基于微米尺度的污水处理菌群中抗生素抗性基因转移特征及扩散机制研究

The transfer and spread of antibiotic resistant genes (ARG) has become a growing concern in the environment. In particular, ARG transfer efficiency is high in wastewater treatment due to the high abundance of microorganisms and complicated conditions involved. Although this has raised wide public concern, the mechanism of ARG transfer and spread in the microbial communities was still poorly understood. In this project, novel microfluidic methods will be developed for in-situ monitoring of the dynamics of ARG transfer in wastewater-treatment microorganisms, thereby allowing the micron-scale analysis of the transfer characteristics and spread mechanism. First, a microfluidic platform will be established for time-lapse tracking of the ARG transfer process within microbial communities on chip. Second, the influence of key environmental factors on ARG transfer will be examined on-chip, and characteristics of the populations containing transferred ARG will be identified using Biolog and high-throughput sequencing. Last, the influence of different environmental factors on horizontal gene transfer and vertical gene transfer will be evaluated at the single-cell level . The results of this project will provide meaningful insights into the transfer and spread processes of ARG in microbial communities under different environmental conditions, thereby providing theoretical basis for ARG control in the aqueous environment.

抗生素抗性基因（ARG）的转移扩散已成为环境研究中的热点问题。其中，污水处理中细菌种类丰富，所处环境条件复杂，ARG转移率高，引起了广泛关注，但目前对污水处理菌群中ARG转移的微观行为特征及扩散机制还缺乏深入了解。本项目建立基于微流控的研究方法，开展ARG转移的微观、原位和动态研究，从微米尺度分析微生物菌群中ARG的转移特征及扩散机制。首先，建立微流控观测系统，研究菌群中ARG转移的过程特征；其次，研究污水处理中主要营养物质、毒性污染物及消毒剂等单一及复合因素对菌群中ARG转移的影响规律，并结合Biolog及高通量测序手段，分析ARG转移细菌的种群特点；最后，在单细胞层面研究ARG的扩散机制，明晰不同环境因素影响下水平转移和垂直转移在ARG扩散中所起的作用。通过本项目的研究，可从微观方面深入理解不同环境下菌群中ARG的转移特征及扩散机制，为控制水环境中ARG的传播提供一定的理论依据。

抗生素抗性基因（ARG）的转移扩散已成为环境研究中的热点问题。其中，污水处理中细菌种类丰富，所处环境条件复杂，ARG转移率高，引起了广泛关注，但目前对污水处理菌群中ARG转移的微观行为特征及扩散机制还缺乏深入了解。本项目建立了基于微流控的ARG转移研究方法，实现了ARG转移的微观、原位和动态过程研究，获得了污泥菌群中ARG转移的动力学特征; 进一步基于单层微流控小室芯片实现单细胞水平上的ARG转移过程研究，明晰了水平转移和垂直转移在ARG扩散中所起的作用，发现固定空间内ARG的水平转移速率变化过程符合流行病传染模型；采用流式分选与高通量测序结合的方法，分析了不同环境条件（温度、营养物浓度、重金属压力等）对ARG转移率的影响，并识别出污泥菌中易接受质粒的菌属。通过本项目的研究，从微观方面深入理解了菌群中ARG的转移特征及扩散机制，为控制水环境中ARG的传播提供一定的理论依据。