Although trans-fatty acid (tFA) intake has been positively associated with coronary heart disease (CHD), the relative effect of consuming industrially produced (IP)- compared with ruminant-produced (RP)-tFA on CHD risk factors is unclear. This study was designed to examine the effects of feeding partially hydrogenated vegetable oil (PHVO), IP-tFA source, and butter oil (BO), RP-tFA source, on the development of atherosclerosis and risk factors associated with CHD. Forty-eight male Hartley guinea pigs were fed a hypercholesterolemic diet containing (9% by weight) PHVO, BO, coconut oil (CO; positive control), or soybean oil (SO; negative control) for 8 or 12 wk (n = 6/group). Morphological analysis revealed that none of the groups developed atherosclerosis. Plasma and hepatic lipids did not differ between the tFA groups, but total and small HDL particles were significantly higher in the BO group than in the PHVO group and mean HDL particle size was significantly smaller in the BO group than in the PHVO group. Compared with the other treatment groups, the SO treatment resulted in significantly lower total cholesterol (TC) and LDL cholesterol in plasma, whereas hepatic TC was significantly higher in the SO group than in the other treatment groups. Plasma and hepatic cholesterol concentrations did not differ between the tFA and CO treatments. These results demonstrate that when fed at a high dose, IP- and RP-tFA had the same effect on established CHD risk factors in male Hartley guinea pigs. The effects of RP-tFA on HDL particle sizes and concentrations warrant further investigation.