Organophosphate (OP) compounds are the most commonly used chemical substances to eliminate pests and insects. Trichlorfon is an OP pesticide used in agriculture against vegetable and fruit pests. In this study, it was aimed to investigate antioxidant responses of Gammarus pulex exposed to trichlorfon. Acetylcholinesterase (AChE), cytochrome P450 (CYP1A1) and glutathione S-transferase (GST) enzyme activities were measured in a microplate reader using the ELISA method.When we compared AChE enzyme activity with control, it increased in all groups after 24 hours (p>0.05) and decreased in 96 hours (p<0.05). When the exposure times were statistically compared, the decrease in the A and B groups was found to be significant only at the end of the 96-hour period (p<0.05). CYP1A1 enzyme activity decreased in all groups compared to control group (p>0.05). GST enzyme activity was increased in both groups after 24 and 96 hours of application compared to control (p<0.05).When we look at the changes in AChE, CYP1A1 and GST enzyme activities in G. pulex against trichlorfon, it was concluded that these enzymes can be used as useful biomarkers. G. pulex has proven to be an effective bioindicator that shows trichlorfon pollution in water.