Effects of breed type and sex on the fatty acid composition of subcutaneous neutral lipid and intramuscular neutral and phospholipids of longissimus lumborum muscle were investigated using 145 steers and 82 heifers that consisted of pure Japanese Black and Holstein and crossbreds among Japanese Black, Holstein, Japanese Brown, and Charolais. Steers and heifers were reared on a high plane of nutrition and were fed the same concentrate diet and rice straw. All animals were slaughtered serially and carcass composition was determined by dissection of the left side of the carcass. Breed type and sex differences of fatty acid percentages of carcass lipids were compared by adjusting the percentages to mean carcass fat percentages. Heifers had higher contents of 18:1 and total monounsaturated fatty acids in subcutaneous and intramuscular neutral lipids than steers (P < .05). The fatty acid composition of intramuscular phospholipids differed between sexes for 16:0, 20:1, and 20:5, but the differences were small. Breed differences were significant (P < .05) in steers for 16:0, 16:1, 18:1, and total saturated and monounsaturated fatty acids in both subcutaneous and intramuscular neutral lipids, and iso-16:0, 16:0, and total saturated fatty acids in phospholipids, respectively. However, in heifers, fewer fatty acids differed (P < .05) among breed types in the neutral lipids. It is suggested that the Japanese Black has a genetic predisposition for producing carcass lipids containing higher concentrations of monounsaturated fatty acids than Holstein, Japanese Brown, or Charolais.