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Impact of coagulation on SARS-CoV-2 and PMMoV viral signal in wastewater solids

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Abstract

Wastewater surveillance (WWS) of SARS-CoV-2 has become a crucial tool for monitoring COVID-19 cases and outbreaks. Previous studies have indicated that SARS-CoV-2 RNA measurement from testing solid-rich primary sludge yields better sensitivity compared to testing wastewater influent. Furthermore, measurement of pepper mild mottle virus (PMMoV) signal in wastewater allows for precise normalization of SARS-CoV-2 viral signal based on solid content, enhancing disease prevalence tracking. However, despite the widespread adoption of WWS, a knowledge gap remains regarding the impact of ferric sulfate coagulation, commonly used in enhanced primary clarification, the initial stage of wastewater treatment where solids are sedimented and removed, on SARS-CoV-2 and PMMoV quantification in wastewater-based epidemiology. This study examines the effects of ferric sulfate addition, along with the associated pH reduction, on the measurement of SARS-CoV-2 and PMMoV viral measurements in wastewater primary clarified sludge through jar testing. Results show that the addition of Fe3+ concentrations in the conventional 0 to 60 mg/L range caused no effect on SARS-CoV-2 N1 and N2 gene region measurements in wastewater solids. However, elevated Fe3+ concentrations were shown to be associated with a statistically significant increase in PMMoV viral measurements in wastewater solids, which consequently resulted in the underestimation of PMMoV-normalized SARS-CoV-2 viral signal measurements (N1 and N2 copies/copies of PMMoV). The observed pH reduction from coagulant addition did not contribute to the increased PMMoV measurements, suggesting that this phenomenon arises from the partitioning of PMMoV viral particles into wastewater solids.

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Acknowledgements

The authors wish to acknowledge the help and assistance of the University of Ottawa, the Ottawa Hospital, the Children’s Hospital of Eastern Ontario, the Children’s Hospital of Eastern Ontario’s Research Institute, Public Health Ontario, and all of their employees who were involved in this project. Their time, facilities, resources, and feedback are greatly appreciated.

Funding

This research was supported by the Province of Ontario’s Wastewater Surveillance Initiative (WSI). It was also supported by a CHEO (Children’s Hospital of Eastern Ontario) CHAMO (Children’s Hospital Academic Medical Organization) grant, which was awarded to Dr. Alex E. MacKenzie.

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Authors and Affiliations

  1. Department of Civil Engineering, University of Ottawa, Ottawa, ON, Canada

    Nada Hegazy, Xin Tian, Patrick M. D’Aoust, Lakshmi Pisharody, Syeda Tasneem Towhid, Élisabeth Mercier, Zhihao Zhang, Shen Wan, Ocean Thakali, Md Pervez Kabir, Wanting Fang, Tram B. Nguyen, Nathan T. Ramsay, Stéphanie Guilherme & Robert Delatolla

  2. Children’s Hospital of Eastern Ontario Research Institute, Ottawa, ON, Canada

    Alex E. MacKenzie & Tyson E. Graber

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  1. Nada Hegazy
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  2. Xin Tian
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Contributions

All authors contributed to the study conception and design as described below. The first draft of the manuscript was written by Nada Hegazy and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Nada Hegazy: experimental work, investigation, formal analysis, writing, review and editing.

Xin Tian: experimental work, review and editing.

Patrick M. D’Aoust: experimental work, review and editing.

Lakshmi Pisharody: investigation, review and editing.

Syeda Tasneem Towhid: experimental work, review and editing.

Élisabeth Mercier: review and editing.

Zhihao Zhang: experimental work, review and editing.

Shen Wan: review and editing.

Ocean Thakali: review and editing.

Md Pervez Kabir: review and editing.

Wanting Fang: review and editing.

Tram B. Nguyen: review and editing.

Nathan T. Ramsay: review and editing.

Alex E. MacKenzie: methodology, validation, review and editing, funding acquisition.

Tyson E. Graber: review and editing.

Stéphanie Guilherme: validation, supervision, review and editing.

Robert Delatolla: methodology, validation, supervision, writing—review and editing, funding acquisition.

Corresponding author

Correspondence to Robert Delatolla.

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Highlights

• Effects of primary coagulation on SARS-CoV-2 and PMMoV measurements are unknown.

• Fe3+ addition to 60 mg/L had no effect on SARS-CoV-2 N1 and N2 measurements.

• Fe3+ addition to 60 mg/L elevated PMMoV measurements due to enhanced liquid-to-solid partitioning.

• Fe3+ addition to 60 mg/L underrepresents PMMoV-normalized N1 and N2 measurements.

• pH change associated with Fe3+ addition did not contribute to the elevation of PMMoV measurements.

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Hegazy, N., Tian, X., D’Aoust, P.M. et al. Impact of coagulation on SARS-CoV-2 and PMMoV viral signal in wastewater solids. Environ Sci Pollut Res 31, 5242–5253 (2024). https://doi.org/10.1007/s11356-023-31444-1

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