In this study the value of subsurface treatment of urban stormwater during Aquifer Storage Transfer Recovery (ASTR) is characterised using quantitative microbial risk assessment (QMRA) methodology. The ASTR project utilizes a multi-barrier treatment train to treat urban stormwater but to date the role of the aquifer has not been quantified. In this study it was estimated that the aquifer barrier provided 1.4, 2.6, >6.0 log10 removals for rotavirus, Cryptosporidium and Campylobacter respectively based on pathogen diffusion chamber results. The aquifer treatment barrier was found to vary in importance vis-à-vis the pre-treatment via a constructed wetland and potential post-treatment options of UV-disinfection and chlorination for the reference pathogens. The risk assessment demonstrated that the human health risk associated with potable reuse of stormwater can be mitigated (disability adjusted life years, DALYs <1 × 10-6) if the aquifer is integrated with suitable post treatment options into a treatment train to attenuate pathogens and protect human health.