The current study aimed to investigate the effects of chlorpyrifos exposure on the activity of acetylcholinesterase (AChE), lipid peroxidation, reduced glutathione, oxidised glutathione, glutathione peroxidase and glutathione S transferase in the kidneys, liver, and gills tissues of Channa punctatus fish. The fish were treated with 3 sub-lethal concentrations of the chlorpyrifos viz. high dose-0.6 pppm, mild dose-0.3 ppm and low dose-0.16 ppm and a control group was kept to compare the results obtained. The results showed a significant decrease in AChE activity in all three tissues of fish, which was dose-dependent and time-dependent. The study also found a significant increase in the concentration of malondialdehyde (MDA), a marker of lipid peroxidation, in all three tissues of fish exposed to chlorpyrifos, which was dose-dependent and time-dependent. The results indicated a significant dose- and time-dependent increase in GSH levels in the kidney, liver, and gills of the fish exposed to chlorpyrifos, while GSSG levels were also elevated, suggesting oxidative stress in the tissues. GPx activity was significantly altered in all three tissues, with higher doses and longer exposure times having greater effects. GST activity was significantly increased in all treatment groups, with the highest activity observed in the high dose group at all-time points. These results suggest that chlorpyrifos exposure can significantly inhibit AChE activity and induce oxidative stress in different tissues of C. punctata fish, which may lead to adverse effects on their health and survival and highlights the importance of monitoring and regulating pesticide use in aquatic environments