Bicycle touring as a modality of physical activity that involves whole-body cardiorespiratory and metabolic functions could be a potential strategy to improve body composition and cardiorespiratory fitness. Therefore, the aim of the current study was to investigate the effects of 7-days bicycle touring activity on body composition and physical fitness. A total of 13 individuals (three women) participated in this quasi-experimental study. The participants were evaluated at baseline and were tracked for up to 12 days after the intervention. The intervention consisted of a 480 km bicycle touring route performed over 8 days. Body weight and body composition (i.e., fat mass and lean mass) were assessed using a bioelectrical impedance analyser. Physical fitness was measured using the International Fitness Scale questionnaire. We conducted a repeated-measures analysis of variance to determine changes in body weight and body composition and paired sample t-tests to analyse changes in physical fitness. Significant differences in fat mass were observed between the baseline and both post-intervention (p = 0.003) and re-test values (p = 0.031). Significant differences were also noted in lean mass between the baseline and post-intervention values (p = 0.003), whereas no significant changes were observed when compared the baseline and re-test values (p = 0.178). Significant differences were obtained in cardiorespiratory fitness when comparing the baseline with the post-intervention values (p = 0.016), whereas no significant differences were noted in general fitness, muscular strength, speed/agility, and flexibility (all p > 0.05). In conclusion, a 7-day bicycle touring intervention can significantly reduce fat mass and increase lean mass and cardiorespiratory fitness in healthy individuals.