Objective-To determine the effects of increases in dietary intake of polyunsaturated and saturated fatty acids on plasma lipid and lipoprotein concentrations and activity of associated enzymes in healthy domestic cats. Animals-16 healthy adult sexually intact female cats. Procedures-A baseline diet (40% energy from fat) and 4 test diets, with increased amounts of fat (51% and 66% energy from fat) from the addition of polyunsaturated and saturated fatty acids, were fed for 6 weeks each. Plasma cholesterol, triglyceride, and lipoprotein cholesterol concentrations, along with activities of lipoprotein lipase, hepatic lipase, and lecithin-cholesterol acyl transferase, were measured at the end of each feeding period. Results-Diet, amount of fat, or ratio of polyunsaturated to saturated fatty acids had no effect on plasma concentrations of cholesterol, triglycerides, and very–low-density or high-density lipoproteins or the activity of lecithin-cholesterol acyl transferase. Low-density lipoprotein concentrations were significantly lower in cats fed a high-fat diet containing polyunsaturated fatty acids. Lipoprotein concentration and hepatic lipase activity were significantly higher in cats fed the fat-supplemented diets, and this was unrelated to whether diets were enriched with polyunsaturated or saturated fatty acids. Conclusions and Clinical Relevance-Diets containing up to 66% of energy from fat were tolerated well by healthy cats and did not affect plasma lipid concentrations. Therefore, high-fat diets probably will not contribute to hypercholesterolemia or hypertriglyceridemia in cats.