Decay of SARS-CoV-2 and surrogate murine hepatitis virus RNA in untreated wastewater to inform application in wastewater-based epidemiology

Warish Ahmed¹, Paul M Bertsch², Kyle Bibby³, Eiji Haramoto⁴, Joanne Hewitt⁵, Flavia Huygens⁶, Pradip Gyawali⁵, Asja Korajkic⁷, Shane Riddell⁸, Samendra P Sherchan⁹, Stuart L Simpson¹⁰, Kwanrawee Sirikanchana¹¹, Erin M Symonds¹², Rory Verhagen¹³, Seshadri S Vasan¹⁴, Masaaki Kitajima¹⁵, Aaron Bivins³

Affiliations

¹ CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Dutton Park, QLD, 4102, Australia. Electronic address: Warish.Ahmed@csiro.au.
² CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Dutton Park, QLD, 4102, Australia.
³ Department of Civil & Environmental Engineering & Earth Sciences, 156 Fitzpatrick Hall, University of Notre Dame, Notre Dame, IN, 46656, USA.
⁴ Interdisciplinary Center for River Basin Environment, University of Yamanashi, 4 - 3 -11 Takeda, Kofu, Yamanashi, 400 -8511, Japan.
⁵ Institute of Environmental Science and Research Ltd (ESR), Porirua, 5240, New Zealand.
⁶ Centre for Immunology and Infection Control, Queensland University of Technology, 300 Herston Road, Herston, QLD, 4006, Australia.
⁷ United States Environmental Protection Agency, Office of Research and Development, 26W Martin Luther King Jr. Drive, Cincinnati, OH, 45268, USA.
⁸ CSIRO Australian Centre for Disease Preparedness, Geelong, VIC, 3220, Australia.
⁹ Department of Environmental Health Sciences, Tulane University, 1440 Canal Street, New Orleans, LA, 70112, USA.
¹⁰ CSIRO Land and Water, Lucas Heights, NSW, 2234, Australia.
¹¹ Chulabhorn Research Institute, 54 Kampangpetch 6 Road, Laksi, Bangkok, 10210, Thailand.
¹² College of Marine Science, University of South Florida, 140 Seventh Avenue South, St. Petersburg, FL, 33701, USA.
¹³ Queensland Alliance for Environmental Health Sciences (QAEHS), University of Queensland, 20 Cornwall Street, Woolloongabba, QLD, 4102, Australia.
¹⁴ CSIRO Australian Centre for Disease Preparedness, Geelong, VIC, 3220, Australia; Department of Health Sciences, University of York, York, YO10 5DD, UK.
¹⁵ Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, North West 8, Kita-ku, Sapporo, Hokkaido, 060-0032, Japan.

PMID: 32861728
PMCID: PMC7451058
DOI: 10.1016/j.envres.2020.110092

Decay of SARS-CoV-2 and surrogate murine hepatitis virus RNA in untreated wastewater to inform application in wastewater-based epidemiology

Warish Ahmed et al. Environ Res. 2020 Dec.

. 2020 Dec:191:110092.

doi: 10.1016/j.envres.2020.110092. Epub 2020 Aug 27.

