云南省普洱市思茅区大面积野鼠自毙疫情实验室检测结果与分析  

Test results and analysis of rodent with unexplained death in a large area in Simao,Pu’er,Yunnan

作　　者：杨珊[2] 宋志忠[1] 高子厚[1] 陈朝年 马顺高 桑卫洪 王鹏[1] 张云智[1] 周济华[1] 左曙青 梁云[1] 杜春红[1] YANG Shan;SONG Zhi-zhong;GAO Zi-hou;CHEN Chao-nian;MA Shun-gao;SANG Wei-hong;WANG Peng;ZHANG Yun-zhi;ZHOU Ji-hua;ZUO Shu-qing;LIANG Yun;DU Chun-hong(Pu’er Center for Disease Control and Prevention,Pu’er,Yunnan 66500,China;不详)

机构地区：[1]云南省地方病防治所,云南大理671000 [2]云南省普洱市疾病预防控制中心 [3]云南省普洱市思茅区疾病预防控制中心 [4]云南省大理白族自治州人民医院 [5]军事科学院军事医学研究院微生物流行病研究所

出　　处：《现代预防医学》2021年第19期3591-3594,3599,共5页Modern Preventive Medicine

基　　金：国家自然科学基金项目(81760607,81360413);病原微生物生物安全国家重点实验室开放基金(SKLPBS1833)。

摘　　要：目的调查引起云南省普洱市思茅区大面积野鼠自毙疫情的病原体,以便为采取相应措施提供科学依据。方法通过现场流行病学调查,采集119只自毙野鼠样本,解剖取脏器进行鼠疫耶尔森菌病原体分离、噬菌体裂解实验、F1抗原、caf1基因等病原学、免疫学及分子生物学检测,以排除鼠疫。同时,脏器样本接种绵羊血琼脂平板、巧克力平板、麦康凯平板、沙氏琼脂培养基等营养培养基和选择性培养基,分离培养细菌、真菌类病原体;选取10只组织腐败较轻的自毙鼠,按肺组织、脑组织、腹腔脏器组织分为3份样本后,研磨、离心,再以0.22μm滤膜过滤,收集离心上清液、滤膜滤出液和滤膜截留残渣共9份样本,腹腔接种雄性、体质量30~40 g昆明小鼠,每组2只,每只0.5 ml。另接种0.5 ml Hank’s液2只鼠作对照。共接种昆明小鼠20只。死亡小鼠解剖观察病理变化,取脏器组织接种上述培养基进行细菌分离培养。结果鼠疫耶尔森菌病原学、免疫学和分子生物学检测均为阴性;自毙野鼠脏器镜检、分离培养及接种实验动物均检出G-短小杆菌,经梅里埃VITEK 2GN、GP卡、16S rRNA扩增鉴定为多杀巴斯德菌(Pasteurella multocida,Pm),概率为99%;分离到的G+粗大杆菌为腊样芽孢杆菌,概率为99%。结论多杀巴斯德菌可能是这次发生野鼠大面积死亡的病原菌。该病原体的生物学特性及危害值得进一步关注。Objective To investigate the pathogens causing the large-scale unexplained deaths of wild rats in Simao,Pu’er,Yunnan,and to provide scientific basis for clarifying the epidemic situation and corresponding measures.Methods Through field epidemiological investigation,119 autopsy rodent samples were collected and dissected to detect pathogenic bacteria,especially Yersinia pestis,via isolation assay,phage lysis test,F1 antigen detection,caf1 gene amplification to verify the infection.The samples of lungs,brains and abdominal visceral organs were collected from 10 self-killed mice with less corruption.After grinding,centrifugation,and then filtering with 0.22μm membrane,9 samples of centrifugal supernatant,membrane filtrate and filter membrane interception residue were collected,0.5 ml of which was inoculated into each male Kunming mice(30~40 g)intraperitoneally,with 2 mice in each group,while another 2 Kunming mice inoculated with 0.5 ml Hank’s solution were adopted as control.A total of 20 Kunming mice were inoculated.The abdominal visceral organs were used for pathogen isolation,by using Sheep Blood Agar,Chocolate Agar,MacConkey Agar and Sabouraud Dextrose Agar.The dead mice were dissected to observe the pathological changes,and the organ tissues were inoculated with the above culture medium for bacterial isolation.Results The etiological,immunological and molecular biological tests of Yersinia pestis were negative.Microscopical examination,isolation,culture of organs of self-killed wild rats,and inoculation of experimental animals all detected G-pumilus,which was identified as Pasteurella multocida by Merier VITEK 2 GN,GP card and 16 S rRNA amplification,with a probability of 99%.The isolated G+crude bacillus was Bacillus cereus,with a probability of 99%.Conclusion Pasteurella multocida may be the main pathogen causing the death of wild rats.The biological characteristics and harm of the pathogen deserve further attention.

关 键 词：云南 野鼠 死亡 鼠疫 多杀巴斯德菌 

分 类 号：R111[医药卫生—公共卫生与预防医学]