v>EPA
A Compend urn of U.S. Wastewater
Surveillance to Support COVID-19
Public Health Response
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
This document was prepared by the United States Environmental Protection Agency (EPA). Neither the
United States government nor any of its employees, contractors, subcontractors, or their employees
make any warrant, expressed or implied, or assume any legal liability or responsibility for any third
party's use of or the results of such use of any information, apparatus, product, or process discussed in
this report, or represent that its use by such party would not infringe on privately owned rights. This
report does not establish guidance or opinions on the best way to establish wastewater surveillance
programs; rather, it catalogues practices by a broad array of organizations and individuals. The objective
is to summarize the information available on conducting these wastewater monitoring programs in the
United States. EPA may update this document in the future.
Questions about this document should be directed to:
U.S. EPA Office of Wastewater Management
1200 Pennsylvania Avenue NW (4201 M)
Washington, D.C. 20460
(202) 564-0748
wastewatertechnologyclearinghouse@epa.gov
U.S. EPA Office of Research and Development
Center for Environmental Solutions and Emergency Response
26 West Martin Luther King Dr. Mail Code 236
Cincinnati, OH 45268
(513) 569-7900
Gutierrez.Sally@epa.gov
Prepared by:
¦ Sally Gutierrez, EPA Office of Research and Development
¦ Kathryn Kazior, EPA Office of Wastewater Management
¦ Smiti Nepal, EPA Office of Wastewater Management
Cover Photos:
¦ Top Left: Katie Watkins, Houston Public Media. The Houston Health Department collects a
wastewater sample as part of the city's wastewater surveillance program supporting COVID-19
public health efforts (Watkins, 2021).
¦ Bottom Left: Ben Siegel, Ohio University. Engineering students prepare to collect a wastewater
sample for SARS-CoV-2 monitoring to support Ohio University's COVID-19 public health efforts
(OHIO News, 2021).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Acknowledgements
EPA Office of Wastewater Management (OWM) and Office of Research and Development (ORD)
would like to acknowledge the efforts of the following people in reviewing and providing feedback:
¦ Leonardo Angelone, National Institutes of Health
¦ Tracy Bone, EPA Office of Science and Technology
¦ Nicole Brinkman, EPA ORD
¦ Jay Garland, EPA ORD
¦ Amy Kirby, United States Centers for Disease Control and Prevention (CDC)
¦ Lisa McFadden, Water Environment Federation
¦ Jeffrey Mercante, CDC
¦ Kevin Oshima, EPA ORD
¦ Jorge SantoDomingo, EPA ORD
EPA would also like to acknowledge the participation of practitioners from the case study wastewater
surveillance programs included in this report who provided information, participated in discussions, and
reviewed their case study write-ups, was valuable and much appreciated. Contributions of the
practitioners are recognized below:
Wastewater Surveillance
Program
Representative
Indiana
¦ Jim McGoff, Indiana Finance Authority
¦ Erica Walker, 120Water
¦ Kyle Bibby and Alex Perkins, University of Notre Dame
Michigan
¦ Rhiannon Bednar, Robert Orellana, Susan Peters, and Mary Grace
Stobierski, Michigan Department of Health and Human Services
¦ Shannon Briggs, Michigan Department of Environment, Great Lakes,
and Energy
¦ Kevin Bakker and Chuanwu Xi, University of Michigan
¦ Erin Dreelin and Joan Rose, Michigan State University
New Mexico
¦ Justin Garoutte, John Rhoderick, and Rebecca Roose, New Mexico
Environment Department
Ohio
¦ Zuzana Bohrerova, Ohio Water Resources Center
Wyoming
¦ Ali Harrist, Franz Fuchs, Stefan Johansson, and Stephanie Pyle,
Wyoming Department of Health
¦ Wanda Manley and Cari Sloma, Wyoming Public Health Laboratory
¦ Bledar Bisha and Sarah Collins, University of Wyoming
Hampton Roads Sanitation
District (Virginia)
¦ Raul Gonzales and Jim Pletl, Hampton Roads Sanitation District
Houston, Texas
¦ Loren Hopkins, Houston Health Department
¦ Lauren Stadler, Rice University
Tempe, Arizona
¦ Wydale Holmes and Rosa Inchuasta, City of Tempe
Clemson University
¦ David Freedman, Clemson University
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
III
Wastewater Surveillance
Program
Representative
University of Arizona
¦ Ian Pepper, University of Arizona
In addition, a formal technical review of the draft document was conducted by professionals with
experience in wastewater surveillance in accordance with EPA Peer Review Guidelines. While every
effort was made to accommodate all the Peer Review comments, the results and conclusions do not
indicate consensus and may not represent the views of all the reviewers. The technical reviewers of this
document included the following:
¦ Jerome Oliver, EPA OWM
¦ Jason Turgeon, EPA Region I
¦ Christobel Ferguson, Water Research Foundation (WRF)
¦ Jonathan Yoder, CDC
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Contents
Acknowledgements ii
Abbreviations viii
Executive Summary I
1 Introduction 3
2 Purpose 5
3 Report Development Approach 6
4 Financial Support 8
4.1 National Science Foundation 9
4.2 Centers for Disease Control and Prevention I 3
4.3 National Institutes of Health 14
4.4 U.S. Department of Health and Human Services 18
4.5 Water Research Foundation 18
4.6 Water Environment Federation 19
4.7 Other Funding Opportunities 19
5 Program Development 21
5.1 Peer-to-peer Communication and Resource Sharing 21
5.1.1 Workshops and Trainings 21
5.1.2 Online Platforms 23
5.2 Researching and Developing Analytical Methods 26
5.3 Ongoing Wastewater Surveillance Support 28
5.4 Inclusion of Rural and Underserved Populations 30
5.5 Ethical and Legal Considerations 31
5.6 Worker Safety 32
6 Implementation of Surveillance Programs 33
6.1 Overview of Surveillance Programs 33
6.2 Wastewater Surveillance Case Studies 42
6.2.1 Indiana 43
6.2.2 Michigan 47
6.2.3 New Mexico 48
6.2.4 Ohio 52
6.2.5 Wyoming 56
6.2.6 Hampton Roads Sanitation District in Hampton Roads, Virginia 59
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6.2.7 Houston, Texas 62
6.2.8 Tempe, Arizona 65
6.2.9 Clemson University in Clemson, South Carolina 68
6.2.10 University of Arizona in Tucson, Arizona 71
7 Wastewater Surveillance Lessons Learned 73
8 References 75
Appendix A. Summary of Wastewater Surveillance Programs
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vi
List of Tables
Table I. List of search terms used to identify wastewater surveillance programs 6
Table 2. NSF-funded research for wastewater surveillance 10
Table 3. NIH wastewater surveillance projects funded under RADx-rad 16
Table 4. Research opportunities identified by WRF to support wastewater surveillance of SARS-
CoV-2 (WRF, 2020j) 22
Table 5. Summary of case study wastewater surveillance programs 42
Table 6. University of Arizona's levels of concern and associated actions for ranges of SARS-
CoV-2 wastewater concentrations 72
List of Figures
Figure I. Jurisdictions using CDC funds to support wastewater surveillance for SARS-CoV-2 as
of April 2021 (WRF, 2021a) 14
Figure 2. COVIDPoopsl9 dashboard (UC Merced, 2021) 24
Figure 3. Utilities performing wastewater surveillance in the United States (WEF, 2021 b) 25
Figure 4. CDC's NWSS DCIPHER analytics dashboard for SARS-CoV-2 wastewater results
(WRF, 2021a) 29
Figure 5. Cumulative samples in the NWSS DCIPHER system (WRF, 2021a) 29
Figure 6. Boise Wastewater Surveillance Dashboard with wastewater SARS-CoV-2 virus copies
per liter on the top graph and confirmed and probable COVID-19 cases on the
bottom graph (Boise, 2021) 36
Figure 7. Utah SARS-CoV-Wastewater Surveillance Dashboard with the service are and recent
wastewater trend on the left, SARS-CoV-2 million gene copies per person, per day
on the top right graph, and daily new cases per 100,000 residents on the bottom
right graph (Utah DEQ, 2021) 37
Figure 8. Missouri Department of Health and Human Services Wastewater Surveillance
Dashboard with color coded symbols to indicate recent SARS-CoV-2 wastewater
trends (Missouri DHSS, 2021) 38
Figure 9. Yale University Wastewater Surveillance Dashboard - Hartford South Meadows
WWTP serving Hartford, West Hartford, Newington, Bloomfield, and
Wethersfield with SARS-CoV-2 wastewater results in copies per militer on the top
graph and daily COVID-19 cases per 100,000 residents on the bottom graph (Yale
University, 2021 a) 40
Figure 10. Athens-Clark County Wastewater Surveillance Dashboard for one of the WWTPs
(Lottetal., 2020) 41
Figure I I. Example plots provided to the utilities including viral gene copies per 100 milliters
(mL) for recent samples, concentrations over time as compared to the other
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
utilities, weekly COVID-19 cases, and mobility metrics from Google and Apple
(I FA, 2020) 45
Figure 12. Plots of viral gene copies (GC) per mL in Indiana communities (Indiana, 2020) 46
Figure 13. Michigan Wastewater Surveillance Program projects (green diamonds) and sampling
locations (blue circles) (Michigan EGLE, 2021) 47
Figure 14. Wastewater surveillance data for the J. Paul Taylor Center (NMED, 2021a) 50
Figure 15. Wastewater surveillance data for Luna County Detention Center (east side) (NMED,
2021a) 51
Figure 16. Ohio Coronavirus Wastewater Monitoring Network Dashboard showing all the
participating utilities on the map and a list of utilities in order of the trend based on
the most recent results (as of April 19, 2021) (Ohio DOH, 2020) 54
Figure 17. Ohio Coronavirus Wastewater Monitoring Network Dashboard showing city-specific
results normalized by WWTP influent flow rate (top graph) and compared to the
number of COVID-19 cases from individual testing (bottom graph) (Ohio DOH,
2020 ) 55
Figure 18. Wyoming State SARS-CoV-2 Wastewater Surveillance Dashboard prevalence ranges
(Wyoming PHL, 2021) 57
Figure 19. Wyoming State SARS-CoV-2 Wastewater Surveillance Dashboard (Wyoming PHL,
2021 ) 58
Figure 20. HRSD's dashboard presents the SARS-CoV-2 wastewater concentration and the new
individual COVID-19 cases overtime (HRSD, 2021a) 60
Figure 21. HRSD's dashboard presents the SARS-CoV-2 wastewater concentration spatially
throughout the collection system overtime (HRSD, 2021a) 61
Figure 22. Areas of Houston covered by the wastewater surveillance program (Houston, 2021) 63
Figure 23. Percent change in wastewater results from July 6, 2020 (Peak) for areas of Houston
covered by the wastewater surveillance program (Houston, 2021) 64
Figure 24. The City of Tempe's wastewater dashboard for SARS-CoV-2 with a map of the areas
sampled (Tempe, 2021a) 67
Figure 25. The Clemson University COVID-19 Wastewater Dashboard color codes the impact
level on a map of each WWTP's collection system area (Clemson, 2021a) 69
Figure 26. The Clemson University COVID-19 Wastewater Dashboard demonstrates the impact
level and virus copies/L in each WWTP influent sampling point over time. The
dashboard also includes variant tracking results (Clemson, 2021a) 70
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VIII
Abbreviations
APHL Association of Public Health Laboratories
ASTHO Association of State and Territorial Health Officials
ASU Arizona State University
CARES Act 2020 Coronavirus Aid, Relief, and Economic Security Act
CDC Centers for Disease Control and Prevention
cDNA complementary deoxyribonucleic acid
COVID-19 coronavirus disease 2019
CT cycle threshold
DCIPHER Data Collation and Integration for Public Health Event Response
DHS United States Department of Homeland Security
EGLE Michigan Department of Environment, Great Lakes, and Energy
ELC Epidemiology and Laboratory Capacity for Prevention and Control of Emerging
Infectious Disease (CDC funding)
EPA United States Environmental Protection Agency
HHD Houston Health Department
HHS United States Department of Health and Human Services
HRSD Hampton Roads Sanitation District
IFA Indiana Finance Authority
L liter
MDHHS Michigan Department of Health and Human Services
mL milliliter
N nucleocapsid gene
NACCHO National Association of County and City Health Officials
NACWA National Association of Clean Water Agencies
NEHA National Environmental Health Association
NIH United States National Institutes of Health
NIST United States National Institute of Standards and Technology
NSF United States National Science Foundation
NSSIL National Sewage Surveillance Interagency Leadership
NWSS National Wastewater Surveillance System
ODH Ohio Department of Health
Ohio EPA Ohio Environmental Protection Agency
Ohio WRC Ohio Water Resources Center
ORD Office of Research and Development (within EPA)
PCR polymerase chain reaction
RADxSM Rapid Acceleration of Diagnostics (NIH initiative)
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
RADx-rad
RADxSM Radical (NIH initiative)
RAPID
Rapid Response Research (NSF funding mechanism)
RNA
ribonucleic acid
RT-ddPCR
reverse transcription digital droplet PCR
RT-PCR
reverse transcription PCR
RT-qPCR
reverse transcription-quantitative PCR
SARS-CoV-2
severe acute respiratory syndrome coronavirus 2
WBE
wastewater-based epidemiology
WDOH
Wyoming Department of Health
WEF
Water Environment Federation
WEST
Water and Energy Sustainable Technology (at the University of Arizona)
WHO
World Health Organization
WPHL
Wyoming Public Health Laboratory
WRF
Water Research Foundation
WWTP
wastewater treatment plant
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Executive Summary
Wastewater surveillance is a community-level approach for monitoring disease or chemical biomarkers
that are excreted in human urine and feces and collected in sewers. Since early 2020, with the start of
the coronavirus disease 2019 (COVID-19) pandemic, scientists and public health practitioners across the
globe have been developing methods and implementing programs to track severe acute respiratory
syndrome coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19, in wastewater. Even though
SARS-CoV-2 is a respiratory virus, wastewater surveillance can be used to track its spread since it can
be shed in the feces of individuals who are symptomatic and asymptomatic (including pre-symptomatic).
Monitoring SARS-CoV-2 levels in untreated wastewater relies on approaches and technologies that have
been and continue to be rapidly deployed and evaluated by federal agencies, non-governmental
organizations, states, wastewater utilities, universities, and industry. Despite the rapidly evolving science
in this field, these entities were able to establish wastewater surveillance programs while developing
sampling and analytical methods. The results of these programs provide useful information to assist
communities in their public health response to the COVID-19 pandemic—highlighting the potential for
wastewater monitoring to serve as a complementary approach to current and future infectious disease
surveillance systems.
The United States Environmental Protection Agency (EPA) created this document to provide
information to those who are interested in implementing wastewater surveillance programs to monitor
SARS-CoV-2 or other pathogenic disease agents and chemical exposures in the future. To support that
goal, this compendium documents the efforts of federal, state, local, and tribal agencies—as well as
associations, universities, and the private sector—throughout 2020 and into early 2021 to explore
federal and other funding sources, develop and implement wastewater surveillance for SARS-CoV-2, and
provide information on how programs were implemented through case studies.
The report describes funding mechanisms from federal agencies, non-governmental organizations,
private foundations, or funding by other means, such as reprogramming of existing funding, that was
used to establish wastewater surveillance programs. The report also discusses how stakeholder
collaboration through workshops, trainings, and other mechanisms allowed the research community to
build upon established protocols for waterborne enteric viruses to advance wastewater surveillance for
SARS-CoV-2. This collaboration was also critical for developing analytical methods for SARS-CoV-2 in
wastewater, refining sampling collection procedures, and interpreting the wastewater testing results.
Tied to the last point, this report documents how some programs communicated the results through
online dashboards and then translated those results into public health responses.
