Organophosphates and synthetic pyrethroid pesticides are used in residential areas and agriculture, discharge into environment, may cause unpredictable toxicity to non-targeted aquatic organisms. The current study aims to assess the effect of these pesticides on the transaminases, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase enzymes activity in brain and gills of Oreochromis niloticus. Fish were exposed for 24 and 48 h (hours) to the LC0 (non-lethal concentrations) of the malathion, chlorpyrifos and λ-cyhalothrin, respectively. For biochemical assay, a spectrophotometer was used. Glutamate oxaloacetate transaminase activity in brain and gill was significantly (p <0.05) elevated and inhibited, respectively, except in 48 h treated samples. The highest (+258%) and lowest (+8.9%) enhancement in this activity was noted in chlorpyrifos and malathion treated gill samples, respectively. The glutamate pyruvate transaminase activity was significantly (p <0.05) enhanced in all treated samples except in 24 h malathion treated brain. The highest (+340.7%) and lowest (+27%) enhancement in this level was recorded in malathion and chlorpyrifos exposed brain and gills samples. Therefore, it is suggested that both these assessments in fish different organs could be used as a useful biomarker to detect environmental pollution. Thus, the alterations in transaminases activity indicated that the tested pesticides are highly toxic to the health of aquatic organisms and should be periodically and regularly monitored in the environment to reduce pollutant stress on non-target biota, particularly fish.