Recently, research has been focusing on high quality and safer animal production by utilizing natural functional materials. The aim of this study was to evaluate the effect of administration of natural Evening Primrose Oil (EPO) on gamma linolenic acid (GLA) levels in milk from Holstein dairy cows. Quality and quantity of milk as well as blood and fatty acids from Holstein cow fed diets supplemented with 2.7-4% calcium-salted EPO coated with palm stearin oil were analyzed. There was no significant difference in yield and composition of milk between control and EPO treatment. However, EPO treatment lowered blood aspartate aminotransferase (AST), somatic cell count (SCC), and cholesterol levels (p < 0.05) compared to untreated control. Blood urea nitrogen (BUN) level was decreased (p < 0.05) in GLA 1 and GLA 2 group when compared with control group. Non-esterified fatty acids (NEFA) concentration was lower (p < 0.05) in GLA 1 and GLA 2 groups than in control group. The level of GLA in milk was increased in EPO group when compared to control. Therefore, results from the present study demonstrate that supplementary EPO has beneficial effects on cow health, showing a decrease in somatic cell count and levels of blood cholesterol, alanine aminotransferase (ALT), and AST. Furthermore, supplementation of EPO improves milk quality with increased amounts of GLA.