Abstract
Due to its antioxidant and immunomodulatory properties, using passion fruit seed oil (PFSO) is a promising strategy to mitigate the effects of heat stress in laying hens, potentially optimizing the absorption of essential minerals such as selenium (Se) and zinc (Zn). Therefore, this study investigated the profile of selenium- and zinc-binding proteins (Se/Zn-BPs) in the hepatic proteome of Lohmann White laying hens (26 weeks old, n = 96) subjected to heat stress and whose diet was supplemented with 0.9% PFSO, using a metalloproteomic approach that combined two-dimensional electrophoresis (2D PAGE), graphite furnace atomic absorption spectrometry (GFAAS), and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The experimental design was a 2 × 2 factorial (temperature: thermoneutral/stress × diet: control/PFSO) design. After 84 days, liver samples were collected and subjected to metalloproteomic analyses. GFAAS analysis showed higher concentrations of Zn and Se in the protein pellets and in 11 specific protein spots of the supplemented groups (thermoneutral/PFSO and stress/PFSO). LC-MS/MS analysis identified 56 Se/Zn-BPs, with a predominance of heat shock chaperones (HSPs) and proteins involved in energy metabolism. In conclusion, PFSO supplementation modulates Se and Zn absorption, promoting a mineral balance that optimizes immune and antioxidant defense processes. This mechanism can lead to a positive impact on the health and productive performance of laying hens under heat stress.