Aim This study was retrospectively designed to evaluate the influence of healthy lifestyle behaviors on the incidence of chronic kidney disease (CKD) during a 5-year follow-up period in middle-aged and older males. Methods The subjects included 252 males without a history of cardiovascular disease, stroke, renal dysfunction and/or dialysis treatment who were not taking any medications. Their lifestyle behaviors were evaluated using a standardized self-administered questionnaire and defined as follows: (1) habitual moderate exercise, (2) daily physical activity, (3) fast walking speed, (4) slow eating speed, (5) no late-night dinner, (6) no bedtime snacking and (7) no skipping breakfast. The participants were divided into four categories, which were classified into quartile distributions according to the number of healthy lifestyle behaviors (7-6, 5, 4 and B3 groups). Results After 5 years, the incidence of CKD [estimated glomerular filtration rate (eGFR)\60 ml/min/1.73 m 2 and/ or proteinuria] was observed in 23 subjects (9.1 %). The Kaplan-Meier survival curves showed that the cumulative incidence of CKD significantly decreased according to an increase in the number of healthy lifestyle behaviors (logrank test: p = 0.003). According to a multivariate analysis, habitual moderate exercise [hazard ratio (HR) 0.20, 95 % confidence of interval (CI) 0.06-0.69, p = 0.011] and no bedtime snacking (HR 0.19,, p = 0.004) were significantly associated with the incidence of CKD. Conclusions These results suggest that the accumulation of healthy lifestyle behaviors, especially those related to habitual moderate exercise and no bedtime snacking, is considered to be important to reduce the risk of CKD.