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Nano‐copper oxides are a versatile inorganic material. As a result of their versatility, the immense applications and usage end up in the environment causing a concern for the lifespan of various beings. The ambiguities surround globally on the toxic effects of copper oxide nanoparticles (CuO‐NPs). Hence, the present study endeavored to study the sub‐lethal acute exposure effects on the developing zebrafish embryos. The 48 hpf LC₅₀ value was about 64 ppm. Therefore, we have chosen the sub‐lethal dose of 40 and 60 ppm for the study. Accumulation of CuO‐NPs was evidenced from the SEM‐EDS and AAS analyzes. The alterations in the AChE and Na⁺/K⁺‐ATPase activities disrupted the development process. An increment in the levels of oxidants with a concomitant decrease in the antioxidant enzymes confirmed the induction of oxidative stress. Oxidative stress triggered apoptosis in the exposed embryos …