天津农学院学报 ›› 2024, Vol. 31 ›› Issue (4): 49-54.doi: 10.19640/j.cnki.jtau.2024.04.010

• 研究与简报 • 上一篇    下一篇

小丑鱼创伤弧菌的分离鉴定及药敏特性研究

霍佳昕, 胡秀彩通信作者, 夏苏东, 吕爱军   

  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 300392
  • 收稿日期:2023-10-29 出版日期:2024-08-31 发布日期:2024-09-11
  • 通讯作者: 胡秀彩(1976—),女,实验师,硕士,主要从事水产动物微生物与免疫学研究。E-mail:huxiucai@126.com。
  • 作者简介:霍佳昕(1997—),女,硕士在读,主要从事水产动物微生物与免疫学研究。E-mail:1009353868@qq.com。
  • 基金资助:
    天津市自然科学基金项目(18JCYBJC29900); 天津市高校重点科研项目(2019ZD14); 天津农学院2021年度校级教材研究项目(2021-A-01)

Isolation, identification and drug sensitivity characteristics of Vibrio vulnificus from Amphiprion percula

Huo Jiaxin, Hu XiucaiCorresponding Author, Xia Sudong, Lü Aijun   

  1. Tianjin Key Laboratory of Aqua-Ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin 300392, China
  • Received:2023-10-29 Online:2024-08-31 Published:2024-09-11

摘要: 鱼类创伤弧菌病是一种流行广泛、危害严重的细菌性传染病。2019年6月,天津某养殖场患病小丑鱼(Amphiprion percula)主要表现为腹部出血、肠道炎症和腹水等临床症状,并导致严重发病死亡。本研究从患病小丑鱼体内分离得到一株致病菌,命名为VA-1菌株,进行细菌形态学观察、生理生化特性分析、16S rDNA基因测序、人工感染试验以及药敏特性等研究,结果显示VA-1菌株为革兰氏阴性细菌,对半乳糖、葡萄糖、纤维二糖、麦芽糖、松二糖、果糖等的试验结果为阳性。Blast检索显示,VA-1菌株的16S rDNA基因序列与Vibrio vulnificus(MH244242.1)序列相似性为100%,系统发育树分析与Vibrio vulnificus亲缘关系最近,自然聚为一支,最终判定该菌株为创伤弧菌(Vibrio vulnificus)。药敏试验结果显示,该菌株对头孢氨苄、阿齐霉素、左氧氟沙星等多种药物敏感,但对四环素和万古霉素耐药。人工感染斑马鱼显示该菌的半数致死量(LD50)为1.33×105 cfu/mL。以上研究结果表明该菌株对鱼类有较强致病性,本研究为小丑鱼创伤弧菌病的诊治奠定了基础。

关键词: 小丑鱼, 创伤弧菌, 分离鉴定, 药敏试验

Abstract: Vibrio vulnificus in fish is one of the widespread and serious bacterial diseases. In June 2019, the diseased clown anemone fish(Amphiprion percula)in Tianjin certain aquaculture farm mainly experienced clinical symptoms such as abdominal bleeding, intestinal enlargement, and fluid accumulation in the body, leading to serious illness and death. In this study, a bacterial strain named VA-1 was isolated from diseased A. percula. Through morphological observation, physiological and biochemical characteristics, 16S rDNA gene sequencing, artificial infection test and drug sensitivity tests, the results showed that VA-1 was Gram-negative bacteria and positive for galactose, glucose, cellobiose, maltose, turanose, fructose and so on. Blast retrieval showed that the similarity between VA-1 strain 16S rDNA gene sequence and Vibrio vulnificus(MH244242.1)was 100%. Phylogenetic tree analysis showed that VA-1 strain was most closely related to Vibrio vulnificus(MH244242.1), and they were naturally clustered into one branch, so the strain was finally identified as Vibrio vulnificus. The results of drug sensitivity test showed that the strain was sensitive to cefalexin, azithromycin, levofloxacin and other drugs, but resistant to tetracycline and vancomycin. Half of the lethal dose(LD50)of healthy Barchydanio rerio var was 1.33×105 cfu/mL. The above results indicate that this strain has strong pathogenicity to fish, laying a foundation for the diagnosis and treatment of V. vulnificus in A. percula.

Key words: Amphiprion percula, Vibrio vulnificus, separation and identification, drug sensitivity

中图分类号: