Soil, geochemical, microbiological and archaeological studies were conducted at eight settlements dating from the Paleolithic to Late Medieval and New Times near the Southern Trans-Urals mountains, Russia. The forest-steppe landscapes, rivers and abundance of mineral resources have attracted people to region since ancient times. Properties of cultural layer (CLs): close to neutral pH, well structured, absence of salinity, enrichment with exchangeable calcium and anthropogenic phosphorus (0.2–0.4%). The majority of CLs start from 3–25 cm and last up to 40–60 cm and contain 6–10% organic carbon (Corg), in a layer of 0–20 cm because modern processes are developing here, at Ishkulovo site (around 14th century) Corg decreases to 1.3%, because CL is below 80 cm and in the absence of fresh organic material it is mineralized. The content of microbial biomass of the 0–10 cm layer is 520–680 μg С/g of soil, microbes cause the emission of 0.2–1.0 μg C–CO2/g soil per h. The proximity of the studied sites to deposits of copper, chromium, zinc, manganese and Magnitogorsk Metallurgical and Uchaly processing plants creates geochemical anomalies of soils. Pollutants are also released into the CLs from bronze products, ore, ceramic fragments and raw materials from their manufacture. It leads to the concentration of pollutants of the 1st (arsenic, lead, zinc) and 2nd (cobalt, copper, nickel, chromium) hazard classes in the studied CLs, often in quantities higher than the clarke and less often – more than the allowable content for human health.