Fine-Scale Temporal Dynamics of SARS-CoV-2 RNA Abundance in Wastewater during A COVID-19 Lockdown

Abstract

Wastewater is a pooled sampling instrument that may provide rapid and even early disease signals in the surveillance of COVID-19 disease at the community level, yet the fine-scale temporal dynamics of SARS-CoV-2 RNA in wastewater remains poorly understood. This study tracked the daily dynamics of SARS-CoV-2 RNA in the wastewater from two wastewater treatment plants (WWTPs) in Honolulu during a rapidly expanding COVID-19 outbreak and a responding four-week lockdown that resulted in a rapid decrease of daily clinical COVID-19 new cases. The wastewater SARS-CoV-2 RNA concentration from both WWTPs, as measured by three quantification assays (N1, N2, and E), exhibited both significant inter-day fluctuations (10 1.2 -10 5.1 gene copies or GC/L in wastewater liquid fractions, or 10 1.4 -10 6.2 GC/g in solid fractions) and an overall downward trend over the lockdown period. Strong and significant correlation was observed in measured SARS-CoV-2 RNA concentrations between the solid and liquid wastewater fractions, with the solid fraction containing majority (82.5%-92.5%) of the SARS-CoV-2 RNA mass and the solid-liquid SARS-CoV-2 RNA concentration ratios ranging from 10 3.6 to 10 4.3 mL/g. The measured wastewater SARS-CoV-2 RNA concentration was normalized by three endogenous fecal RNA viruses (F + RNA coliphages Group II and III, and pepper mild mottle virus) to account for potential variations that may occur during the multi-step wastewater processing and molecular quantification, and the normalized abundance also exhibited similar daily fluctuations and overall downward trend.

On the Critical Role of Human Feces and Public Toilets in the Transmission of COVID-19: Evidence from China

“…Up till now, the viral RNA of SARS-CoV-2 has been detected in sewage with wastewater in many countries and regions around the globe, and even in surface waters in some places. Some notable examples are listed alphabetically here, but are not limited to: Canada ( D'Aoust et al, 2021 ), Finland ( Hokajrvi et al, 2021 ), Germany ( Agrawal et al, 2021 ; Westhaus et al, 2021 ), Hong Kong ( Cheung et al, 2020 ; Xu et al, 2020b ), Hungary ( Roka et al, 2021 ), India ( Chakraborty et al, 2021 ; Kumar et al, 2021 ), Iran ( Tanhaei et al, 2021 ), Israel ( Bar-Or et al, 2021 ), Italy ( La Rosa et al, 2021 ), Mexico ( Coronado et al, 2021 ), Netherlands ( Medema et al, 2020 ), Pakistan ( Haque et al, 2021 ), Qatar ( Saththasivam et al, 2021 ), Saudi Arabia ( Alahdal et al, 2021 ), Serbia ( Kolarevic et al, 2021 ), Spain ( Randazzo et al, 2020 ), Switzerland ( Fernandez-Cassi et al, 2021 ), the United Kingdom ( Hillary et al, 2021 ; Martin et al, 2020 ), the United States ( Li et al, 2021c ; Sherchan et al, 2021 ; Wang et al, 2020b ), and Venezuela ( Chacin-Bonilla & Chacon, 2021 ).…”

High frequency, high throughput quantification of SARS-CoV-2 RNA in wastewater settled solids at eight publicly owned treatment works in Northern California shows strong association with COVID-19 incidence

“…Even after mixing with wastewater liquid, coronaviruses have a stronger affinity to the solid fraction of wastewater, even higher than the affinities of nonenveloped viruses 13 . Studies comparing SARS-CoV-2 RNA concentrations in the liquid and solid fractions of real wastewater have demonstrated that the solids harbor 3-4 orders of magnitude higher concentrations of SARS-CoV-2 RNA on a per mass basis than liquid influent 3,14 . As a result, the settling of wastewater solids that happens during primary treatment at most wastewater treatment plants serves as a built-in concentration step for wastewater monitoring.…”

Development of a rapid pre-concentration protocol and a magnetic beads-based RNA extraction method for SARS-CoV-2 detection in raw municipal wastewater

“…5 As such, the monitoring of wastewater for SARS-CoV-2 occurrence to investigate the prevalence of COVID-19 infections in a given population is an approach that has been rapidly adopted worldwide as the pandemic continues to spread. [6][7][8][9][10][11][12][13] Reliable tests for SARS-CoV-2 infections target the viral genome through quantitative reverse transcription PCR (RT-qPCR). 14,15 The use of RT-qPCR led to the first report of SARS-CoV-2 RNA detection in untreated wastewater in a proof-of-concept study that demonstrated the applicability of wastewater surveillance for COVID-19 as a tool to understand infection rates within communities.…”