We studied the effect of mefenoxam on soil biochemical properties. Soil was mixed with three rates of mefenoxam (0.5, 1 and 2 L ha(-1)) and incubated for 83 days. Fungicide was applied to the soil four times during the experiment, according to the manufacturer's instructions. Soil ergosterol, dehydrogenase, urease, β-glucosidase, and phosphatase activities were measured during the experiment. Compared to controls, soils with the highest doses of mefenoxam demonstrated decreased ergosterol and dehydrogenase activities by 81 and 27 %, respectively; whereas, urease, β-glucosidase, and phosphatase activities increased. These results suggest that mefenoxam may possibly have consequences for agronomic crop production due to the negative effect on soil fungal populations and stimulation of the growth of soil bacterial activity.