Solid-liquid distribution of SARS-CoV-2 in primary effluent of a wastewater treatment plant
Femi F. Oloye,
Yuwei Xie,
Mohsen Asadi,
Jonathan K. Challis,
Charles A. Osunla,
Pu Xia,
Jenna Cantin,
Oluwabunmi P. Femi-Oloye,
Markus Brinkmann,
Kerry N. McPhedran,
Mike Sadowski,
Sudhir Pandey,
Paul D. Jones,
Chand Mangat,
Mark R. Servos,
John P. Giesy
Affiliations
Femi F. Oloye
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada; Corresponding author at: Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada.
Yuwei Xie
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada; Nanjing Institute of Environmental Science, Ministry of Ecology and Environment of China, Nanjing 210042, China; Key Laboratory of Pesticide Environmental Assessment and Pollution Control, Ministry of Ecology and Environment of China, Nanjing 210042, China
Mohsen Asadi
Department of Civil, Geological and Environmental Engineering, College of Engineering, University of Saskatchewan, Saskatoon, SK, Canada
Jonathan K. Challis
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada
Charles A. Osunla
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada
Pu Xia
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada
Jenna Cantin
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada
Oluwabunmi P. Femi-Oloye
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada
Markus Brinkmann
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada; School of Environment and Sustainability, University of Saskatchewan, Saskatoon, SK, Canada; Global Institute for Water Security, University of Saskatchewan, Saskatoon, SK, Canada
Kerry N. McPhedran
Department of Civil, Geological and Environmental Engineering, College of Engineering, University of Saskatchewan, Saskatoon, SK, Canada
Mike Sadowski
Saskatoon Water Department, Wastewater Treatment Plant, City of Saskatoon, Saskatoon, SK, Canada
Sudhir Pandey
Saskatoon Water Department, Wastewater Treatment Plant, City of Saskatoon, Saskatoon, SK, Canada
Paul D. Jones
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada; School of Environment and Sustainability, University of Saskatchewan, Saskatoon, SK, Canada
Chand Mangat
Antimicrobial Resistance and Nosocomial Infections, National Microbiology Laboratory / Public Health Agency of Canada, Canada
Mark R. Servos
Department of Biology, University of Waterloo, Waterloo, ON, Canada
John P. Giesy
Toxicology Centre, University of Saskatchewan, Saskatoon, SK, Canada; Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK, Canada; Department of Environmental Sciences, Baylor University, Waco, TX, USA; Department of Zoology and Center for Integrative Toxicology, Michigan State University, East Lansing, MI, USA
Distributions of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and fecal viral biomarkers between solid and liquid phases of wastewater are largely unknown. Herein, distributions of SARS-CoV-2, Pepper Mild Mottle Virus (PMMoV), and F-RNA bacteriophage group II (FRNAPH-II) were determined by viral RNA RT-qPCR. Comparison of viral recovery using three conventional fractionation methods included membrane filtration, a combination of mid-speed centrifugation and membrane filtration, and high-speed centrifugation. SARS-CoV-2 partitioned to the solids fraction in greater abundance compared to liquid fractions in a combination of mid-speed centrifugation and membrane filtration and high-speed centrifugation, but not in membrane filtration method in a particular assay, while fecal biomarkers (PMMoV and FRNAPH-II) exhibited the reciprocal relationship. The wastewater fractionation method had minimal effects on the solids-liquids distribution for all viral and phage markers tested; however, viral RNA load was significantly greater in solid–liquid fractions viral RNA loads compared with the than whole-wastewater PEG precipitation. A RNeasy PowerWater Kit with PCR inhibitor removal resulted in greater viral RNA loads and lesser PCR inhibition compared to a QIAamp Viral RNA Mini Kit without PCR inhibitor removal. These results support the development of improved methods and interpretation of WBE of SARS-CoV-2. • Distribution of SARS-CoV-2 to liquid and solid portions was addressed. • Addressing PCR inhibition is important in wastewater-based epidemiology. • Fraction methods have minimal effect.
