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Mice were fed conjugated linoleic acid (CLA) with or without fish oil or aspirin, which deplete tissue arachidonic acid and block arachidonic acid metabolism, respectively. Mice fed fish oil did not lose body fat when supplemented with CLA but mice fed soy oil did. Aspirin did not alter CLA-induced body fat loss. CLA may be metabolized to an isomer of arachidonic acid to induce a loss of body fat. However, this body fat loss is apparently not mediated via alteration of prostaglandin synthesis. Understanding the regulation of body fat by CLA may offer insight into the mechanisms of body fat regulation in cattle.