There is a significant inverse relationship between high density lipoprotein cholesterol (HDL-C) and cardiovascular events. Previous studies have reported increased HDL-C in response to 4 weeks of pharmacological doses of omega-3 fatty acids (n-3 FA) (3-5g/day). Our hypothesis was that a low dose of n-3 FA (eicosapentaenoic and docosahexaenoic acid), one achievable in the diet, ingested in the form of one fish oil capsule three times per day by college-aged and middle-aged men, over a short period of 2 weeks, preceded and followed by a single bout of exercise, would increase HDL-C levels. We determined serum lipid profile (triglyceride, VLDL-C, LDL-C, HDL-C, total cholesterol) in response to a single bout of exercise (60 min, 55-60% HRmax) after consuming a low dose of 0.9 g n-3 FA/day for 14 days in eight middle-aged men (age 48.9 ± 1.4) and seven college-aged men (age 21 ± 2.5). There were no significant changes with blood lipids except in HDL-C. In middle-aged and college-aged men HDL-C significantly increased (P<0.005). The college-aged men’s TC/HDL-C ratio significantly increased (P<0.05). Our findings suggest that two weeks of ingesting dietary achievable doses of n-3 FA followed by a single bout of exercise increase HDL-C, which is associated with a decreased risk for coronary heart disease (CHD).