Through its research conducted in early 2021, EPA identified 14 states with large-scale SARS-CoV-2
wastewater surveillance programs, along with 160 local communities or academic institutions conducting
wastewater surveillance. While these groups developed and relied on different methods, their efforts
were ultimately successful in detecting SARS-CoV-2 and initiating action to prevent continued spread of
COVID-19. From these programs, EPA selected 10 case studies that highlight different approaches for
implementing programs and analyzing wastewater data to track the presence of SARS-CoV-2. These
examples can inform how to establish and implement wastewater surveillance throughout the COVID-
19 pandemic or for future pandemics and public health crises. For example, the case study programs
highlighted that much of their success was dependent on effective collaboration with multiple partners,
flexibility to adapt their programs as the science evolved, support from within their organization,
transparent communication with stakeholders on how wastewater testing results could be used to help
their communities respond to the pandemic, and adequate funding. These programs highlighted how
wastewater surveillance can be an important and effective tool for early detection of SARS-CoV-2,
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
especially among disadvantaged or vulnerable populations where clinical testing may not be widely
available.
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
3
Introduction
Wastewater surveillance is a community-level approach for monitoring chemical metabolites, bacteria,
and viral pathogens that are excreted in human urine and feces and collected in sewers. It has been
successfully used to detect various agents of diseases (e.g., poliovirus, hepatitis B, norovirus) in
populations for decades and more recently to understand community-level drug use (e.g., opioids). A
major advantage of wastewater-based methods is that they are not subject to the same reporting and
recall biases that can occur when epidemiologic data are collected from individual community members
or health care providers. For example, wastewater samples can be used to identify the true spectrum of
drugs being consumed by a population rather than relying on individual self-reported information. In
addition, wastewater-based methods can produce near real-time data that represent an entire
community or smaller subsets of a community (e.g., at the sub-county level, at the individual facility or
building level).
Beginning in early 2020, wastewater surveillance received renewed attention in light of the COVID-19
pandemic. Because individuals with symptomatic or asymptomatic infection can shed the SARS-CoV-2
virus in their feces, quantitative measures of SARS-CoV-2 in wastewater can provide useful information
on changes in total COVID-19 infection in the community contributing to the wastewater (CDC,
2021a). Scientists and public health practitioners across the globe therefore quickly mobilized to develop
methods and programs to track SARS-CoV-2 in wastewater. For example, researchers in the
Netherlands began testing sewage for SARS-CoV-2 from six cities and the Schiphol airport in February
2020, before the Netherlands reported their first COVID-19 case. A month after the wastewater
program began, the Netherlands detected low levels of the virus in sewage from several sites.
Subsequent increases in sewage viral concentrations correlated with increases in reported COVID-19
prevalence in the community (Medema et al„ 2020). Italian researchers also began testing for SARS-
CoV-2 in wastewater in areas of high (e.g., Milan) and low (e.g., Rome) COVID-19 prevalence in
February 2020, with first detections of SARS-CoV-2 in wastewater observed in late February (La Rosa et
al„ 2020). Studies conducted in the United States in March and April 2020 had similar success detecting
the presence of SARS-CoV-2 in wastewater (e.g., Massachusetts) (Wu et al„ 2020). Early efforts such as
these highlighted the potential for sewage surveillance to serve as a complementary measure for
monitoring COVID-19 spread in a community.
surveillance can serve as an early warning system of
increased COVID-19 spread. These programs can also provide data at the smaller sewershed or facility
level and in communities where timely COVID-19 individual testing is underutilized or unavailable.
There are numerous documented instances where SARS-CoV-2 wastewater surveillance data have been
Federal, state, public health, and environment
departments throughout the United States, as well
as academic institutions, tribes, utilities, and others
have developed wastewater monitoring programs
for SARS-CoV-2 to monitor trends within a
sewershed or at a targeted site (e.g., a facility or
building). According to the Centers for Disease
Control and Prevention (CDC), the virus has in
some cases been detected in wastewater prior to
reported cases in the community and trends in virus
concentrations in wastewater have preceded trends
in newly reported cases by multiple days (CDC,
2021 a). These findings suggest that wastewater
Wastewater Analysis for SARS-CoV-2
Multiple testing methods are used to quantify
SARS-CoV-2 in wastewater. These laboratory
tests typically quantify ribonucleic acid (RNA)
—or the genetic signature of SARS-CoV-2 in
wastewater. This means that wastewater
surveillance programs for SARS-CoV-2 are not
measuring the infectious virus directly, but
instead are measuring viral RNA as an indicator
of virus concentrations. Note that where this
report refers to measurements of the virus in
wastewater, the SARS-CoV-2 genetic material
is what is being measured.
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
used to prioritize individual testing resources, inform other community mitigation strategies, and
monitor effectiveness of interventions over the past year. For example, a state or local public health
department may increase individual testing or ramp up public health messaging and outreach following a
rise in SARS-CoV-2 measurements in wastewater.
For most community-level wastewater surveillance programs, samples are typically collected at the
influent of a wastewater treatment plant (WWTP). Other smaller scale programs sample at locations
throughout the collection system, including at lift stations or from manholes that carry sewage from
individual buildings. Wastewater samples are then analyzed for the presence of SARS-CoV-2 using a
nucleic acid-based reverse transcription polymerase chain reaction (RT-PCR) assay for gene markers
that are unique to the virus. Results provide insight on COVID-19 among the population served by the
utility's collection system and/or subsections within a community.
It is important to note that wastewater surveillance for SARS-CoV-2 is a developing field and
researchers are still learning about the dynamics of viral shedding in feces and viral persistence in
wastewater. More data on the prevalence and concentrations of SARS-CoV-2 shed in the feces of
infected individuals are needed to better understand the relationship between SARS-CoV-2
concentrations in wastewater and the number of individuals infected with COVID-19. Furthermore, low
levels of infection in a community may not be captured by sewage surveillance (CDC, 2021a). Other
complexities that arise when interpreting wastewater data include the mobility of the population
contributing to the wastewater, industrial wastewater contribution, stormwater, other factors (e.g.,
cleaning, dilution), and variability in wastewater flow and fecal load. It is also important to note that
community-level monitoring at a WWTP does not capture residences and businesses on a septic-based
system or facilities that have their own system (e.g., correctional facilities); however, some separate
wastewater surveillance efforts are also monitoring community septic tanks.
Because of these limitations, SARS-CoV-2 wastewater data are often considered in tandem with other
COVID-19-related data to inform public health. Data from wastewater surveillance programs are not
meant to replace other COVID-19 surveillance systems, but rather to offer complementary data.
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5
2 Purpose
EPA compiled this report to capture some of the notable and unique efforts to develop and use
wastewater surveillance to detect and monitor the SARS-CoV-2 virus genetic material in untreated
wastewater throughout 2020 and into early 2021. As a result, this report does not include
advancements in wastewater surveillance that may have occurred after February 2021. The report
includes details on developing and establishing wastewater monitoring programs throughout the United
States and summarizes details such as funding mechanisms, sampling approaches used, sample analysis
methods development, data interpretation, and public health responses. The report also describes the
robust collaboration between organizations to share knowledge and optimize the wastewater
surveillance process that occurred during this period.
Through discussions with some key programs, EPA compiled lessons learned for the development and
implementation of wastewater surveillance programs, in the face of rapidly evolving science and a global
pandemic. These examples and the many success stories can inform wastewater surveillance efforts for
future pandemics and public health crises.
This report documents SARS-CoV-2 wastewater surveillance efforts throughout the country by federal
agencies, non-governmental stakeholders, state agencies, tribal agencies, local agencies, utilities,
academic institutions, and private entities. The report begins by describing how EPA gathered
information (Section 3) and then summarizes funding mechanisms that have provided financial support to
researchers and other groups implementing wastewater surveillance (Section 4). Next, the document
provides examples for how entities worked collaboratively to advance wastewater surveillance practices
(Section 5.1), develop and research analytical methods (Section 5.2), provide ongoing support for
wastewater surveillance strategies (Section 5.3), serve rural and underserved populations (Section 5.4),
consider ethical and legal concerns (Section 5.5), and develop resources to protect wastewater utility
workers (Section 5.6). The report also provides an overview of some of the SARS-CoV-2 wastewater
surveillance programs that were established by various entities (Section 6.1) and presents 10 case
studies, which offer unique examples of program leadership, collaboration, funding, sampling design, data
presentation, and data interpretation (Section 6.2), with a summary of lessons learned from wastewater
surveillance programs (Section 7). Appendix A presents a summary of SARS-CoV-2 wastewater
surveillance programs, based on a review of publicly available information in January 2021.
This report is not intended to serve as a comprehensive summary of all wastewater surveillance efforts
for SARS-CoV-2 in the U.S. or as a guidance/framework document. Rather, it documents and provides
examples and perspective on the wide variety of activities conducted by diverse entities to develop and
implement ways to use wastewater surveillance to complement existing public health measures for the
COVID-19 pandemic.
It is important to note that this report and EPA do not endorse or make any judgement or provide
guidance on a specific model for wastewater surveillance programs. This report is intended to provide
information, lessons learned, and examples to assist communities in designing, developing, and
implementing these programs as needed.
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3 Report Development Approach
To summarize programs providing financial support and methods development research/guidance for
wastewater surveillance of SARs-CoV-2 over the past year (see Sections 4 and 5), EPA searched for and
reviewed federal agency, non-governmental organization, and other stakeholder (e.g., professional
associations) websites with a focus on wastewater and public health. EPA also relied on information
from press releases, publicly available grant/contracting documents, webinars/virtual conference
presentations, research project updates from funding agencies, and news articles. Except for the 10 case
studies presented in Section 6.2, EPA did not meet with the federal agencies, non-governmental
organizations, or other stakeholders described throughout this report to collect additional information
on their wastewater surveillance programs. Additionally, EPA did not conduct a comprehensive
literature review in support of this report.
As part of this report, EPA also compiled details on existing wastewater surveillance programs in the
United States using publicly available information (see Section 6 and Appendix A). EPA's goal in this
effort was to identify and understand the breadth of state, local (e.g., city, county), tribal, and university
wastewater surveillance programs. To gather this information, EPA conducted keyword searches in
January 2021 using various combinations of wastewater, COVID-19, and program-specific search terms
presented in Table I and an internet search engine. For example, a combination of "wastewater,"
"monitoring," "COVID-19," and "university" keywords used to identify surveillance efforts on
college/university campuses.
Table I. List of search terms used to identify wastewater surveillance programs.
Wastewater Search Terms
COVID-19 Search Terms
Program-Specific Terms
Wastewater
COVID-19
University
Sewage
COVID
College
Manhole
SARS-CoV-2
School
WWTP
Outbreak
City
Surveillance
County
Monitoring
Local
Dashboard
State
Wastewater Testing
State-wide
Tribe
Territories
EPA also reviewed websites of known collaborative initiatives (e.g., the COVID-19 Wastewater Based
Epidemiology [WBE] Collaborative [COVID-19 WBE Collaborative, 2021]) to identify additional
surveillance programs. EPA also reviewed the COVIDPoopsl9 website (UC Merced, 2021), which
compiles wastewater surveillance programs for SARS-CoV-2 across the globe, and a Slack workspace
website for informal communication regarding SARS-CoV-2 wastewater surveillance (see additional
discussion in Section 5.1).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Once the wastewater surveillance programs were identified, EPA compiled information on the programs
from program websites and dashboards, news articles and press releases, program summaries and
reports, and more. EPA searched for and reviewed information across all state wastewater surveillance
programs. However, EPA's ability to summarize information about all the local, tribal, and university
programs identified was limited given the number of such programs identified (more than 150). For all of
the state programs and a subset of the local and university programs, EPA recorded publicly available
details, including:
¦ Program location (e.g., city, state).
¦ Organization leading the program.
¦ Partners or collaborating organizations.
¦ Wastewater surveillance program website link.
¦ Sampling start date and end date, if applicable.
¦ Summary of sampling locations (e.g., WWTP influent, manholes at correctional facilities or
dormitories).
¦ Number of sampling locations.
¦ Analytical methods (e.g., reverse transcription-quantitative PCR [RT-qPCR]).
¦ Wastewater testing results presentation (e.g., tables, graphs, normalization, units).
¦ Public health actions/outcomes as available.
EPA then reviewed the identified state, local, and university-led programs to select a diverse subset with
unique aspects to highlight as case studies. EPA selected programs that demonstrate the wide variety of
ways that programs were financed (e.g., self-funded versus using Coronavirus Aid, Relief, and Economic
Security [CARES] Act funding) and implemented (e.g., sampling locations, analytical methods), as well as
how the programs interpreted the wastewater results (e.g., compared to individual cases, normalized by
wastewater flow rate or population size) and the public health measures the programs took based on
the wastewater results (e.g., individual testing, public education and outreach). Once EPA identified the
programs for case studies, EPA met with the practitioners and researchers leading each program in
order to gather additional information beyond what was found in the search of publicly available sources
listed above. EPA used this information to develop the case study summaries presented in Section 6.2.
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8
4 Financial Support
As more information became available on the ability to detect SARS-CoV-2 in untreated wastewater,
federal agencies, non-governmental organizations, and various stakeholders quickly recognized the need
to develop and support wastewater surveillance programs.
On March 27, 2020, the federal government enacted the CARES Act to address the health, economic,
and societal impacts of the COVID-19 pandemic. Federal agencies distributed CARES Act funds through
grants, contracts, and other mechanisms to interested researchers and other groups. The CARES Act
funded many different types of programs (e.g., individual testing, unemployment relief), including funds to
support wastewater surveillance programs. Some federal agencies, non-governmental organizations, and
other stakeholders also provided financial support from their non-CARES Act annual funding to support
wastewater surveillance programs.
Early in the COVID-19 response, numerous federal agencies and non-governmental stakeholders
initiated independent programs tied to wastewater surveillance, including providing funding to support
the COVID-19 response. To coordinate multiple initiatiatives across interested groups, the U.S.
Department of Health and Human Services (HHS) and CDC created the National Sewage Surveillance
Interagency Leadership (NSSIL) Committee in June 2020. Since late July of 2020, the NSSIL Committee
has held monthly meetings "to exchange information and discuss federal agency-specific missions, roles,
activities, and stakeholder engagement related to wastewater surveillance of SARS-CoV-2" (CDC,
2020a). The NSSIL Committee includes involvement from the following federal agencies:
¦ HHS
¦ CDC
¦ EPA
¦ Department of Homeland Security (DHS)
¦ Department of Defense
¦ United States Geological Survey
¦ National Institutes of Health (NIH)
¦ National Science Foundation (NSF)
¦ Department of Veterans Affairs
The NSSIL Committee also includes the following non-governmental stakeholders:
¦ Association of Public Health Laboratories (APHL)
¦ Association of State and Territorial Health Officials (ASTHO)
¦ Council of State and Territorial Epidemiologists
¦ National Association of County and City Health Officials (NACCHO)
¦ National Environmental Health Association (NEHA)
¦ Water Environment Federation (WEF)
¦ WRF
The NSSIL Committee originally included three federal interagency workgroups: Implementation and
Planning Workgroup, Science and Technology Evaluation for Practice Research, and Wastewater Testing
Surge Capacity (CDC, 2020a). The Implementation and Planning Workgroup develops and implements
the National Wastewater Surveillance System (NWSS), sewage sampling and testing capacity, and
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9
guidance documents for sewage sampling, testing, and data interpretation for public health action. The
NWSS links to a real-time, public health data platform known as DCIPHER (Data Collation and
Integration for Public Health Event Response) developed by CDC and designed to store, analyze, and
display public health data, including SARS-CoV-2 wastewater data collected throughout the country
(CDC, 2021a). See Section 5.3 for more information on CDC's NWSS. The Science and Technology
Evaluation for Practice Workgroup coordinates the exchange of information on wastewater sampling,
testing, and data interpretation throughout federal agencies. This workgroup also coordinates forums to
connect and inform the public and partner organizations (CDC, 2020a). The Wastewater Testing Surge
Capacity workgroup was originally designed to utilize federal laboratories to provide timely as-needed
wastewater analyses (CDC, 2020a). However, NSSIL decided this workgroup was no longer necessary
because of the rapid increase in wastewater analytical support from commercial and state laboratories
(CDC, 2021b).
