phylogenetic analysis revealed that the strain exhibited a close genetic relationship with P.vulgaris,which shared over 99% genetic homology.Consequently,strain B20200824 was identified as P.vulgaris.The results of the drug susceptibility test showed that the strain was resistant to norfloxacin,amoxicillin,enrofloxacin,and 3 other drugs;it showed intermediate resistance to doxycycline and erythromycin,and susceptible to azithromycin,cefotaxime,florfenicol,and 2 other drugs.The MIC of florfenicol was 0.25 μg/mL,while the MICs for imipenem,levofloxacin,and azithromycin were 0.125 μg/mL,and for cefotaxime it was 0.061 25 μg/mL,respectively.These results were consistent with those obtained from the susceptibility assay.Taken together,these findings demonstrate that P.vulgaris can cause disease in P.nigromaculatus,and florfenicol is recommended for controlling infections caused by P.vulgaris in clinical practice.

关 键 词：黑斑蛙 普通变形杆菌 药敏试验 耐药性 

分 类 号：S966.32[农业科学—水产养殖]