致半滑舌鳎烂尾病的创伤弧菌的分离鉴定及中草药对其抑菌效果研究  

Isolation and identification of Vibrio vulnificus causing rotting tail disease in half-smooth tongue sole and its inhibition by Chinese herbs

作　　者：刘勃兴 崔舒畅 任海 靳晓敏[1] 韩红生 史秋梅[1] 高桂生[1] LIU Bo-xing;CUI Shu-chang;REN Hai;JIN Xiao-min;HAN Hong-sheng;SHI Qiu-mei;GAO Gui-sheng(Hebei Normal University of Science&Technology,Hebei Key Laboratory of Preventive Veterinary Medicine,Qinhuangdao 066004,China;Qinhuangdao Liangfeng Marine Ecological Technology Development Co.,Ltd.,Qinhuangdao 066600,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 [2]秦皇岛粮丰海洋生态技术开发有限公司,河北秦皇岛066600

出　　处：《中国预防兽医学报》2021年第5期489-494,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：河北省科技厅项目(2028701Z);河北科技师范学院海洋专项(2018HU014);河北科技师范学院海洋专项(2018HU007)。

摘　　要：为检测秦皇岛市昌黎县某渔场养殖的致半滑舌鳎烂尾病的致病原,检测分离菌对抗生素的敏感性以及中草药对其的体外抑菌效果,本研究从患病鱼尾部肌肉分离纯化到一株细菌,采用TCBS鉴定培养基培养,结果可见绿色菌落,边缘整齐,表面光滑,轻微凸起;经革兰氏染色后镜检可见,分离菌为革兰氏阴性菌,两端钝圆,形状似月牙;经ATB全自动微生物鉴定仪判定,分离菌为创伤弧菌,符合率为99%;通过16S rDNA序列分析进一步鉴定分离菌为创伤弧菌,命名为CLVV-1。采用动物回归试验检测分离菌株的致病性,结果显示CLVV-1菌株可致半滑舌鳎出现烂尾病征;通过KB纸片法检测分离菌株对14种抗生素的药物敏感性,结果显示,CLVV-1菌株对氨苄西林等6种抗生素高度敏感,对阿米卡星等共4种抗生素中介,对土霉素等共4种抗生素耐药;经牛津杯法和二倍稀释法检测15味中草药水煎提取液对分离菌株的体外抑菌效果,结果显示,石榴皮等8味中草药液对CLVV-1菌株具有抑菌效果,而白头翁等7味中草药液对CLVV-1菌株无抑菌效果。本研究为创伤弧菌感染所致的半滑舌鳎烂尾病的防治提供参考,并首次为相关中草药制剂防治该菌的应用奠定基础。To detect the pathogen causing the tail rot of half-smooth tongue sole(Cynoglossus semilaevis)cultured in a fishing ground of Changli County,Qinhuangdao City,test the sensitivity of the isolate to antibiotics,and explore the in vitro bacteriostatic effect of herbs on the isolate,a bacterial strain was isolated and purified from the diseased tail muscle,and cultured in TCBS identification medium,the results showed that green colonies formed on the plate,with neat edges,smooth,and slight projections.Microscopic examination after gram staining showed that the isolated bacteria were Gram-negative bacteria with distant ends and shaped like crescent moon.The automatic microbial identification instrument determined that the isolate was Vibrio vulnificus,with a compliance rate of 99%.The isolate was identified by 16S rDNA sequence analysis,and the results further showed that it was Vibrio vulnificus,which was subsequently named CLVV-1.The pathogenicity of the isolate was detected by animal regression test,the results showed that CLVV-1 caused tail rot of half-smooth tongue sole.The drug sensitivity of the isolate to 14 antibiotics was detected by KB disk method,it showed that CLVV-1 strain was highly sensitive to ampicillin and other 6 antibiotics,moderately sensitive to amikacin and other four antibiotics,and resistant to oxytetracycline and other four antibiotics.The in vitro bacteriostatic effect of 15 Chinese herbal medicine decoction extracts on the isolate was detected by Oxford cup method and double dilution method,it showed that eight kinds of extracts such as Pomegranate peel had bacteriostatic effect on CLVV-1,while other seven kinds of extracts including the root of Chinese pulsatilla had no antibacterial effect.This study provides a reference for the prevention and treatment of the tail rot disease of half-smooth tongue sole caused by Vibrio vulnificus infection and lays a foundation for the research of related Chinese herbal medicine preparations.

关 键 词：半滑舌鳎 烂尾病 创伤弧菌 耐药性 中草药 

分 类 号：S852.61[农业科学—基础兽医学]