The purpose of this study was to compare blood lipid responses to continuous versus accumulated exercise. Nine participants completed the following conditions on separate occasions by treadmill walking/jogging at 70% of VO2max: 1) one 500-kcal session and 2) three 167 kcal sessions. Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TG) concentrations were measured from serum samples obtained 24 h prior to and 24 and 48 h after exercise. All blood lipid responses were analyzed in 2 (condition) x 3 (time) repeated measures ANOVAs. HDL-C increased by 7 mg/dL over baseline at 48 h post-exercise with three accumulated sessions versus 2 mg/dL with continuous exercise (P < 0.05). Triglyceride concentrations were unchanged in both conditions. These findings suggest that three smaller bouts accumulated on the same day may have a modestly greater effect for achieving transient increases in HDL-C compared to a continuous bout of similar caloric expenditure.