ABSTRACT: Postprandial glucose (PPG) is an independent predictor of cardiovascular events and death, regardless of diabetes status. Whereas changes in physical activity produce changes in insulin sensitivity, it is not clear whether changes in daily physical activity directly affect PPG in healthy free-living persons.We used continuous glucose monitors to measure PPG and PPG excursions (?PPG, postmeal - premeal blood glucose) at 30-min increments after meals in healthy habitually active volunteers (n = 12, age = 29 ± 1 yr, body mass index = 23.6 ± 0.9 kg·m(-2), VO2max = 53.6 ± 3.0 mL·kg(-1)·min(-1)) during 3 d of habitual (?10,000 steps per day) and reduced (<5000 steps per day) physical activity. Diets were standardized across monitoring periods, and fasting-state oral glucose tolerance tests (OGTT) were performed on the fourth day of each monitoring period.During 3 d of reduced physical activity (12,956 ± 769 to 4319 ± 256 steps per day), PPG increased at 30 and 60 min after a meal (6.31 ± 0.19 to 6.68 ± 0.23 mmol·L(-1) and 5.75 ± 0.16 to 6.26 ± 0.28 mmol·L(-1), P < 0.05 relative to corresponding active time point), and ?PPG increased by 42%, 97%, and 33% at 30, 60, and 90 min after a meal, respectively (P < 0.05). Insulin and C-peptide responses to the OGTT increased after 3 d of reduced activity (P < 0.05), and the glucose response to the OGTT did not change significantly.Thus, despite evidence of compensatory increases in plasma insulin during an OGTT, ?PPG assessed by continuous glucose monitoring systems increased markedly during 3 d of reduced physical activity in otherwise healthy free-living individuals. These data indicate that daily physical activity is an important mediator of glycemic control, even among healthy individuals, and reinforce the utility of physical activity in preventing pathologies associated with elevated PPG.