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Western diets are dominated by the consumption of chemically modified foods, characterized by a deficiency of n-3 polyunsaturated fatty acids (PUFAs). Lack of n-3 PUFAs is also present in livestock feed, which negatively affects livestock health, including immune response, and results in a low content of n-3 PUFAs in animal products. The goal of this experiment was to study the effect of the addition of feed containing PUFAs produced by the fungus <i>Mortierella alpina</i> on immune parameters (IgA, MUC-2, IGF-2, phagocytoses and selected lymphocyte subsets) and the composition of the intestinal microbiota of hens and egg fatty acids profile. Hens were divided into groups (control, F10—supplemented with 10% of fermented feed, F15—supplemented with 15% of fermented feed). The relative expression of all genes was markedly upregulated, mainly in the F15 group. Likewise, in F15, a significant increase in both phagocytes engulfing capacity and the level of oxidative burst was observed. Neither CD T cell subpopulations nor the CD4/CD8 ratio were significantly affected. A significant increase in small intestinal enterobacteria was observed in the F15. The fatty acid profile of eggs in both experimental groups showed an increased proportion of n-3 PUFAs and decreased n-6/n-3 PUFAs ratio. The results of this work show that the addition of 15% omega-3 acids enriched fermented feed positively affected the immune response of laying hens and improved the fatty acid composition of eggs.