Rice can be the contaminating with soil-borne bacteria. Furthermore, the contaminated bacteria can be grown during immersion process for produce wet-milled rice flour. Therefore, disinfectants can be added during the immersion process. Antibacterial activities of the natural disinfectant, fermented rice spent water (FRSW), and the chemical disinfectants, chlorine dioxide (CD) and sodium benzoate (SB), were respectively determined when added in pure cultures of target bacteria such as Salmonella typhimurium, Escherichia coli, and Bacillus cereus or when added to immersion water in the immersion process. In addition, rinsing effects for removing bacteria were determined when rice was rinsed with water before and after the immersion process. Antibacterial activities were rapidly increased as increasing amounts of the disinfectants are added to the pure cultures of the target bacteria. Antibacterial activity of CD was the most effective among the three tested disinfectants when added to the pure cultures of the target bacteria, respectively. Those of the same disinfectants were increased when they were increasingly added to the immersion water. However those of the disinfectants were less effective when added to the immersion water. On the other hand, rinsing effects for removing bacteria were the most effective when rice was rinsed only with water without the immersion process. Collectively, rinsing rice with water only was more effective than using disinfectants in the immersion water during rice flour production.