1株大黄鱼源金黄杆菌的分离鉴定和耐药性分析

doi:10.16431/j.cnki.1671-7236.2022.03.036

摘要/Abstract

摘要： 【目的】探究引发浙江省某养殖场大黄鱼发病的病原及其致病性和耐药情况。【方法】无菌条件下剖检发病大黄鱼，取其肝脏和肾脏组织，划线分离培养细菌，通过形态学观察、生理生化试验、16S rRNA基因序列分析鉴定分离菌的种属特性，通过人工感染和组织切片制备评价分离菌对大黄鱼的致病性，最后通过药敏纸片法检测分离菌的耐药性。【结果】分离菌在培养基上外观为表面凸起、光滑和边缘整齐的黄色圆形菌落；革兰氏染色后光学显微镜下呈现出单个分散或成对分布的红色短棒状杆菌；该分离菌可在麦康凯琼脂上生长，可利用葡萄糖产酸；16S rRNA基因序列相似性分析及系统进化树显示，该分离菌与金黄杆菌相似性高达97.0%以上并聚为一类。动物回归试验结果显示，分离菌可引起大黄鱼的肝脏、肾脏和脾脏发生明显病变，半数致死量为6.32×10⁴ CFU/mL，表明其对大黄鱼具有较强致病性；药敏结果显示，分离菌对阿米卡星和左氧氟沙星敏感，对庆大霉素、新霉素和红霉素中度敏感，对头孢克肟、氨苄西林、卡那霉素、呋喃唑酮等耐药。【结论】本研究分离了大黄鱼源致病株金黄杆菌，并通过其致病性和耐药性研究为金黄杆菌引发水产动物发生病害的防控和治疗提供参考。

关键词: 金黄杆菌; 分离鉴定; 人工感染; 序列分析; 耐药性

Abstract: 【Objective】 The study was aimed to explore the pathogens, pathogenicity and drug resistance of Pseudosciaena crocea in a farm in Zhejiang province.【Method】 The diseased Pseudosciaena crocea was dissected under sterile conditions, the liver and kidney tissues were taken, and the bacteria were isolated and cultured by scribing.The species characteristics of the isolated bacteria were identified by morphological observation, physiological and biochemical test and 16S rRNA gene sequence analysis.The pathogenicity of the isolated bacteria to Pseudosciaena crocea was evaluated by artificial infection and tissue section.Finally, the drug resistance of the isolated bacteria was detected by drug sensitive paper method.【Result】 The appearance of the isolated bacteria on the culture medium was yellow round colonies with convex, smooth surface and neat edges.After Gram staining, there were single scattered or paired distribution of Corynebacterium red under an optical microscope.The isolated strain could grow on MacConkey agar and produce acid by glucose.16S rRNA gene sequence similarity analysis and phylogenetic tree showed that the isolated strain had more than 97.0% similarity with Chryseobacterium sp. and clustered into one class.The results of animal regression test showed that the isolated bacteria could cause obvious lesions in the liver, kidney and spleen of Pseudosciaena crocea, and the median lethal dose was 6.32×10⁴ CFU/mL, indicating strong pathogenicity to Pseudosciaena crocea.The results of drug sensitivity showed that the isolated bacteria were sensitive to amikacin and levofloxacin, moderately sensitive to gentamicin, neomycin and erythromycin, and resistant to cefixime, ampicillin, kanamycin and furazolidone.【Conclusion】 In this study, the pathogenic strain of Chryseobacterium isolated from diseased Pseudosciaena crocea was reported, and its pathogenicity and drug resistance were studied to provide references for the prevention, control and treatment of aquatic animal diseases caused by Chryseobacterium sp..

Key words: Chryseobacterium sp.; isolation and identification; artificial infection; sequence analysis; drug resistance

中图分类号:

S941.42

蔡杰, 吴佳怡, 潘苑霞, 呼高伟, 张林州. 1株大黄鱼源金黄杆菌的分离鉴定和耐药性分析[J]. 中国畜牧兽医, 2022, 49(3): 1135-1143.

CAI Jie, WU Jiayi, PAN Yuanxia, HU Gaowei, ZHANG Linzhou. Isolation, Identification and Drug Resistance Analysis of Chryseobacterium sp. from Pseudosciaena crocea[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1135-1143.

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