In five short-term experiments conducted in Victoria in 1997 and 1998, grazing dairy cows were given either pasture alone or pasture supplemented with high-energy concentrates, and the fatty acid profiles of milk fat were measured. We established the effects of these feeds on some aspects of milk fat of importance for human nutrition, but we specifically focused on the hypothesis that conjugated linoleic acid (CLA) concentrations in milk fat increase as pasture intake increases, and decrease as more concentrates are fed. In agreement with previous research, feeding fresh pasture alone resulted in high concentrations (1·0–1·8 g/100 g milk fat) of CLA. When the effect of level of pasture consumption on CLA content was examined, a significant positive relationship (r2=0·35; P<0·05) was obtained. When cereal grain concentrates were used to supplement pasture intake, the CLA content of milk fat generally declined (P<0·05), except where the amount of concentrates given led to a marked reduction in total milk fat concentration. The use of cereal grain concentrates also generally resulted in significant (P<0·05) increases in medium-chain saturated fatty acids, but always reduced the contribution of butyric acid to milk fat, from 4·5 to 3·9 g/100 g milk fat, on average.