畜牧与饲料科学 ›› 2017, Vol. 38 ›› Issue (6): 12-12.doi: 10.12160/j.issn.1672-5190.2017.06.005

• 基础科学 • 上一篇    下一篇

仔猪黄痢病原菌的分子生物学鉴定及多重耐药基因检测

路振香;郭伟娜;龚争;刘畅;汪训磊   

  1. 安徽科技学院动物科学学院,安徽凤阳233100
  • 出版日期:2017-06-20 发布日期:2017-06-20
  • 通讯作者: 路振香
  • 作者简介:路振香(1964-),女,副教授,硕士,主要从事动物传染病学的教学和科研工作。
  • 基金资助:
    安徽科技学院兽医学重点学科建设项目(AKZDXK2015A04); 安徽省科技攻关计划项目(1301ZC03088)

Molecular Identification and Detection of Multidrug Resistance Gene of Pathogenic Bacteria Associated with Piglets Diarrhea

LU Zhen-xiang, GUO Wei-na, GONG Zheng, LIU Chang, WANG Xun-lei (College of Animal Science, Anhui Sciences and Technology University, Fengyang 233100, China)   

  • Online:2017-06-20 Published:2017-06-20

摘要: 旨在确定引起安徽地区猪场仔猪黄痢发生的病原菌种类,并从分子水平上揭示其多重耐药机制。利用麦康凯培养基对从仔猪黄痢病死猪肝脏样本中初步分离到的9株细菌进行选择性培养;提取分离菌株的基因组DNA,利用PCR法扩增其16SrDNA序列并进行测序;将测序结果提呈至NCBI数据库,与GenBank中已发表的序列进行BLAST比对,对分离菌株进行种水平鉴定;采用纸片扩散法开展分离菌株对抗菌药物的敏感性试验;利用PCR法检测分离菌株携带多重耐药基因AcrA的情况。结果表明,9株分离菌在麦康凯琼脂培养上均呈中等大小的玫瑰红色菌落,符合大肠杆菌的菌落特征;9株分离菌的16SrDNA序列与GenBank中公布的大肠杆菌16SrDNA序列的相似性均在99.0%以上,提示分离菌株均为大肠杆菌;9株分离菌对12种抗菌药物呈现不同程度的耐药性,对阿米卡星、大观霉素和氯霉素的耐药性较强;9株分离菌的AcrA基因携带率为100%。综上提示,多重耐药性大肠杆菌是引起该地区猪场仔猪黄痢发生的致病菌,在临床治疗中应引起足够重视。

Abstract: The aim of the present study was to identify the pathogenic bacteria associated with piglets diarrhea prevailing in pig farms of Anhui Province, and to reveal the molecular mechanism of their muhidrug resistance. Nine strains isolated from the liver samples of dead piglets with diarrhea were subjected to selective culture using MacConkey medium; the genomic DNA of the isolates was extracted and used to amplify 16S rDNA by PCR, and the amplified product was sequenced; in order to identify the isolates to genus, the 16S rDNA sequences of the isolates were deposited to NCBI and compared with the published sequences in GenBank; the susceptibility of the isolates against antimicrobial agents was assessed by disk diffusion method; the presence of muhidrug resistance gene AcrA among the isolates were screened by PCR. The results showed that the rose-coloured colonies with moderate size were observed in MacConkey medium, which was in accord with the growing characteristics of Escherichia coli (E . coli) in MacConkey medium; the 16S rDNA sequences of the nine isolates had I〉99.0% similarity with the previously reported 16S rDNA sequences of E. coli in GenBank, indicating that the isolates were identified as E. coli; the nine isolates exhibited different degrees of resistance to twelve antimicrobial agents, and the higher resistance to amikacin, spectinomycin and chloramphenicol was observed; all of the isolates were AcrA gene positive. The combined data suggest that the muhidrug resistant E. coli was the etiological agent of piglets diarrhea in pig farms of this region and enough attention should be paid to the infectious diseases caused by the isolates in clinical therapy.

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