In a 2×2 factorial design, 12 Thai Native and 12 Holstein bulls were fed ad libitum a total mixed ration (20 : 80; roughage : concentrate) with whole cottonseed (WCS) or sunflower seed (SFS) as oilseed sources. The rations contained 7% crude fat and were fed for 90 days. Plasma was taken at three times during the experiment, and at slaughter the longissimus dorsi and subcutaneous fat were sampled for fatty acid analysis. Ration did not affect rumen fermentation parameters. The plasma fatty acid profile was not affected by ration. In subcutaneous fat, a ration×breed interaction for the saturated fatty acid (SFA) and c9t11 CLA proportions was observed, resulting from larger differences between the rations in Thai Native compared with Holstein bulls. The WCS ration resulted in higher proportions of SFA and lower proportions of monounsaturated fatty acids and c9,t11 CLA compared with the SFS ration (P<0.01). In the intramuscular fat, the WCS ration was also associated with a lower c9t11 CLA proportion (P<0.01) and higher SFA proportion (P<0.05). The intramuscular proportion of polyunsaturated acids was higher and the proportion of SFA was lower in Thai Native compared with Holstein bulls (P<0.05), irrespective of ration.