Slow sand filters (SSFs) have been widely used in the construction of water plants in rural areas. It is necessary to find river sand of suitable particle size to improve SSF treatment of micro-polluted water so as to ensure the effective and long-term operation of these plants. In this study, SSF1# (particle size of 0.1-0.5 mm), SSF2# (particle size of 0.5-1 mm), and SSF3# (particle size of 1-1.5 mm) were selected. The physical absorption, CODMn and NH4+-N removal effect, and microbial community were analyzed. According to Langmuir and Freundlich adsorption model fitting, the smaller the particle size of the river sand, the more pollutants are adsorbed under the same conditions. SSF1# has the shortest membrane-forming time, highest CODMn and NH4+-N removal rate, and highest Shannon estimator, indicating that there are more abundant microbial species in the biofilm. Mesorhizobium, Pannonibacter, Pseudoxanthomonas, Aquabacterium, Devosia, and other bacteria have different proportions in each system, each forming its own stable biological chain system. The effluent quality of the three SSFs can meet drinking water standards. However, river sand with a particle size range of 0.1-0.5 mm is easily blocked, and thus the recommended size range for SSF is 0.5-1 mm. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.