The influence of anthropogenic pollution, particularly with polycyclic aromatic hydrocarbons (PAHs) on soil toxicity and spread of antibiotic resistance genes (ARGs) is extremely important nowadays. We studied 20 soil samples from a technogenically polluted site, municipal solid wastes (MSW) landfills, and rural settlements in the southwestern part of the Rostov Region of Russia. A close correlation was established between the results of biosensor testing for integral toxicity, the content of genes for the biodegradation of hydrocarbons, and the concentration of PAHs in soils. The relation between the quantitative content of ARGs and the qualitative and quantitative composition of PAHs has not been registered. Soils subjected to different types of the anthropogenic pressure differed in PAHs composition. The technogenic soils are the most polluted ones. These soils are enriched with 5 ring PAHs and carry the maximum variety of assayed ARGs, despite the fact that they do not receive household or medical waste. •At biotesting soils contaminated with industrial PAHs are more toxic than landfills.•Technogenically polluted soils are enriched with 5 ringPAHs compared to landfills.•Technogenically polluted soils contain more diverse ARGs.