AbstractSpatially distributed properties of the subsurface result in varying water saturation and preferential flow paths, which lead to heterogeneous solute transport patterns and heterogeneous microbial environments. This, in turn, influences the distribution of nutrients and energy gradients, microbial biomass, and activity thereof. By their very nature, current field sampling techniques do not resolve subsampling scale heterogeneities in microbial biomass and activity, resulting in inaccurate estimates of microbially mediated carbon and nitrogen turnover in the heterogeneous subsurface. Thus, in this study, we undertook a numerical modeling approach to study the impact of spatial heterogeneity on microbially mediated carbon and nitrogen turnover in the vadose zone. We adapted an established biogeochemical process network that captures a variety of respiration pathways, carbon decomposition strategies, and microbial life processes to simulate microbially mediated carbon and nitrogen turnover in variably saturated spatially heterogeneous settings, using an established numerical tool (OGS#BRNS). The fractionation of microbial communities into active and inactive states, as well as immobile and mobile states followed could be linked to the bulk average saturation. Lastly, we identified three reactive systems, distinguished by the rate ratio of aerobic respiration and transfer of oxygen from the air to the water phase, to evaluate the impact of spatial heterogeneity on carbon and nitrogen removal in subsurface heterogeneous domains. Specifically, when this ratio is approximately 1, there is no impact on carbon removal, while when this ratio is very high, then carbon removal decreases as the domain tends to be oxygen limited.