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Objective The aim of this study was to evaluate the effects of occupational exposure of Federal Highway Police (PRF) officers to cholinesterase-inhibiting insecticides (CII). Methods We evaluate erythrocyte and plasma cholinesterase activity, pulse rate, systolic and diastolic pressure, and clinical evaluation through the Mini Mental State Examination (MMSE) and the Diagnostic and Statistical Manual of Mental Disorders (DSM-V). Results All PRF officers evaluated were male, between 22 and 49 years of age. Pulse rate of the subjects were statistically superior in the post-exposure moment when compared to pre-exposure moment. Inhibition of acetylcholinesterase and butyrylcholinesterase was significant in the post-exposure moment when compared to pre-exposure moment. Conclusions In conclusion, the results of the present work show that there are significant biochemical changes which can be the beginning of serious deleterious effects to the health of PRF officers.