This study was conducted to examine the effects of dietary supplementation with vegetable oils on performance of high-yielding lactating cows and conjugated linoleic acid (CLA) content in milk fat. Twelve lactating Holstein cows in early lactation (30 to 45 d postpartum) were used in a triple 4 x 4 Latin square design. In each period, the cows in each group were fed the same basal diet and received one of the following treatments: 1) control (without oil), 2) 500 g of cottonseed oil, 3) 500 g of soybean oil, and 4) 500 g of corn oil. Each experimental period lasted for 3 wk, with the first 2 wk used for adaptation to the diet. Supplementation with vegetable oils tended to increase milk yield, with the highest milk yield in the cottonseed oil group (35.0 kg/d), compared with the control (34.4 kg/d). Milk fat percentage was decreased, but there were few effects on percentage and yield of milk protein as well as milk fat yield. The cows fed added soybean oil produced milk with the highest content of trans-11 C(18:1) (23.8 mg/g of fat), which was twice that of the control (12.6 mg/g of fat). Content of cis-9, trans-11 CLA in milk fat increased from 3.5 mg/g in the control to 6.0, 7.1, and 10.3 mg/g for the cows fed oils from cottonseed, corn, and soybean, respectively. A significant linear relationship existed between trans-11 C(18:1) and cis-9, trans-11 CLA. Supplementation with oils doubled the content of total fatty acids in blood plasma, with little difference between different vegetable oil sources. Octadecenoic acid content was significantly higher in blood plasma of animals fed added oils from cottonseed and soybean than those fed with corn oil and control. The plasma trans-11 C(18:1) content was significantly higher in the oil-added animals than in control. Supplementation of vegetable oils tended to improve milk production of lactating cows, and the CLA content in milk fat was significantly increased. Soybean oil seemed to be the optimal source to increase CLA production.