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Selenium nanoparticles (SeNPs) are fast becoming a key instrument in several applications such as medicine or nutrition. Questions have been raised about the safety of their use. Male rats were fed for 28 days on a monodiet containing 0.5, 1.5, 3.0 and 5.0 mg Se/kg. Se content in blood and liver, liver panel tests, blood glucose, total antioxidant capacity (TAC), the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were analysed. Liver and duodenum were subjected to histopathology examination. The weight gain of rats showed no differences between tested groups. Se content in blood was higher in all treated groups compared to the control group. The liver concentration of Se in the treated groups varied in the range from 222 to 238 ng/g. No differences were observed in the activity of AST (aspartate aminotransferase), ALP (alkaline phosphatase) and TAS (total antioxidant status). A significant decrease in ALT activity compared to the control group was observed in the treated groups. GPx activity varied from 80 to 88 U/mL through tested groups. SOD activity in liver was decreased in the SeNP-treated group with 5 mg Se/kg (929 ± 103 U/mL). Histopathological examination showed damage to the liver parenchyma and intestinal epithelium in a dose-dependent manner. This study suggests that short-term SeNP supplementation can be safe and beneficial in Se deficiency or specific treatment.