华南虎源产碱普罗威登斯菌的分离鉴定及致病性研究  

Isolation,Identification and Pathogenicity of Providencia alcalifaciens from South China Tiger

作　　者：李雨琪 朱智豪 郑欣 许癸宗 蔡思思 周娴静 靳雨欣 刘璐 许静茹 陈芳婷 薛梦如 康娅莉 薛羽希 陈腾腾 林锡潘 林开雄 谭聪灵 张敏 范克伟 LI Yuqi;ZHU Zhihao;ZHENG Xin;XU Guizong;CAI Sisi;ZHOU Xianjing;JIN Yuxin;LIU Lu;XU Jingru;CHEN Fangting;XUE Mengru;KANG Yali;XUE Yuxi;CHEN Tengteng;LIN Xipan;LIN Kaixiong;TAN Congling;ZHANG Min;FAN Kewei(Engineering Research Center for the Prevention and Control of Animal Original Zoonosis of Fujian Province University,Fujian Provincial Key Laboratory for the Prevention and Control of Animal Infectious Diseases and Biotechnology,College of Life Sciences,Longyan University,Longyan 364000,China;College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Fujian Meihuashan Institute of South China Tiger Breeding,Longyan 364201,China;Chongqing Nanchuan District Animal Husbandry and Veterinary Fishery Center,Chongqing 408499,China)

机构地区：[1]龙岩学院生命科学学院,福建省家畜传染病防治与生物技术重点实验室,动物源性人兽共患病防控福建省高校工程研究中心,龙岩364000 [2]福建农林大学动物科学学院,福州350002 [3]福建梅花山华南虎繁育研究所,龙岩364201 [4]重庆市南川区畜牧兽医渔业中心,重庆408499

出　　处：《中国畜牧兽医》2024年第3期1320-1328,共9页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金青年科学基金项目(31602048);福建省高校产学合作项目(2020N5012);福建省自然科学基金项目(2023J01990);国家级大学生创新创业训练计划项目(202011312012、202111312006、202311312009);新罗区产学研科技联合创新项目(2022XLXYZ009)。

摘　　要：【目的】分离鉴定导致华南虎急性死亡的病原菌,确定其致病性及药物敏感性,为今后该病的防控提供科学依据。【方法】利用形态学、分子生物学确定分离菌株种类,并对其进行小鼠致病性试验、药敏试验。【结果】从死亡华南虎幼虎肠道中分离到1株细菌。分离菌在鲜血琼脂平板上呈现灰白色菌落,在麦康凯平板上呈现白色菌落;镜检为革兰阴性杆菌,命名为FJ/Tiger201802。结合16S rRNA测序结果,确定其为产碱普罗威登斯菌。遗传进化分析显示,FJ/Tiger201802与鱼源分离株HXM38(MZ734428.1)位于同一进化分支,亲缘关系较近,16S rRNA基因核苷酸相似性为99.5%。分离菌株接种试验小鼠,发病小鼠出现精神萎靡、被毛松乱、扎堆、眼角出现分泌物等症状,分离菌株对小鼠的半数致死量(LD50)为1×108.2CFU,病理剖检及病理组织学结果表明,FJ/Tiger201802的致病性较强,主要侵害小鼠的肝脏、肺脏、肠等组织器官。药敏试验结果表明,氨苄西林、氨曲南、头孢呋辛、头孢哌酮、头孢曲松、头孢吡肟、头孢他啶、头孢西叮、氧氟沙星、链霉素、妥布霉素和庆大霉素对分离菌株具有较强的体外生长抑制作用,对头孢唑林、头孢噻吩不敏感。【结论】本研究分离到1株华南虎源产碱普罗威登斯菌,可引起小鼠感染发病。研究结果可为产碱普罗威登斯菌病的临床诊断和治疗提供参考依据。【Objective】The aim of this experiment was to isolate and identify the pathogenic bacteria causing the acute death of South China tiger,determine its pathogenicity and drug sensitivity,and provide scientific basis for the prevention and control of the disease in the future.【Method】Morphological and molecular biology were used to determine the types of isolated strains,and the pathogenicity test and drug sensitivity test of the isolated strains were carried out.【Result】A strain of bacteria was isolated from the intestine of dead South China tiger cubs.The isolated bacteria showed gray-white colonies on the fresh blood agar plate and white colonies on the MacConkey plate.Microscopically,it was Gram-negative bacilli,named FJ/Tiger201802.Combined with the results of 16S rRNA sequencing,it was identified as Providencia alcalifaciens.Phylogenetic analysis showed that FJ/Tiger201802 was closely related to HXM38(MZ734428.1)in the same evolutionary branch,and the nucleotide similarity of 16S rRNA gene was 99.5%.The mice inoculated with the isolated strain showed symptoms such as listlessness,loose hair,clumping,and secretions in the eyes.The median lethal dose(LD 50)of the isolated strain to mice was 1×108.2 CFU.The results of pathological anatomy and histopathology showed that the pathogenicity of FJ/Tiger201802 was strong,mainly affecting the liver,lung,intestine and other tissues and organs of mice.The results of drug sensitivity test showed that ampicillin,aztreonam,cefuroxime,cefoperazone,ceftriaxone,cefepime,ceftazidime,cefoxitin,ofloxacin,streptomycin,tobramycin and gentamicin had a strong inhibitory effect on the growth of the isolated strain in vitro,and the isolate were not sensitive to cefazolin and cefalotin.【Conclusion】In this study,a strain of Providencia alcalifaciens from South China tiger was isolated,which could cause infection in mice.The results provided a reference for the clinical diagnosis and treatment of Providencia alcaligenes.

关 键 词：华南虎 产碱普罗威登斯菌 分离鉴定 致病性 

分 类 号：S852.61[农业科学—基础兽医学]