华南虎源铜绿假单胞菌的分离鉴定及其生物学特性研究  

Isolation,identification and biological characteristics of Pseudomonas aeruginosa from South China tiger

作　　者：李雨琪 吴林悦 黄林洁 朱智豪 许静茹 蔡思思 周娴静 邓莹滋 靳雨欣 陈芳婷 林开雄 陈腾腾 林锡潘 林俊宏 黄雨欣 许癸宗 俞国华 范克伟 LI Yu-qi;WU Lin-yue;HUANG Lin-jie;ZHU Zhi-hao;XU Jing-ru;CAI Si-si;ZHOU Xian-jing;DENG Ying-zi;JIN Yu-xin;CHEN Fang-ting;LIN Kai-xiong;CHEN Teng-teng;LIN Xi-pan;LIN Jun-hong;HUANG Yu-xin;XU Gui-zong;YU Guo-hua;FAN Ke-wei(Fujian Provincial Key Laboratory for the Prevention and Control of Animal Infectious Diseases and Biotechnology,Engineering Research Center for the Prevention and Control of Animal Original Zoonosis of Fujian Province University,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)

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,福建龙岩364000 [2]福建农林大学动物科学学院,福建福州350002 [3]福建梅花山华南虎繁育研究所,福建龙岩364201

出　　处：《中国兽医科学》2023年第6期715-726,共12页Chinese Veterinary Science

基　　金：国家自然科学基金青年科学基金项目(31602048);福建省农业科技重大专项(2019NZ09005);福建省高校产学合作项目(2020N5012);福建省高校新世纪优秀人才支持计划项目(闽教科〔2018〕47号);福建省自然科学基金项目(2019J01801);国家级大学生创新创业训练计划项目(202011312012,202111312010,202111312006);龙岩学院校级科技创新团队建设项目(岩学院科研[2018]13号);新罗区产学研科技联合创新项目(2022XLXYZ009)。

摘　　要：为探明福建省梅花山华南虎繁育研究所1只3岁华南虎公虎的死亡原因,无菌采集死亡虎的肝、脾、肺等组织样本,并接种于鲜血琼脂培养基分离优势细菌,利用表型特性及16S rRNA基因序列分析鉴定分离菌株;通过毒力基因检测、小鼠致病性试验、药敏试验及耐药基因检测对分离菌株进行致病性和耐药性研究。结果显示,从死亡华南虎肝组织中分离到1株铜绿假单胞菌,命名为FJ/Tiger-201801;该分离菌与26株铜绿假单胞菌16S rRNA的核苷酸序列同源性为97.1%~99.6%,其中与水貂源CS-20株、林麝源FMDP002株、人源Kasamber2株的同源性高达99.6%;毒力基因检测结果显示,该分离菌携带exoT、exoS、exoY这3种毒力基因;动物致病性试验结果显示,该分离菌对小鼠具有较强的致病性,试验小鼠在攻毒后13 h内全部死亡,并对死亡小鼠的肝、肺、脾等组织器官造成了严重的病理组织损伤;对11类30种常用抗菌药物的药敏试验结果显示,该分离菌株对氨苄西林等其中8类18种抗菌药物均表现耐药,对多黏菌素B等6种抗菌药物表现中度敏感,仅对头孢吡肟、氧氟沙星、左氧氟沙星、环丙沙星、妥布霉素和哌拉西林这6种抗菌药物表现敏感;对8类35种耐药基因检测结果显示,该分离菌同时携带有喹诺酮类、β-内酰胺类和四环素类抗菌药物的6种耐药基因。综上,本研究从死亡华南虎肝组织中分离获得了1株具有较强致病力且耐药严重的铜绿假单胞菌,为华南虎等野生动物铜绿假单胞菌病的防控提供了重要的参考依据。In order to investigate the dead cause of a 3-year-old male South China tiger from the Fujian Meihuashan Institute of South China Tiger Breeding.The liver,spleen,lung and other tissues of dead tiger were collected aseptically,and the dominant bacteria were isolated on fresh blood agar medium.Phenotypic characteristics and 16S rRNA gene sequence analysis were used to identify the isolated strains.Pathogenicity and drug resistance of the isolated strains were studied through virulence gene detection,mouse pathogenicity testing,drug sensitivity testing,and drug resistance gene assay.The results showed that a strain of Pseudomonas aeruginosa was isolated from the liver tissue of the dead South China tiger,named FJ/Tiger-201801.The nucleotide sequence homology between this isolate and the 16S rRNA of 26 strains of Pseudomonas aeruginosa was 97.1%—99.6%,among which the homology with the CS-20 strain from mink,FMDP002 strain from forest musk,and Kasamber 2 strain from human was as high as 99.6%.The results of virulence gene testing showed that the isolated strain carried exoT,exoS,and exoY virulence genes.The results of animal pathogenicity tests showed that the isolated strain had a strong pathogenicity to mice.All the mice died within 13 hours after the attack,and caused severe pathological tissue damage to the liver,lungs,spleen,and other tissues and organs of the dead mice.The results of drug sensitivity tests on 30 commonly used antimicrobial agents in 11 categories showed that the isolated strain was resistant to 18 antimicrobial agents in 8 categories,including ampicillin,and moderately sensitive to 6 antimicrobial agents,including polymyxin B.It was only sensitive to 6 antimicrobial agents,including cefepime,ofloxacin,levofloxacin,ciprofloxacin,tobramycin,and piperacillin.The detection results of 35 drug resistance genes in 8 categories showed that the isolated bacteria simultaneously carried 6 resistance genes for quinolones,β-lactams and tetracyclines.In summary,this study isolated a strain of Pseudomonas a

关 键 词：华南虎 铜绿假单胞菌 分离鉴定 致病性 

分 类 号：S852.614[农业科学—基础兽医学]