Dissecting horizontal and vertical gene transfer of antibiotic resistance plasmid in bacterial community using microfluidics
Dissecting horizontal and vertical gene transfer of antibiotic resistance plasmid in bacterial community using microfluidics
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使用微流控技术剖析细菌群落中抗生素抗性质粒的水平和垂直基因转移
DOI：
10.1016/j.envint.2019.105007
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发表时间：
2019-10-01
影响因子：
11.8
通讯作者：
Yin, Huabing
中科院分区：
文献类型：
--
作者：
Li, Bing;Qiu, Yong;Yin, Huabing
抗生素耐药基因（ARGs）的传播已成为全球健康的新威胁。虽然水平基因转移（HGT）被认为是主要途径之一，但越来越多的证据表明，垂直基因转移（VGT）也参与其中。然而，传统的基于培养的方法无法区分HGT和VGT，导致结论往往相互矛盾。在这里，单细胞微流体与延时成像已经成功地用于剖析质粒介导的HGT和VGT对环境社区中ARG传播的贡献。以具有甲氧苄啶耐药性的ARG编码质粒pKJK5的大肠埃希菌为供体，定量了三种代表性抗生素（甲氧苄啶、四环素和阿莫西林）对活性污泥细菌群落中ARG转移过程的影响。研究发现，HGT受到抗生素及其靶点（供体、受体单独或共同）的抑制机制的影响，而VGT则对转偶联物的形成和ARG的传播起着重要作用。甲氧苄啶对供体和转缀合物具有高度的抗性，其存在显著增加了HGT和VGT率。虽然四环素和阿莫西林都对供体有抑制作用，但由于抑制机制不同，对HGT率的影响也不同。此外，我们还证明了HGT在社区中的传播动力学可以用流行病感染模型来描述，该模型与HGT和VGT的芯片定量测量相结合，为研究和预测现实社区中ARG的传播动力学提供了一个有前途的工具。
The spread of antibiotic resistance genes (ARGs) has become an emerging threat to the global health. Although horizontal gene transfer (HGT) is regarded as one of the major pathways, more evidence has shown the significant involvement of vertical gene transfer (VGT). However, traditional cultivation-based methods cannot distinguish HGT and VGT, resulting in often contradictory conclusions. Here, single-cell microfluidics with time-lapse imaging has been successfully employed to dissect the contribution of plasmid-mediated HGT and VGT to ARG transmission in an environmental community. Using Escherichia coli with an ARG-coded plasmid pKJK5 with trimethoprim resistance as the donor, we quantified the effects of three representative antibiotics (trimethoprim, tetracycline and amoxicillin) on the ARG transfer process in an activated sludge bacterial community. It was found that HGT was influenced by the inhibitory mechanism of an antibiotic and its targets (donor, recipient alone or together), whereas VGT contributes significantly to the formation of transconjugants and consequently ARG spreading. Trimethoprim is highly resisted by the donor and transconjugants, and its presence significantly increased both the HGT and VGT rates. Although tetracycline and amoxicillin both inhibit the donor, they showed different effects on HGT rate as a result of different inhibitory mechanisms. Furthermore, we show the kinetics of HGT in a community can be described using an epidemic infection model, which in combination with quantitative measure of HGT and VGT on chip provides a promising tool to study and predict the dynamics of ARG spread in real-world communities.