Objective: Accumulating evidence suggests that, in addition to various psychosocial parameters, affective responses to exercise play an important role in subserving future exercise behavior. This study comprehensively evaluated whether acute exercise duration and recovery period influenced the relationship between moderate-intensity walking exercise and mood profile. Method: We employed a randomized controlled cross-over trial. Participants completed two laboratory visits, separated by one-week. One of the visits involved a mood profile assessment with no exercise, while the other visit involved a mood profile assessment after an acute bout of exercise. Participants (N = 352; 22 per group; young [ M-age = 21 yrs] healthy adults) were randomized into one of 16 experimental groups: 10, 20, 30, 45 or 60 min bout of exercise coupled with either a 5, 15 or 30 min recovery period. The exercise bout was of moderate-intensity (40-59% of HRR). Mood profile was assessed from the POMS survey, considering subscales of depression, anger and hostility. Results: For all three mood profile parameters, there was no evidence of a group x time interaction effect. However, the main effect for time was statistically significant for each mood parameter. These significant results demonstrate that, generally, exercise had a favorable effect on each of the mood profile, regardless of exercise duration and recovery period. In addition to the significant main effects for time, we also observed a significant main effect for group for the mood parameter hostility. With the exception of the group 13 (60 min of exercise with 5 min recovery) and the 3 groups that employed a 10-min bout of exercise (groups 1-3), all other experimental groups had a lower (better) hostility score after the exercise visit. Conclusions: Generally, exercise had a favorable effect on various mood profiles, regardless of exercise duration (between 10 and 60 min) and recovery period (between 5 and 30 min).