Flow-driven biomarker movement in gravitational sewers for wastewater-based epidemiology and public health monitoring

The movement of biological (genetic viral, fungal or bacterial) and chemical indicators (BCIs) within sewer networks is critical to wastewater-based epidemiology (WBE) enabling accurate calculation of chemical and pathogen loads within a community. These quantified BCIs, which include genetic material from pathogens as well as pharmaceuticals, from a range of classes, serve as proxies for community-wide health and behaviour patterns. However, a critical knowledge gap exists in understanding how different BCIs move within complex sewer systems, which could lead to misinterpretation of community-level data. This study aims to address this gap by investigating the transport behaviour of 5 common BCIs (carbamazepine, metoprolol, naproxen, venlafaxine and PMMoV) in a real-world gravitational sewer network. In addition, we also spiked the wastewater with deuterated caffeine-d9, allowing discrimination from native caffeine present in the network and therefore, enabling an accurate assessment of recovery due to no public use. Our results revealed that all targets travelled with limited dispersion throughout the first stage of the gravitational sewer, 0.8 km after introduction into the network. It was observed that carbamazepine (logD = 2.8 at sewer pH), exhibited more dispersion throughout the remaining 2.3 km of the gravitational system, showing a broader, more asymmetric trace with increased tailing, which potentially indicates sorption to the solid phase, impacting its movement through the network. All other chemical targets had similar movement patterns, indicating a lower tendency to bind to the solid phase (logD < 1, at average sewer pH). Loads were calculated using dye-predicted flow rates and normalized to caffeine-d9. Carbamazepine loads were under-predicted by 74 %, attributed to losses to the solid phase throughout the sewer system. Conversely, metoprolol, naproxen, and venlafaxine loads were over-predicted (146 %, 32 %, and 129 %, respectively), likely due to additional public inputs. Our results demonstrate that more hydrophilic chemicals move throughout the sewer network with limited dispersion while hydrophobic compounds may experience significant losses. These findings have important implications for the accurate interpretation of WBE data, future BCI tracing studies and the selection of appropriate chemical markers for community health monitoring.