Reclamation of domestic wastewater for agricultural irrigation is viewed as a sustainable option to create an altative water source and address water scarcity.Free-living amoebae (FLA),which are amphizoic protozoa,are widely distributed in various environmental sources.The FLA could cause considerable environmental and health risks.However,little information is available on the risk of these protozoa.In this study,we evaluated the feasibility using rural domestic wastewater for agricultural irrigation,and analyzed dynamic changes of the microbial community structure and FLA populations in raw and treated wastewater,as well as the phyllosphere and rhizosphere of lettuce production sites that were irrigated with different water sources.The bacterial community dynamics were analyzed by terminal restriction fragment length polymorphism (T-RFLP).The bacterial community structures in the influent were similar to that in the effluent,while in some cases relative abundances varied significantly.The populations of Acanthamoeba spp.and Hartmannella vermiformis in the anaerobically treated wastewater were significantly higher than in the raw wastewater.The vegetables could harbor diverse amoebae,and the abundances of Acanthamoeba spp.and H.vermiformis in the rhizosphere were significantly higher than in the phyllosphere.Accordingly,our studies show insight into the distribution and dissemination of amoebae in wastewater treatment and irrigation practices.