The reuse of treated wastewater (e.g. for irrigation) is a common practice to combat water scarcity problems world-wide. However, the potential spread of opportunistic pathogens and fecal contaminants like Enterococci within the subsoil could pose serious health hazards. Additional sources (e.g., leaky sewer systems, livestock farming) aggravate this situation. This study contributes to an understanding of pathogen spread in the environment, using a combined modelling and experimental approach. The impact of quartz sediment and certain wastewater characteristics on the dissemination of Enterococcus faecalis JH2–2 is investigated. The transport processes of advection-dispersion and straining were studied by injecting conservative saline tracer and fluorescent microspheres through sediment packed columns, and evaluating resulting breakthrough curves using models. Similarly, simultaneously occurring reactive processes of microbial attachment, decay, respiration and growth were studied by injecting Enterococcus faecalis JH2–2 suspended in water with or without dissolved oxygen (DO) and nutrients through sediment, and evaluating resulting inlet and outlet concentration curves. The processes of straining, microbial decay and growth, were important when DO was absent. Irreversible attachment was important when DO was present. Sensitivity analysis of each parameter was conducted, and field scale behavior of the processes was predicted, to facilitate future work.