Authors

Affiliations

¹ CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Dutton Park, QLD, 4102, Australia. Electronic address: Warish.Ahmed@csiro.au.
² CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Dutton Park, QLD, 4102, Australia.
³ Department of Civil & Environmental Engineering & Earth Sciences, 156 Fitzpatrick Hall, University of Notre Dame, Notre Dame, IN, 46656, USA.
⁴ Interdisciplinary Center for River Basin Environment, University of Yamanashi, 4 - 3 -11 Takeda, Kofu, Yamanashi, 400 -8511, Japan.
⁵ Institute of Environmental Science and Research Ltd (ESR), Porirua, 5240, New Zealand.
⁶ Centre for Immunology and Infection Control, Queensland University of Technology, 300 Herston Road, Herston, QLD, 4006, Australia.
⁷ United States Environmental Protection Agency, Office of Research and Development, 26W Martin Luther King Jr. Drive, Cincinnati, OH, 45268, USA.
⁸ CSIRO Australian Centre for Disease Preparedness, Geelong, VIC, 3220, Australia.
⁹ Department of Environmental Health Sciences, Tulane University, 1440 Canal Street, New Orleans, LA, 70112, USA.
¹⁰ CSIRO Land and Water, Lucas Heights, NSW, 2234, Australia.
¹¹ Chulabhorn Research Institute, 54 Kampangpetch 6 Road, Laksi, Bangkok, 10210, Thailand.
¹² College of Marine Science, University of South Florida, 140 Seventh Avenue South, St. Petersburg, FL, 33701, USA.
¹³ Queensland Alliance for Environmental Health Sciences (QAEHS), University of Queensland, 20 Cornwall Street, Woolloongabba, QLD, 4102, Australia.
¹⁴ CSIRO Australian Centre for Disease Preparedness, Geelong, VIC, 3220, Australia; Department of Health Sciences, University of York, York, YO10 5DD, UK.
¹⁵ Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, North West 8, Kita-ku, Sapporo, Hokkaido, 060-0032, Japan.

PMID: 32861728
PMCID: PMC7451058
DOI: 10.1016/j.envres.2020.110092

Abstract

Wastewater-based epidemiology (WBE) demonstrates potential for COVID-19 community transmission monitoring; however, data on the stability of SARS-CoV-2 RNA in wastewater are needed to interpret WBE results. The decay rates of RNA from SARS-CoV-2 and a potential surrogate, murine hepatitis virus (MHV), were investigated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in untreated wastewater, autoclaved wastewater, and dechlorinated tap water stored at 4, 15, 25, and 37 °C. Temperature, followed by matrix type, most greatly influenced SARS-CoV-2 RNA first-order decay rates (k). The average T₉₀ (time required for 1-log₁₀ reduction) of SARS-CoV-2 RNA ranged from 8.04 to 27.8 days in untreated wastewater, 5.71 to 43.2 days in autoclaved wastewater, and 9.40 to 58.6 days in tap water. The average T₉₀ for RNA of MHV at 4 to 37 °C ranged from 7.44 to 56.6 days in untreated wastewater, 5.58-43.1 days in autoclaved wastewater, and 10.9 to 43.9 days in tap water. There was no statistically significant difference between RNA decay of SARS-CoV-2 and MHV; thus, MHV is suggested as a suitable persistence surrogate. Decay rate constants for all temperatures were comparable across all matrices for both viral RNAs, except in untreated wastewater for SARS-CoV-2, which showed less sensitivity to elevated temperatures. Therefore, SARS-CoV-2 RNA is likely to persist long enough in untreated wastewater to permit reliable detection for WBE application.

Keywords: COVID-19; Decay; Enveloped virus; Murine hepatitis virus; SARS-CoV-2; Temperature; Wastewater.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

**Fig. 1**
Mean decay curves of SARS-CoV-2 and MHV RNA over time (days) in untreated wastewater, autoclaved wastewater and tap water microcosms. The measurements were linearized premised on first-order decay, in which the natural log (ln)-transformed measured concentration at each time point was divided by the concentration at time zero. In some instances, the error bars (SD) are too small to illustrate.

**Fig. 2**
Log₁₀-transformed mean first-order decay rate constants (log₁₀k; k in units per day) of SARS-CoV-2 and MHV RNA as observed at 4, 15, 25, and 37 °C in each matrix. Each marker represents one mean log₁₀k value as observed in the current study.

See this image and copyright information in PMC

References

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Decay of SARS-CoV-2 and surrogate murine hepatitis virus RNA in untreated wastewater to inform application in wastewater-based epidemiology

Affiliations

Decay of SARS-CoV-2 and surrogate murine hepatitis virus RNA in untreated wastewater to inform application in wastewater-based epidemiology

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Miscellaneous