Rural toilet reform in cold areas is the focus and difficulty of China’s toilet revolution. In this paper, by carrying out the septic tank gradient burial depth test and regular seasonal monitoring of the operation effect of rural septic tanks in cold areas, the frost protection effect and treatment effect of rural toilets in cold areas are clarified, which provide important references for the rural toilet revolution and its practical significance in cold areas of China. Based on controlled tests and correlation analysis methods, different gradient burial depths of septic tanks (40, 60, and 80 cm) and different seasonal operation effects (spring, summer, autumn, and winter) were studied to analyze the influence of factors such as burial depths and temperatures on the effectiveness of anti-freezing and treatment of septic tanks. The results found that the septic tank effluent indexes of the six cold-zone test households could not meet requirements of Class Ⅰ-B in the discharge standard of pollutants for municipal wastewater treatment plants (GB 18918-2002). (The criteria for Class I-B are as follows: COD emission limit of 60 mg/L; SS emission limit of 20 mg/L; TN emission limit of 20 mg/L; TP emission limit of 1 mg/L; NH3-N emission limit of 8 mg/L); The average removal of suspended solids (SSs), chemical oxygen demand (COD), total nitrogen (TN), total phosphorus (TP), NH3-N, and NO3-N by septic tanks was 79.7%, 21.8%, 10.5%, 13.5%, 9.76%, and 2.38%, respectively; the water temperature was significantly positively correlated with SS, TN, and TP; the double-layer insulated septic tank with medium burial depth (60 cm) could operate normally in cold areas in winter. The treatment effect of septic tanks on various indicators basically shows that summer > autumn > spring > winter. Temperature is the main limiting factor affecting the treatment effect of septic tanks. Capsule: By carrying out septic tank gradient burial depth tests and regular seasonal monitoring of the operating effect of rural septic tanks in cold areas, we can clarify the anti-freezing effect of rural toilets in cold areas and the effectiveness of treatment.