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Our study focused on the toxicity evaluation of copper oxide nanoparticles (CuO-NPs) on a bioindicator; the land snail Helix aspersa. Their effects were studied by a targeted approach in the laboratory, by evaluating the oxidative stress biomarkers (GSH, GST, GPx, CAT, and LPO) in hepatopancreas and kidney. The snails were exposed to increasing concentrations (50, 100, 150, and 200 mg/kg) of CuO-NPs mixed in wheat flour during a sub-chronic treatment period of 45 days. Our results show that: CuO-NPs can induce oxidative stress, by producing reactive oxygen species (ROS), which was confirmed by the decrease in glutathione (GSH) level and reduction of its metabolizing enzyme the glutathione-s-transferase (GST) in both organs, as they trigger the detoxification system resulting in increased activity of the glutathione peroxidase (GPx) and catalase defense enzyme and lipide peroxidation indices within the hepatopancreas.