1. Laying hen performance, yolk fat fatty acid concentrations and firmness of eggs were evaluated with respect to the inclusion in the diet of conjugated linoleic acid (CLA) and fish oil. 2. Nine diets were arranged factorially, with three levels of supplementation of CLA (1, 3 and 5 g/kg) and fish oil (0, 14 and 20 g/kg). 3. Type of diet did not affect egg production traits. 4. CLA addition increased yolk weight and yolk fat concentrations of CLA, saturated and total long-chain n-3 fatty acids, but decreased those of monounsaturated and total long-chain n-6 fatty acids. 5. Fish oil addition increased long-chain n-3 fatty acids yolk fat concentrations but decreased those of CLA, saturated and long-chain n-6 fatty acids. 6. Effects of CLA addition on yolk fat concentrations of C22:4 n-6 and C20:5 n-3 were greater when no fish oil was added to the diet. 7. CLA supplementation increased linearly yolk moisture and firmness and altered albumen and yolk pH.