Federal agencies and non-governmental stakeholders of the NSSIL prioritized making resources available
for SARS-CoV-2 wastewater surveillance programs that would support public health efforts to decrease
the spread of the virus. One identified resource was financial support for COVID-19-related wastewater
surveillance research, development, and implementation.
This section summarizes some of the funding mechanisms provided by federal agencies, non-
governmental organizations, and other stakeholders; it is not an exhaustive list of all funding provided to
wastewater surveillance programs. There are many wastewater surveillance programs that used or are
using funding from other sources (e.g., private foundations, reallocations of existing funding). See the
case studies in Section 6.2 for several detailed examples of how specific entities funded their wastewater
surveillance programs.
4.1 National Science Foundation
On April 3, 2020, NSF announced a request for proposals with a focus on "non-medical, non-clinical-
care research" to better understand the spread of COVID-19, educate on the science of virus
transmission and prevention, and develop processes to address the pandemic. NSF used their Rapid
Response Research (RAPID) funding mechanism, which allows the agency to quickly review proposals
and award grants for projects that have a critical urgency with regard to identified circumstances (NSF,
2020a). Traditional grant programs often take months between an agency's releases of a request for
proposal and project funding, which can be ineffective during emergencies such as the COVID-19
pandemic (NSF, 2020b). NSF's RAPID grant proposal requirements are also more streamlined than
traditional grant programs, requiring no more than five pages explaining why the proposed research is
urgent and why RAPID is the most appropriate grant program (NSF, 2020c). NSF's announcement
indicated up to $200,000 in funding for projects conducted over a period of up to one year. However,
NSF noted that they would consider projects of a longer duration with sufficient justification (NSF,
2020a).
NSF received $76 million from the CARES Act, with $75 million allocated for grants and $1 million for
the administration of those grants (NSF, 2020d). As of January 2021, NSF granted a total of 1,229
awards for COVID-19 projects using the CARES Act funding and $208 million from their fiscal year
2020 budget (NSF, 2020e). As of April 2021, 16 of these grants supported research on SARS-CoV-2 in
wastewater, as shown in Table 2 (NSF, 2020f).
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Table 2. NSF-funded research for wastewater surveillance.
Project Title
Award
Number
Description
Collaborative Research: RAPID:
Wastewater Informed
Epidemiological Monitoring for
SARS-CoV-2
2027752
and
2027758
University of Notre Dame and Georgia Tech to develop methods to monitor SARS-CoV-2 in wastewater
and connect those measurements with epidemiological data to model outbreak dynamics and estimate the
overall prevalence of the virus in communities over time. The issued project budget was $153,000 to
Notre Dame and $45,503 to Georgia Tech (NSF, 2020g; NSF, 2020h).
Collaborative Research: RAPID:
Coronavirus Persistence,
Transmission, and Circulation in
the Environment
2023057
and
2022877
University of Michigan and Stanford University to monitor SARS-CoV-2 in WWTPs in the San Francisco
Bay Area to determine whether WWTP monitoring can be used to catch outbreaks early. This is part of
a larger project funded with $68,591 issued to University of Michigan and $ 130,000 issued to Stanford
University (NSF, 2020i; NSF, 2020j).
RAPID: Tracking the Coronavirus
in Municipal Wastewater
2027679
Oregon State University to analyze SARS-CoV-2 from samples collected at WWTPs and sewer access
points throughout Oregon to identify infected communities, determine virus levels within each
community, and recognize possible underlying contributing factors of the continued spread of the virus.
The total project budget was issued at $100,000 (NSF, 2020k).
RAPID: Tribal Capacity to
Evaluate COVID-19 using
Wastewater-based Epidemiology
2038372
Arizona State University to investigate the feasibility of using SARS-CoV-2 wastewater surveillance to
monitor COVID-19 in tribal communities and to develop culturally appropriate research training and
educational materials for wastewater utility operators, health professionals, and tribal leaders. The total
project budget was issued as $200,000 (NSF, 20201).
RAPID: COVID-19's Impact on
the Urban Environment, Behavior,
and Wellbeing
2028564
Arizona State University to assess how public health interventions in response to the COVID-19
pandemic impact the environment, human behavior, and wellbeing of the public. Researchers will analyze
urban wastewater samples collected daily in Tempe, Arizona, in an effort to characterize various
biomarkers of environmental health and wellbeing (e.g., air pollutants, medications, allergy suppressants,
stimulants and depressants, drugs of abuse, dietary markers). The project budget is $199,998 (NSF,
2020m).
RAPID: Viral Structure-Function-
activity in the Engineered
Wastewater Cycle
2026599
Columbia University to research the fate of SARS-CoV-2 and other viruses during WWTP processes,
with a total funding of $ 198,388 (NSF, 2020n).
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Table 2. NSF-funded research for wastewater surveillance.
Project Title
Award
Number
Description
RAPID: Monitoring for SARS-
CoV-2 to Elucidate Infection
Dynamics Across Major
Metropolitan Areas of the U.S.
2029025
North Carolina State University, the University of Southern California, Rice University, and Howard
University to monitor SARS-CoV-2 in wastewater in four cities, one each in California, North Carolina,
Texas, and Washington, D.C. to address knowledge gaps in the use of wastewater surveillance as a public
health monitoring tool for a variety of communities. The project budget was issued as $200,000 (NSF,
2020o). See additional details in the "Monitoring SARS-CoV-2 in Major Metropolitan Areas of the U.S."
call-out box below.
RAPID: Determine Community
Disease Burden of COVID-19 by
Probing Wastewater Microbiome
2027059
University of Hawaii to develop a highly efficient concentration and detection method for enveloped
viruses (like SARS-CoV-2) in wastewater and to collect time-sensitive wastewater samples from
communities impacted by the disease to determine the abundance, diversity, and temporal dynamics of
SARS-CoV-2 and other enveloped viruses. The total project budget was $ 151,956 (NSF, 2020p).
RAPID: Determination of Health
Risks and Status from SARS-CoV-
2 Presence in Urban Water Cycle
2029515
University of Utah to develop efficient techniques to extract and monitor SARS-CoV-2 in wastewater and
to understand human health risks associated with the presence of SARS-CoV-2 in influent and treated
effluent at WWTPs. Total funding issued for this project was $123,706 (NSF, 2020q).
Research Coordination Network
for Wastewater Surveillance of
SARS-CoV-2
2038087
University of Notre Dame, Howard University, Stanford University, and Arizona State University to
create a Research Coordination Network to connect researchers from across the country for quick and
efficient knowledge transfer on SARS-CoV-2 wastewater surveillance with a project budget of $299,995
(NSF, 2020r). See additional details in the "Creating the Research Coordination Network for SARS-CoV-
2" call-out box below.
RAPID: Identifying Geographic
and Demographic Drivers of Rural
Disease Transmission for
Improved Modeling and Decision
Making
2029866
University of North Carolina to examine drivers of disease transmission in rural areas and explore
differences with urban areas, with the ultimate goal of improving pandemic management in rural areas.
Researchers will use data (e.g., health surveillance data, cellphone-based mobility data, land use features,
commuting patterns) from three rural and three urban counties in North Carolina to develop a
susceptible-exposed-infected-recovered model. They will also collect wastewater samples to quantify the
prevalence of SARS-CoV-2 and to provide a complementary non-clinical metric to validate their model.
Total funding issued for this project was $ 135,593 (NSF, 2020s).
SBIR Phase 1: Automated In-Situ
High-Resolution COVID-19
Wastewater-Based Epidemiology
2041400
FLUIDION US, Inc. to develop an optimized RT-qPCR approach for in-situ sampling and analysis of
different viruses (including SARS-CoV-2, its variants, and other emerging viruses) and an instrumentation
platform that is applicable to the early detection of viruses. As part of this work, researchers will explore
the latest advances in molecular biology protocols, such as an extraction-free single step RT-qPCR. Total
funding issued for this project was $256,000 (NSF, 2020t).
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Table 2. NSF-funded research for wastewater surveillance.
Project Title
Award
Number
Description
RAPID COVID-19 DCL response:
Wastewater Pathogen Tracking
Dashboard
2033137
Battelle Memorial Institute to evaluate wastewater data from four locations to determine the prevalence
of SARS-CoV-2 and other viral pathogens, as well as to detect and quantify viral mutations through the
use of gene sequencing. Additionally, researchers will develop a predictive risk model to identify
neighborhoods to initiate contact tracing due to high SARS-CoV-2 in the wastewater relative to the
number of confirmed COVID-19 cases. Total funding issued for this project was $ 197,375 (NSF, 2020u).
RAPID-REU Site: Mitigating the
Impact of COVID-19 Pandemic on
Undergraduate Research Training
in the Biosciences
2034045
Montana State University to quantify SARS-CoV-2 in wastewater from a Montana WWTP using a CDC
test kit protocol as an indicator of community spread of the virus for use by public health officials. This is
just one of several projects covered by the $75,042 project budget, all of which will engage
underrepresented minority students in research through projects that can be conducted remotely (NSF,
2020v).
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Monitoring SARS-CoV-2 in Major Metropolitan Areas of the U.S.
In May 2020, NSF awarded a $200,000 RAPID grant (award number 2029025) to a collaboration
between North Carolina State University, the University of Southern California, Rice University, and
Howard University to create SARS-CoV-2 wastewater surveillance programs in four major
metropolitan areas with varying sewershed sizes/populations, infection rates, and required COVID-19
prevention strategies (e.g., mask mandates). Each university received 25 percent of the NSF grant to
purchase analytical equipment and materials and to collectively develop an analytical method for SARS-
CoV-2 in wastewater. Each university was responsible for optimizing a portion of the analytical method
(e.g., RNA extraction, sample storage, virus concentration) using samples taken from their respective
cities (USC, 2021). The program collected samples from WWTPs in Los Angeles, California; the
District of Columbia; Raleigh, North Carolina; and Houston, Texas, to address knowledge gaps in the
use of wastewater surveillance as a public health monitoring tool. Researchers acknowledged that the
largest hurdles with the project were deciding between analytical approaches, considering how much
virus each method recovers, the associated expenses for the supplies, and the required duration of the
methods. Additionally, the project team initially had to overcome supply and equipment shortages due
to the pandemic, limiting their ability explore certain analytical methods (NSF, 2020o; Shah, 2020; USC,
2021).
Creating the Research Coordination Network for SARS-CoV-2
In July 2020, NSF provided nearly $300,000 to researchers collaborating from the University of Notre
Dame, Howard University, Stanford University, and Arizona State University to create a Research
Coordination Network to connect interested research groups studying SARS-CoV-2 in wastewater
across the United States (award number 2038087). The Research Coordination Network addresses
knowledge gaps in the development of wastewater analysis methods by initiating virtual activities such
as conferences, workshops, training videos, and seminars. Additionally, the researchers support sharing
knowledge globally by connecting with international wastewater surveillance networks and efforts. The
goal of the Research Coordination Network is to exchange of knowledge in an effort to accelerate
program optimization and facilitate the ongoing development of wastewater surveillance (NSF, 2020r).
4.2 Centers for Disease Control and Prevention
In place since 1995, CDC's Epidemiology and Laboratory Capacity for Prevention and Control of
Emerging Infectious Diseases (ELC) Cooperative Agreement has provided funding to "all 50 states,
several large health departments, and U.S. territories, and affiliates to detect, respond to, control, and
prevent infectious diseases" (CDC, 2021 b). On April 23, 2020, HHS announced that CDC1 received
$631 million in CARES Act funding to support state and local COVID-19 response efforts for the 64
health department recipients (i.e., 50 states, six large cities, and eight territories2) already supported in
the current five-year funding period (CDC, 2020c). This funding was intended to support "key activities
related to COVID-19 in the areas of epidemiology, laboratory, and informatics" (CDC, 2020c). None of
this initial CARES Act funding was used to support wastewater surveillance programs (CDC, 2021 c). On
May 18, 2020, CDC announced that it would distribute an additional $10.25 billion in CARES Act
1 While CDC is an agency within HHS, CDC distributed the CARES Act funding independently through existing
CDC financial mechanisms.
2 The six large cities are: Chicago, IL; Houston, TX; Los Angeles County, CA; New York City, NY; Philadelphia,
PA; and Washington, D.C. The eight territories are: American Samoa, Federated States of Micronesia, Guam,
Mariana Islands, Marshall Islands, Palau, Puerto Rico, and U.S. Virgin Islands (CDC, 2021c).
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14
funding to states, territories, and local jurisdictions through the ELC cooperative agreements (CDC,
2020d). Six states and two local jurisdictions allocated some of this funding to support wastewater
surveillance efforts (CDC, 2021c). In September 2020, CDC distributed a supplemental ELC award
totaling $2.5 million to eight states3 to establish the early implementer network for NWSS. This initial
funding was used to support public health department capacity to coordinate sample collection and
testing, along with data submission to NWSS (CDC, 2021c).
In January 2021, ELC awarded an additional $19 billion in 2021 Coronavirus Response and Relief
Supplemental Appropriations Act funding to support an expanded COVID-19 response, which could
include further development, implementation, and expansion of wastewater surveillance programs
(CDC, 2021 d). Thirty states, one city, and two territories opted to allocate a total of $203 million to
wastewater surveillance efforts (CDC, 2021c). This included support to Ohio and Houston, Texas (see
case studies for Ohio in Section 6.2.4 and Houston, Texas, in Section 6.2.7). As of August 2021, 43
jurisdictions are using CDC-distributed funds to support wastewater surveillance, including 37 state
health departments, four local health departments, and health departments from two U.S. territories
(see Figure I) (CDC, 2021g).
Figure I. Jurisdictions using CDC funds to support wastewater surveillance for SARS-CoV-
2 as of August 2021 (CDC, 2021 g).
4.3 National Institutes of Health
NIH4 launched the Rapid Acceleration of Diagnostics (RADxSM) initiative on April 29, 2020, to "speed
innovation in the development, commercialization, and implementation of technologies for COVID-19
3 The eight states are: California, North Carolina, Ohio, South Carolina, Utah, Virginia, Washington, and Wisconsin
(CDC, 2021c).
4 While NIH is an agency within HHS, NIH established the RADxSM initiative independently from HHS.
Puerto Rico
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15
testing" (NIH, 2021a). The initiative has four programs, each with a unique focus area and budget to
fund projects (NIH, 2020a):
1. RADxSM Tech
2. RADxSM Advanced Technology Platforms
3. RADxSM Underserved Populations
4. RADxSM Radical
The RADxSM Radical (RADx-rad) supports new nontraditional approaches to address gaps in COVID-19
testing, including wastewater analysis to identify the virus and measure the spread of infection,
particularly among high-risk populations (NIH, 2020a). RADx-rad has a total budget of $200 million,
with grants and supplements supported by I I NIH institutes and centers (NIH, 2020b; NIH, 2020c).
In August 2020, as part of the RADx-rad program, NIH issued a series of funding opportunities for
wastewater surveillance through notices of special interest and emergency awards for researchers to
apply for project funding. NIH issues notices of special interest to support high-priority and high-
opportunity areas of science. NIH uses emergency awards to provide expedited funding in response to
public health emergencies such as the one declared for COVID-19. NIH announced emergency awards
to support wastewater detection of SARS-CoV-2, with $ 19 million available to fund five to ten
wastewater surveillance initiatives (NIH, 2020d). In December 2020, NIH issued seven awards to
support wastewater surveillance and one award to support data coordination across grantees (NIH,
2021 b). All listed wastewater surveillance projects have a start date of January 2021 and are projected
to end in December 2022; the data coordination effort is expected to continue through November
2024. A brief summary of the awards related to wastewater surveillance is provided in Table 3 and a
summary of the data coordination center is presented in the following call-out box.
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Table 3. NIH wastewater surveillance projects funded under RADx-rad.
Project Title
Project Number
Description
Development and Proof-of-
Concept Implementation of the
South Florida Miami RADx-rad
SARS-CoV-2 Wastewater-Based
Surveillance Infrastructure
1UO1DA053941 -01
The University of Miami (partnering with Weill Cornell Graduate School of Medical
Sciences) to develop data standards and infrastructure for wastewater surveillance program
data, optimize wastewater surveillance sampling and analytical methods, and integrate the
results with public health data to develop predictive models for community spread utilizing
total project funding of $2.7 million (NIH, 2021c).
