1株草龟维氏气单胞菌的分离鉴定及药敏谱分析

doi:10.11843/j.issn.0366-6964.2020.10.021

摘要/Abstract

摘要： 旨在确定河北昌黎县某花鸟鱼店宠物草龟发病死亡的病因，笔者从发病草龟体内分离纯化出1株优势菌CLW-1，通过解剖观察、细菌分离培养、革兰染色镜检、生化鉴定和16S rDNA基因测序对其分析鉴定，并通过药物敏感性测试检测菌株的耐药性，通过斑马鱼接种试验和毒力基因检测其致病性。结果显示，该菌在血琼脂平板形成湿润、表面光滑、呈β溶血的灰白色菌落，在RS琼脂培养基上长出光滑湿润的黄色菌落；革兰染色镜检为阴性短小球杆菌；16S rDNA基因序列同源性分析表明，分离菌株CLW-1与维氏气单胞菌为同一族，相似性均达99%以上；斑马鱼接种试验结果显示，该菌具有较高的致病力，其LD₅₀为1.26×10⁶CFU；毒力基因扩增结果表明，分离菌株CLW-1具有aerA、Exu、Lip和Ser毒力基因；药敏试验结果显示，分离菌株CLW-1对哌拉西林、头孢唑林和恩诺沙星等敏感；对呋喃唑酮、苯唑西林和红霉素等中敏；对麦迪霉素、克林霉素和万古霉素等耐药。通过试验结果确定此分离菌株CLW-1为维氏气单胞菌，携带毒力基因，具有较高致病力，应用对应的抗生素治疗效果较好，为维氏气单胞菌病的防治奠定基础。

关键词: 草龟, 维氏气单胞菌, 分离鉴定, 药敏试验, 毒力基因

Abstract: To determine the cause of death of Chinemys reevesii in a pet shop in Changli county, Hebei province, a dominant strain CLW-1 was isolated and purified from the diseased Chinemys reevesii. It was analyzed and identified through anatomical observation, bacterial isolation and culture, Gram staining and microscopic examination, biochemical identification and 16S rDNA gene sequencing, the drug resistance of the strain was detected through drug sensitivity test, and its pathogenicity was detected through zebrafish inoculation test and virulence gene experiment. The results showed that the bacteria formed moist, smooth surface, gray colony with β hemolysis on blood agar medium, and grew smooth and moist yellow colony on RS agar medium; Gram staining and microscopic examination showed negative coccidiosis; Homology analysis of 16S rDNA gene sequence showed that the isolated strain CLW-1 belongs to the same group with Aeromonas veronii, and the homology is above 99%; Inoculation test results of zebrafish showed that the strain had high pathogenicity, and its LD₅₀ was 1.26×10⁶ CFU; The virulence gene amplification results showed that the isolated strain CLW-1 had virulence genes of aerA, Exu, Lip and Ser; Drug sensitivity test results showed that the isolated strain CLW-1 was sensitive to piperacillin, cefazolin and enrofloxacin, etc; Susceptibility to furazolidone, oxacillin and erythromycin, etc; Resistance to midecamycin, clindamycin and vancomycin, etc. According to the test results, the isolated strain CLW-1 is confirmed to be Aeromonas veronii, which carries virulence genes and has high pathogenicity, it has great therapeutic effect by applying corresponding antibiotics, thus laying a foundation for the prevention and treatment of Aeromonas veronii.

Key words: Chinemys reevesii, Aeromonas veronii, isolation and identification, drug sensitivity test, virulence gene

中图分类号:

S852.61

王苗, 张雪佳, 杜万年, 于秀剑, 刘勃兴, 吴同垒, 史秋梅, 张志强. 1株草龟维氏气单胞菌的分离鉴定及药敏谱分析[J]. 畜牧兽医学报, 2020, 51(10): 2528-2535.

WANG Miao, ZHANG Xuejia, DU Wannian, YU Xiujian, LIU Boxing, WU Tonglei, SHI Qiumei, ZHANG Zhiqiang. Isolation, Identification and Drug Sensitivity Test of a Strain of Aeromonas veronii from Chinemys reevesii[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(10): 2528-2535.

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