Abstract: In order to prevent rice cake from mould and spoilage, while to greatly keep its flavor and quality, bactericidal effect of pickling combined with water-bath sterilization process was studied. The factors including pickling solution volume fraction, pickling time, water-bath temperature and water-bath time were investigated by mean of orthogonal design, with the total number of initial colonies and sensory evaluation as indicators. Traditional isolation and identification methods were applied to analyze the microbial flora of rice cake. Results showed that the influences of factors affecting the total number of initial colonies followed the order, from high to low, water-bath time, water-bath temperature, pickling time, pickling solution volume fraction; the influences of factors affecting sensory evaluation followed the order, from high to low pickling solution volume fraction, water-bath temperature, pickling time, water-bath time. Variance analysis results indicated that only pickling solution volume fraction affected the sensory evaluation significantly. The optimized sterilization conditions were pickling solution volume fraction of 2%, pickling time of 15 min, water-bath temperature of 95 ℃ and water-bath time of 40 min. By isolating and identifying microbial flora from rice cake sample, bacteria as Bacillus, Micrococcus, and Klebsiella oxytoca, and fungus as Penicillium, Aspergillus, and Talaromyces were mainly detected in the fresh-keeping rice cake. Meanwhile, the main bacteria and fungus existing in the sterillized rice cake were Bacillus, Penicillium and a small amount of Talaromyces, other bacteria were killed or had low concentration which was lower than the detection limit. The results indicated that sterilization process combining pickling with water-bath sterilization gave rice cake a significant antibacterial effect that could effectively prevent microorganism from growing and reproducting. The shelf life of rice cake could reach 6 months under room temperature.