Agricultural land faces pressures from intensive farming and climate change, leading to soil quality degradation. Identifying suitable biotic and abiotic indicators of agricultural soil status is crucial. This study investigates microbial and physicochemical soil parameters in four diverse agricultural areas in Central Poland: two at the Pilica and two at the Bzura catchments. Samples were collected from three zones in a gradient of distance from local streams: ecotone (1 m), transitional (6 m) and agricultural (30 m), observing spatial differences in the aforementioned parameters. This novel - spatial approach highlighted relationships among soil physiochemistry and microbial parameters (16S rRNA, heterotrophic communities, metabolic activity, bacterial nutrient-transforming potential and antibiotic resistance genes - ARGs). The Bzura agricultural areas, with better quality of soil, had remarkedly higher microbial metabolic activity (~1.58 AWCD index) and abundance of nitrifying (~6.77 x 10 5 amoA gene copy g -1 of soil) and denitrifying (~5.58 x 10 5 nosZ gene copy g -1 of soil) bacteria compared to Pilica (1.22 AWCD, 2.56 x 10 5 amoA and 4.18 x 10 5 nosZ, respectively). High levels of K, P and N-NH 4 significantly differentiated Bzura to Pilica, suggesting these elements increased microbial parameters. Nitrifying bacteria decreased in high soil organic matter and increased in high N-NH 4, making them more sensitive and better soil quality indicators than denitrifying bacteria. Nevertheless, detection of intI and ARGs ( mcr3-6, tetM, ermC and sul1) was higher in Bzura than Pilica, indicating that increase microbial parameters in good agricultural quality soils could pose a risk for the transmission of emerging contaminants.