多价噬菌体精准控制污水处理厂耐药菌的策略及微生态机制研究

WWTPs的污水和污泥中高丰度的ARB和ARG对生态环境和人类健康构成严重威胁。噬菌体是一种选择性强、高效安全的杀菌剂。然而，已报道的用于控制问题细菌的噬菌体大多具有窄宿主范围，无法克服污水处理系统中目标ARB浓度过低的局限性。本项目将采用多价噬菌体对污水处理系统中的ARB进行精准控制。通过epicPCR技术结合传统分离培养的方法，分析活性污泥中几种典型的ARB种群结构多样性，获取优势ARB菌株；并以其为宿主菌，通过连续多宿主法筛选可靶向裂解多“种”ARB的多价噬菌体；挑选感染力强、宿主范围稳定的多价噬菌体，考察单一噬菌体和噬菌体混合制剂分别对活性污泥中ARB的削减能力，获得多价噬菌体控制污水处理系统中ARB的最佳策略；探索多价噬菌体控制活性污泥中ARB的微生态机制，评估其生态风险。本项目对于推动噬菌体在污水处理系统ARB控制中的应用具较重要意义。

The high content of antibiotic-resistant bacteria (ARB) and antibiotic-resistant gene (ARG) in sewage and sludge of WWTPs pose serious threat to the ecological environment and human health. Bacteriophage is a highly selective, highly effective and safe antibacterial agent. However, most of the phages reported for controlling problem bacteria have a narrow host range, which cannot overcome the limitation of low ARB concentration in the wastewater treatment systems. In this project polyvalent bacteriophages will be used to precisely control ARB in wastewater treatment systems. With epicPCR technology combined with traditional culture-dependent methods, the diversity of ARB in activated sludge will be analyzed to obtain the dominant ARB strains. Then the dominant ARB will be used as host bacteria to screen for the polyvalent bacteriophages that can target multiple species of ARB. The polyvalent bacteriophages with strong infectivity and stable host range will be selected to investigate the ability of single phage and phage cocktail to reduce ARB in activated sludge, and then the best strategy for polyvalent bacteriophages to control ARB in wastewater treatment systems will be obtained. The micro-ecological mechanism of ARBs in activated sludge controlled by polyvalent bacteriophages will be explored, and the ecological risks will be evaluated. This project is of great significance to promote the application of phage in the control of ARBs in wastewater treatment systems.