The present study was conducted in order to evaluate the effects of calf presence during milking and cow breed on milk fatty acid (FA) profile. 30 Prim'Holstein (H) and Salers (S) primiparous lactating cows were milked in the presence (CP) or in the absence (CA) of their calves, during the 9 months of lactation. Milk FA profile was significantly influenced by breed: 4:0, 12:0, 18:2 n-6 and cis-18:1 had higher concentrations in milk from H cows, whereas 14:0, 18:3 n-3, cis-9, trans-11-CLA and trans-18:1 had higher concentrations in milk from S cows. The presence of the calf during milking also had an important effect: it decreased the milk fat content, as well as 18:0, 18:2 n-6, 18:3 n-3 and trans-18:1 levels and it increased 16:0 level. Cis-9, trans-11-CLA concentration increased in the presence of the calf, for H cows, whereas it decreased for S cows.