吲哚放线杆菌AIFJ1607株全基因组测序及耐药特性分析  

Analysis of whole genome sequence and drug resistance of Actinobacillus indolicus AIFJ1607 strain

作　　者：车勇良[1] 陈秋勇[1] 陈如敬[1] 吴学敏[1] 王隆柏[1] 刘玉涛[1] 周伦江[1] CHE Yongliang;CHEN Qiuyong;CHEN Rujing;WU Xuemin;WANG Longbai;LIU Yutao;ZHOU Lunjiang(Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fujian Animal Diseases Control Technology Development Center,Fuzhou 350013,China)

机构地区：[1]福建省农业科学院,畜牧兽医研究所,福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《中国兽医学报》2021年第12期2377-2384,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31872502);福建省公益类科研院所专项资助项目(2019R1026-13);福建省农业科学院科研资助项目(AGY2018-6)。

摘　　要：研究旨在探究吲哚放线杆菌AIFJ1607株的多重耐药机制。首先采用生化试验和16S rRNA的测序分析来鉴定细菌种类,然后应用药敏和MIC试验来分析20种抗生素的敏感性,再次采用全基因组测序和生物信息学方法来分析细菌的全基因组序列特征和耐药特性,最后分离纯化细菌质粒并测序来分析该质粒特征。结果显示,生化鉴定和16S rRNA测序分析结果显示该菌生长需要NAD或X因子的参与,过氧化氢酶和吲哚为阳性,尿素酶和cAMP试验为阴性,且16S rRNA序列与吲哚放线杆菌具有99%的同源性。药敏试验结果显示该菌对庆大霉素、妥布霉素、大观霉素、链霉素、新霉素、卡那霉素、青霉素、头孢、氯霉素、氟苯尼考、磺胺、四环素、博莱霉素、甲硝唑和恩诺沙星等15种抗生素耐药。全基因组序列分析结果显示了13种耐药基因,分别为ble、NimC/NimA、tetB、norA、bla;、sul2、Cat3、aph33ib、aac(3)-Ⅱa、aph33ib、aph6id、aph3ia、ant3ia,另外还有一种砷抑制剂arsR基因。并且发现了多种插入序列(IS),分别为IS6、IS10、IS30、ISApl1和ISApl4。质粒序列分析结果显示质粒pAIFJ1含有氟苯尼考耐药基因floR,且与副猪嗜血杆菌质粒pHPSGC具有很高的同源性。结果表明,该分离菌株AIFJ1607为吲哚放线杆菌,药敏试验结果与全基因组序列分析的耐药基因结果一致,分离质粒与副猪嗜血杆菌中的质粒高度同源预示着该质粒pAIFJ1可能在吲哚放线杆菌和副猪嗜血杆菌之间进行水平转移。The study aimed to explore antibiotic resistant mechanism of the multidrug-resistant Actinobacillus indolicus(A.indolicus)strain AIFJ1607.Bacterial species was identified through biochemical assays and 16 S rRNA gene sequencing.Antibiotic resistance phenotype for 20 antibiotics was determined using K-B method and MIC determination.The whole-genome sequence of AIFJ1607 strain was determined using the PacBio Sequel and was analyzed using bioinformatics.Antibiotic resistance gene determinants were identified using ResFinder.The genetic characterization of the plasmid pAIFJ1 carrying floR gene from AIFJ1607 strain was analyzed by sequencing,annotating and phylogenetic tree.Biochemical test showed NAD or X factor was required for the growth of the AIFJ1607 strain,catalase and indole were positive,urease and cAMP tests were negative.The sequence of 16 S rRNA of the AIFJ1607 strain had 99%homology to A.indolicus.Antibiotic sensitivity test showed that the AIFJ1607 strain was resistant to 15 antibiotics,such as gentamicin,tobramycin,spectinomycin,streptomycin,neomycin,kanamycin,penicillin,cephalosporin,chloramphenicol,florfenicol,sultanilamide,tetracycline,bleomycin,metronidazole and enrofloxacin.Whole-genome sequence analysis showed that there were 13 antibiotic resistance genes,which were ble,NimC/NimA,tetB,norA,blaROM-1,sul2,cat3,aph33 ib,aac(3)-Ⅱa,aph33 ib,aph6 id,aph3 ia and ant3 ia.Moreover,ArsR gene,an arsenic inhibitor,was also found in the whole genome.In the meantime,a variety of insertion sequences(IS)were also found,including IS6,IS10,IS30,ISApl1 and ISApl4.The results of plasmid sequencing showed that the pAIFJ1 containing floR gene,which was highly homologous to the plasmid pHPSGC in Haemophilus parasuis.The results revealed that the bacteria AIFJ1607 strain was A.indolicus.The results of antibiotic sensitivity test were consistent with the results of whole-genome sequence analysis.The high homology between the isolated plasmid and the plasmid in Haemophilus parasuis indicated that the plasmid pAIFJ1 migh

关 键 词：吲哚放线杆菌 全基因组测序 耐药特性 插入序列 

分 类 号：S855.1[农业科学—临床兽医学]