Milk with an increased content of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and conjugated linoleic acid (CLA) was obtained by incorporating fish oil into the feed of cows. The 4 feed treatments used were a control diet of 57% forage and 43% concentrate mix with EnerGII fat supplement at 1.65% of dietary DM, or EnerGII in the basal diet was partially replaced with 1) 0.21% partially ruminally inert calcium salts of 71% fish oil given at 0.41% of DM; 2) 0.41% inert calcium salts of 71% fish oil given at 0.83% of DM; or 3) 0.83% inert calcium salts of 43% fish oil given at 0.83% of DM. The cows were milked after 5 and 8 wk and the EPA, DHA, and CLA contents in the pasteurized whole milk were determined. The presence of off-flavors in the milk was investigated after 3 and 10 d of storage. Twelve judges were trained to evaluate the presence of grassy, fishy, oily, oxidized, and rancid off-flavors. Although levels of EPA, DHA, vaccenic acid, and CLA increased for all 4 treatments, a trained sensory panel detected no difference in milk flavor between treatments and the control, with little or no intensity of off-flavors. Results suggest that feeding fish oil and EnerGII at varying levels enhanced CLA, EPA, DHA, and total n-3 fatty acids in milk over the length of the experiment without negatively affecting milk flavor. This creates the potential for a more marketable and healthful product.