Wastewater Analysis of SARS-
CoV-2 in Tribal Communities
1U01DA053976-01
Arizona State University to develop a WBE Tribal Coordination Center and measure
SARS-CoV-2 in wastewater across U.S. reservations, with a project budget of $ 1.5 million.
Another goal of this project is to "show that WBE is a non-invasive, culturally appropriate
biomonitoring strategy that can be adopted and implemented by tribal communities" (NIH,
2021 d).
Improved Scalability, Sensitivity,
and Interpretability of Pathogen
Detection, Including SARS-CoV-
2, in Wastewater Using High-
throughput, Highly Multiplexed
Digital Array PCR Technology
1 UO 1DA053899-01
University of North Carolina Chapel Hill to explore limitations of molecular technologies
like RT-qPCRto quantify SARS-CoV-2 (e.g., lack of streamlined pre-analytical processing
steps) and to demonstrate other comprehensive and low-cost approaches that could be
used to rapidly address novel pathogen threats in the future, with a project budget of $ 1
million (NIH, 202 le).
Wastewater Assessment for
Coronavirus in Kentucky:
Implementing Enhanced
Surveillance Technology
1 UO 1DA053903-01
University of Kentucky to develop analytical methods for rapid quantification in the field
(i.e., "create a sensitive, robust, and field-friendly platform for testing wastewater for SARS-
CoV-2 RNA" rather than a laboratory setting), to validate the approach with side-by-side
comparisons to conventional wastewater surveillance, and then establish wastewater
surveillance programs by training WWTP operators to use these methods in remote areas
of Appalachian Kentucky. The issued project budget was $1.8 million (NIH, 202 If).
Wastewater Detection of
COVID-19
1 UO 1DA053893-01
Missouri State Department of Health and Senior Services to support wastewater
surveillance at congregate facilities and to use these data to help estimate the per patient
contribution and longevity of SARS-CoV-2 RNA in wastewater. This will be done by
increasing the number of facilities sampled, adjusting the sampling frequency, and comparing
the wastewater results to individual testing. Missouri is also using these funds to evaluate
and define factors that contribute to SARS-CoV-2 signal suppression in wastewater, with a
total funding of $2 million (NIH, 202 Ig).
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Table 3. NIH wastewater surveillance projects funded under RADx-rad.
Project Title
Project Number
Description
Optimizing SARS-CoV-2
Wastewater-based Surveillance
in Urban and University Campus
Settings
1UO1DA053949-01
Columbia University Health Sciences to optimize wastewater surveillance conducted at a
diverse, urban university (including dorms, research buildings, and medical facilities) and the
surrounding sewersheds and WWTPs. Researchers will also model case counts using
normalized wastewater data. The team was awarded $2.5 million for this effort (NIH,
202 Ih).
Bioinformatics Framework for
Wastewater-based Surveillance
of Infectious Diseases
301LMO13129-02S2
Arizona State University (Tempe campus) to develop and implement a near real-time
wastewater surveillance bioinformatics framework for SARS-CoV-2 at the national, city,
and intra-sewershed or neighborhood level, and to translate SARs-CoV-2 data from RT-
qPCR analysis and high-throughput sequencing into information for monitoring population
health. The goal of this work is to "lead to a better understanding of how WBE can
support population-level monitoring of SARS-CoV-2 and similar infectious disease threats"
(NIH, 202 li). As part of this work, researchers will also incorporate findings into and
expand the online dashboard Arizona State University developed in collaboration with
Tempe, Arizona (see case study in Section 6.2.8). The project was funded at $571,000 in
funding (NIH, 202li).
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Establishing a RADx-rad Data Coordination Center
On August 6, 2020, NIH released an emergency award under RADx-rad to fund the development of a
RADx-rad Data Coordination Center to provide support to RADx-rad awardees with administration
operations and logistics; data collection, integration, and sharing; and data management and use.
Beyond connecting RADx-rad programs, the Data Coordination Center also provides data to the NIH-
based data center that supports other NIH initiatives (NIH, 2020e).
On December 21, 2020, NIH awarded the University of California San Diego and the University of
Texas Health Science Center at Houston $5,954,423 in funding from the Paycheck Protection Program
and Health Care Enhancement Act of 2020 for initial development of the Data Coordination Center in
2021 (project number I U24LM0I 3755-01) (NIH, 2020e; NIH 2021 j). The project team designed the
RADx-rad Discoveries and Data Coordination Center to include three main functions (NIH, 2021 k):
1. Administration Core: To focus on logistics and communications for the Data Coordination
Center and facilitate data sharing and data use agreements to collect and distribute data
generated by the RADx-rad awardees.
2. Data Core: To develop tools and approaches to assist the RADx-rad awardees with data
collection, harmonization, and sharing (e.g., common metadata requirements, mapping
standards, data hosting).
3. Discovery and Diagnostic Core: To support RADx-rad awardees in ensuring the data they
collect is usable for the Data Coordination Center by providing expert consultation,
developing best practices, and more.
The Data Coordination Center includes information on all NIH RADx-rad awardees, a list of
resources for the awardees that includes topics such as data flow and guidance for common data
elements, and training and networking events for researchers (NIH, 2021 k).
4.4 U.S. Department of Health and Human Services
In November 2020, HHS (with support from CDC) awarded a competitive contract of $ 1.55 million in
CARES Act funding to AquaVitas, a commercial wastewater testing and analytics company, to complete
a six-week wastewater testing pilot study of WWTPs serving about 10 percent of the U.S. population
for a period of six weeks (CDC, 2020b; FPDS, 2021). The results from this pilot test were provided to
state and local public health agencies and incorporated into CDC's NWSS (CDC, 2020b).
In February 2021, HHS issued a follow-on request for proposals to collect wastewater samples, analyze
samples, and transfer results from at least 320 WWTP throughout the United States, including tribes
and territories (the total representing approximately 30 percent of the U.S. population), into CDC's
NWSS (CDC, 2020b; HHS, 2021). Biobot Analytics was awarded the contract on May 24, 2021, with
sample collection scheduled anticipated to begin on June 14, 2021, and anticipated run for a period of
nine weeks (Biobot, 2021a; Biobot, 2021 b). Under this contract, Biobot Analytics will expand on the
previous HHS-led wastewater epidemiology program, while incorporating genomic sequencing in an
effort to track COVID-19 variants (Biobot, 2021 a).
4.5 Water Research Foundation
WRF initiated and funded multiple research projects to advance wastewater surveillance for SARs-CoV-
2 based on the research needs identified at their first International Water Research Summit in April
2020 (WRF, 2020a) (see additional details in Section 5.1). Three research areas were identified for
additional funding and research: (I) interlaboratory and methods assessment, (2) stability of SARS-CoV-
2 genetic signal in wastewater, and (3) impact of storage and pre-treatment methods on signal strength
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(WRF, 2020b). Funds were awarded to address the first two topic areas to research teams following a
competitive contract process.
In June 2020, WRF published a request for qualifications to identify a research team to evaluate existing
laboratory methods for sensitivity and precision to detect the SARS-CoV-2 in untreated wastewater
(WRF, 2020c). In July 2020, WRF named Trussell Technologies as the recipient of $200,000 to lead the
"Interlaboratory and Methods Assessment of the SARS-CoV-2 Genetic Signal in Wastewater" study
(WRF, 2020d). The work started in early Fall 2020 and involved collecting composite wastewater
samples tested by the research team's laboratory and submitted to other participating laboratories for
analysis using their own methods. WRF funded the project using internal funds as well as supplemental
funding from the Bill and Melinda Gates Foundation (WRF, 2020e).
In August 2020, WRF announced another request for qualifications to find a research team that could
identify ways to optimize SARS-CoV-2 sewage sampling designs in order to quickly detect COVID-19
spread through communities contributing to sewersheds of various sizes (WRF, 2020f). In October
2020, WRF named the University of California at Irvine as the recipient of $300,000 for a study titled
"Understanding the Factors that Affect the Detection and Variability of SARS-CoV-2 in Wastewater"
(WRF, 2020g). Using wastewater samples (sewage and septage) collected in Los Angeles County and
analyzed for SARS-CoV-2, the research team is developing recommendations for optimal sample design
at three different scales: large urban sewersheds, medium-sized regional sewersheds, and small regional
systems (WRF, 2020h). Funding for this project was provided by WRF as well as the Bill and Melinda
Gates Foundation, and work is anticipated to be completed in 2021 (WRF, 2020h).
WRF has also funded at least one other study related to wastewater monitoring for SARS-CoV-2. In
September of 2019, WRF awarded $295,000 to the University of Notre Dame to evaluate the
persistence and disinfection of Lassa virus in WWTPs (WRF, 2020i). The scope was later expanded to
include SARS-CoV-2. The goal of this study is to develop a user-friendly model capable of estimating
environmental releases and worker exposures to these two viruses to help prepare the wastewater
industry for the next epidemic or pandemic. Work is anticipated to be completed in 2022 (WRF, 2020i).
4.6 Water Environment Federation
WEF has entered into a cooperative agreement with CDC to support CDC in providing information on
SARS-CoV-2 guidance to the water and wastewater utility sector, which comprises the majority of
WEF's membership. WEF is developing a network for information sharing within the sector. The
Network for Wastewater-based Epidemiology will host the utility community of practice for wastewater
surveillance (WEF, 2021 b). In addition, WEF will provide training on wastewater surveillance to utilities
and public health personnel (WEF, 2021a). WEF is also working with CDC to evaluate use of
wastewater surveillance in correctional facilities as a supplement to case surveillance data and as a
possible early warning for COVID-19 in these facilities. For this effort, WEF will be working with
multiple states, the first of which is Oklahoma (WEF, 2021c).
4.7 Other Funding Opportunities
Beyond funding opportunities through federal agencies and non-governmental organizations, wastewater
surveillance programs are utilizing funds from private foundations, existing program funding, and pro-
bono resources. Two examples of private foundations funding wastewater surveillance programs or
research on SARS-CoV-2 analytical methods include:
¦ The Bill and Melinda Gates Foundation, which provided almost $390,000 to the California
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Institute of Technology to develop a rapid quantification method for detecting SARS-CoV-2 in
wastewater (BMGF, 2021).
¦ The Foundation for a Healthy Kentucky, which provided funding to pay for the equipment needed
to analyze wastewater samples collected from the Mayfield WWTP for SARS-CoV-2, as part of a
larger $60,000 project. The project team includes Graves County Health Department, Mayfield
Electric and Water Systems, Murray State University, and the University of Louisville (Healthy KY,
2020).
Some wastewater surveillance programs were supported by funds from other existing programs. For
example, Hampton Roads Sanitation District (HRSD) used available funds from ratepayers to conduct
SARS-CoV-2 wastewater sampling and analysis (see case study in Section 6.2.6 for additional details)
(HRSD, 2021 b). Wyoming also reprogrammed existing funding they received from CDC's ELC to
support their wastewater surveillance program (see case study in Section 6.2.5) (Wyoming, 2021).
An example of pro-bono wastewater surveillance support comes from BioBot Analytics, a private
company based out of Boston, Massachusetts. BioBot initiated a nationwide pro-bono wastewater
sampling and analysis program in April 2020 to help establish their SARS-CoV-2 testing protocols and
expand wastewater surveillance efforts. From March 25 through May 31, 2020, BioBot Analytics
received over 1,800 wastewater samples from 360 WWTPs across 43 U.S. states and Canadian
providences (Biobot, 2020). Results highlighted the utility of wastewater surveillance as an early warning
of a potential COVID-19 outbreak. For example, wastewater data from New Castle County, Delaware,
spiked between three and seven days before a spike in COVID-19 cases was identified through individual
testing (BioBot, 2020). Biobot was able to quickly develop this program by leveraging some of their
previous work monitoring wastewater for opioids.
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5 Program Development
Numerous international efforts have demonstrated that wastewater surveillance can be used to detect
outbreaks of other viruses (e.g., poliovirus, norovirus) earlier than through clinical surveys (Brouwer et
al„ 2018; Deshpande et al„ 2003; Ivanova et al„ 2019; Kazama et al„ 2017; Lago et al„ 2003). At the start
of the COVID-19 pandemic, researchers and practitioners needed to quickly develop and/or adapt
analytical methods, sample collection procedures, and data interpretation approaches specific to SARS-
CoV-2. Much of this work began in early 2020 with collaborative efforts among international experts
from utilities; universities; and local, state, and federal governments. Many of these efforts were held
virtually and recorded for future reference. As the science rapidly evolved, teams leading surveillance
programs throughout the United States and in other countries continually learned from one another via
frequently revised online forums that documented research findings, lessons learned, best practices,
guidelines, and recommendations.
As the field continued to evolve over the past year, new technologies emerged that improved the speed
and accessibility of wastewater testing and analysis for implementation at various scales. Over time,
multiple analytical methods and program frameworks were proven successful and were shared widely
across the wastewater surveillance community. The field has matured by having reliable methods,
programs, safety measures, and ethical considerations, many of which are being constantly refined and
updated with the latest research and information. This section discusses some of the key areas of
program development.
5.1 Peer-to-peer Communication and Resource Sharing
Early in the COVID-19 pandemic, there were few guidelines and recommendations available on sampling
and analysis methods for SARS-CoV-2 in wastewater. The global research community worked in
partnership to build upon established protocols for waterborne enteric viruses and further progress the
field of wastewater surveillance. This international collaboration among experts was a crucial step in the
success of wastewater surveillance programs for SARS-CoV-2 during the pandemic. Federal, state, tribal,
and local agencies; utilities; universities; and other groups have all participated in and benefited from the
continued collaborative spirit and partnership across the field. Some of these efforts are discussed
below.
5.1.1 Workshops and Trainings
WRF released a 42-question survey via emails campaigns, Linkedln, Instagram, Twitter, and Facebook
from April 16-24, 2020, to gather information on sampling design (e.g., locations, frequency, composite
vs. grab, shipping, storage), sample processing (e.g., preliminary treatment, concentration, purification),
extraction, detection, sample volume requirements, and analytical methods validation and controls
(Zhou et al„ 2020). WRF used the results from the survey to inform the content for their virtual multi-
day International Water Research Summit on COVID-19 held in late April 2020. WRF convened over
50 international experts from utilities, academia, consulting, and government to advance the work
researchers were performing throughout the world (WRF, 2020a). The International Water Research
Summit included discussions on the following topics in an effort to identify best practices and
recommended approaches:
¦ Sample collection and preservation
¦ Analytical methods for identifying SARS-CoV-2 in wastewater
¦ SARS-CoV-2 concentrations in wastewater in comparison to community COVID-19 cases
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¦ Communication of results
During the International Water Research Summit, participants also developed a "near-term research
roadmap" that identified critical knowledge gaps requiring immediate research (WRF, 2020a).
Participants categorized the necessary research into four themes: (I) analytical methods variability,
reproducibility, and reliability; (2) viral shedding rate of SARS-CoV-2 in feces and the associated genetic
signal; (3) interpretation of results to support public health efforts; and (4) risk assessment of potential
exposure pathways for wastewater workers (WRF, 2020j). Participants also identified 12 research
opportunities to advance wastewater surveillance for SARS-CoV-2 across these themes, as shown in
Table 4 (WRF, 2020j). Based on these findings, WRF sought funding partners and began soliciting
applications for qualified organizations to conduct research on these topics. See Section 4.5 for details
on some of the projects WRF funded.
Table 4. Research opportunities identified by WRF to support wastewater
surveillance of SARS-CoV-2 (WRF, 2020j).
Priority
Theme
Research Opportunity
High
Methods
Intra- and interlaboratory assessments on sampling regimes and
molecular methods.
High
Shedding rate and
genetic signal
Effect of wastewater pre-treatment on genetic signal.
High
Shedding rate and
genetic signal
Dilution and persistence of the genetic signal in the sewer collection
system. Targeted integrated study (in well-characterized systems that
have good hydraulic models).
