The aim of the study was to evaluate the effect of organic solvents, commonly used in ecotoxicity studies on the activity of soil microorganisms. Two solvents, often applied in the studies of the effects of hydrophobic organic contaminants in the environment, acetone and dichloromethane, were employed in the experiments. The evaluation of the effects of the solvents was based on the measurements of two parameters describing the overall activity of soil microorganisms (activity of dehydrogenases) and the activity of the specific group of nitrification bacteria (potential of nitrification). The experiments included different laboratory conditions (soil material properties, time of contact, methods of solvent amendment). The results show that the solvents introduced to soils in the amount commonly used in the ecotoxicity studies (10-20 cm3 kg(-1)) may inhibit the activity of soils microorganisms: the effect was particularly visible in the case of potential of nitrification determinations. Employment of the method of gradual application of the solvents to soils led to significant decrease of those negative effects.