Patrick Breadner, Nivetha Srikanthan, Hadi Dhiyebi, Samina Hayat, Leslie Bragg, Yuwei Xie, John Giesy, and Mark Servos (2022). Partitioning behaviour of SARS-CoV-2 in wastewater. Proceedings of the GWF Annual Open Science Meeting, May 16-18, 2022.

The field of wastewater-based epidemiology has experienced rapid expansion and development since the onset of the COVID-19 pandemic. It has become a vital resource for tracking the spread of COVID-19 across communities. However, the absence of standardized methods across labs and remaining methodological questions can impact the interpretation of data for public health efforts. In particular, characterizing how RNA fragments of SARS-CoV-2 partition in wastewater is a central part of understanding its fate and behaviour in the wastewater. Additionally, various labs analyze either the solid or liquid fraction, and this has implications for the interpretation of analytical results and trends. The partitioning of SARS-CoV-2 was examined in a series of experiments that were conducted using centrifugation with varied spin time and force settings, polyethylene glycol precipitation, and filtration of wastewater. The endogenous pepper mild mottled virus (PMMoV) that is ubiquitous in wastewater was also included as it is commonly used to normalize the SARS-CoV-2 signal for trend analysis. It was demonstrated that SARS-CoV-2 splits evenly between the liquid and solid fractions under most conditions, whereas PMMoV was measured predominantly in the liquid fraction. When centrifugation conditions were changed, a longer and harder spin resulted in a shift toward the solid fraction. Developing a better understanding of the fate of SARS-COV-2 in wastewater and creating a foundation of best practices is key to supporting the current pandemic response but also to prepare for future potential infectious diseases.

AOSM2022 Next generation solutions to ensure healthy water resources for future generations First Author: Patrick Breadner (University of Waterloo) Additional Authors: Nivetha Srikanthan (University of Waterloo), Hadi Dhiyebi (University of Waterloo), Samina Hayat (University of Waterloo), Leslie Bragg¬ (University of Waterloo), Yuwei Xie (University of Saskatchewan), John Giesy (University of Saskatchewan), and Mark Servos (University of Waterloo)