High
Risk
Evaluation of potential for infectious virus in wastewater and generation
of aerosols.
High
Interpretation of
results
Correlations to clinical data for the assessment of community
prevalence. How to leverage wastewater surveillance to provide useful
data to public health stakeholders.
High
Interpretation of
results
Define partnership opportunities.
High
Shedding rate and
genetic signal
Viral shedding rate, duration, and demographics in symptomatic and
asymptomatic infections (RNA copies per gram of feces).
Medium
Methods
Impacts of sample collection method (composite vs. grab, duration of
composite, time of day).
Medium
Methods
Distribution of virus (or RNA copies) in liquid and solid phase.
Medium
Interpretation of
results
How to effectively translate COVID-19 research into pandemic
preparedness and wastewater surveillance for future needs.
Low
Methods
Which spike organism to use for quality assurance/quality control
purposes.
Low
Methods
Comparative methods review for enveloped viruses, focusing on the
concentration from wastewater matrix.
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23
WRF continues work on wastewater surveillance and most recently in April 2021 held a second
International Symposium on COVID-19 Wastewater Surveillance (WRF, 2021a).
As another example, the National Institute of Standards and Technology (NIST) held a virtual workshop
in June 2020 to discuss challenges with measuring and detecting SARS-CoV-2 in wastewater with five
expert presenters.5 The main goals for the workshop included discussing the merits of applying NIST's
expertise in producing standards and reference materials to the most pressing monitoring issues (NIST,
2020).
Tailored to applying surveillance on a more local level, NACCHO and CDC hosted a webinar in June
2020 titled "Water, Sanitation, and Hygiene During the COVID-19 Pandemic" (NACCHO, 2020a;
NACCHO, 2020b). The webinar featured three speakers, two from CDC and one from a local health
department.6 The topics included general information on proper cleaning and disinfection, minimizing
risk of Legionella in drinking water before reopening unoccupied buildings, options for disaster shelters,
along with wastewater surveillance for SARS-CoV-2. Panelists reviewed sampling and analysis techniques
for wastewater testing of SARS-CoV-2, discussed the benefits of early detection of SARS-CoV-2, and
raised concerns regarding common misconceptions about the field (e.g., the virus is always infectious in
sewage) (NACCHO, 2020a; NACCHO, 2020b). NACCHO included wastewater surveillance for SARS-
CoV-2 as one of the sessions in their Preparedness Summit annual conference in April 2021 (NACCHO,
2021).
5.1.2 Online Platforms
In addition to collaboration through workshops and webinars, the wastewater surveillance community
has developed numerous interactive online resources. One example is the COVIDPoopsl9 dashboard,
created and managed by the University of California at Merced. This dashboard provides a global map of
reported SARS-CoV-2 wastewater surveillance efforts, including the number of public dashboards,
universities, countries, and surveillance programs (referred to as testing sites), all of which is updated
daily. For each identified program, the dashboard provides a link to the program website, relevant news
articles, and/or associated publications for the surveillance program (UC Merced, 2021). The University
of California at Merced published the methods for developing the COVIDPoops 19 dashboard
(Naughton et al„ 2021).
As of early May 2021, the COVIDPoops 19 dashboard includes 80 dashboards, 256 universities, 54
countries, and 2,216 surveillance programs, as shown in Figure 2 (UC Merced, 2021). This dashboard is
part of the larger COVID-19 WBE Collaborative that encourages partnership and discussions among
those in the wastewater surveillance field (COVID-19 WBE Collaborative, 2021). The COVID-19 WBE
Collaborative website provides a list of resources, including a COVID-19 WBE Slack workspace that is
an organized web-based channel for groups to collaborate and talk about their programs; a Protocols.io
group that houses information on sample collection techniques, analytical methods, quality control, and
results interpretation; and a link to the website for Research Coordination Network on wastewater
surveillance. The Research Coordination Network website, funded by NSF and designed by researchers
from the University of Notre Dame, Howard University, Stanford University, and Arizona State
University, offers networking opportunities to increase collaboration among wastewater surveillance
programs (University of Notre Dame, 2021). Information is provided through webinars, workshops,
5 Bharat Ramakrishan, OpenBiome; Manoj Dadlani, CosmosID; Katerina Papp, Southern Nevada Water Authority;
Dr. Kyle Bibby, University of Notre Dame; and Aparna Keshaviah, Mathematica.
6 Amy E. Kirby, PhD, MPH, CDC; Jasen Kunz, MPH, CDC; Rob Blake, MPH, Transylvania County Department of
Public Health.
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
training videos, and links to other resources (e.g., open data repositories) (see additional details on this
NSF project in Section 4.1).
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Another example of an interactive online resource is the international Water Action Platform, created
and managed by Isle Utilities and supported by various sponsors. While the platform was initiated in
March of 2020 in response to the COVID-I9 pandemic, it has since expanded to cover a wide range of
wastewater-related topics (e.g., asset management, communications, sustainability). The Water Action
Platform encourages peer-to-peer collaboration by hosting regular webinars to share lessons learned
and best practices from utilities around the world, summarizing news updates and press releases, and
supporting a series of "knowledge hubs" via WhatsApp. As of September 2020, more than 1,300 people
from 758 organizations across 92 countries are participating in the Water Action Platform (Water
Action Platform, 2021).
WEF is also compiling articles, research, news updates, and links to other sites for up-to-date
information on best practices for wastewater surveillance on a weekly basis. WEF's coronavirus website
breaks out resources into sections by topics such as vaccine resources, water sector-specific
information, training and education resources, and water sector coronavirus assistance. The water
sector-specific information includes recent articles, podcasts, webcasts, and fact sheets that were
published by WEF or other sources (WEF, 202Id). WEF has also developed a Network of Wastewater-
Based Epidemiology website focusing on U.S. utilities performing wastewater surveillance. As of May I 3,
2021, WEF's dashboard included 5I7 utilities, 159 academic institutions, eight laboratories, and one
other program performing or supporting wastewater surveillance programs, as shown in Figure 3 (WEF,
202 lb).
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25
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Figure 3. Utilities performing wastewater surveillance in the United States (WEF, 202lb).
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26
5.2 Researching and Developing Analytical Methods
Beginning early in the COVID-19 pandemic, many federal agencies, state agencies, non-governmental
stakeholders, and the academic research community worked both independently and together to quickly
develop and refine methods to support wastewater surveillance for SARS-CoV-2. This section presents
several examples of such efforts.
Laboratory analytical methods for the detection of
SARS-CoV-2 RNA in wastewater have continually
evolved based on new science and resources. The
NWSS website (CDC, 2021a) outlines commonly
used wastewater surveillance methods for:
¦ Sample preparation (e.g., storage,
homogenization, sample clarification).
¦ Sample concentration (e.g., ultrafiltration,
polyethylene glycol precipitation,
ultracentrifugation).
¦ RNA extraction.
¦ SARS-CoV-2 RNA detection and
quantification (e.g., RT-qPCR and reverse
transcription digital droplet PCR [RT-
ddPCR]).
SARS-CoV-2 RNA Measurement Methods
Once the SARS-CoV-2 RNA is extracted from
the wastewater sample, laboratories quantify the
amount of RNA using RT-qPCR, RT-ddPCR, or
other less common forms of RT-PCR. The viral
RNA is enzymatically transcribed to
complementary deoxyribonucleic acid (cDNA) in
a process called reverse transcription (RT). The
cDNA is then used in a second enzymatic
reaction (PCR) to make many copies of a small
fragment of the transcribed viral RNA. RT-PCR
(qPCR or ddPCR) can occur in two steps, where
these enzymatic reactions take place sequentially
in separate tubes, or as a one-step reaction
where the two enzymatic processes occur in
sequence in the same tube (CDC, 202If).
CDC has also developed an online resource that lists reverse transcription PCR (RT-PCR) primers and
probes for two N gene assays of SARS-CoV-2, last updated in June 2020 (CDC, 2020e). The APHL has
compiled a list of selected COVID-19 molecular testing resources for individual testing and their key
characteristics (e.g., test name, manufacturer, complexity, testing throughput capacity) (APHL, 2020).
APHL also provides a downloadable Excel file summarizing a longer list of test methods for individual
testing, with additional details on performance and limits of detection (APHL, 2021a), and a dashboard
summarizing laboratory testing capacity and capabilities based on results of a weekly survey of 99 state,
local, and territorial health laboratories, focusing on individual testing (APHL, 2021 b). While the APHL
resources are focused on individual testing, some of the information may be useful to consider for
wastewater samples (e.g., buffer supply limitations).
One example of a common laboratory analytical
method is that developed by IDEXX Reference
Laboratories. IDEXX developed a novel, highly
sensitive RT-qPCR test to detect NI and N2 gene
targets of SARS-CoV-2 (IDEXX, 2021).
SARS-CoV-2 N Gene Targets
There are two distinct assays targeting the
SARS-CoV-2 nucleocapsid gene, NI and N2,
that can be measured using RT-qPCR or RT-
ddPCR (CDC, 202If).
In the summer of 2020, NIST developed reference materials consisting of synthetic fragments of the
SARS-CoV-2 virus RNA to aid in the analysis and development of RT-qPCR assays for SARS-CoV-2
(360dx, 2020; NIST, 2020). Researchers and laboratories can use the reference materials to assess
detection limits for SARS-CoV-2 assays or calibrate other in-house or commercial SARS-CoV-2
controls. This allowed researchers and laboratories to further refine their analytical methods for SARS-
CoV-2. NIST provided the reference material free of charge through funding from the CARES Act in
exchange for user feedback to further improve and develop the materials (360dx, 2020; NIST, 2020).
NIST is continuing to work with the DHS Science and Technology Directorate and the University of
Louisville School of Medicine to further develop and refine reference materials and standards for
quantifying SARS-CoV-2 with RT-qPCR assays, which will allow for additional development in the
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27
analytical methods (DHS, 2021). The new standards will allow SARS-CoV-2 wastewater surveillance
results to be more readily shared and compared across surveillance programs, which will in turn inform
more useful healthcare decisions (DHS, 2021).
As mentioned in Section 4.5, WRF funded several studies to further advance the wastewater
surveillance field based on research needs identified at their International Water Research Summit
(WRF, 2020h; WRF, 2020e). One such study was a methods assessment and interlaboratory comparison
of wastewater results for SARS-CoV-2, in which WRF assessed the reliability of various SARS-CoV-2
testing methods to produce accurate and repeatable results (WRF, 2020c). For this, WRF invited
laboratories to participate and analyze the same composite samples using their own analytical methods.
The study ultimately compared distinct RNA extraction and quantification methods used at 32 U.S.-
based laboratories, which included a combination of commercial; city, state, and federal government;
academic research; and wastewater utility laboratories, as well as some commercial manufacturers.
Study results provided critical information on the performance of various methods and their use when
conducting laboratory analyses for wastewater surveillance studies and programs. The results are
summarized in an executive summary available on WRF's website and a journal article (WRF, 2020k;
Pecson et al„ 2021).
In addition to WRF's interlaboratory comparison, in June 2020, EPA's Office of Research and
Development (ORD) began researching and developing a method for concentrating and then quantifying
the amount of SARS-CoV-2 virus in wastewater using samples from WWTPs in Ohio (see Section 6.2.4
for details on Ohio's wastewater surveillance program). Some of ORD's research also evaluated the
ability to concentrate and detect a surrogate coronavirus called OC43 from larger sample volumes of
wastewater (McMinn et al„ 2021). EPA is continuing research on applying this approach to detect SARS-
CoV-2 and developing culture-based methods to detect SARS-CoV-2 in wastewater, similar to the
culture-based methods for detecting coliforms where bacteria grow on media. EPA has also collaborated
with Australia's Commonwealth Scientific and Industrial Research Organization on several review and
research activities on SARS-CoV-2 wastewater surveillance ranging from sampling design (Ahmed et al„
2021), stability of molecular targets (Ahmed et al„ 2020a), and recoveries using surrogate coronavirus
(Ahmed et al„ 2020b). All these research activities further support the refinement of SARS-CoV-2
wastewater methods, with a long-term goal of developing "sensitive, standardized methods to detect
and quantify SARS-CoV-2 in raw sewage, including infectious virus" (EPA, 2021).
Other laboratory quantification methods have since evolved to allow experts and nonexperts alike to
quickly extract viral RNA from wastewater and quantify viral RNA in the field rather than in a
laboratory setting, potentially offering same-day SARS-CoV-2 wastewater results. For example,
LuminUltra—in collaboration with Halifax Water and Dalhousie University Halifax, Nova Scotia—
created a portable and rapid SARS-CoV-2 wastewater testing solution. This new technology is described
by the company as being able to run multiple samples within 90 minutes, without the need for extensive
laboratory expertise (LuminUltra, 2020; LuminUltra, 2021). LuminUltra's quantification kit and others,
such as those developed by BioRad, QIAGEN, and Zymo, have been developed to support cost-effective
and rapid expansion of wastewater testing methods for SARS-CoV-2 (Bio-Rad, 2021; Qiagen, 2021;
Zymo, 2021).
In early 2021, the research community also began exploring methods for detecting and quantifying
SARS-CoV-2 variants (e.g., the United Kingdom variant, B. 1.1.7 [Alpha]; the South African variant,
B. 1.351 [Beta]). This work is being done through genomic sequencing of wastewater samples and the
development of primers and probes capable of detecting individual mutations that characterize specific
variants via RT-qPCR. Some examples are Rice University sequencing the whole SARS-CoV-2 genome
(see Houston Case Study in Section 6.2.7) and Clemson University quantifying the B. 1. 1.7 in the
wastewater (see Clemson University Case Study in Section 6.2.9) (Houston, 2021; Clemson, 2021b).
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5.3 Ongoing Wastewater Surveillance Support
As the field of SARS-CoV-2 wastewater surveillance continues to grow, organizations have developed
and shared best practices and guidance with their stakeholders. The organizations include large
international groups, such as the World Health Organization (WHO) that published recommendations
on the ethics of wastewater surveillance, to smaller nonprofits such as WEF that published safety
information for WWTP workers. This section describes the work of organizations like these that have
or are providing ongoing support for wastewater surveillance.
Various federal agencies have contributed to the field of SARS-CoV-2 wastewater surveillance on a
national level. For example, and as described in Section 4, CDC and HHS, in collaboration with other
federal agencies, established the NWSS in September 2020 to, in part, provide support and coordination
for wastewater surveillance of SARS-CoV-2. The NWSS website offers technical support for sampling
methods (e.g., location and frequency), laboratory methods, and public health interpretation of
wastewater testing data (CDC, 2021a). The NWSS DCIPHER platform is a national database of
wastewater testing results for SARS-CoV-2 collected by state, tribal, local, and territorial health
departments in coordination with wastewater utilities. Data are submitted to NWSS through the
DCIPHER platform by state, tribal, local, and territorial health departments using a standard collection
instrument that defines the minimum set of reporting guidelines (CDC, 2021a). CDC analyzes the data
submitted to the system in real time and reports results to health departments via the DCIPHER data
reporting analytics dashboard (CDC, 2021a).
The NWSS DCIPHER analytics system corrects wastewater testing results for laboratory method
performance, as well as the composition of wastewater and the number of people living in the
community under surveillance—ensuring comparability across labs, across communities, and through
time (WRF, 2021a). The data reporting dashboard presents maps of WWTPs sewersheds under
surveillance, time series plots of wastewater results, and information on SARS-CoV-2 trend status (e.g.,
sustained increase, increasing, plateaued, decreasing, sustained decrease) at each sampling location for
use by public health departments (WRF, 2021a). A screen shot of part of the data reporting dashboard
is shown in Figure 4 (CDC, 2021 g). As of the end of August 2021, SARS-CoV-2 wastewater results for
almost 20,000 samples have been submitted to NWSS (see Figure 5) (CDC, 2021 g).
