The effects of level of fish oil inclusion in the diet on grass silage intake, and milk yield and composition of dairy cows offered either 5 or 10 kg concentrates/d were evaluated in a ten treatment, partly balanced, changeover design experiment involving 50 cows in early lactation. Concentrates were prepared to provide 0, 150, 300 or 450 g fish oil/cow per d or 300 g fish oil/cow per d from a premix when each animal was offered 5 kg/d. The fish oil was predominantly from herring and mackerel caught in the North Atlantic while the fish oil premix was obtained from a commercial source and used palm kernel expeller as a carrier. Increasing fish oil supplementation decreased silage dry matter intake and the concentrations of milk fat and protein, and increased milk yield and diet digestibility. There were significant interactions between concentrate feed level and level of fish oil for silage intake and milk yield. Other than for the concentrations of milk fat and protein, and 20[ratio ]4n−6 fatty acids, the source of fish oil did not affect forage intake or animal performance. Fish oil supplementation also decreased the concentrations of milk protein by 0·9 g/kg for each 100 g increase in fish oil supplementation, the depression being similar at each level of concentrate feeding. Supplementing the feed of dairy cows with 450 g fish oil/cow per d decreased the concentration of milk fat by 15 g/kg. This study also showed that feeding dairy cattle with fish oil is an efficient method of increasing eicosapentaenoic acid in the human diet through transfer into milk.