Polyunsaturated fatty acids are important membrane components that influence membrane integrity and fluidity. In the present study, the effect of oral supplementation for 60 days with essential fatty acids (omega 3, 6 and 9) and vitamin E on canine semen quality was evaluated. Sixteen dogs were selected for the experiment; eight were used as the control group and eight received the fatty acid supplemented diet for 60 days. Semen samples were taken every 15 days during the entire experimental period and were analyzed for volume (ml), motility (%), vigour (0-5), concentration (x10(6)/ml), morphology of spermatozoa (%), plasma membrane integrity (%; using the hyposmotic swelling test) and thermoresistance (motility and vigour after 4 h at 38 degrees C). We concluded that, daily supplementation with omega 3, omega 6 and omega 9 fatty acids, together with vitamin E, for a period of 60 days, significantly increased the semen volume of the treated group after 15 days of supplementation; the vigour and concentration of spermatozoa were superior after the first month of supplementation, while the percentage of morphologically abnormal spermatozoa decreased and the cells were protected against thermal stress.