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29
ANALYTICS DASHBOARD
NATIONAL WASTEWATER SURVEILLANCE SYSTEM
ALERTS LIST
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O STEWERSHED TREND CLASSIFICATION MAP © WASTEWATER EPI CURVES
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| UNKNOWN | SUSTAINED INCREASE INCREASE PLATEAU DECREASE | SUSTAINED DECREASE
TRENDS CLASSIFICATION GRID
Sustained Increase
14
(37%)
Increase
0
(0%)
Plateau
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0
(0%)
Sustained Decrease
(3%)
Expand Collapse
16,000
-5. 12,000
E
CO
tn
8,000
4,000
0
January March May July
Figure 5. Cumulative samples in the NWSS DCIPHER system (CDC, 2021g).
Figure 4. CDC's NWSS DCIPHER analytics dashboard for SARS-CoV-2 wastewater results
(CDC, 2021g).
20,000
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30
As mentioned in Section 4.4, HHS (with support from CDC) funded wastewater testing from WWTPs
serving 10 percent of the U.S. population (estimated at 100 WWTPs) in late 2020 for a period of six
weeks. HHS is continuing to support this nationwide pilot study to test the scalability of an early
warning system and public health response framework through a second phase that is anticipated to
cover 30 percent of the U.S. population. The results of this work are especially relevant in smaller
communities with the potential for underfunded individual testing programs that may be underreporting
cases of COVID-19 (CNBC, 2020).
As mentioned in Section 4.5, WRF funded a research project to develop specific implementation and
sample design recommendations at different types of sewersheds: large urban sewersheds, medium-sized
regional sewersheds, and small regional systems (WRF, 2020h).
On a more local level, multiple organizations have and continue to provide general guidance about
sampling and analysis methods to support programs. For example, in June of 2020, the National
Association of Clean Water Agencies (NACWA) released a guidance document for utilities to aid in the
development of wastewater surveillance programs for SARS-CoV-2. The document is based on lessons
learned from NACWA member utilities who were already sampling for SARS-CoV-2 in wastewater.
NACWA's document includes a recommendation for utilities to engage and work with their local
and/or state health departments for the wastewater surveillance program. NACWA prepared this
document in response to concerns that SARS-CoV-2 wastewater surveillance and communication of
results are outside the purview of a wastewater utility; however, utilities are receiving requests to
conduct surveillance from community members, private companies, and other organizations. Utilities
were also concerned about assessing costs and data validation since there is no consistent analytical
method. NACWA indicated that the resource is a "living document" and will be updated as guidance
and information evolves (NACWA, 2020).
As another example, the Environmental Research Institute of the States and ASTHO, under a
Memorandum of Agreement with EPA ORD, published an issue brief on the detection of SARS-CoV-2 in
wastewater in November 2020. This brief summarizes early studies conducted by states, identifies
research gaps, outlines common practices for collecting wastewater samples (e.g., sample location, type,
and frequency; documentation; analytical methods; reporting), and provides recommendations to
scientists and public health and environmental officials moving forward (ERIS and ASTHO, 2020).
Based on lessons learned at their April 2020 International Water Research Summit, WRF has shared
multiple resources for surveillance programs. One such document is a detailed summary of
recommendations from global experts that participated in the International Water Research Summit,
covering "potential uses of wastewater surveillance for tracking COVID-19, sampling design, analytical
tools, and communication of results to public health decision-makers, the public, and other key
stakeholders" (WRF, 2020j). WRF also published an appendix on best practices for sample collection
and storage and developed a field sample collection form that outlines the information that should be
recorded during sample collection (e.g., wastewater flow rate, type of sewer system, air temperature,
sample water characteristics such as temperature, total suspended solids, chlorine residual, pH) (WRF,
20201). The field collection form also provides space to record information about sample storage and
processing at the laboratory (e.g., temperature of sample upon receipt at the laboratory). WRF's
website includes links to their press releases on current projects and funding opportunities (WRF,
2021b).
5.4 Inclusion of Rural and Underserved Populations
As wastewater surveillance efforts expanded across the country, federal, state, tribal, and local agencies
and other non-governmental stakeholders have recognized the importance of selecting sites with
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31
consideration for potential vulnerabilities of the community (e.g., at-risk populations, rural areas,
communities underserved by health care systems). Ohio's COVID-19 Populations Needs Assessment
acknowledged that COVID-19 would likely disproportionally impact people of color, rural populations,
and individuals with disabilities since these groups already deal with less access to healthcare and more
negative social determinants of health than others (Nemeth et al„ 2020). Wastewater surveillance can
provide insight into COVID-19 spread without biases that may be associated with individual testing, such
as race, economic status, age, testing site hours and locations, and many other factors.
For example, NEHA published an article outlining the benefits and best practices of wastewater
surveillance in rural areas. The article, "Sewage Monitoring in Rural Communities: A Powerful Strategy
for COVID-19 Surveillance," explains how wastewater surveillance is a strong supplement to individual
testing programs in these areas because rural and underserved areas face challenges for detection and
management of the outbreak. As an example, individual testing may not be feasible or cost-effective in
these areas, and wastewater surveillance can provide greater coverage and broader detection in smaller
towns. However, the article notes that wastewater surveillance in rural communities may be challenging
due to the number of people (estimated at 20 percent) that live in areas without WWTPs and instead
use septic systems. Additionally, rural areas may not have trained individuals to conduct the sampling
and perform the laboratory analyses, so the article suggests rural wastewater surveillance will require
coordination between WWTPs, laboratories (commercial and academic), and public health departments
(NEHA, 2020).
One thing to consider when establishing wastewater surveillance programs is the number of people
contributing to the wastewater. If the population is too small, robust individual testing and contact
tracing programs may be more effective at evaluating the spread of COVID-19 than wastewater
surveillance. However, if individual testing and contract tracing are not available, then wastewater
surveillance may be useful for public health officials.
Multiple case study programs considered vulnerable and underserved communities when creating or
executing the sampling program. The Ohio wastewater surveillance program prioritized sampling at
WWTPs that service large and/or vulnerable populations. The previously mentioned vulnerability report
that ODH and Ohio State University developed strongly guided Ohio's sampling site selection (Ohio,
2021; Nemeth et al„ 2020). Other programs used community relations to institute public health
measures more effectively within vulnerable populations. Tempe, Arizona partnered with Guadalupe,
Arizona, to monitor the Guadalupe's wastewater, primarily comprised of low-income and minority
families. The Mayor and City Council of Guadalupe used the wastewater results to enforce COVID-19
public health messaging and guide future decision-making. Tempe also created a multi-lingual door-to-
door education campaign about COVID-19 and how to protect oneself while distributing masks and
care packages for use for vulnerable populations within Tempe (Tempe, 2021 d). The Houston Health
Department (HHD) used the wastewater results to identify zip codes most in need of public health
interventions and used leaders within the local communities to connect with residents. HHD'S
community partnership was to conduct COVID-19 prevention outreach, such as scheduling
appointments and organizing secure transportation to testing and vaccination sites more effectively to
vulnerable and difficult to reach areas (Houston, 2021).
5.5 Ethical and Legal Considerations
Beyond guidance for sampling and methods, the wastewater surveillance community has considered and
developed guidance on related ethical concerns. The Canadian Water Network (CWN) convened
experts in a Public Health Advisory Group to discuss public health application and communication and
the ethical issues posed by wastewater monitoring (CWN, 2020a). The Canadian Water Network also
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32
released ethics and communication guidance in September 2020, applying WHO's guidelines on ethical
issues in public health surveillance (WHO, 2017) to wastewater-based SARS-CoV-2 data (CWN,
2020b). WHO released a document in August 2020 outlining the uses of wastewater surveillance
programs and considerations when creating a program, including ethical and legal considerations. WHO
specifically addressed concerns about potential stigmatization of the community but also noted that
wastewater surveillance would be less likely than individual testing to be stigmatizing due to the
wastewater characteristic as a pooled sample. WHO also acknowledged that these concerns may be
heightened because wastewater surveillance is often done without community consent. WHO cautioned
programs to avoid disproportionately targeting already-stigmatized communities with public health and
social measures (WHO, 2020). Researchers from multiple universities also published an academic paper
on the legal and ethical considerations with wastewater surveillance (Gable et al„ 2020).
5.6 Worker Safety
Guidance has also been developed regarding utility worker safety when supporting wastewater
surveillance efforts for SARS-CoV-2. For example, WEF compiled information about the viability of
SARS-CoV-2 in wastewater and the related safety concerns for workers at the WWTP and throughout
the collection system into a web-based "Water Professional's Guide to COVID-19" (WEF, 2020a). This
provides resources for safety based on the hierarchy of controls, the most effective control solutions
for the most employees, and guidance from the Occupational Safety and Health Administration (WEF,
2020a). WEF also published a document titled "Protecting Wastewater Professionals from COVID-19
and Other Biological Hazards" based on the findings of an expert panel of academics and practitioners
(WEF, 2020b). The document summarizes the most current, evidence-based information on protecting
worker health and safety with respect to exposure to biological hazards associated with wastewater
from agencies and authorities external to WEF, as well as the peer-reviewed literature. Occupational
Safety and Health Administration also has recommended guidance for solid waste and wastewater
management workers and employers during the COVID-19 pandemic (OSHA, 2020).
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6 Implementation of Surveillance
Programs
As wastewater surveillance for SARS-CoV-2 gained traction in the United States, many state and local
agencies, utilities, and universities, along with a few tribal agencies, developed and implemented
monitoring programs. Section 6.1 briefly summarizes publicly available information on programs initiated
by 14 states and 161 tribes, municipalities, and universities. Details are provided for each program in
Appendix A. Section 6.2 provides additional information for a subset often diverse programs, gathered
through open-ended interviews with program leads.
This discussion is not meant to serve as a comprehensive summary of all wastewater surveillance efforts
across the country. EPA identified the programs in this document in January and February of 2021
through publicly available resources, such as the COVIDPoopsl9 dashboard (see Section 5.1), and by
searching the internet for program websites, data dashboards, research papers, reports, news articles,
and press releases that provided general information on SARS-CoV-2 monitoring programs (UC
Merced, 2021). EPA included programs based on the amount of publicly available information found at
the time. EPA recognizes that there are many other ongoing wastewater surveillance programs for
SARS-CoV-2 at both the state and local level, including tribes. Refer to Section 3 for additional
information on data collection methods.
6.1 Overview of Surveillance Programs
Most early monitoring efforts for SARS-CoV-2 in wastewater focused on wastewater influent (i.e.,
untreated sewage that enters a WWTP). As wastewater monitoring gained traction throughout the
pandemic, however, programs expanded to collect wastewater from sewer manholes, sewer lines, lift
stations, and sewer cleanouts to target smaller isolated communities or specific facility populations (e.g.,
correctional facilities, long-term care facilities, K-12 schools). Universities also developed programs to
monitor SARS-CoV-2 in sewage from student housing on campus, and in some cases, within the
surrounding community. Universities were either able to quickly initiate programs by leveraging
longstanding research programs that were already in place involving pathogens in wastewater (e.g.,
Michigan State University) or by pivoting from previous research and adapting their expertise to
wastewater monitoring (e.g., Clemson University) (Michigan, 2021a; Clemson, 2021b). The Berkeley
Water Center at the University of California Berkeley summarized the experience of 25 college and
university wastewater surveillance programs in the United States from the Fall 2020 semester (Harris-
Lovett et al„ 2021). Many universities also provided laboratory support and technical expertise to local
and state agencies (e.g., Rice University), as well as individual utilities (e.g., University of Arizona)
(Houston, 2021; University of Arizona, 2021).
Wastewater surveillance programs have been implemented at the national, state, tribal, and local level
using both commercial and academic research laboratories. At the national level, for example, Biobot
Analytics initiated a nationwide pro bono sampling and analysis program in April 2020, and HHS started
wastewater testing at 100 WWTPs across the country for a period of six weeks beginning in November
2020 (see Section 4) (BioBot, 2020; FPDS, 2021). HHS, with support from CDC, has also established a
nationwide wastewater surveillance program evaluating SARS-CoV-2 in WWTP influent (see Section 4.4
for details). At the state level, many state health and environment departments have developed
statewide surveillance programs (e.g., Ohio) or funded them on a more local level (e.g., Michigan) (Ohio
DOH, 2020; Michigan, 2021b). Local agencies and individual utilities have initiated monitoring programs
as well, typically with funds from local government or ratepayers. In some cases, states and local
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government have worked together to execute programs (e.g., Vermont) (Burlington, 2021). In other
cases, states worked directly with utilities (e.g., Indiana) (Indiana, 2020). Some wastewater surveillance
programs receive support from consulting firms. For example, the wastewater surveillance program in
Bergen County, New Jersey, is supported by AECOM and Columbia University (AECOM, 2020), while
CDM Smith is supporting the U.S. Department of Veterans Affairs with their wastewater surveillance
pilot project (CDM Smith, 2020).
Methods employed in early programs vary greatly with respect to sample location, sample type,
frequency and time of collection, laboratory processing and analysis, and data interpretation. Regarding
sample collection, the number of sampling locations varies from small, localized programs with less than
ten unique sampling locations (e.g., Boise, Idaho; Burlington, Vermont) to larger statewide programs
with upwards of 300 locations (e.g., Michigan) (Boise, 2021; Burlington, 2021; Michigan, 2021a). Sampling
frequency similarly varies across programs, with most programs collecting samples once or twice weekly
and a few collecting samples daily.
Sample Collection Types
Wastewater samples are collected as either composite
samples or one-time grab samples. While composite
samples are often considered more representative of Grab samPles consist of a sin§le discrete
community fecal contributions, grab samples are samPle that is representative of the
sometimes advantageous since they can be collected wastewater at the time of collection,
quickly and at a much lower cost (CDC, 2021 e). Composite samples are collected over time
Composite samples are most frequently collected at the to represent the average wastewater over
influent of WWTPs, while grab samples are collected that time. A common composite sampling
when access is limited (e.g., no electric hookup) or there approach is to collect a set volume every
are security concerns at a given sampling location. In 15 minutes for 24-hours using a
general, grab samples are collected in the morning hours programmed automatic composite sampler
when a greater percentage of the population is assumed (EPA, 201 3).
to use the restroom, as indicated by the daily peak in
wastewater flow. During the early months of the pandemic, automatic composite samplers were in
limited supply and many programs had no choice but to collect grab samples. Depending on the
program, samples are collected by utility staff, private contractors, or students.
State and local programs typically send wastewater samples to commercial or university laboratories for
analysis, while most universities analyze their program samples in on-campus laboratories. In some cases,
programs send samples to multiple laboratories as part of an interlaboratory comparison to validate and
improve analytical methods. Several programs started by using a commercial laboratory and then later
switched to a local university or state laboratory to decrease costs and reduce turnaround time
between sample collection and receiving the results (e.g., Boise, Idaho; Houston, Texas) (Boise, 2021;
Houston, 2021). Most programs analyze their wastewater samples with RT-qPCR, while a smaller
percentage rely on the RT-ddPCR method. A handful of programs use both technologies. Programs
typically analyze samples for both the NI and N2 gene targets of SARS-CoV-2, while several also analyze
fecal indicator organisms such as the pepper mild mottle virus.
Many state, local, and university wastewater surveillance programs communicate their results to the
public in real time through the use of online data dashboards. Several have also produced more detailed
summary reports (e.g., Indiana) (IFA, 2020) or published their methods and preliminary results in peer-
reviewed journals (e.g., HRSD in Virginia) (Indiana, 2020; Gonzales et al„ 2020).7 Within these
resources, programs present the concentration of the virus measured in wastewater (e.g., "genome
7 As of February 2021, many articles on this topic are only available in preprint version and have therefore not yet
been certified by a formal peer review process.
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35
copies/liter [L]"), normalized virus concentrations (e.g., normalized by population size, wastewater flow),
or both concentrations. Normalized concentrations are used to account for changes in wastewater
dilution (e.g., due to water other than wastewater entering the sewer system) and differences in relative
human waste input over time (CDC, 2021 e).
