半滑舌鳎源海藻希瓦氏菌的分离鉴定及药敏试验  

Isolation,identification and drug sensitivity test of Cynoglossus semilaevis-derived Shewanella algae

作　　者：白和平 刘畅 张鹿 张志强[1] 吴同垒[1] 高桂生[1] 史秋梅[1] BAI Heping;LIU Chang;ZHANG Lu;ZHANG Zhiqiang;WU Tonglei;GAO Guisheng;SHI Qiumei(Hebei Province Key Laboratory of Preventive Veterinary Medicine,Hebei Normal University of Science and Technology,Qinhuangdao 066004,China)

机构地区：[1]河北科技师范学院河北省预防兽医学重点实验室,河北秦皇岛066004

出　　处：《黑龙江畜牧兽医》2022年第18期81-84,142,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：河北省科技重大专项(20286701Z);秦皇岛市科学技术研究与发展计划项目(201803B006)。

摘　　要：河北省昌黎县某半滑舌鳎鱼养殖场于2022年6月中旬出现发病,为了从死亡半滑舌鳎腹水中分离优势感染菌,试验采用了细菌分离纯化、革兰氏染色、生化鉴定、16S rRNA序列分析进行病原鉴定,并采用纸片扩散法检测分离菌对22种抗生素的敏感性,琼脂扩散法检测分离菌对22种中药的抑菌效果。结果表明:分离菌在血琼脂培养基上培养可见光滑圆润、灰色凸起状,呈β溶血的菌落,革兰氏染色镜检为阴性小杆菌,生化特性与海藻希瓦氏菌生化特性符合率为96%,分离菌16S rRNA基因序列与GenBank中海藻希瓦氏菌同源性达98.94%;分离菌对头孢曲松、头孢噻呋、大观霉素敏感,对氨苄西林、阿莫西林、卡那霉素等13种抗生素耐药;中药五味子、乌梅、诃子对分离菌的抑菌作用较强,秦皮、黄柏、紫花地丁等11味中药无抑菌效果。说明分离菌为海藻希瓦氏菌,对抗生素头孢曲松、头孢噻呋、大观霉素和中药五味子、乌梅、诃子6种药物敏感。A Cynoglossus semilaevis fish farm in Changli County,Hebei Province,developed a disease in mid-June 2022.In order to isolate the dominant infection bacteria from the ascites of the dead Cynoglossus semilaevis,bacterial isolation and purification,Gram staining,biochemical identification,and 16 S rRNA sequence analysis were used in the experiment.The Kirby-Bauer test paper method was used to detect the sensitivity of the isolated bacteria to 22 kinds of antibiotics,and the agar diffusion method was used to detect the bacteriostatic effect of 22 kinds of traditional Chinese medicines.The results showed that the isolated bacteria cultured in blood agar medium were smooth,round,gray and convex,β-hemolytic colony;Gram staining and microscopic examination showed negative bacilli.The coincidence rate of biochemical characteristics with that of Shewanella algae was 96%.The homology of 16 S rRNA gene sequence of the isolated bacteria with that of Shewanella algae in GenBank was 98.94%.The isolates were sensitive to ceftriaxone,ceftiofur and spectinomycin,and resistant to ampicillin,amoxicillin and kanamycin.Schisandra chinensis,Fructus chebulae and Terminalia chebula had the strongest bacteriostatic effect on the isolated bacteria,while 11 traditional Chinese medicines such as Cortex fraxini,Cortex phellodendri and Herba violae had no bacteriostatic effect.The results suggested that the isolated strain was Shewanella algae,which was sensitive to antibiotics ceftriaxone,ceftiofurol,megamycin,Chinese traditional medicine Schisandra chinensis,Fructus chebulae and Terminalia chebula.

关 键 词：半滑舌鳎 海藻希瓦氏菌 分离 鉴定 16S rRNA基因 抗生素药敏试验 中药药敏试验 

分 类 号：S852.61[农业科学—基础兽医学]