1株鸭源非O1/O139群霍乱弧菌的分离鉴定与致病性分析

doi:10.16431/j.cnki.1671-7236.2021.04.034

摘要/Abstract

摘要： 为探明引起贵州省某鸭场雏鸭发病的病原及其致病性和耐药情况，本研究对该鸭场送的疑似细菌感染病鸭进行剖检，取鼻黏膜、心脏和肝脏等组织器官接种于培养基中进行细菌分离鉴定，通过对分离菌进行药敏试验、动物回归试验和毒力基因检测研究其耐药情况和致病性。结果显示，分离菌在血琼脂培养基上生长16 h后呈现为边缘整齐、有光泽的乳白色菌落，伴有β-溶血现象，经革兰氏染色后在生物显微镜下呈两端钝圆、弧状、排列无规则的革兰氏阴性短小杆菌，与霍乱弧菌相符；16S rDNA基因序列同源性及系统进化树显示，该分离菌与霍乱弧菌同源性高达99.6%~99.7%聚为一支；药敏试验结果显示，分离菌对大部分药物都表现为耐药，其中对氨苄西林、克林霉素、复方新诺明、苯唑西林和克林霉素等抗菌药耐药性较强，对头孢哌酮和头孢曲松敏感；动物回归试验显示，分离菌可导致试验组雏鸭5 d内全部发病死亡，表明该分离菌对雏鸭具有较强的致病性；毒力基因PCR检测结果显示，检测的霍乱弧菌相关毒力基因hlyA、ompW和chxA为阳性，而检测的O1群rfb、O139群rfb、tcpA及ctxA基因为阴性，表明本次分离的霍乱弧菌携带有致病基因，但不属于O1和O139血清群。结果表明，该鸭场雏鸭发病的疫情病原为非O1/O139血清群霍乱弧菌，该菌致病性强且对多种抗菌药物耐药。本试验结果为贵州省鸭霍乱弧菌病的防控提供了参考依据。

关键词: 鸭; 非O1/O139群霍乱弧菌; 药敏试验; 毒力基因

Abstract: In order to find out the pathogen,pathogenicity and drug resistance of ducklings in a duck farm in Guizhou province,we dissected the ducks suspected of bacterial infection in the duck farm,tissues and organs such as nasal mucosa,heart and liver were inoculated in the culture medium for bacterial isolation and identification,and studied the drug resistance and pathogenicity of the isolate by drug sensitivity test,animal regression test and virulence gene detection.The results showed that the isolated bacteria grew on the blood agar medium for 16 h and showed a smooth and glossy milky white colony with β-hemolysis.The isolated bacteria were Gram-negative short bacilli with blunt round ends and irregular arrangement under the biological microscope,which was consistent with Vibrio cholera.The homology and phylogenetic tree of 16S rDNA gene sequence showed that the homology of the isolate and Vibrio cholerae were as high as 99.6% to 99.7% and were clustered into one branch.The results of drug sensitivity showed that the isolate was resistant to most drugs,including ampicillin,clindamycin,compound trimethoprim,oxacillin,clindamycin and so on,and the isolate was highly sensitive to cefoperazone and ceftriaxone.The results of animal regression test showed that the isolated strain could cause all ducklings to die within 5 days,which indicated that the isolate had strong pathogenicity to ducklings.PCR detection of virulence genes showed that hlyA,ompW and chxA genes of Vibrio cholerae were positive,while the O1 group rfb,O139 group rfb,tcpA and ctxA genes were negative,indicating that the Vibrio cholerae isolated in this study carried pathogenic genes,but did not belong to O1 and O139 serogroups.The results showed that the pathogen was non-O1/O139 Vibrio cholerae,which was highly pathogenic and resistant to many antibiotics.The results provided a reference for the prevention and control of duck Vibrio cholerae in Guizhou province.

Key words: duck; non-O1/O139 Vibrio cholerae; drug susceptibility test; virulence gene

中图分类号:

S852.61

张天天, 吴征卓, 陈国权, 王娜, 周碧君, 程振涛, 文明, 王开功. 1株鸭源非O1/O139群霍乱弧菌的分离鉴定与致病性分析[J]. 中国畜牧兽医, 2021, 48(4): 1457-1465.

ZHANG Tiantian, WU Zhengzhuo, CHEN Guoquan, WANG Na, ZHOU Bijun, CHENG Zhentao, WEN Ming, WANG Kaigong. Isolation,Identification and Pathogenicity Analysis of a Strain of Non-O1/O139 Vibrio cholerae from Duck[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1457-1465.

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