Monitoring and Testing for SARS-CoV-2
(Covid-19 virus) in Sewage wastewater
Ching-Yi Tsai, Ph.D. and Chin S. Yang, Ph.D.
Since the
initial outbreak of Covid-19 pneumonia in China in late 2019, the disease has become
a pandemic. Over the last several months, we have slowly but surely learned more
about the causative agent, SARS-CoV-2. We now know that both symptomatic and
asymptomatic carriers of SARS-CoV-2 shed the virus in their fecal matter, which
is eventually discharged into the sewage wastewater system. Many scientists
have proposed using the so-called wastewater-based epidemiology (WBE) to
monitor the prevalence of infections within a population.
(WBE or wastewater-based
surveillance) is an area of epidemiology that use results or
concentrations of pathogen or biomarker testing in wastewater for the
determination of the likely exposure and disease outbreaks in a population.
This is achieved by measuring biological entities (often biomarkers) in
wastewater generated by the people contributing to a wastewater treatment plant
catchment. Wastewater-based epidemiology have been used to estimate illicit drug (e.g., opioids) use
in communities or populations, but can also be used to measure the consumption
of caffeine, various pharmaceuticals and other compounds. Wastewater-based epidemiology has also been
adapted to measure the load of pathogens such as SARS-CoV-2
in a community. The community can be as large as a small to mid-size cities to
as small as a nursing home or a college dormitory. It differs from SARS-CoV-2 patient testing in that the results reflect
the viral load in a population. Wastewater-based epidemiology is an
interdisciplinary endeavor that draws on input from specialists such as wastewater treatment plant operators, microbiologists and epidemiologists.
In
the current
pandemic, WBE employing RT-PCR was
used in various countries as a complementary method to assess the load of SARS-CoV-2 in populations. Reports of SARS-CoV-2 WBE have been published in studies of
populations within countries such as Canada,
China,
the Netherlands, Singapore, Spain and
the United States, although the studies were
limited to small to mid-size communities. WBE of SARS-CoV-2 potentially offers a very useful
source of information on the prevalence and temporal trends of COVID-19 in
communities and a potent tool to combat SARS-CoV-2
spread within a community.In
a couple practical uses, we have found SARS-CoV-2
WBE can be a useful tool for Universities (dormitories) to determine whether to
close the dormitory entirely, to perform COVID-19 testing for all residents or
both and for all communities that have COVID-19 carriers that are asymptomatic.
It is also important to point out at present time that there is insufficient
evidence to indicate whether SARS-CoV-2 in wastewater is infectious or not.
Planning to sample and test SARS-CoV-2 in wastewater
To be
able to use the test results, it is important to be able to identify individual
wastewater and sewage collection system, its distribution and access points for
sample collection in a community, whether it is a nursing home, a school, or a
University dormitory.
Because
generation rates in the influent wastewater can fluctuate during the day, it is
recommended that time composite samples are collected on a daily basis. A
24-hour flow proportional sampling is preferred (EPA, 2017). There are several
factors to consider when planning to sample.
1.
Communications: you may have to
work closely with facility managers or building engineers to prepare and
identify suitable access points for setting up sampling equipment and for
collecting samples. It is easier if you prepare in advance.
2.
Sample collection equipment and
disposables: there are wastewater samplers available commercially. Contact the
manufacturer or distributor for information (or perform an on-line search).
Obtain your disposables, such as tubing, collection bottles, gloves, from your
suppliers.
3.
Collection frequency: decide your
collection frequency, whether it is once weekly or daily. It is always better to
have more data points but more sampling means higher costs.
4.
Days (weekdays v. weekend;
holidays v. workdays) of collection; decide your collection day(s). It may be
more meaning to collect samples during weekdays, e.g., office buildings. One
the other hand, there is little difference whether samples are collected during
weekdays or weekend, e.g., nursing homes or dormitories.
5.
Sample collection and handling:
all sample collection devices must be clean and sterile before use. Disposables
are used only once to avoid cross contamination. Samples, once collected, must
be kept at approximately 4°C. SARS-CoV-2
is stable at 4°C and is sensitive to warmer
temperature. Samples must be properly labeled and documented on a
chain-of-custody.
6.
Sample storage and shipping: all
samples must be stored at or close to 4°C and shipped with ice packs. No ice cubes or dry ice. Ice packs and coolers
must be washed, cleaned and disinfected between uses. 70% rubbing alcohol or
diluted bleach solutions can be used for the disinfection. Deliver your samples
to the lab ASAP, either in person or by curriers or overnight shipping.
