Wastewater-based surveillance (WBS) is increasingly used for monitoring disease targets in wastewaters around the world. This study, performed in Ottawa, Canada, identifies a decrease in SARS-CoV-2 wastewater measurements during snowmelt-induced sewer flushing events. Observations first revealed a correlation between suppressed viral measurements and periods of increased sewage flowrates, air temperatures above 0 °C during winter months, and solids mass flux increases. These correlations suggest that high sewage flowrates from snowmelt events or intense precipitation events lead to the scouring of previously settled solids in sewers and the subsequent entrainment of these solids into the transported wastewaters. Collection of WBS samples during flushing events hence contains a heterogeneous mixture of solids, including resuspended solids with varying degrees of decay. Therefore flushing events can present a challenge for accurately measuring disease target viral signals when using solids-based analytical methods. This study demonstrates that resuspended solids entrained in the wastewaters during flushing events retain PMMoV signal while the SARS-CoV-2 signal is significantly reduced due to the slower decay rate of pepper mild mottle virus (PMMoV) compared to SARS-CoV-2 within wastewaters. Hence current normalization methods using PMMoV are shown to be ineffective in correcting for flushing events and the associated resuspension of settled solids, as the PMMoV signal of settled solids within sewers does not account for the differential decay rates experiences by SARS-CoV-2 signal in settled solids. Instead, this study identifies RNA to PMMoV correction factor as an effective approach to correct for flushing events and to realign SARS-CoV-2 signal with COVID-19 hospital admission rates within communities. As such, the study highlights the key physicochemical parameters necessary to identify flushing events that affect SARS-CoV-2 WBS measurements and introduces a novel RNA to PMMoV correction factor approach for solids-based analysis of SARS-CoV-2 during flushing events, enhancing the accuracy of WBS data for public health decision-making.
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