One common design for public dashboards used by multiple jurisdictions is to have a map showing the
location of the WWTPs that are sampled along with geographic boundaries of the collection system
served by each WWTP. Users can then click a specific sampling location to view a line graph of virus
concentrations measured in wastewater through time. Some dashboards also present individual COVID-
19 testing results, either as a separate graph (e.g., Utah, Wisconsin, Colorado) or overlaid with the
wastewater data (e.g., Ohio) (Utah DEQ, 2021; Wisconsin DOHS, 2020; Colorado DPH, 2021; Ohio
DOH, 2020). This allows users to compare trends between SARS-CoV-2 wastewater results and
COVID-19 case counts based on individual testing.
Boise, Idaho, is one example of a city with a public dashboard that displays wastewater testing results
along with individual COVID-19 testing results. Boise has been sampling and testing influent to its two
WWTPs since May 2020. Boise presents the average of the daily SARS-CoV-2 wastewater results from
the two WWTPs (weighted based on the WWTP's flow) on their data dashboard, as presented in
Figure 6. The top graph shows the wastewater results in virus copies/L and the bottom graph shows
new cases of COVID-19 over the same timeframe for the entire county (Boise, 2021).
In some cases, dashboards indicate recent trends in wastewater results either with descriptive text or
with a specific symbol or color on their sampling location maps. One example is the Utah SARS-CoV-2
Sewage Monitoring Program, which collects samples weekly at 42 WWTPs representing approximately
80 percent of the state's population. Utah's dashboard includes a map depicting each WWTP's service
area with color-coded symbols to indicate the recent sewage trend based on the virus concentrations
from the four most recent samples (see Figure 7). Utah combines wastewater flow and service area
population data to estimate viral concentrations in units of SARS-CoV-2 gene copies per person, per
day, for each WWTP, shown in the top graph. Utah's dashboard also shows the daily new cases per
100,000 residents for each WWTP sewershed, shown in the bottom graph (Utah DEQ, 2021).
Various other state, local, and university programs have started to define trends in wastewater data,
albeit using different methods. The Wisconsin COVID-19 Wastewater Surveillance dashboard shows a
map of the WWTP service areas along with wastewater SARS-CoV-2 results presented in gene copies
per person, per day, along with the seven-day average COVID-19 case rate for the sewershed for each
of the 65 participating WWTPs (as of April 2021). Wisconsin defines trends in SARS-CoV-2
concentrations and individual COVID-19 testing results using a linear regression model and the most
recent two weeks of data. Increasing trends are when the change from the prior seven-day period to
the most recent seven-day period is greater than or equal to 10 percent and statistically significant
(Wisconsin DOHS, 2021). The Ohio Coronavirus Wastewater Monitoring Network dashboard
compares the average of the most recent two samples with the average of the prior third and fourth
sample to determine the percent change between those averages; less than 50 percent change is defined
as stable, 50 to 100 percent change is defined as increasing or decreasing, and more than 100 percent
change is defined as substantially increasing or decreasing (Ohio DOH, 2021). The Missouri Sewershed
Surveillance Project's COVID-19 Tracking Tool uses a story map to summarize wastewater data from
more than 50 participating WWTPs. Missouri presents trends in the SARS-CoV-2 wastewater results
over the past two weeks using map symbology (see Figure 8) (Missouri DHSS, 2021).
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Wastewater Test Results (virus copies per liter)
Ada County New Cases (confirmed and probable)
Jul 2020 Sep 2020 Nov 2020 Jan 2021 Mar 2021
Figure 6. Boise Wastewater Surveillance Dashboard with wastewater SARS-CoV-2 virus copies per liter on the top graph and
confirmed and probable COVID-19 cases on the bottom graph (Boise, 2021).
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Salt Lake CityWRF
Estimated population served: 209,645
Monitoring results and case counts Sewage non-detections plotted at 1. MGC = million gene copies. Sewersheds with 1-5 cases plotted at rate of2.
~ All data Past month Lo8 y-axes J 3
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Figure 7. Utah SARS-CoV-Waste water Surveillance Dashboard with the service are and recent wastewater trend on the left,
SARS-CoV-2 million gene copies per person, per day on the top right graph, and daily new cases per 100,000 residents on the
bottom right graph (Utah DEQ, 2021).
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38
Peoria
Increasing (triangle): Viral load either
increased by 40% or more from last week,
or increased by 25% or more for the last
two weeks.
No Change (circle): Viral load did not
change or the changes did not meet the
criteria for Increasing or Decreasing status.
Decreasing (inverted triangle): Viral load
decreased by the three consecutive >25%
decreases or >30% decreases in two of the
previous three weeks.
At locations symbolized with larger
symbols, viral load may not have
changed significantly over the past two
weeks but is high compared to previous
measurements in that location.
Trends are based on EWMA of number of
viral shedding in fences may occur for up
to 30 days after infection.
Trends may be unavailable if there is
insufficient data to perform calculations.
These will be shown in Gray (square).
Figure 8. Missouri Department of Health and Human Services Wastewater Surveillance Dashboard with color coded symbols
to indicate recent SARS-CoV-2 wastewater trends (Missouri DHSS, 2021).
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39
Due to the recent emergence of SARS-CoV-2, there is no standardized way to predict the number of
infected individuals reliably and accurately from wastewater sample results. However, some programs
have used wastewater data to make predictions about the number of people with COVID-19 based on
the SARS-CoV-2 wastewater results. For example, Yale University collects and analyzes daily
wastewater samples for SARS-CoV-2 from seven WWTPs across Connecticut, covering nearly one
million residents (Yale University, 2021 b). For each participating WWTP, Yale graphs the wastewater
SARS-CoV-2 results over time along with a seven-day moving average. Yale also graphs the available
daily COVID-19 cases. For the most recent approximately two weeks Yale predicts the daily COVID-19
cases using the wastewater results. Yale replaces the predicted data with individual COVID-19 testing
results as that information becomes available. Figure 9 presents an example of the graphs Yale generates
for the Hartford South Meadows WWTP (Yale University, 2021a). Another example is summarized in
the Wyoming case study in Section 6.2.5.
In some cases, wastewater surveillance programs have tied trends in wastewater data to events that
occurred in the local communities where the wastewater samples are collected. For example, the
University of Arizona consistently observed spikes in SARS-CoV-2 wastewater results collected from
the WWTP a week after major events and holidays; when this occurred, they often saw similar spikes in
individual COVID-19 cases the following week. The University of Arizona was also able to track the
effectiveness of public health interventions through the wastewater sampling. For example, one week
after Arizona issued the stay-at-home policy, the SARS-CoV-2 wastewater results decreased, with the
individual COVID-19 cases decreasing the subsequent week (University of Arizona, 2021).
Some public-facing dashboards indicate events, like these, in an effort to help explain trends observed in
SARS-CoV-2 measurements from wastewater. For example, the dashboard for Athens-Clarke County,
Georgia, indicates significant events on the timeseries plot with vertical dashed lines. These dashed lines
refer to events such as the day that Athens-Clarke County's mask mandate was put into place, the first
day of the fall semester at the University of Georgia, and the day of Georgia's broad reopening (see
Figure 10) (Lott et al„ 2020).
In general, wastewater surveillance programs have been initiated to complement individual testing or to
support more comprehensive surveillance for COVID-19 in areas with limited resources. Documented
public health decisions triggered by or directly linked to wastewater monitoring results range from mask
mandates to canvassing neighborhoods with educational materials. In some cases, wastewater data have
served as an early warning of a potential COVID-19 outbreak, providing information ahead of individual
testing data and allowing leadership to quickly intervene (e.g., increase individual testing to quarantine
infected individuals, initiate targeted educational outreach efforts). This is most often seen at the local-
or facility-1 eve I, where wastewater data represent smaller segments of the population (e.g., sub-
sewershed monitoring) or a specific facility (e.g., correctional facility, university dorm). For example, in
September 2020, the University of San Diego detected SARS-CoV-2 in wastewater from the Revelle
College area. Within 14 hours of these results and out of an abundance of caution, the university sent
targeted messages to members of the campus community to notify them of the results and to encourage
them to get tested as soon as possible and monitor themselves for symptoms (UC San Diego, 2020).
The university immediately expanded individual testing to enable campus employees and students to get
tested over the following two days. The university tested more than 650 individuals and ultimately
identified two positive cases. These individuals immediately self-isolated, potentially preventing a larger
outbreak (Clark, 2020). Additional examples are provided in the case studies included in Section 6.2.
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40
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41
Date
~ 95% Confidence Interval ^Total Viral Copies (predicted) Observed Total Viral Copies
Figure 10. Athens-Clark County Wastewater Surveillance Dashboard for one of the WWTPs (Lott et al., 2020).
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42
6.2 Wastewater Surveillance Case Studies
EPA conducted a series of interviews with a subset of practitioners and researchers leading wastewater
monitoring efforts at the state and local level, as well as on university campuses. For this, and as
described in Section 3, EPA focused on ten programs that demonstrate the wide variety of ways that
programs were financed and implemented. The goal of this effort was to learn more about the diverse
history and use of wastewater surveillance for SARS-CoV-2 among participants, including what
prompted each initiative, initial and ongoing funding mechanisms, collaboration and organization with
other entities, and sample collection and laboratory analytical methods. EPA also gathered insight on
how wastewater data have been used to support decision-making and critical lessons learned along the
way. EPA interviewed technical leads for the following wastewater surveillance programs listed in Table
5. These programs are also included in Appendix A summarizing the collaborations, sampling details,
methods, and dashboards.
Table 5. Summary of case study wastewater surveillance programs.
Wastewater Surveillance
Program
Unique Aspects of the Program Highlighted in the Case Study
Indiana
The program was led by the Indiana Finance Authority (IFA).
Participating utilities conducted sample collection at individual facilities
and WWTPs. The program focused on communities with
universities/colleges due to the transient nature of student populations
and published a report documenting their decision-making process. See
details in Section 6.2.1.
Michigan
The statewide program included 20 different projects led by various
universities and utilities. It was built off the existing state laboratory
network and workflow used to support the beach water monitoring
program. See details in Section 6.2.2.
tate Programs
New Mexico
The program focuses sampling efforts entirely at individual facilities (e.g.,
correctional facilities, youth shelters). At some facilities, samples are
collected at multiple locations that divide the facility into smaller
resident populations. The state developed a public-facing dashboard
summarizing results and published a press release highlighting a success
story at a facility. See details in Section 6.2.3.
V)
Ohio
The program involves large-scale collaboration between multiple state
agencies, U.S. EPA ORD, and numerous universities. WWTPs are
sampled throughout the state. The program involves an analytical
methods group—which includes eight laboratories—to foster SARS-
CoV-2 methods development. The state developed a public-facing
dashboard that depicts trends in wastewater results. See details in
Section 6.2.4.
Wyoming
The state provided financial incentives to participating utilities. A public-
facing dashboard shows wastewater results as well as modeled
predictions for the percent of the population infected with COVID-19,
based on wastewater data. See details in Section 6.2.5.
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Table 5. Summary of case study wastewater surveillance programs.
Wastewater Surveillance
Program
Unique Aspects of the Program Highlighted in the Case Study
l/l
Hampton Roads
Sanitation District in
Hampton Roads, Virginia
The utility collects samples in the local community and analyzes them for
SARS-CoV-2 at their own laboratory. The utility also analyzes
wastewater samples from other utilities and organizations throughout
the state and has documented their approach in a peer-reviewed journal
article. See details in Section 6.2.6.
Local Program
Houston, Texas
The city collects wastewater samples from all WWTPs within the city, as
well as at individual facilities and other locations within the sewershed.
The city uses the wastewater data, along with other data sources such as
COVID-19 individual testing results and vaccination rates, to identify
"hot spots" for targeted public health intervention. Members of the local
community support outreach efforts. See details in Section 6.2.7.
Tempe, Arizona
The city quickly developed and implemented a wastewater surveillance
program for SARS-CoV-2 by building off its existing opioid wastewater
monitoring program. The city compares local events to the wastewater
results. See details in Section 6.2.8.
'rograms
Clemson University in
Clemson, South Carolina
The university categorizes wastewater results by "impact level" on a
public-feeing dashboard. This dashboard also includes results for variants.
The university uses the wastewater data to support its COVID-19
response and the city of Clemson used the wastewater results to
support its mask mandate. See details in Section 6.2.9.
hi-
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University of Arizona in
Tucson, Arizona
The university first analyzed samples for SARS-CoV-2 from utilities
across the country and then began analyzing samples collected on
campus. The university has developed action levels for their campus
wastewater surveillance program and used the wastewater data to
prevent an outbreak in a dorm—a story that garnered national attention.
See details in Section 6.2.10.
6.2.1 Indiana
The I FA recognized the possibility of accessing CARES Act funding to develop a wastewater surveillance
program for SARS-CoV-2. I FA made a request to the state and was subsequently provided the CARES
funding. IFA is a state agency that oversees Indiana's debt issuance and provides efficient and effective
financing solutions for state, local, and business interests (IFA, 2021). In addition, the IFA manages the
state's federally funded State Revolving Fund Loan Programs. As a result, IFA already had mechanisms in
place to distribute the CARES Act funding in support of the wastewater surveillance program, as
opposed to the Indiana State Department of Health or the Indiana Department of Environmental
Management. Indiana established a wastewater surveillance program for SARS-CoV-2 from August to
December of 2020 in 14 communities across the state. IFA chose to focus exclusively on communities
with college and university campuses due to the transient nature of student populations and the
potential increased spread of infectious diseases (Indiana, 2021).
IFA selected l20Water to manage the initiative and work directly with participating utilities (i.e., eligible
utilities included those that served public or private universities with at least 1,000 enrolled students and
student housing; all eligible utilities were invited to participate). l20Water hosted virtual training
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44
sessions on sample collection procedures, sent weekly sampling kits, coordinated between participating
labs, leveraged technology for data collection and results management and collaboratively developed a
final report. Utility partners collected their own samples and saw themselves as "stewards of the data",
which many shared with public health decision-makers. Utilities identified sampling locations within the
university collection systems (e.g., influent to WWTPs on-campus, manholes or lift stations that
represent specific dormitory populations, Greek housing) and then collected the samples. Some utilities
also chose to collect samples at other locations in the community (e.g., off-campus housing apartment
complexes, correctional facilities, long-term care facilities). IFA and l20Water encouraged utilities to
collect wastewater samples at additional locations that would provide useful information to their
communities. In return for their participation, utilities received sample test kits, site selection guidance,
and expert review and interpretation of the analytical results (Indiana, 2021).
For this program, utilities sent the wastewater samples to either Microbac Laboratories or the
University of Notre Dame for SARS-CoV-2 analysis. Indiana included two laboratories to explore and
better understand the precision of the novel analytical methods being used (Indiana, 2021). Utilities sent
portions of the same wastewater sample to both laboratories in support of an interlaboratory
comparison study. Microbac Laboratories analyzed samples via RT-PCR while the University of Notre
Dame used RT-ddPCR. Staff from the two laboratories met weekly to discuss the results and identify
ways to further refine and improve their methods, fostering a strong partnership. In general, wastewater
results were comparable across laboratories (Indiana, 2020).
After sending samples to the laboratories, utilities received a weekly summary report that included
graphs of wastewater results through time along with other relevant information (see Figure I I). In
some cases, utilities communicated these results to and worked directly with local health departments
(Indiana, 2021).
IFA released a public report on the program in late 2020 that summarized the purpose of the
wastewater surveillance, the interlaboratory study, and trends in wastewater results (IFA, 2020). The
report also included a detailed analysis of temporal trends in SARS-CoV-2 in wastewater compared to
temporal trends in COVID-19 cases at both the community and campus level (see Figure 12). The
report notes that temporal trends in wastewater mirrored trends in individual COVID-19 testing results
for most communities, but that the trends appeared to occur at the same time (Indiana, 2020). In
general, the wastewater data were not found to serve as a leading indicator of new cases for the Indiana
program due to extended sample turnaround time and difficult-to-interpret fluctuations in the results.
However, the wastewater results offered useful complementary information to other health data on
COVID-19 (e.g., individual testing results) (Indiana, 2021).
