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The article presents a study examining the effects of ozone treatment on soil microflora. The impact of ozonation on microbial development and soil properties is analyzed. Chernozem soil from spring wheat grown using organic farming techniques was used in laboratory studies. It was found that ozone treatment has different effects on various soil microflora depending on the treatment regime. The effectiveness of ozone depends on the metabolic characteristics of microorganisms, their antioxidant defense mechanisms, the physiological structure of their cells, etc. Small doses act as a stress factor, exerting a therapeutic effect and stimulating the development of microorganisms in the soil. At an ozone treatment dose of 504 mg min/m³, populations of Bacillus bacteria increased, the content of imperfect fungi of the genus Fusarium increased, and the number of micromycetes increased by 20-60%, compared to non-ozonized soil. Moreover, the favorable environment contributed to an increase in the abundance of not only beneficial soil microflora but also the development of pathogens. Research has established that the optimal ozone treatment dose under the given conditions is 907 mg min/m³. This dose provides a balance between maintaining the abundance of beneficial Bacillus bacteria and suppressing pathogenic Fusarium fungi. Consequently, ozone treatment, when properly administered, can regulate the composition of soil microflora. The experiment provides a scientific basis for managing the composition of the soil microbiome through ozonation. This will help maintain the fertility of agricultural land, increasing its agronomic value and improving its microbiological composition, thereby increasing crop yields in an environmentally friendly manner.