The effect of an atrazine formulation on microbial biomass, microbial respiration, ATP content and dehydrogenase and urease activity in a semiarid soil and the influence of time on the response of soil microbial activity to the herbicide treatment were assessed. The atrazine formulation was added to soil as aqueous solutions of different concentrations of active ingredient to obtain a range of concentrations in the soil from 0.2 to 1000 mg kg−1. Microcosms of soil with the different herbicide concentrations and untreated control soil were incubated for 6 h, 16 and 45 days. In general, an increase in the measured microbiological and biochemical parameters with atrazine concentration in soil was observed. The increase in microbial activity with atrazine pollution was noticeable after lengthy incubation.