Effects of diazinon, at different concentrations and exposure times, were investigated in freshwater fish, Cyprinus carpio , to elucidate the possible mode of action on lipid peroxidation together with the inhibitory effect of diazinon on acetylcholinesterase activity and changes in tissue protein levels. Cholinesterase inhibition is considered to be a specific biomarker of exposure to organophosphorus pesticides. Fish were exposed to 0.0036 mu g/L, 0.018 mu g/L, and 0.036 mu g/L (sublethal) concentrations of diazinon for 5, 15, and 30 days, and biochemical measurements were carried out spectrophotometrically. Brain was chosen as an indicator tissue because it is a target system for the organophosphorus action. More than 20% decline in acetylcholinesterase activity relative to mean activity of the controls was observed in the diazinon-exposed groups. Protein content decreased significantly after 15 days of exposure to 0.018 mu g/L and 0.036 mu g/L diazinon and after 30 days of exposure to 0.036 mu g/L. Malondialdehyde level declined markedly compared with the control levels. This study showed that prolonged exposures of C. carpio to diazinon had significant effects on brain acetylcholinesterase activity and that environmentally relevant concentrations of diazinon can significantly inhibit brain acetylcholinesterase activity. Altered protein content was probably due to the high energy demand under pesticide stress or inhibition of de novo enzyme synthesis. The decreased malondialdehyde content may reflect the possibility of better protection against oxidative stress.