Monoculture of Eucalyptus urophylla × E. grandis negatively impacts soil and forest ecosystems. Mixed plantations with thinning practices are proposed to improve soil fertility, though their effects on soil carbon (C), nitrogen (N), and microbial functions are not well known. We conducted an experiment on Eucalyptus forests (Planted in 2008) with varying thinning intensities (D1=no-thinning, D2=60%, D3=70%, D4=85%) and stand types (Pure Forest-PF) and Mixed ForestMF) to assess soil quality at depths of 0-20 and 20-40 cm. Results showed significant improvements in soil physical properties, including total, dissolved, particulate, and resistant organic carbons, light fraction carbon, and microbial biomass carbon, with increases of 65%, 97%, 149%, 111%, 87%, and 30% in D3 and MF treatments compared to PF. Additionally, total nitrogen (TN), nitrate nitrogen (NO3 --N), ammonium nitrogen (NH4 +-N), and microbial biomass N significantly increased in D3 under MF treatment. Thinning up to 70% enhanced soil enzyme activities and populations of cellulose-decomposing and ammonifying bacteria in MF. Positive correlations were found between soil physiochemical properties and these bacteria. Our findings suggest that mixed plantations in Eucalyptus forests enhance soil functions, structure, biochemical attributes, and nutrient cycling, informing forest management in subtropical regions.