Microbial populations are primarily responsible for the decomposition of organic residues, the nutrients cycle, and the flow of energy inside of soil. The present study was undertaken to link soil microbiological and soil biochemical parameters with soil- and litter-quality conditions in the surface layer from 5 sites differing in plant cover, in stand age, and in land-use history. The aim was to see how strongly these differences affect the soil microbial attributes and to identify how microbiological processes and structures can be influenced by soil and litter quality. Soil and litter samples were collected from 5 sites according to different land use: preserved forest, nonpreserved forest, secondary forest, pasture, and eucalyptus plantation. Soil and litter microbial biomass and activity were analysed and DNA was extracted from soil. The DNA concentrations and soil microbial C and N correlated positively and significantly, suggesting that these are decisive nutrients for microbial growth and time required for microbial biomass renewal. The litter microbial biomass represented a source of C and N higher than soil microbial biomass and can be an important layer to contribute to tropical soil with low C and N availability. The litter quality influenced the litter and soil microbial biomass and activity and the soil bacterial diversity. The chemical and nutritional quality of the litter influenced the structure and microbial community composition in the eucalyptus plantation.