This study quantifies the effects of extruded linseed and soybean (ELS) dietary supplementation on milk yield, composition, and fatty acid profiles, as well as first-service conception rate in Holstein dairy cows. Seventy-eight open Holstein dairy cows were divided into two groups: (1) a control, which received a basal diet; and (2) a test group, which received a basal diet supplemented with the ELS (650 g/kg of extruded linseed and 150 g/kg of extruded soybean) at a rate of 100 g/kg. In the ELS group, milk yield per day and solid not fat (SNF) yield increased by 3.26% and 0.88%, respectively, in relation to the control. Percentage milk fat decreased significantly by 1.4% in the ELS group when compared with the control. The ELS supplement resulted in a decrease in saturated fatty acids (SFAs) and an increase in monounsaturated (MUFAs) and polyunsaturated fatty acids (PUFAs) in milk. In conclusion, the supplementation of dairy cow feed with 100 g/kg of ELS increases milk yield and milk unsaturated fatty acids (especially MUFAs and PUFAs). ELS supplementation also causes a decrease in percentage fat and SFA levels but does not affect the first-service conception rate or the incidence rate of mastitis.