Excessive sedentary time is related to poor mental health. However, much of the current literature uses crosssectional data and/or self-reported sedentary time, and does not assess factors such as sedentary bout length. To address these limitations, the influence of objectively measured sedentary time including sedentary bout length (i.e. < 30 min, ≥30 min) on mood, stress, and sleep, was assessed in 271 healthy adults (49% women; age 27.8 ± 3.7) across a 1-year period between 2011 and 2013 in Columbia, SC. Participants completed the Profile of Mood States and the Perceived Stress Scale, and wore a Sensewear Armband to assess sedentary time, physical activity, and sleep for ten days at baseline and one year. A series of fixed-effects regressions was used to determine the influence of both baseline levels and changes in daily sedentary time (total and in bouts) and physical activity on changes in mood, stress, and sleep over one year. Results showed that across the year, decreases in total sedentary time, and time in both short and long bouts, were associated with improvements in mood, stress and sleep (p < 0.05). Increases in physical activity were only significantly predictive of increases in sleep duration (p < 0.05). Thus, reductions in sedentary time, regardless of bout length, positively influenced mental wellbeing. Specifically, these results suggest that decreasing daily sedentary time by 60 min may significantly attenuate the negative effects of high levels of pre-existing sedentary time on mental wellbeing. Interventions manipulating sedentary behavior are needed to determine a causal link with wellbeing and further inform recommendations.