The purpose of this study was to update the evidence on the health benefits of cycling. A systematic review of the literature resulted in 16 cycling-specific studies. Cross-sectional and longitudinal studies showed a clear positive relationship between cycling and cardiorespiratory fitness in youths. Prospective observational studies demonstrated a strong inverse relationship between commuter cycling and all-cause mortality, cancer mortality, and cancer morbidity among middle-aged to elderly subjects. Intervention studies among working-age adults indicated consistent improvements in cardiovascular fitness and some improvements in cardiovascular risk factors due to commuting cycling. Six studies showed a consistent positive dose-response gradient between the amount of cycling and the health benefits. Systematic assessment of the quality of the studies showed most of them to be of moderate to high quality. According to standard criteria used primarily for the assessment of clinical studies, the strength of this evidence was strong for fitness benefits, moderate for benefits in cardiovascular risk factors, and inconclusive for all-cause mortality, coronary heart disease morbidity and mortality, cancer risk, and overweight and obesity. While more intervention research is needed to build a solid knowledge base of the health benefits of cycling, the existing evidence reinforces the current efforts to promote cycling as an important contributor for better population health. © 2011 John Wiley & Sons A/S.