Older adults spend more time engaged in sedentary behavior (SB) than any other segment of the population. Interrupting sedentary time with short bouts of walking improves 24-hour glycemic control compared to uninterrupted sitting. However, it is not known if short-walking bouts are as beneficial to 24-hour glycemia as a single bout of continuous exercise. PURPOSE: To compare the effectiveness of multiple, short intermittent walking bouts and one, calorically equivalent continuous bout of walking on 24-hour glycemia in older adults. METHODS: Healthy, overweight/obese older adults (N=18, 67 ± 5 y, BMI = 32.2 ± 4,3 kg/m2) completed two, 24h conditions in a whole room indirect calorimeter; 1) Intermittent walking (IW): 1.5 min of moderate intensity treadmill walking (36 min total) every 30 minutes and 2) continuous walking (CW): 36 min continuous, moderate intensity treadmill walking performed in the morning (~8AM). Outside of the prescribed walking times, subjects remained in SB for the remainder of the waking day. Continuous glucose monitoring was used to measure interstitial glucose concentrations every 5 minutes. Energy and macronutrient intake was standardized between conditions. RESULTS: 24-hour energy expenditure (2257 ± 329 vs. 2165 ± 302 kcal, mean ± SD) and RQ (0.84 ± 0.03 vs. 0.84 ± 0.03) were similar during IW and CW, respectively. Peak postprandial glucose following dinner was lower (p<0.05) during IW (120.4 ± 10.7 mg/dl) compared to CW (135.3 ± 15.3 mg/dl). No differences were observed in any other 24 hour glycemia variables, including 24 hour area under the glucose curve (IW = 154862 ± 12724 mg/dl, CW = 158096 ± 15156 mg/dl), glycemic variability (standard deviation of 24 hour glucose concentrations) (IW = 12.2 ± 4.4 mg/dl, CW = 12.2 ± 4.2 mg/dl), and peak postprandial glucose concentrations following breakfast (IW = 144.0 ± 22.7 mg/dl, CW = 144.8 ± 27.2 mg/dl) and lunch (IW = 137.9 ± 17.1 mg/dl, CW = 139.2 ± 17.8 mg/dl). CONCLUSION: These results suggest IW had similar effects as CW on 24-hour glycemia, although the postprandial glucose response to meals consumed later in the day may be lower with IW. IW may improve cardiometabolic health in older adults.