It is well known that the consumption of trans fatty acids (TFA) from industrial sources (iTFA) increases the risk of cardiovascular disease (CVD), but little is known about the effect of TFA from ruminants (rTFA) on CVD risk factors. This study compared the effect of 4 diets (high in rTFA, moderate in rTFA, high in iTFA, and low in TFA from any source) on plasma LDL cholesterol concentrations and other CVD risk factors in 38 healthy men. Results showed that the consumption of iTFA was associated with unfavourable changes in CVD risk factors (higher concentrations of total cholesterol, apolipoprotein B, LDL cholesterol, and lower concentrations of HDL cholesterol) that were significantly greater than changes observed with the moderate intake of rTFA. However, high intake of rTFA also led to negative changes in cholesterol concentrations, whereas moderate intake of rTFA had no effects on studied CVD risk factors. In summary, this study showed that high intake of rTFA may lead to detrimental changes in lipid CVD risk factors, similar to those that have been attributed to iTFA. However, moderate intake of rTFA through consumption of large quantities of dairy products appeared to have a minimal impact on CVD risk factors. The authors proposed that the general population’s current rTFA intake, which corresponds to approximately one-third of that in the moderate rTFA diet of the present study, is unlikely to cause negative changes in CVD risk.