一株牛源溶酪巨型球菌致病性及全基因组特征分析  

Pathogenicity and draft genome characteristics analysis of a bovine-derived Macrococcus caseolyticus

作　　者：薛嘉熹 范文雨 曹鑫艳 高之煜 顾兰英 张秦川 李世义 盛金良[1] 孙延鸣[1] 张彦兵[1] XUE Jiaxi;FAN Wenyu;CAO Xinyan;GAO Zhiyu;GU Lanying;ZHANG Qinchuan;LI Shiyi;SHENG Jinliang;SUN Yanming;ZHANG Yanbing(College of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang 832000,China)

机构地区：[1]石河子大学动物科技学院,新疆石河子832003

出　　处：《石河子大学学报（自然科学版）》2024年第4期438-448,共11页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(32360901);新疆维吾尔自治区青年科学基金项目(2022D01B197);石河子大学青年创新培育人才项目(CXPY202211);石河子大学高层次人才科研启动项目(RCZK202043)。

摘　　要：目的本研究旨在研究1株牛源溶酪巨型球菌(Macrococcus caseolyticus,M.caseolyticus)的致病性和全基因组特征,挖掘该菌株的毒力和耐药相关基因。方法在新疆石河子市某牛场中采集1例病死牛肺脏,从牛肺脏灌洗液中分离鉴定了1株牛源溶酪巨型球菌,在此基础上进行了药敏试验、动物感染试验;并通过Illumina的二代测序技术对该菌株进行全基因组框架图构建,构建多个管家基因同源进化树,对编码基因、重复序列、非编码RNA等进行预测,并在GO、KEGG、COG、NR和CAZy数据库对基因组的基本功能特征进行注释,结合NR数据库的注释和PHI、CARD数据库注释,明确其基因组中毒力因子、耐药性等相关基因。结果从牛肺中成功分离到一株M.caseolyticus,通过感染兔子试验证实该菌株具有较强的致病性,病理切片结果显示兔子肺泡数量减少,大量炎性细胞渗出,肺泡囊增大,肺泡囊中充满渗出物,红细胞渗出,细支气管管壁塌陷。该菌经平均核苷酸相似度分类和系统发育树确定为溶酪巨型球菌,全基因组大小为2209314 bp,CG%含量为37.22%,平均长度为855 bp,编码2304个基因;其中tRNA有58个、rRNA有8个、sRNA有1个;含有3条CRISPRs序列。在GO数据库中有1028个基因得到注释,分别注释到分子功能、细胞组分和生物过程3大类;在KEGG数据库中共有1420个编码基因得到注释;在COG数据库中,共注释有23种功能的1961个基因;同时,在全基因组测序结果中发现51个毒力因子和对氨基糖苷类、β-内酰胺类和多肽类等抗生素的多种耐药基因,药物敏感性试验中菌株对测试的12类抗生素仅对磺胺类(复方新诺明)、林可霉素类(林可霉素)、青霉素类(氨苄西林)耐药。结论本研究从病死牛肺脏中分离得到1株牛源溶酪巨型球菌并命名为SHZXbov7,该菌具有较强的致病性,含有多种毒力因子和耐药基因,菌株SHZXbov7的基因组测序和注释将丰富溶酪巨�Objective In this study,we investigated the pathogenicity and genome-wide characteristics of a strain of Macrococcus caseolyticus(M.caseolyticus),and explored the virulence and drug resistance-related genes of this strain.Methods A strain of bovine-derived M.caseolyticus was isolated and identified from the dead bovine lungs collected at a cattle farm in Shihezi City,Xinjiang.Subsequently,drug sensitivity tests,animal tests,and pathological and histological observations were conducted.The draft genome sequencing of this strain was then performed using second-generation sequencing technology of illumina.Further,species affinities were further evaluated based on the sequencing results.The composition of the target genome,including coding genes,repetitive sequences,and non-coding RNAs,was determined,and basic functional features of the genome were annotated in databases such as GO,KEGG,COG,NR,and CAZy.Furthermore,a combination of annotations from the NR database and the PHI and CARD databases was utilized to identify genome virulence factors,drug resistance,and other related genes.Results Pathological section results revealed various abnormalities in the rabbit,including a decrease in the alveoli count,exudation of inflammatory cells,enlargement and filling of alveolar sacs with exudate,thinning of vascular walls,decreased permeability,exudation of red blood cells,and collapse of the fine bronchial tube walls.The pathogen was identified as M.caseolyticus through mean nucleotide similarity classification and phylogenetic tree analy-sis.Its draft genome size is 2209314 base pairs,with a CG%content of 37.22%and an average gene length of 855 base pairs.The genome encodes 2304 genes,including 58 tRNAs,8 rRNAs,and 1 sRNA,as well as 3 sequences of CRISPRs.Additionally,1028 genes were annotated in the GO database,covering three major categories:molecular functions,cellular components,and biological processes.Furthermore,1420 coding genes were annotated in the KEGG database,and 1961 genes were annotated with 23 functions in

关 键 词：溶酪巨型球菌 耐药基因 毒力基因 基因组草图测序 

分 类 号：R943[医药卫生—药剂学]