BACKGROUND: Omega-3 fatty acids can be conducive toreducing the harmful effects of physiological stress through reducing the stimulatory activity of glutamatergic neurons.OBJECTIVES: The aim of this study was to investigate the effects of omega-3 fatty acids on performance, blood parameters, and the profile of fatty acids in the egg yolks of laying hen under stress.METHODS: This study was performed using 96 LSL-Lite laying hens in a 2 × 3 factorial experiment with 2 factors, namely stress (no dexamethasone and 1.5 mg/kg of diet dexamethasone) and omega-3 fatty acids (the levels of 0, 0.24 and 0.48 % of diet), in a completely randomized design for 70 days from 35 to 44 weeks of age.RESULTS: Physiological stress reduced the feed intake, the percentage of egg production, and egg mass in laying hens (P<0.05). Omega-3 fatty acid supplements had no effects on these traits. The groups receiving 0.48 % of omega-3 fatty acids had the lowest heterophil:lymphocytes (P<0.05), and there was no difference between stressed and non-stressed groups. The ratio of omega-6 to omega-3 fatty acids was higher in birds previously affected by stress (P<0.05). Feeding with omega-3 fatty acids in birds increased omega-3 fatty acids in egg yolk and reduced the ratio of omega-6 fatty acids to omega-3 in laying hens (P<0.05).CONCLUSIONS: Based on the results of this experiment, physiological stress had a negative effect on performance traits, and the use of omega-3 fatty acids in laying hens under stress had no effect on performance traits. Physiological stress altered the profile of fatty acids and increased the proportion of omega-6 to omega-3 fatty acids in egg yolk. The use of omega-3 fatty acids reduced the negative effects of stress and improved the profile of fatty acids in egg yolk.