7.
Biosafety: individuals who
collect and handle the samples must observed biosafety. Prepare and use PPE as
well as disinfectants or sanitizers before, during and after sampling and
handling of the samples.
Sampling
in each building or facility is different. If you have any questions concerning
SARS-CoV-2 sampling in general or need to discuss SARS-CoV-2 sampling and
testing whether indoors or in wastewater systems, please feel free to contact
us at 856-767-8300 or by email chingyi.tsai@prestige-em.com or
Theresa.lehman@prestige-em.com.
References:
Warish Ahmed, Nicola Angel, Janette Edson, Kyle Bibby, Aaron
Bivins, Jake W. O'Brien, Phil M. Choi, Masaaki Kitajima, Stuart L. Simpson, Jiaying Li, Ben Tscharke, Rory Verhagen, Wendy J.M. Smith, Julian Zaugg,
Leanne Dierens, Philip Hugenholtz,
Kevin V. Thomas, Jochen F. Mueller. 2020. First confirmed detection of
SARS-CoV-2 in untreated wastewater in Australia: A proof of concept for the
wastewater surveillance of COVID-19 in the community. Science of the
Total Environment. https://doi.org/10.1016/j.scitotenv.2020.138764
Aaron
Bivins, Justin Greaves, Robert Fischer, Kwe Claude Yinda,
Warish Ahmed, Masaaki Kitajima, Vincent J. Munster,
and Kyle Bibby. 2020. Persistence of SARS-CoV‑2 in Water and Wastewater. Environ. Sci. Technol.
Lett. https://dx.doi.org/10.1021/acs.estlett.0c00730
EPA, 2017.
Wastewater Sampling. Number: SESDPROC-306-R4 (February 13, 2017).
Paola Foladori, Francesca Cutrupi,
Nicola Segata, Serena Manara,
Federica Pinto, FrancescaMalpei, Laura Bruni, Giuseppina La Rosa. 2020. SARS-CoV-2 from faeces to wastewater treatment: What do we know? A review. Science of the Total Environment 743 (2020) 140444.
https://doi.org/10.1016/j.scitotenv.2020.140444
Jordan Peccia, Alessandro Zulli, Doug E.
Brackney, Nathan D. Grubaugh,
Edward H. Kaplan, Arnau Casanovas-Massana , Albert I. Ko, Amyn A. Malik,
Dennis
Wang, Mike Wang, Joshua L. Warren, Daniel M. Weinberger, Wyatt Arnold, and Saad
B. Omer. 2020. Measurement of SARS-CoV-2 RNA in wastewater tracks community
infection dynamics. Nature Biotechnology, Vol. 38:
1164–1167. https://doi.org/10.1038/s41576-020-0684-z.
Walter Randazzo, Pilar Truchado, Enric Cuevas-Ferrando, Pedro Simon, Ana Allende, Gloria Sanchez. 2020. SARS-CoV-2 RNA in wastewater anticipated COVID-19
occurrence in a low prevalence area. Water Research 181: 1-8. https://doi.org/10.1016/j.watres.2020.115942
Samendra P. Sherchan,
Shalina Shahin, Lauren M.Ward, Sarmila Tandukar, Tiong G. Aw, Bradley Schmitz , Warish Ahmed, Masaaki Kitajima. 2020. First detection of SARS-CoV-2 RNA in
wastewater in North America: A study in Louisiana, USA. Science
of the Total Environment. 743:1-6. https://doi.org/10.1016/j.scitotenv.2020.140621
Sims, N., B. Kasprzyk-Hordern. 2020. Future perspectives of wastewater-based
epidemiology: Monitoring infectious
disease spread and resistance to the community level. Environment
International 139 (2020) 105689. https://doi.org/10.1016/j.envint.2020.105689
Wu FQ;
Xiao A; Zhang JB; Gu XQ; Lee WL; Kauffman K ; Hanage
WP; Matus M ; Ghaeli N;
Endo N; Duvallet C; Moniz K; Erickson TB; Chai PR;
Thompson J; Alm EJ. 2020. SARS-CoV-2 titers in wastewater are higher than expected from clinically
confirmed cases. medRxiv preprint doi: https://doi.org/10.1101/2020.04.05.20051540
Date: October 27,
2020