Acetylcholinesterase activity was measured in workers potentially exposed to pesticides that are frequently used in agriculture in 7 provinces in Colombia between 1998 and 2001.During this period, local health centers in the Departments (provinces) of Boyacá, Caldas, Huila, Meta, Norte de Santander, Santander and Valle del Cauca monitored a total of 25,242 workers for acetylcholinesterase activity. The Limperos and Ranta method, modified by Edson, was used to detect levels of pesticide exposure.The worker sample consisted of 78.9% men and 21.1% women. Thirty-nine percent of the workers were between 26 and 40 years of age, and 66% had social security. The most common work activities were use of spray applicators (39.1%) and harvesters (24.9%). Of the 25,356 tests for acetylcholinesterase activity, 7.6% showed abnormal activity levels.In the zones investigated, organophosphosphates were the most commonly used pesticides (42.4%), followed by carbamates (17.8%), organochlorines (8.4%) and chlorinates (6.6%). The diversity of pesticides in use underlines the need to increase the variety of biomarkers for monitoring exposed workers.