两株SPF级实验小鼠病原细菌的分离、鉴定及其基因组特征分析  

Isolation,identification,and genomic characterization of two strains of pathogenic bacteria from SPF mice

作　　者：石伟雄# 高飞 苏磊 SHI Weixiong;GAO Fei;SU Lei(National Human Diseases Animal Model Resource Center,Institute of Laboratory Animal Science,Chinese Academy of Medical Sciences,Beijing 100021,China;College of Animal Sciences(Laboratory Animal Center),Jilin University,Changchun 130062,Jilin,China)

机构地区：[1]中国医学科学院医学实验动物研究所国家人类疾病动物模型资源库,北京100021 [2]吉林大学动物科学学院(实验动物中心),吉林长春130062

出　　处：《微生物学通报》2025年第2期859-874,共16页Microbiology China

基　　金：国家重点研发计划(2021YFF0702900);吉林省科技发展计划(20230203145SF);国家科技资源共享服务平台项目(国家病原微生物资源库-NPRC-32)。

摘　　要：【背景】实验动物中不断发现干扰科学研究的新病原体,库氏棒杆菌(Corynebacterium kutscheri)和铜绿假单胞菌(Pseudomonas aeruginosa)作为影响实验动物健康的病原微生物,是国标中要求的必检微生物。本研究从吉林大学实验动物中心饲养的无特定病原体(specific pathogen-free,SPF)级小鼠体内分离到1株疑似库氏棒杆菌和1株疑似铜绿假单胞菌。【目的】鉴定这些病原微生物的种类、生物学特性、系统发育关系及基因组特征,为该病原微生物标准物质的研制及检测评价技术体系的建立提供生物资源并奠定基础。【方法】采用革兰氏染色、药敏试验、生理生化等方法对病原生物学特征进行鉴定;结合PCR鉴定、系统发育分析、全基因组测序、生物信息学等技术明确该菌的遗传学特征。【结果】形态观察、16S rRNA基因序列、基因组分析及系统发育分析证实,该分离菌株分别被鉴定为库氏棒杆菌CCPM-B-B-019-2402-1和铜绿假单胞菌CCPM-B-B-013-2402-3;药敏试验表明,库氏棒杆菌CCPM-B-B-019-2402-1对苯唑西林、呋喃妥因和苄青霉素耐药,铜绿假单胞菌CCPM-B-B-013-2402-3对替卡西林/克拉维酸和替加环素耐药,对头孢他啶、环丙沙星、左氧氟沙星等9种抗生素敏感;全基因组测序结果表明,库氏棒杆菌CCPM-B-B-019-2402-1携带有fagA、fagB、fagC、fagD等重要毒力基因,铜绿假单胞菌CCPM-B-B-013-2402-3携带有plcB、tse6、tse5等毒力基因。【结论】对SPF级小鼠气管组织和盲肠内容中分离出的2株病原菌进行了鉴定,并对2株病原菌进行了基因组特征分析,该研究为实验动物病原细菌标准株的建立,以及实验动物病原标准物质的研制和实验动物的质量控制提供了重要资源和支撑。[Background]New pathogens that interfere with scientific research have been constantly discovered in laboratory animals.Corynebacterium kutscheri and Pseudomonas aeruginosa as pathogenic microorganisms affecting the health of laboratory animals are required to be monitored in the national standard.In this study,suspected strains of C.kutscheri and P.aeruginosa were isolated from specific pathogen-free(SPF)mice reared at the Laboratory Animal Center of Jilin University.[Objective]To identify the species and reveal the biological characteristics,phylogenetic relationship,and genomic characteristics of these pathogens,providing valuable biological resources and laying a foundation for developing reference materials and establishing an evaluation system for these pathogens.[Methods]The biological characteristics of the pathogens were studied by Gram staining,drug sensitivity test,and physiological and biochemical methods.PCR,phylogenetic analysis,whole genome sequencing,and bioinformatics analysis were performed to clarify the genetic characteristics of the strains.[Results]Through morphological observation,16S rRNA sequencing,genome analysis,and phylogenetic analysis,the isolates were identified as C.kutscheri CCPM-B-B-019-2402-1 and P.aeruginosa CCPM-B-B-013-2402-3,respectively.C.kutscheri CCPM-B-B-019-2402-1 was resistant to oxacillin,nitrofurantoin,and benzylpenicillin.P.aeruginosa CCPM-B-B-013-2402-3 showed resistance to ticarcillin/clavulanic acid and tigecycline,while it demonstrated sensitivity to 9 antibiotics including ceftazidime,ciprofloxacin,and levofloxacin.The results obtained from whole genome sequencing indicated that C.kutscheri CCPM-B-B-019-2402-1 carried key virulence genes such as fagA,fagB,fagC,and fagD,whereas P.aeruginosa CCPM-B-B-013-2402-3 harbored virulence genes like plcB,tse6,and tse5.[Conclusion]In this study,two strains of pathogenic bacteria isolated from the tracheal tissue and cecum of SPF mice were identified,and their genomic characteristics were analyzed.This study provides import

关 键 词：库氏棒杆菌 铜绿假单胞菌 生物学特征 基因组特征 系统发育分析 

分 类 号：S858.91[农业科学—临床兽医学]