Abstract Objective Increasing omega-3 fatty acid (FA) intake, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), is associated with numerous health benefits; however, the benefits on inflammation appear to vary depending on the study population examined. While improvements in inflammatory status have been reported in the elderly, there is less evidence regarding the effects of fish oil supplementation on inflammation in young adults. The goal of the present study was to examine the influence of fish oil supplementation on lipid metabolites and the inflammatory status of young healthy men. Materials/Methods Fasted serum samples were collected from 10 young healthy males (23.4±1.7years) before and after a 3-month supplementation of fish-oil containing 2.0g EPA and 1.0g DHA. Samples were analyzed to investigate changes in FA profiles, bioclinical parameters (e.g. triglyceride and hs-CRP), and a panel of 26 eicosanoids. Paired t-tests were used to evaluate changes between the time points. Results Serum triglycerides decreased (P=0.0006) while the proportion of HDL-c (relative to total cholesterol) increased significantly (P=0.0495) after fish oil supplementation. Specific monounsaturated and polyunsaturated FA levels were changed following supplementation, including reductions in palmitoleic and oleic acid, and, as expected, increases in EPA and DHA. We also observed increases in eicosanoids, namely prostaglandin-F2α (P<0.0001) and thromboxane-B2 (P=0.0296), after fish oil supplementation. Conclusions A 3-month fish oil supplementation in young healthy men improved circulating triglyceride levels and the HDL-c ratio while, concomitantly, increasing the concentrations of two eicosanoids (prostaglandin-F2α and thromboxane-B2). This suggests that fish oil supplementation does have significant benefits in young healthy adults and that specific omega-6-derived eicosanoids can help to further our understanding regarding the beneficial link between omega-3 FA and inflammation.