key: cord-328686-5ik5em5a
authors: Zhao, L.; Atoni, E.; Du, Y.; Zhang, H.; Donde, O.; Huang, D.; Xiao, S.; Ma, T.; Shu, Z.; Yuan, Z.; Tong, L.; Xia, H.
title: First study on surveillance of SARS-CoV-2 RNA in wastewater systems and related environments in Wuhan: Post-lockdown
date: 2020-08-21
journal: nan
DOI: 10.1101/2020.08.19.20172924
sha: 
doc_id: 328686
cord_uid: 5ik5em5a

Wastewater-based epidemiology (WBE) has emerged as an effective environmental surveillance tool in monitoring fecal-oral pathogen infections within a community. Congruently, SARS-CoV-2 virus, the etiologic agent of COVID-19, has been demonstrated to infect the gastrointestinal tissues, and be shed in feces. In the present study, SARS-CoV-2 RNA was concentrated from wastewater, sludge, surface water, ground water, and soil samples of municipal and hospital wastewater systems and related environment in Wuhan during the COVID-19 middle and low risk periods, and the viral RNA copies quantified using RT-qPCR. From the findings of this study, during the middle risk period, one influent sample and three secondary treatment effluents collected from Waste Water Treatment Plant 2 (WWTP2), as well as two influent samples from wastewater system of Hospital 2 were SARS-CoV-2 RNA positive. One sludge sample collected from Hospital 4; which was obtained during low risk period, was positive for SARS-CoV-2 RNA. These study findings demonstrate the significance of WBE in continuous surveilling and monitoring of SARS-CoV-2 at the community level, even when the COVID19 prevalence is low. Therefore, the application of WBE is principally useful in tracking the level of infections in communities and the risk assessment of the secondary environment.

of this study, during the middle risk period, one influent sample and three secondary treatment effluents 23 collected from Waste Water Treatment Plant 2 (WWTP2), as well as two influent samples from wastewater 24 system of Hospital 2 were SARS-CoV-2 RNA positive. One sludge sample collected from Hospital 4; which was 25 obtained during low risk period, was positive for SARS-CoV-2 RNA. These study findings demonstrate the 26 significance of WBE in continuous surveilling and monitoring of SARS-CoV-2 at the community level, even 27 when the COVID19 prevalence is low. Therefore, the application of WBE is principally useful in tracking the 28 level of infections in communities and the risk assessment of the secondary environment. 29 30 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted August 21, 2020. . https://doi.org/10.1101/2020.08.19.20172924 doi: medRxiv preprint Letter/Short report 31 Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the causative agent for coronavirus disease 32 COVID-19, a current public health crisis of global concern (Zhou et al. 2020). COVID-19 was declared a 33 pandemic by World Health Organization (WHO) on 11 th March 2020 (Cucinotta and Vanelli 2020), after its first 34 report in Wuhan to other cities in China and thereafter spread to many other countries. COVID-19 is primarily 35 transmitted via respiratory droplets that people cough, sneeze or exhale, and may also be spread via fomites 36 Shanghai. In their study findings, SARS-CoV-2 RNA positive samples were detected from wastewater 57 discharges of the designate hospitals (http://www.water8848.com/news/202003/30/123415.html). 58

Due to the outbreak of SARS-CoV-2 in Wuhan, stringent quarantine measures were enforced throughout the 59 city on January 23, 2020 by the local Hubei government. After 11 weeks of lockdown, traffic control measures 60 begun to be officially lifted on April 8, and life started getting back to normal. As of 7 th April, there were 574 61 confirmed cases and 673 cases under medical surveillance in Wuhan (http://wjw.wuhan.gov.cn/sy/). Indeed, 62 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted August 21, 2020. . https://doi.org/10.1101/2020.08.19.20172924 doi: medRxiv preprint Wuhan, and it was lowered from high to medium risk on 25 th March 2020, and thereafter lowered it from 64 medium to low risk on 8 th April. Currently there is no published report on tracking of SARS-CoV-2 in 65 wastewater and related environment in Wuhan during the epidemic or after the epidemic. 66

