This study was designed to evaluate the effects of two feeding methods on milk yield, composition and fatty acid (FA) profile obtained from dual‐purpose cattle. Twenty‐four Aosta Red Pied cows beyond peak of lactation were assigned to two groups and fed hay and concentrates in the proportions 0.69 and 0.31 on a dry matter basis for 10 weeks. Concentrates were offered separately from forages 6 times a day (separate ration, SR) or as a total mixed ration (TMR). The feeding method did not significantly influence dry matter intake (16.8 vs. 16.9 kg/head/day for SR‐ and TMR‐fed cows, respectively), milk yield (17.4 vs. 17.5 kg/head/day), milk fat, protein and lactose contents (36.4 vs. 35.2, 33.5 vs. 32.8, and 47.3 vs. 47.4 g/kg) and yields (607.9 vs. 613.4, 567.4 vs. 572.7 and 805.5 vs. 829.7 g/head/day). The overall milk FA profile was very similar between groups. Milk concentrations of FA used as indirect markers of rumen function (C18:2 t10c12, odd‐ and branched‐chain FA) and the extent of ruminal biohydrogenation were comparable (P > 0.05) between SR‐ and TMR‐fed cows, suggesting that ruminal pH did not vary considerably as a consequence of the feeding strategy applied.