ABSTRACT: Cycling is a recreational activity and mode of commuting with substantial potential to improve public health in many countries around the world. The aim of this study was to examine prospective associations between recreational and commuter cycling, changes in cycling habits, and risk of type 2 diabetes (T2D) in Danish adults from the Diet, Cancer and Health cohort study.At baseline from 1993 to 1997, 24,623 men and 27,890 women from Denmark, 50-65 y of age and free of T2D and other chronic diseases, underwent a number of assessments, including completing a lifestyle questionnaire also addressing cycling habits. Approximately 5 y later, at a second examination, participants completed a new, updated lifestyle questionnaire. Cox regression was used to estimate hazard ratios (HRs) of incident T2D registered in the Danish National Diabetes Registry, according to recreational and commuter cycling and changes in cycling habits, with adjustment for a priori known T2D risk factors. During 743,245.4 person-years of follow-up (mean follow-up 14.2 y), 6,779 incident cases of T2D were documented. Multivariable adjusted HRs (95% confidence interval [CI]) were 1, 0.87 (0.82, 0.93), 0.83 (0.77, 0.89), 0.80 (0.74, 0.86) and 0.80 (0.74, 0.87) (p for trend = <0.001) for 0, 1-60, 61-150, 151-300, and >300 min/wk of total cycling (recreational and commuter cycling), respectively. In analysis of seasonal cycling, multivariable adjusted HRs (95% CI) were 1, 0.88 (0.83, 0.94), and 0.80 (0.76, 0.85) for non-cyclists, seasonal cyclists (those cycling only in summer or winter), and those cycling during both summer and winter, respectively. How changes in total cycling from baseline to the second examination affected risk was also investigated, and multivariable adjusted HRs (95% CI) were 1, 0.88 (0.78, 1.01), 0.80 (0.69, 0.91), and 0.71 (0.65, 0.77) for non-cyclists and for those who ceased, initiated, or continued cycling between baseline and the second examination, respectively. Lastly, in the analysis of commuter cycling, multivariable HRs (95% CI) were 1, 0.72 (0.60, 0.87), 0.83 (0.69, 1.00), and 0.70 (0.57, 0.85) (p for trend = <0.001) for cycling 0, 1-60, 61-150, and >150 min/wk to work, respectively. The main limitation of the current study is the use of self-reported physical activity.Commuter and recreational cycling was consistently associated with lower risk of T2D in Danish adults. Our results also provide evidence that late-in-life initiation of or continued engagement in cycling lowers risk of T2D.