In the present study, a total of 216 samples that covered middle and low risk periods were collected from 67 various points including wastewater treatment plants, designated hospitals for COVID-19, lake and river that 68 are close to the COVID-19 designated hospitals, between the months of April and May, 2020. Except for the 69 ground water samples, the collection sites were mainly distributed in two districts; with the first one being CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted August 21, 2020. . https://doi.org/10.1101/2020.08.19.20172924 doi: medRxiv preprint From the findings of this study, during the middle risk period, positive samples were detected both in 83 municipal and hospital wastewater systems. One of the three influent samples (7.4 ×10 3 copies/L), three of 84 six secondary treatment effluents (5.3 × 10 3 and 1.0 ×10 4 copies/L) of WWTP2, and two samples from CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted August 21, 2020. . https://doi.org/10.1101/2020.08.19.20172924 doi: medRxiv preprint Although SARS-CoV-2 RNA surveillance in wastewaters is a useful WBE drive, the public health risk associated 109 with water cycle is unclear since viral particles infectivity in sewage and faeces is yet to be determined in 110 addition to its probable fecal-oral transmission. Indeed, a recently conducted study has inferred that risk of 111 infection from wastewater and river is insignificant due to the low success rate in cell culture of SARS-CoV-2 112 from water samples in spite of the high RNA copies (Rimoldi et al. 2020) . is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted August 21, 2020. . https://doi.org/10.1101/2020.08.19.20172924 doi: medRxiv preprint surveillance and detection of SARS-CoV-2 was a prime priority due to the peak in COVID-19 infection in 127

Wuhan. Moreover, there was no regulation that guided WWTP on periodic collection and storage of water 128 samples over a period of time. 129

In conclusion, the detection of SARS-CoV-2 RNA in various wastewater systems and related environmental 130 samples in Wuhan shows the significance of WBE in continuous surveilling and monitoring SARS-CoV-2 at the 131 community level, under low prevalence record of human illnesses, in contrast to clinical surveillance. This 132 application is principally useful in remote communities and confined populations where mass sampling for 133 the entire population may not be easily achievable at the onset due to inadequate resources or occurrence of 134 asymptomatic patients, though effective sampling techniques is of great essence for achieving accurate 135 results. 136

First Confirmed Detection of SARS-CoV-2 in Untreated Wastewater in Australia: A 145 Proof of Concept for the Wastewater Surveillance of COVID-19 in the Community

Monitoring Human Enteric Viruses in Wastewater and Relevance to Infections 148

Encountered in the Clinical Setting: A One-Year Experiment in Central France

Sentinel Surveillance of SARS-CoV-2 in Wastewater Anticipates the 151 Occurrence of COVID-19 Cases

Wastewater-Based Epidemiology Biomarkers: Past, Present and Future

WHO Declares COVID-19 a Pandemic

Presence of SARS-Coronavirus-2 in Sewage

Presence and Vitality of SARS-CoV-2 Virus in Wastewaters and Rivers

First Detection of SARS-CoV-2 in Untreated Wastewaters in Italy

Through Monitoring of Sewer Systems

COVID-19 May Transmit through Aerosol

A New Coronavirus Associated with Human Respiratory Disease in China

SARS-CoV-2 Titers in Wastewater Are Higher than Expected from Clinically Confirmed 194 Cases

Evaluation of Lockdown Impact on SARS-CoV-2 Dynamics through Viral Genome 197 Quantification in Paris Wastewaters

Wastewater-Based Epidemiology for Early Detection of Viral 200 Outbreaks

Molecular and Serological Investigation of 2019-NCoV Infected Patients: Implication of

We sincerely thank Prof. Hongping Wei and the entire team at the National Biosafety Laboratory in Wuhan, 138China for the support they extended to us. This work was supported by the Wuhan Bureau of Science and 139Technology (202002020101010022). 140

The authors declare no conflict of interest. 142 143 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review)The copyright holder for this preprint this version posted August 21, 2020. 173 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted August 21, 2020. 

. CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted August 21, 2020. 207 208 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review)The copyright holder for this preprint this version posted August 21, 2020. . https://doi.org/10.1101/2020.08.19.20172924 doi: medRxiv preprint