The long-term use of vineyards and their renewal causes numerous chemical changes in the bioavailable copper (Cu), zinc (Zn) levels, organic carbon and microbial properties of the soil. However, there are few works with an approach to the use of vineyards in subtropical soils. Thus, the objective of this work was to evaluate the relationship between physical-chemical properties, microbial basal respiration, activity, richness, and diversity from vineyards and nearby forest reference soils in three different areas in southern Brazil. Each area underwent three treatments: old cultivation, i.e., a well-established vineyard; renewed vineyard; and reference, a native forest located adjacent to the vineyards. The physical-chemical properties clay content, moisture, pH, organic matter (OM), carbon (C), nitrogen (N), phosphorus (P), potassium (K), copper (Cu), and zinc (Zn) level; the basal respiration (BR), microbial biomass Carbon (MBC), microbial biomass Nitrogen (MBN), hydrolysis of fluorescein diacetate (FDA), microbial quotient (qMic), and metabolic quotient (qCO2); and microbial richness and Shannon diversity index were determined. The organic C level differently influenced the microbial activity and richness in each of the three experimental areas. The change in microbial diversity indexes impacted soil biological activity in two locations. Vineyard renewal promotes the reduction of Cu (- 48.8%) and zinc (- 20%) bioavailability and increases microbial diversity in subtropical soils in southern Brazil.