Increasing the nutritional quality of fat in dairy products is a rising consumers claim motivated by the potential benefits of long chain n-3 fatty acids and low n-6/n-3 ratio on human health. Camelina is a sustainable crop and a good source of n-3 fatty acids. The goal of the trial was to study the effects of camelina cake inclusion in the diet of milking dairy cows on production and milk composition. Twenty-two primiparous and multiparous Holstein Friesian dairy cows housed at the Experimental Station (Centro Zootecnico Didattico Sperimentale) of the University of Milan in Lodi were divided in two homogeneous groups for milk production, parity and DIM (24.62 kg, 1.95, 160d). Diets consisted of a TMR corn silage-based, supplemented with 800g/h/d of camelina cake (CAME) or an isonitrogenous and isoenergetic soybean-based premix (CTR). Performances were recorded weekly. Milk composition were assessed weekly on fresh samples. Milk samples for milk fatty acid compositional analysis were collected on day 0, 21, and 42 of the trial. Data were analyzed by MIXED procedure by SAS. No differences were detected for milk production, feed intake, live body weight and body condition score. CAME group had a reduced milk fat percentage (4.27 vs. 4.72%; P < 0.05) compared to CTR but not a reduced production of fat (1.11 vs. 1.17 kg). Saturated milk fatty acids were also reduced in CAME compared with CTR (2.8 vs. 3.14%; P < 0.05). Linolenic acid and rumen biohydrogenation intermediates were increased in CAME whereas stearic acid decreased. Camelina cake could be useful to improve nutrition value of milk, but its impact on rumen metabolism and possible interferences on biohydrogenation steps need further attention.