Soil carbon is intimately related to the living part of the organic matter, as represented by the soil microbial biomass, which mediates the decomposition, mineralization, and immobilization of organic carbon available in soils under different land-use systems. Forest-to-agriculture conversion and land-use change often lead to a loss in microbial biomass carbon (MBC) and shifts in microbial activity, directly influencing the soil carbon dynamics. The main aim of this study was to evaluate the effects of land-use change and geographical distribution on the microbial and environmental patterns related to soil C-dynamics. We evaluated MBC and microbial respiration in soils under five different land-use systems and two contrasting seasons, at a regional scale in Santa Catarina State, Southern Brazil. At the west mesoregion, changes in the MBC were correlated to sampling season in forest and grassland systems. Yet at the plateau mesoregion, we observed a land-use effect, as MBC decreased in no-till and crop-livestock integration systems. At the two mesoregions, forest and grassland had presented the highest values of MBC and microbial activity, as represented by microbial respiration. The grassland sites have presented lower values of the metabolic quotient (qCO2) and higher values of the microbial quotient (qMic). The qCO2 was lower in winter for all land-use systems. The forest sites have shown the highest total and particulate organic carbon values. The chemical-physical characteristics have shown correlations with microbiological variables related to the soil microbial C-dynamics. The land-use intensity, season, and geographic location were the main drivers of changes in microbial C-dynamics.