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The use of pesticides saved millions of lives by killing the insects which are known to be potential vectors of disease, however, those pesticides have harmful effects on human health and on the environment public health and environmental problems caused using pesticides outweigh the benefits. The objective of this study is to assess the level of poisoning with the AChE test due to cotton producer’s pesticide exposure in the biggest cotton production townships of northern Benin (Banikoara and Kandi), and to identifying the risk factors.Through a cross sectional study, we recruited 190 pesticides sprayers for at least 5 years. They were submitted to a questionnaire and to acetylcholinesterase (AChE) test using the Test Mate model 400 devices (EQM Research Inc) with main component of the device: photometric sensor and principle based on the works by Ellman. By a non-probabilistic sampling, we recruited 190 farmers from the two townships fulfilling all the inclusion criteria and available to participate to all stages of the study.The studied population is essentially young: 83.16% were under 45 years old, with 75% illiterates. 70.3% of the farmers have more than 10 years of spraying experience. We noted that 2.06% of the farmers still used domestic containers to prepare the pesticides. As precautions to prevent poisoning after spraying, 10.31% of the surveyed farmers drink milk. There was a significant AChE decrease between pre-exposure (AChE 3.08±2.3 UI/ml) and post-exposure (AChE 2.65±0.52 IU/ml); p=0.009. 73.1% of the famers were concerned by that inhibition. Those who could read the pictograms faced less inhibition of AChE (p<0.05). The age variables, level of education and experience of pulverisation do not have any influence on AChE inhibition.AChE monitoring is needed for the surveillance of farmers.