杂交鲟类志贺邻单胞菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2019.01.031

摘要/Abstract

摘要：

试验旨在对鲟体内分离菌进行鉴定、致病力探究和耐药性分析。本研究从濒死鲟体内分离到1株细菌（NZ-2017），通过形态学观察、生理生化特性鉴定、16S rRNA基因分析及系统进化树构建等方法对分离菌种属进行确定，并采用纸片扩散法进行药敏试验，PCR方法检测耐药基因，人工感染小鼠试验确定分离菌的致病力。结果显示，NZ-2017为革兰氏阴性杆菌，其16S rRNA基因经测序及BLAST比对，显示其与类志贺邻单胞菌的同源性达99.70%，结合生理生化鉴定结果确定分离菌为类志贺邻单胞菌。药敏试验结果显示，分离菌对氧氟沙星、诺氟沙星、环丙沙星、多西环素4种药物敏感，对卡那霉素、阿莫西林、泰乐菌素、复方新诺明4种药物耐药；5种耐药基因检测结果显示，该菌携带3种耐药基因：Sul1、Sul2和Intl1，与药敏表型相符。人工感染试验结果显示，该菌对小鼠有致病力。本研究对贵州地区鲟发病流行的病因作出了准确诊断，为该地区细菌性鱼病预防和合理用药提供依据。

关键词: 杂交鲟; 类志贺邻单胞菌; 分离鉴定; 16S rRNA基因; 药敏试验; 耐药基因

Abstract:

This paper was aimed to study on isolation,identification and drug resistance analysis of a bacterial (NZ-2017),which was isolated from local sick hybrid sturgeons.Morphology,physiological and biochemical characteristics and phylogenetic analysis of 16S rRNA gene were used to identify its species.The filter paper diffusion method test,antimicrobial resistance genes test and animal regression test were carried out to analyze its pathogenicity.The results showed that NZ-2017 was a gram-negative bacillus,which was 99.70% homologous with Plesiomonas shigelloides 16S rRNA gene fragment by sequencing and BLAST analysis.With the results of physiological and biochemical identification,NZ-2017 was confirmed as Plesiomonas shigelloides.The drug sensitivity test results showed that NZ-2017 was sensitive to ofloxacin,norfloxacin,ciprofloxacin,doxycycline,and resistance to kanamycin,amoxicillin,tylosin and sulfamethoxazole.The antimicrobial resistance genes test results showed that there were three antimicrobial resistance genes (Sul1,Sul2 and Intl1),which was compatible with phenotype.The animal regression test result showed that NZ-2017 had pathogenicity to mouse.This study conducted an accurate diagnosis of the cause of the epidemic of sturgeons in Guizhou,and provided a basis for the prevention and controls to this bacterial disease of fish in this region.

Key words: hybrid sturgeon; Plesiomonas shigelloides; isolation and identification; 16S rRNA gene; antibiotic sensitivity test; antimicrobial resistance genes

中图分类号:

S852.61⁺2

张明洋, 胡安东, 程振涛, 姜海波, 文明. 杂交鲟类志贺邻单胞菌的分离鉴定及耐药性分析[J]. 《中国畜牧兽医》, 2019, 46(1): 264-270.

ZHANG Mingyang, HU Andong, CHENG Zhentao, JIANG Haibo, WEN Ming. Isolation,Identification and Drug Resistance Analysis of Plesiomonas shigelloides from Hybrid Surgeon[J]. , 2019, 46(1): 264-270.

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