Soil samples from the upper 10-cm-thick layer of the humus horizon (without forest litter) were taken in Podol’sk and Serpukhov districts (1130 and 1080 km2, respectively) of Moscow oblast. At each sampling site, ecosystem (forest, plowland, or fallow), soil (soddy-podzolic, soddy-gley, bog-podzolic, meadow alluvial, gray forest, and anthropogenically transformed soils of lawns and industrial zones), predominant vegetation, and topography (floodplain and low, medium, and upper parts of watersheds) were determined. The carbon content of the microbial biomass (Cmic) was determined by the method of substrate-induced respiration; we also determined the rate of basal (microbial) respiration (BR) and the organic carbon content, pH, and particle-size distribution. Overall, 237 samples from Serpukhov district and 45 samples from Podol’sk district were analyzed. The BR/Cmic ratios (respiration quotient qCO2) and Cmic/Corg ratios were calculated. The Cmic content in the soils ranged from 43 to 1394 μg C/kg; the BR varied from 0.06 to 25 μg CO2-C/g per h, qCO2, from 0.34 to 6.52 μg CO2-C/mg Cmic per h; and the Cmic/Corg ratio, from 0.19 to 10.65%. It was found that the most significant factors affecting the variability of the Cmic and BR are the parameters of ecosystem (50% and 80%, respectively) and soil (30% and 9%, respectively). The most significant variability of these indices was found in forest soils; it was mainly controlled by the soil texture (33 and 23%) and the Corg content (19 and 24%). The Cmic parameter made it possible to differentiate the soils of the territory for the purposes of their evaluation, monitoring, and biological assessment more clearly than the BR value and the soil chemical characteristics.