The purpose of this study was to compare 1) the preferred cadences and 2) the aerobic demand response to cadence manipulation of highly fit, experienced cyclists and equally fit noncyclists. Eight cyclists (C) and eight non-cyclists (NC) pedaled at 200 W under six randomly ordered cadence conditions (50, 65, 80, 95, 110 rpm and preferred cadence) on a Velodyne trainer. The JOURNAL/mespex/04.02/00005768-199311000-00011/ENTITY_OV0312/v/2017-07-20T222401Z/r/image-pngO2 responses of C and NC to cadence manipulation were similar. Both groups displayed lower JOURNAL/mespex/04.02/00005768-199311000-00011/ENTITY_OV0312/v/2017-07-20T222401Z/r/image-pngO2 values at lower cadences. JOURNAL/mespex/04.02/00005768-199311000-00011/ENTITY_OV0312/v/2017-07-20T222401Z/r/image-pngO2 differences between C and NC across cadences were not significant. Mean preferred pedaling cadence surprisingly was somewhat higher for NC (91.6 ± 10.5 rpm) than C (85.2 ± 9.2 rpm), but the difference was not significant. The most economical cadence was significantly lower for C (56.1 ± 6.9 rpm) than NC (62.9 ± 4.7 rpm). Thus, cycling experience did not substantially influence preferred cadence nor economy during moderate intensity cycling by highly fit athletes. We speculate that preferred cadence and economy similarities between C and NC are associated with similarities in the dynamic muscular training of the groups.