Department of Microbiology and Parasitology, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand.
Centre of Excellence in Medical Biotechnology, Naresuan University, Phitsanulok, Thailand.
PLoS One. 2018 Dec 6;13(12):e0208468. doi: 10.1371/journal.pone.0208468. eCollection 2018.
Conjugation is a type of horizontal gene transfer (HGT) that serves as the primary mechanism responsible for accelerating the spread of antibiotic resistance genes in Gram-negative bacteria. The present study aimed to elucidate the mechanisms underlying the conjugation-mediated gene transfer from the extensively drug-resistant Acinetobacter baumannii (XDR-AB) and New Delhi Metallo-beta-lactamase-1-producing Acinetobacter baumannii (NDM-AB) to environmental isolates of Acinetobacter spp. Conjugation experiments demonstrated that resistance to ticarcillin and kanamycin could be transferred from four donors to two sodium azide-resistant A. baumannii strains, namely, NU013R and NU015R. No transconjugants were detected on Mueller-Hinton Agar (MHA) plates containing tetracycline. Plasmids obtained from donors as well as successful transconjugants were characterized by PCR-based replicon typing and S1-nuclease pulsed-field gel electrophoresis (S1-PFGE). Detection of antibiotic resistance genes and integrase genes (int) was performed using PCR. Results revealed that the donor AB364 strain can transfer the blaOXA-23 and blaPER-1 genes to both recipients in association with int1. A 240-kb plasmid was successfully transferred from the donor AB364 to recipients. In addition, the aphA6 and blaPER-1 genes were co-transferred with the int1 gene from the donor strains AB352 and AB405. The transfer of a 220-kb plasmid from the donors to recipient was detected. The GR6 plasmid containing the kanamycin resistance gene (aphA6) was successfully transferred from the donor strain AB140 to both recipient strains. However, the blaNDM-1 and tet(B) genes were not detected in all transconjugants. Our study is the first to demonstrate successful in vitro conjugation, which indicated that XDR-AB contained combination mechanisms of the co-transfer of antimicrobial resistance elements with integron cassettes or with the plasmid group GR6. Thus, conjugation could be responsible for the emergence of new types of antibiotic-resistant strains.
结合是一种水平基因转移(HGT),是导致革兰氏阴性菌中抗生素耐药基因传播的主要机制。本研究旨在阐明广泛耐药鲍曼不动杆菌(XDR-AB)和新德里金属β-内酰胺酶-1 产生的鲍曼不动杆菌(NDM-AB)与环境分离株鲍曼不动杆菌之间通过接合介导基因转移的机制。接合实验表明,来自四个供体的对替卡西林和卡那霉素的耐药性可以转移到两种对叠氮化钠耐药的鲍曼不动杆菌菌株 NU013R 和 NU015R 上。在含有四环素的 Mueller-Hinton 琼脂(MHA)平板上未检测到转导子。从供体获得的质粒和成功的转导子通过基于 PCR 的复制子分型和 S1-核酸酶脉冲场凝胶电泳(S1-PFGE)进行了表征。使用 PCR 检测抗生素耐药基因和整合酶基因(int)。结果表明,供体 AB364 株可以将 blaOXA-23 和 blaPER-1 基因与 int1 一起转移到两个受体中。成功地从供体 AB364 转移了一个 240-kb 的质粒到受体。此外,来自供体 AB352 和 AB405 的 aphA6 和 blaPER-1 基因与 int1 一起被共同转移。从供体到受体的 220-kb 质粒转移被检测到。携带卡那霉素耐药基因(aphA6)的 GR6 质粒成功地从供体 AB140 转移到两个受体株。然而,所有转导子中均未检测到 blaNDM-1 和 tet(B)基因。我们的研究首次证明了体外结合的成功,这表明 XDR-AB 包含了抗生素耐药元件与整合子盒或质粒群 GR6 共同转移的组合机制。因此,结合可能是导致新型抗生素耐药菌株出现的原因。