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The objective of this study was to evaluate the impact of three specific ruminant (R) milk fats resulting from modification of the cow's diet on cardiovascular risk factors in healthy volunteers. R-milk fats were characterized by increased content in total trans fatty acids (R-TFAs) and parallel decrease in saturated fatty acids (SFAs).A total of 111 healthy, normolipemic men and women have been recruited for a monocentric, randomized, double-blind and parallel intervention, 4-week controlled study. Volunteers consumed three experimental products (butter, dessert cream and cookies) made with one of the three specific milk fats (55 g fat per day). During the first week (run-in period), the subjects consumed on a daily basis dairy products containing 72% SFA/2.85% R-TFA (called ‘L0’). For the next 3 weeks of the study (intervention period), the first group continued to consume L0 products. The second group received dairy products containing 63.3% SFA/4.06% R-TFA (called ‘L4’), and the third group received dairy products containing 56.6% SFA/12.16% R-TFA (called ‘L9’).Plasma concentrations of high-density lipoprotein (HDL)-cholesterol were not significantly altered by either diet (P=0.38). Compared to L0 diet, L4 diet contributed to reduce low-density lipoprotein (LDL)-cholesterol (−0.14±0.38 mmol/l, P=0.04), total cholesterol (−0.13±0.50 mmol/l, P=0.04), LDL-cholesterol/HDL-cholesterol (−0.14±0.36, P=0.03) and total cholesterol/HDL-cholesterol (−0.18±0.44, P=0.02).Different milk fat profiles can change cardiovascular plasma parameters in human healthy volunteers. A limited increase of the R-TFA/SFA ratio in dairy products is associated with an improvement in some cardiovascular risk factors. However, a further increase in R-TFA/SFA ratio has no additional benefit.