| Summary: | The applications of nanoparticles (Nps) as food additives, health enhancers, and antimicrobials in animal production are increasing. The aim of this study was to evaluate the effect of selenium (Se) nanoparticles (Se<sup>0</sup>Nps) stabilized with L-cysteine (Se<sup>0</sup>Nps/L-Cys), as a nutritional supplement, on immunological, oxidative status, and productive parameters in <i>O. mykiss.</i> TEM and SEM-EDS showed the accumulation of spherical Se<sup>0</sup>Nps entirely composed by elemental selenium (Se<sup>0</sup>) as intracellular and extracellular deposits in <i>Pantoea agglomerans</i> UC-32 strain. The in vitro antioxidant capacity of Se<sup>0</sup>Nps/L-Cys was significant more efficient ROS scavengers than Se<sup>0</sup>Nps and Na<sub>2</sub>SeO<sub>3</sub>. We also evaluate the effect of Se<sup>0</sup>Nps/L-Cys on cell viability and oxidative stress in RTgill-W1, RTS-11, or T-PHKM <i>Oncorhynchus mykiss</i> cell lines. Se<sup>0</sup>Nps/L-Cys showed less toxic and high antioxidant activity than Se<sup>0</sup>Nps and Na<sub>2</sub>SeO<sub>3</sub>. Finally, the dietary Se<sup>0</sup>Nps/L-Cys had a significant better effect on both plasma lysozyme and respiratory burst activity (innate immune response), on tissular Gpx activity (oxidative status), and on well-being (productive parameter) of <i>O. mykiss</i> when it is compared to Se<sup>0</sup>Nps and Na<sub>2</sub>SeO<sub>3</sub>. Se<sup>0</sup>Nps/L-Cys is a promising alternative for nutritional supplement for <i>O. mykiss</i> with better performance than Na<sub>2</sub>SeO<sub>3</sub> and Se<sup>0</sup>Nps, ease to implementation, and reduced environmental impact.
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