The increasing use of plant-based ingredients in aquafeeds for Atlantic salmon has led to increasing feed concentrations of plant sterols (phytosterols) combined with decreasing concentrations of cholesterol. Questions have been raised regarding how this may affect performance, health and lipid metabolism in a carnivorous fish species adapted to a high cholesterol diet in nature, and whether phytosterols could be involved in the liver lipid accumulation commonly observed in salmon fed diets with high content of vegetable oils.A feeding trial with 9 diet groups with varying additions of phytosterols and cholesterol was conducted at two temperatures (6 and 12°C). Fish grew from an initial weight of ~250g to a final weight of ~900g. Overall, the addition of phytosterols and/or cholesterol within the range tested (1353–3147mgkg−1 phytosterols, 867–3153mgkg−1 cholesterol and phytosterol:cholesterol ratios of 0.7–3.2) exhibited few effects on fish performance, lipid metabolism and health. The main hypothesis regarding phytosterols or phytosterol:cholesterol ratio affecting liver lipid accumulation appears to be false. However, dietary cholesterol did increase muscle pigmentation at both temperatures, while fillet dry matter and hepatosomatic index were also increased by dietary cholesterol at 12°C. Dietary cholesterol also had some minor effects on the fatty acid composition of liver polar lipids, probably mediated by LXR. In general, more effects were seen from variations in dietary cholesterol compared to phytosterols.The current trial is the first to feed salmon graded levels of phytosterols, cholesterol and their ratio without changing the dietary fatty acid composition, and thereby isolating the effects of dietary sterols.