Microplastics (MPs) are a big and growing environmental concern, with studies showing sublethal to acute biological impacts on typical aquatic organisms. However, little is known about the biological effects of naturally weathered MPs, particularly focusing on mitochondria dysfunction as the key trigger of the biological effects. Therefore, in this study, naturally weathered MPs were produced from day-to-day life products, characterized, and chronically exposed (21 days) to adult zebrafish at the concentration of 0.1 and 1 mg/L. Locomotion and unconditioned anxiety-like behaviour was assessed. Mitochondrial respiration, membrane potential, mitochondrial complex activity and oxidative-related parameters were evaluated in the brain and liver. The results revealed the weathered MPs as a copolymer of propylene and ethylene that induced anxiety-like behaviour. There was an increase in brain catalase activity while the brain lactate dehydrogenase activity was inhibited after exposure to 1 mg/L. Brain glutathione levels were increased while their ratio was not affected. Mitochondrial respiratory chain complex Ⅱ and IV were also significantly decreased in the brain, although not compromising mitochondrial function. On the other hand, exposure to 1 mg/L caused a deficiency in liver mitochondrial respiration and decreased mitochondrial membrane potential, which were associated with the mitochondrial respiratory chain inhibition. An increase in hepatic superoxide dismutase and catalase activity was noticed, supporting the occurrence of ROS-induced ROS release as the potential trigger for the mitochondrial dysfunction. Overall, these findings highlight the potential indirect and cumulative environmental effects these particles may pose to aquatic ecosystems.