Supplementation with eicosapentaenoic and docosahexaenoic acids (EPA and DHA) through the addition of fish oils to mammal diets during lactation benefits milk production, litter growth and the litter immune system, but there is little evidence supporting the use of oils that have been dried using a spray-drying method designed to cool and crystalize fat particles. The objective of this study was to evaluate the effect of a spray-dried dietary fat enriched with EPA and DHA on the fatty acid composition of sow milk. Fifteen pregnant sows were assigned to three dietary treatments from day 100 of gestation until weaning (day 28). Control sows (CONT) were fed an unsupplemented basal diet, and supplemented sows were fed the basal diet plus 20 g (FOPF20) or 40 g (FOPF40) of a spray-dried powdered fat enriched with EPA and DHA. Milk and colostrum compositions and milk yield were similar among diets, and the main milk fatty acids (FA) were C14:0, C16:0, C18:0, C18:1 n-9 and C18:2 n-6. Supplementation with EPA and DHA (FOPF20 and FOPF40) decreased the saturated FA contents and increased the amount of polyunsaturated FA. In summary, the results indicated that supplementing sow diets with EPA and DHA could decrease the saturated fatty acid content and increase the polyunsaturated fatty acid content of milk.