猪源副猪链球菌的分离鉴定及全基因组测序分析  

Identification and whole genome sequencing analysis of swine origin and Streptococcus parasuis

作　　者：王云飞 陆童 董文龙 沙万里 闫满 李国江 WANG Yunfei;LU Tong;DONG Wenlong;SHA Wanli;YAN Man;LI Guojiang(College of Animal Science and Technology,Jilin Agricultural University,Changchun130118,China;College of Animal Science and Technology,Jilin Institute of Agricultural Science and Technology,Jilin City,Jilin 132109,China;Jilin Province Key Laboratory of Preventive Veterinary Medicine,Jilin City,Jilin 132109,China;Jilin Province Pig Ecological Breeding and Disease Prevention and Control Science and Technology Innovation Center,Jilin City,Jilin 132109,China)

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]吉林农业科技学院动物科技学院,吉林吉林132109 [3]吉林省预防兽医学重点实验室,吉林吉林132109 [4]吉林省猪生态养殖及疫病防控科技创新中心,吉林吉林132109

出　　处：《中国兽医学报》2024年第10期2159-2164,共6页Chinese Journal of Veterinary Science

基　　金：吉林省科技创新平台资助项目(YDZJ202302CXJD038)。

摘　　要：从我国吉林地区患有肺炎的病猪分离到优势菌株,对分离菌株进行16S rRNA鉴定,确定为副猪链球菌(Streptococcus parasuis,S.parasuis),命名为WYF-8B。药敏纸片扩散法检测结果显示,WYF-8B对杆菌肽、磺胺间甲氧苄啶、林可霉素、红霉素、磷霉素、阿奇霉素、青霉素、多西环素、复方新诺明、磺胺异恶唑、环丙沙星、恩诺沙星、四环素、苯唑西林、庆大霉素、头孢唑林、氨苄西林均耐药;对氟苯尼考,氯霉素高度敏感。生化鉴定结果显示,WYF-8B可发酵水杨酸、麦芽糖、乳糖、蔗糖、七叶苷、葡萄糖;不发酵甘露醇、尿素、棉子糖、木糖、阿拉伯糖、山梨醇。利用第二代全基因组测序技术对WYF-8B耐药基因型进行研究,结果发现其携带对大环内酯类抗生素的抗性基因Mef(E)、ErmB,对β-内酰胺类抗生素的抗性基因pbp2b、pbp1a、pbp2x,对氟喹诺酮类抗生素的抗性基因gyrA、parC、grlB,对氨基糖苷类抗生素的抗性基因ANT(6)-Ia、AAC(6')-Ie-APH(2'')-Ia、ANT(9)-Ia、APH(3')-Ⅲa,对林可酰胺类抗生素的抗性基lnuB、lmrC、lmrD,对四环素类抗生素的抗性基因tetM,对杆菌肽的抗性基因bcrA,结果显示耐药表型与测序结果的耐药基因型相符。WYF-8B毒力因子研究结果显示,WYF-8B共携带13个毒力因子(pavA、fbp54、clpP、cps4I、wbtB、cpsI、tufA、galE、hasC、groEL、wbtL、gndA、wbtF),其作用机制主要为促进细菌的扩散、转移、定居、抗吞噬及不被宿主免疫细胞识别等。药物敏感试验结果为研究副猪链球菌的临床用药提供参考,耐药表型与毒力因子为新型药物及疫苗的研发奠定基础。Streptococcus parasuis(S.parasuis)is a close relative of S.suis and can cause meningitis,pneumonia or systemic symptoms in affected animals.In this paper,the dominant strain was isolated from diseased pigs with pneumonia in Jilin,China,and the isolated strain was identified as S.parasuis by 16S rRNA,it was named WYF-8B.Resistance was tested by the susceptibility tablet diffusion method,and WYF-8B was resistant to bacitracin,sulfamethoxoprim,lincomycin,erythromycin,fosfamycin,azithromycin,penicillin,doxycycline,cotrimotrixazole,sulfamisoxazole,ciprofloxacin,enrofloxacin,tetracycline,benzoxacillin,gentamicin,efazolin,and ampicillin.It was highly sensitive to florfenicol,chloramphenicol.Iochemical identification revealed that WYF-8Bcould ferment salicylic acid,maltose,lactose,sucrose,oxaesin,and glucose;nonfermentable mannitol,urea,raffinose,xylose,arabinose,and sorbitol.Using the second-generation whole-genome sequencing technology to study the WYF-8Bresistant genotype,it was found that it carried the macrolide antibiotic resistance genes Mef(E)and ErmB;resistance genes pbp2b,pbp1a,and pbp2xtoβ-lactam antibiotics;resistance genes gyrA,parC,and grlBto fluoroquinolones;resistance genes ANT(6)-Ia,AAC(6′)-Ie-APH(2′)-Ia,ANT(9)-Ia,APH(3′)-Ⅲato aminoglycoside antibiotics;resistance groups lnuB,lmrC,lmrD against lincomide antibiotics;resistance gene tetM to tetracycline antibiotics;the resistance gene of bacitracin,bcrA.The results found that the resistance phenotype matches the resistance genotype of the sequencing results.The study of WYF-8Bvirulence factors found that WYF-8Bcarried 13virulence factors(pavA,fbp 54,clpP,cps 4,cps 4I,wbtB,cps,sI,tufA,galE,hasC,groEL,wrbtL,wbbtF),which mainly acted to promote bacterial diffusion,transfer,settlement,antiphagocytosis and immune evasion.The results of drug sensitivity test provide a reference for the study of clinical medication of S.parasuis,and drug resistance phenotype and virulence factors lay the foundation for the research and development of new drugs and vaccin

关 键 词：副猪链球菌 全基因组测序 耐药基因 猪 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]