Objective. To establish the influence of soil treatment methods on the number of earthworms and microbial biomass in dark-chestnut heavy loam soil with observance of the following crop rotation: soybean – winter wheat – corn; verify the hypothesis of the negative influence of traditional soil treatment technologies on the number of earthworms. Methods. Analytical, field, laboratory, system analysis, statistical. Results. During the studies conducted in 2018–2021 at Askanian State Agricultural Experimental Station of the Institute of Irrigated Agriculture of the NAAS, the positive influence of zero soil treatment technologies on the studied parameters versus with traditional technologies was established. A 2.5-fold increase in the number of earthworms and a 1.1-fold increase in microbial biomass was reported with zero soil treatment technologies. The interaction between soil treatment factors and cover crops in the structure of crop rotation was 23.7 %. Soil treatment technologies had a negligible influence on the size of microbial biomass — at the level of 1.4 %. The significance of the action of cover crops in crop rotation was 10.0 %. The experiment also showed a higher number of earthworms in the soil with zero soil treatment technologies with cover crops and traditional technologies for control, due to better live environment and favourable conditions for reproduction and distribution of the population depending on soil treatment technologies. Changes in the density of the soil layer 0–30 cm were within the measurement error (for zero soil treatment technologies the studied parameter was 1.23 g/cm3, for traditional — 1.24 g/cm3). Compaction of 0–20 cm soil layer with zero and traditional soil treatment technologies was reported. The content of soil organic matter by the years of studies under zero soil treatment technologies increased to 3.0 %, which is 1.2 times higher than with traditional technologies. Conclusion. Under the conditions of stationary field experiment the positive influence of zero soil treatment technologies on early changes of biological properties of soil, which were characterized by increase in number of earthworms and size of microbial biomass, has been established. The positive influence of cover crops on soil quality was determined, which was characterized by signs of biota activation in the 0–30 cm layer and improvement of biological and physical properties of the soil.