The aim of this study was to evaluate the effect of a Herbal product (Nutrition Bio-Shield Superfood® (NBS)) supplementation in diets varying in metabolizable energy (ME) and crude protein (CP) contents on the internal egg quality and fatty acid profile of eggs in commercial laying hens. A total of 420 layer hens aged from 63-74 weeks were assigned into 10 treatments with 6 replicates of in a completely randomized design with a 5×2 factorial arrangement with 5 dietary levels of NBS (0, 0.5, 1, 1.5, 2 g/kg diet) and 2 concentration of nutrients (standard and 5% diluted for ME and CP). The diluted diet significantly (P < 0.05) increased the shell thickness and shape index compared to standard diet; however, it decreased shell weight and concentrations of margaric, elaidic, linoleic, docosahexaenoic, poly unsaturated fatty acids (PUFA), n3, n6, and the ratio of n3:n6 fatty acids in egg yolk compared to the standard diet (P ≤ 0.05). NBS supplementation decreased egg yolk myristic, margaric, palmitic and ginkgolic acids concentrations, linearly (P < 0.05). In addition, significant interaction effect was found between diet dilution and NBS supplementation on the Haugh unit (HU), shape index, albumen percentage and egg yolk margaric acid concentration. In general, reduction of energy and protein in the diet of laying hens by 5% of the standard had no adverse effect on egg quality parameters.

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