金鱼腐皮病病原菌的分离鉴定及药敏试验

天津农学院学报 ›› 2017, Vol. 24 ›› Issue (2): 53-57.

金鱼腐皮病病原菌的分离鉴定及药敏试验

康雪微¹, 胡秀彩¹, 孙金辉¹, 孙敬锋¹, 吕爱军^{1,通信作者}, 宋亚娇², YEONG Yiksung¹, 石洪玥¹, 郭立¹

1. 天津农学院水产学院,天津市水产生态及养殖重点实验室,天津 300384;
2. 河南师范大学水产学院,河南新乡 453007

收稿日期:2016-09-14 出版日期:2017-06-30 发布日期:2019-10-15
通讯作者: 吕爱军（1973-）,男,内蒙古太仆寺旗人,教授,博士,主要从事水产动物微生物与免疫学方面研究。E-mail：lajand@126.com。
作者简介:康雪微（1994-）,女,黑龙江海伦人,本科在读,主要从事水族科学与技术方面研究。E-mail：2425173584@qq.com。
基金资助:
国家自然科学基金项目“弹状病毒诱导斑马鱼皮肤和鳃免疫应答的蛋白质组研究”（31272692）; 天津市自然科学基金重点项目“黏蛋白在半滑舌鳎皮肤免疫应答过程中的功能研究”（16JCZDJC33500）及“半滑舌鳎肠道微生态与免疫力关系研究”（15JCZDJC34000）; 天津市水产产业技术体系创新团队“虾、贝类疾病防治”（ITTFRS2017009）

Isolation, Identification and Antibiotics Susceptibility of the Pathogenic Bacteria Isolated from the Putrid Skin Diseased Carassius Auratus

KANG Xue-wei¹, HU Xiu-cai¹, SUN Jin-hui¹, SUN Jing-feng¹, LÜ Ai-jun^{1,Corresponding Author}, SONG Ya-jiao², YEONG Yiksung¹, SHI Hong-yue¹, GUO Li¹

1. Tianjin Key Lab of Aqua-Ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin 300384, China;
2. College of Fisheries, Henan Normal University, Xinxiang 453007, Henan Province, China

Received:2016-09-14 Online:2017-06-30 Published:2019-10-15

摘要/Abstract

摘要： 从患腐皮病金鱼肝脏中分离获得1株细菌,编号为KL-1,通过细菌形态观察、生理生化特性、16S rDNA克隆测序及构建发育树分析等系统鉴定,并进行动物感染试验、药敏特性研究。结果表明,KL-1菌株为革兰氏阴性短杆菌,可发酵葡萄糖产气,氧化酶、接触酶、赖氨酸脱羧酶等为阳性;进一步采用PCR方法扩增16S rDNA序列,将其亚克隆至pMD18-T载体中,测序获得长度为1 507 bp,与Aeromonas veronii模式菌株ATCC 35624（X74684）序列相似性为99.66%,构建系统进化树分析与其聚为一支,鉴定KL-1菌株为维氏气单胞菌（Aeromones veronii）。动物感染试验表明,对斑马鱼半数致死量LD₅₀为1.4×10⁷cfu/mL。药敏试验结果显示,该病原菌对头孢克肟、头孢哌酮、头孢噻肟、头孢氨苄、萘啶酸、恩诺沙星、左氟沙星、诺氟沙星、氟苯尼考和氯霉素等药物敏感。

关键词: 金鱼, 维氏气单胞菌, 致病性, 药敏试验

Abstract: A bacterial strain was isolated from the liver of the Putrid Skin Diseased Carassius auratus, named KL-1. The morphological observation, physiological and biochemical characteristics, the cloning and sequencing of 16S rDNA and phylogenetic tree analysis, animal infection test and the antibiotic susceptibility analysis were carried out in this study. Results showed that KL-1 strain was a rod-shaped, gram-negative bacillus, it can produce acid and gas fromglucose, oxidase, catalase and lysine decarboxylase showed positive reaction; Furthermore, the 16S rDNA sequences was amplified by PCR method, and subcloned into pMD18-T vector, a product of sequencing was 1 507 bp in length. It had a 99.66% similarity of 16S rDNA sequence with Aeromonas veronii type strain ATCC 35624（X74684）, and clustered into one group with A. veronii, so the KL-1 strain was identified as A. veronii. Artificial infection test showed that the median lethal（LD₅₀）to zebrafish was 1.4×10⁷ cfu/mL. The susceptibility testing with different antibiotics showed that it was sensitive to cefixime, cefoperazone, cefotaxime, cephalexin, clindamycin, nalidixic acid, enrofloxacin, difloxacin, norfloxacin, and florfenicol, chloramphenicol etc.

Key words: goldfish, Aeromonas veronii, pathogenicity, drug sensitivity test

中图分类号:

S943

康雪微, 胡秀彩, 孙金辉, 孙敬锋, 吕爱军, 宋亚娇, YEONGYiksung, 石洪玥, 郭立. 金鱼腐皮病病原菌的分离鉴定及药敏试验[J]. 天津农学院学报, 2017, 24(2): 53-57.

KANG Xue-wei, HU Xiu-cai, SUN Jin-hui, SUN Jing-feng, LÜ Ai-jun, SONG Ya-jiao, YEONG Yiksung, SHI Hong-yue, GUO Li. Isolation, Identification and Antibiotics Susceptibility of the Pathogenic Bacteria Isolated from the Putrid Skin Diseased Carassius Auratus[J]. Journal of Tianjin Agricultural University, 2017, 24(2): 53-57.

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http://xuebao.tjau.edu.cn/CN/Y2017/V24/I2/53

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