An increase in the intake of the n-3 series polyunsaturated fatty acids (PUFA) is recommended by nutritionists for the human diet and beef is a significant source of these fatty acids. Enhancing the n-3 PUFA content of beef is important in view of the generally saturated nature of fatty acids in ruminant meats and the potentially negative effect this can have on human health. This study examined the effects of breed and diet on the fatty acid composition of beef M. longissimus. Ninety-six steers were used, 48 Aberdeen Angus cross (AA) and 48 Holstein-Friesian (HF). At 6months of age, 3 groups were identified, to be slaughtered at 14, 19 and 24months, respectively. Each group consisted of eight steers of each breed fed on a concentrate or a grass silage diet, rich in n-6 and n-3 PUFA, respectively. The intake of the concentrate diet was restricted so that steers of each breed grew at a similar rate on each diet. The early maturing AA produced heavier, fatter carcasses with better conformation. Animals fed grass silage had higher carcass fatness and conformation scores and higher levels of neutral lipid and total lipid in muscle than those fed concentrate. When all animals were pooled, a decline in PUFA% as total muscle lipid increased was evident. Feeding a grass silage diet rich in α-linolenic acid (18:3n-3) increased levels of this fatty acid in muscle neutral lipid by a factor of about 3.0 compared with the concentrate diet, as well as enhancing the synthesis of the n-3 series long-chain C20-22 PUFA in the phospholipid fraction, including docosahexaenoic acid (DHA, 22:6n-3). In contrast, both levels and proportions of linoleic acid (18:2n-6) and the n-6 series C20-22 PUFA were higher in animals fed the concentrate diet. The proportions of 18:1trans and conjugated linoleic acid (CLA) in muscle neutral lipid were higher in animals fed concentrate compared with silage in all 3 groups. This was partly due to increased consumption of 18:2n-6. The ratio of PUFA to saturated fatty acids (P:S) in muscle was reduced by feeding grass silage, partly as the result of increased fat deposition. However, the increase in levels of n-3 series fatty acids with silage-feeding resulted in beneficially low n-6:n-3 ratios in muscle in all age groups (approximately 1.2 compared with 12.0 in the concentrate diet). Subtle breed differences in PUFA amounts and proportions were noted. Holstein-Friesians had higher proportions of PUFA and higher P:S ratios compared with AA, partly due to a higher proportion of phospholipid in total lipid. In phospholipid itself, HF in the 19 and 24months groups had higher proportions of most n-3 PUFA. In all age groups the ratio of DHA to its precursor, 18:3n-3 was higher in HF.