鲤源嗜水气单胞菌分离鉴定、耐药性及毒力基因检测

doi:10.16431/j.cnki.1671-7236.2019.07.035

摘要/Abstract

摘要：

为查明贵州某鲤鱼养殖厂鲤鱼死亡的病因，本试验进行了细菌分离培养、菌体形态观察、细菌生理生化鉴定、16S rRNA基因测序分析、药敏试验、耐药基因检测、动物感染试验、毒力基因检测等。结果显示，从死亡鲤鱼病变组织中分离到的细菌呈圆形凸起、表面光滑的灰白色菌落，为革兰氏阴性短杆菌；生化鉴定结果显示，分离菌具有运动性，能发酵葡萄糖产酸产气，精氨酸双水解酶、赖氨酸脱羧酶、M-R和V-P试验阳性；应用细菌16S rRNA基因通用引物进行PCR扩增获得大小为1 421 bp的片段，测序发现，该分离菌与16株嗜水气单胞菌相应序列同源性达75.3%~100.0%；药敏试验结果显示，该分离菌对氟苯尼考、头孢曲松、妥布霉素、氧氟沙星、恩诺沙星5种药物呈现高度敏感，对复方新诺明、磺胺异噁唑、羧苄西林等药物呈现耐药，经耐药基因PCR检测显示，该分离菌携带Sul1、Sul2和Intl1 3种耐药基因，与药敏表型相符；人工感染试验显示，该分离菌有致病力，经毒力基因检测显示，该分离菌携带aer、hly和act 3种毒力基因。上述试验结果表明，本研究分离得到了一株耐磺胺类药物的鲤源嗜水气单胞菌，为该鲤鱼养殖厂的鲤鱼疾病防治与合理用药提供了科学依据。

关键词: 鲤鱼; 嗜水气单胞菌; 分离鉴定

Abstract:

In order to find out the cause of death of carp in a carp farm in Guizhou,this experiment carried out bacterial isolation and culture,morphological observation of bacteria,physiological and biochemical identification of bacteria,16S rRNA gene sequencing,bacterial drug sensitivity test,drug resistance gene detection,animal infection test,virulence gene detection and so on.The results showed that the bacteria isolated from the diseased tissues of dead carp were round,gray-white colony with smooth surface and were Gram-negative brevibacterium.Biochemical identification showed that the isolated bacteria were motile,could ferment glucose to produce acid and gas,and were positive for arginine dihydrolase,lysine decarboxylase,M-R and V-P tests.The bacterial 16S rRNA gene universal primers were used for PCR amplification,and obtained a 1 421 bp fragment.Sequencing results showed that the isolate had 75.3% to 100.0% homology with 16 strains of Aeromonas hydrophila.Drug susceptibility test results showed that the isolate was highly susceptible to florfenicol,ceftriaxone,tobramycin,ofloxacin and enrofloxacin,and was resistant to compound sulfamethoxazole,sulfamethoxazole and carbenicillin.PCR result showed that the isolate carried Sul1,Sul2 and Intl1 resistance genes,which were consistent with the drug sensitivity phenotype.Artificial infection test showed that the isolate was pathogenic.Virulence gene test results showed that the isolate carried aer,hly and act 3 virulence genes.In conclusion,a sulfonamide-resistant strain of Aeromonas hydrophila from carp was isolated in this study,which provided a scientific basis for disease control and rational drugs use for carp in this farm.

Key words: carp; Aeromonas hydrophila; isolation and identification

中图分类号:

S917.1

胡安东, 张明洋, 张飘, 杨霞, 程振涛, 姜海波, 文明. 鲤源嗜水气单胞菌分离鉴定、耐药性及毒力基因检测[J]. 《中国畜牧兽医》, 2019, 46(7): 2167-2175.

HU Andong, ZHANG Mingyang, ZHANG Piao, YANG Xia, CHENG Zhentao, JIANG Haibo, WEN Ming. Isolation, Identification,Drug Resistance and Virulence Genes Detection of Aeromonas hydrophila from Carp[J]. , 2019, 46(7): 2167-2175.

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