The project's overall goal was to provide university communities with a tool for and knowledge about
identifying COVID-19 outbreak risks, and to evaluate ways that communities could utilize wastewater
surveillance in the future. Utilities were eager to contribute to this area of research, though they were
unsure how to interpret results accurately (Indiana, 2021).
Although Indiana has used all the CARES Act funding allocated to this program, IFA sees wastewater
surveillance as a valuable tool for the future. IFA is exploring other funding mechanisms for new
surveillance opportunities (Indiana, 2021).
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SARS-CoV-2 observed concentration
(gene copies/1 OOmL) by site, Angola
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Angola Vs Steuben in red
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Figure 12. Plots of viral gene copies (GC) per mlL in Indiana communities (Indiana, 2020).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
47
6.2.2 Michigan
The Michigan wastewater surveillance program, which is led by the Michigan Department of Health and
Human Services (MDHHS) and the Michigan Department of Environment, Great Lakes, and Energy
(EGLE), began in September 2020 and ended February 2021, To quickly stand up a wastewater testing
pilot project, EGLE relied on their established sampling program and laboratory network used to
support the beach water monitoring program. As part of the beach program, EGLE had previously
provided $500,000 to the laboratory network, led by Michigan State University, to procure RT-qPCR
machines that the laboratories used to establish RT-qPCR analytical methods for E. coll As a result,
these laboratories were able to easily transition to measuring SARS-CoV-2 in wastewater (Michigan,
2021a).
Michigan funded the SARS-CoV-2 wastewater surveillance pilot project using CARES Act funding.
MDHHS received CARES Act funding and provided these funds to EGLE for the statewide wastewater
surveillance pilot project. EGLE used their existing grant process to distribute funding for local
wastewater surveillance programs throughout the state. The grant application required a partnership
with a wastewater utility and local health department, and ideally a laboratory to analyze the wastewater
samples (Michigan, 2021 b). EGLE distributed $10 million in grants to 20 projects that included over 270
sampling locations and community-scale samples from WWTPs and building samples from colleges and
universities, assisted living and long-term care facilities, and K-12 schools with in-person learning (shown
in Figure 13) (Michigan, 2021a).
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sampling locations (blue circles) (Michigan EGLE, 2021).
The 19 public, private, and academic laboratories participating in Michigan's program worked collectively
to quickly develop a SARS-CoV-2 analytical method (Michigan, 2021a; Michigan, 2021 b). All laboratories
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48
purchased the PCR primers and probes identified by CDC. Due to potential supply issues with RT-
qPCR machines and supplies, most of the laboratories shifted to using RT-ddPCR. Michigan State
University lead the RT-ddPCR analytical methods development. To ensure the SARS-CoV-2 wastewater
results were consistent between laboratories, EGLE developed a Quality Assurance Project Plan that
identified the control requirements and calculation methods, such as converting the cycle threshold
(CT) score into virus copies/L (Michigan, 2021a). Michigan State University and Saginaw Valley State
University participated in the WRF's interlaboratory methods evaluation study (WRF, 2020e).
Collaboration was a strength of Michigan's wastewater surveillance program. Michigan State University
led weekly calls with the whole project team and hosted a Microsoft Teams page that included details on
methods and quality assurance procedures. Michigan State University also hosted virtual office hours for
all participants to answer questions and troubleshoot efforts (Michigan, 2021a).
Local projects provided the results to their local communities in real time to support public health
actions. For example, the University of Michigan shared the SARS-CoV-2 wastewater results with the
local health department and the campus COVID-19 response team. The campus COVID-19 response
team conducted individual testing on students living in dormitories that had high levels of the virus in
wastewater. In addition, the campus closed public locations, such as gyms, if high levels were detected
from those buildings (Michigan, 2021a). As another example, one of the Michigan project teams was
sampling wastewater from a Native American-owned casino, and the community decided to close the
casino based on a combination of the wastewater surveillance data and community testing.
The Michigan wastewater surveillance pilot project ended in February 2021 after spending all the
allocated CARES Act funding. Some of the participating sites continued wastewater surveillance using
other funding sources. As of April 2021, MDHHS is reestablishing the statewide SARS-CoV-2
wastewater surveillance program using allocated funding from CDC's ELC Emerging Detection
Enhancement award, with tentative plans to start in June 2021 (Peters, 2021).
6.2.3 New Mexico
The New Mexico wastewater surveillance program, led by the state's Environment Department, began
in late 2020. Funded in part by the CARES Act, the program tracks the SARS-CoV-2 virus in wastewater
from congregate facilities (e.g., youth shelters, detention centers). New Mexico chose to focus on these
facilities to support direct public health action and because residents of congregate facilities are at higher
risk of exposure to and illness from COVID-19. The wastewater results serve as an early indicator
that—along with personal nasal or saliva testing and other public health interventions—is used to
prevent or minimize a potential COVID-19 outbreak (NMED, 2021 b).
As of early April 2021, New Mexico sampled 17 facilities in various regions of the state once or twice a
week. Some facilities have multiple sampling points that divide the facility into smaller resident
populations and/or designate separate COVID-19 quarantine areas (NMED, 2021a). This generates
wastewater results that are more actionable for the facilities and public health professionals (NMED,
2021b).
New Mexico publishes the wastewater sampling results on a dashboard, categorizing the results by
current level of concern and current trend, similar to other program dashboards (e.g., Clemson
University) (NMED, 2021a). The current level of concern is defined as:
• less than 5,000 virus copies/L = no/minimal impact
• 5,000 to 10,000 virus copies/L = low
• 10,000 to 100,000 virus copes/L = moderate
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49
• 100,000 to 1,000,000 virus copies/L = high
• greater than 1,000,000 virus copies/L = very high
The current trend is described as no trend, increasing, or decreasing based on the three most recent
samples (NMED, 202Id).
New Mexico's wastewater surveillance program has helped multiple facilities successfully identify
COVID-19 positive residents or staff through follow-up testing. One example is at J. Paul Taylor Center,
a juvenile detention facility, where SARS-CoV-2 levels in the wastewater were consistently non-
detectable. Then, on December 23, 2020, the surveillance program detected increased SARS-CoV-2
levels in the wastewater sample and notified the facility (see Figure 14) (NMED, 2021a). In response, the
J. Paul Taylor Center conducted individual testing of all its residents and staff and found three positive
staff members, one of whom was at the facility on the day that the sample was collected. The positive
staff members quarantined off site, and the subsequent wastewater results returned to non-detectable
levels (NMED, 2021c).
Another example is from Luna County Detention Center, which has two sampling points: one on the
west side of the facility, which includes a COVID-19 quarantine area, and one on the east side of the
facility, which should not have any residents with COVID-19. The wastewater sample from the east side
was in the no/low impact level of concern until January 5, 2021, when the sample showed SARS-CoV-2
levels of over 1,000,000 virus copies/L (see Figure 15) (NMED, 2021a). Once again, the program notified
the facility, which conducted 100 percent individual testing and identified 27 residents and 10 staff
members with COVID-19. The inmates were transferred to the quarantine area on the west side of the
facility and the staff quarantined off site, resulting in decreased SARS-CoV-2 levels in the wastewater
(NMED, 2021b).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
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Figure 14. Wastewater surveillance data for the J. Paul Taylor Center (NMED, 2021a).
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Figure 15. Wastewater surveillance data for Luna County Detention Center (east side) (NMED, 2021a).
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52
6.2.4 Ohio
The Ohio wastewater surveillance program, led by the Ohio Department of Health (ODH) and Ohio
Environmental Protection Agency (Ohio EPA), began in July 2020 after Ohio Governor DeWine became
aware of the utility of wastewater surveillance and wanted to develop a program. ODH and Ohio EPA
coordinated with the Ohio Water Resources Center (Ohio WRC) to implement the surveillance
program using CARES Act funding. Ohio WRC had existing relationships with universities in Ohio, along
with mechanisms to distribute funding to project teams. The U.S. EPA ORD in Cincinnati, Ohio also
supported the program using internal funding (Ohio, 2021; Ohio DOH, 2020).
To begin the program, Ohio WRC invited researchers from about 10 universities and U.S. EPA to
participate in a statewide monitoring program kickoff. The team established four subgroups to quickly
develop every aspect of the program: a leadership group, site selection group, analytical methods group,
and statistical modeling group. The site selection group had a goal to sample influent to WWTPs in as
many counties to include as many people as possible throughout Ohio. The site selection group solicited
partnerships with utilities directly and through state organizations, prioritizing WWTPs that served large
or vulnerable populations and were willing to participate (Ohio, 2021). The site selection group used the
vulnerability report that ODH and Ohio State University developed to support Ohio's COVID-19
efforts. The vulnerability report identified populations that are at risk for disproportionate burdens of
illness and death from SARS-CoV-2 overlaid with social factors (e.g., median household income,
occupation, education level) and healthcare resources (e.g., hospitals) (Nemeth et al„ 2020). As of
March 2021, Ohio's program included 65 WWTPs throughout the state, as presented in Figure 16
(Ohio DOH, 2020).
The analytical methods group included a network of university laboratories, ORD's laboratory, and out-
of-state commercial laboratories. Each of the eight participating laboratories developed their own
analytical method to foster SARS-CoV-2 methods development. Due to the pandemic creating supply
chain issues for components of the analytical methods (e.g., buffers), some of the labs also had to adjust
their method based on availability. Once a month, the program performs an interlaboratory validation
whereby each lab receives a sample from the same WWTP for analysis. Once results are available, the
participating laboratories meet to discuss the results and troubleshoot any items. For example, none of
the laboratories pasteurize the samples due to lessons learned that pasteurization reduced the RNA
PCR signal. Ohio WRC and U.S. EPA ORD also developed quality assurance and quality control
requirements using positive spikes and human marker comparisons (Ohio, 2021). Additionally, U.S. EPA
ORD and Ohio State University participated in the WRF's interlaboratory methods evaluation study
(WRF, 2020e).
The statistical modeling group's focus was to evaluate whether the SARS-CoV-2 wastewater results
could serve as an early warning system for potential COVID-19 outbreaks. The number of samples per
week and analytical turnaround time were critical to the program's success. The program originally
began with weekly sampling, but shifted to sampling twice per week to get more frequent results for the
local health departments and enable more reliable trend analysis. The labs provide the results to the
programs within three days of sample collection (Ohio, 2021). The statistical modeling group evaluated
different trend analysis methods and modeling approaches to estimate the number of people shedding
SARS-CoV-2 in sampled areas. The statistical modeling group decided to perform linear regression of
the last ten wastewater results (normalized by flow). Ohio WRC said these linear regressions are not
yet on the dashboard as of April 2021 (Bohrerova, 2021).
Ohio WRC compiles the results from all the labs, normalizes the results by WWTP influent flow rate to
account for variations in flow rate due to rainfall, and uploads daily the results to ODH to be published
on the program's dashboard (shown in Figure 17). ODH together with Ohio WRC and other
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53
stakeholders determined thresholds based on the results for the dashboard by comparing the average of
the last two samples with the average of the prior third and fourth samples (Ohio, 2021; Ohio DOH,
2020). The thresholds are:
¦ Substantial increase in SARS-CoV-2 levels (purple) (greater than 100% increase)
¦ Increase in SARS-CoV-2 levels (red) (50% to 100% increase)
¦ Steady SARS-CoV-2 levels (gray) (49% decrease to 49% increase)
¦ Decrease in SARS-CoV-2 levels (blue) (greater than 50% decrease)
For utilities with significant increases in SARS-CoV-2 in the wastewater, ODH contacts the local health
district and provides toolkits to assist with messaging about public health measures. ODH also
coordinates with the local health districts to provide additional testing and contact tracing capabilities if
necessary (Ohio, 2021).
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54
Wastewater Treatment Plant Locations and Boundaries
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and a list of utilities in order of the trend based on the most recent results (as of April 19, 2021) (Ohio DOH, 2020).
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55
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influent flow rate (top graph) and compared to the number of COVID-19 cases from individual testing (bottom graph) (Ohio
DOH, 2020).
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56
6.2.5 Wyoming
The Wyoming wastewater surveillance program, led by the Wyoming Public Health Laboratory
(WPHL), Wyoming Department of Health (WDOH), and University of Wyoming, began in May 2020
using CARES Act funding and CDC's ELC funding. The program sampled both individual facility
wastewater and community wastewater, including WWTP influent and lift stations within the
communities' wastewater collection systems (Wyoming, 2021).
Wyoming incentivized community participation in the sampling efforts. First, Wyoming provided the
wastewater sampling kits and coordinated sample shipping for each of the participating communities. In
addition, Wyoming paid each community $300 per sample for their time and efforts and provided
bonuses to communities that were consistently sampling. Most communities sample the WWTP influent
twice a week. Next, since Wyoming required composite samples for participation in the program,
Wyoming purchased composite samplers for the communities and reimbursed up to $4,000 for each
composite sampler that communities purchased on their own. Wyoming also provided technical
assistance to communities through the Wyoming Association of Rural Water Systems. Wyoming paid
the Wyoming Association of Rural Water Systems for each community that participated in the program
to further incentivize community participation (Wyoming, 2021). As a result, the Wyoming program
currently includes 31 communities (Wyoming PHL, 2021). However, barriers to participation included
concern from some communities that the detection of SARS-CoV-2 in the wastewater could lead to
more restrictive public health measures, such as business shutdowns or gathering restrictions. Other
communities were concerned a detection of SARS-CoV-2 in the wastewater would further stigmatize
the community (Wyoming, 2021).
In addition to the WWTP influent or collection system sampling conducted by communities, the
Wyoming program currently samples from 15 community living facilities with high-risk populations.
These include Wyoming Department of Corrections facilities, Wyoming Department of Family Services
housing, and dorms at the University of Wyoming and Laramie County Community College. Wyoming
hired a contractor to collect composite samples from each facility to minimize the burden on the
facilities (Wyoming, 2021).
Initially, WPHL analyzed all the wastewater samples, but as the program grew, WDOH began
coordinating with the University of Wyoming to analyze some of the samples. The cooperation was
enhanced because WPHL had previously established relationships with University of Wyoming faculty
and had plans for other collaborative activities. WPHL and the University of Wyoming shared the same
testing methods to ensure consistency through the testing process. Additionally, certain laboratory staff
had been trained and worked at WPHL and then transferred to work at the university. WDOH then
combines the data from both laboratories into one file for further statistical analysis (Wyoming, 2021).
WDOH also models the estimated prevalence rates and uncertainty using the wastewater result as a
proxy estimate for the percent of people infected with COVID-19 in each community (i.e., not for
individual facilities) (Wyoming PHL, 2021). WDOH felt the prevalence rate was important to
communicate the results to the participating communities and the general public, since CT and
copies/mL are harder to understand (Wyoming, 2021). WDOH relied on information from a German
study on fecal shedding rates for nine individuals and then ran data simulations to develop the estimation
model (Wyoming PHL, 2021; Wolfel et al„ 2020).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Wyoming's online wastewater surveillance dashboard includes a map
of each community participating in the program. The color of the icon
corresponds to the prevalence level of the virus (see Figure 18) with
gray dots indicating communities without samples in the last 14 days.
In addition, icons on the map outlined in red indicate an increasing
prevalence level in the community. Once a user selects a community,
Wyoming's dashboard adds three additional graphs of estimated
prevalence rate, CT count (i.e., raw data from qPCR), and virus gene
copies/mL all over time (see Figure 19). Wyoming's dashboard also
includes graphs of virus gene copies/mL for each of the facility
monitoring locations.
Wyoming is comparing the community-level wastewater results to
the state individual testing results to identify potential gaps in
individual testing. As vaccines are available, Wyoming plans on using
the SARS-CoV-2 wastewater program to monitor the spread of
COVID-19 in communities with more limited access to vaccines. The
facilities are also using the wastewater results to make public health
decisions. For example, Honor Farm, a Wyoming Department of
Corrections facility, conducted individual testing of all inmates and
staff due to an increase in SARS-CoV-2 in the wastewater (Wyoming,
2021).
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ranges (Wyoming PHL,
2021).
Beyond COVID-19, Wyoming is evaluating options for future wastewater surveillance use, such as in
detection of other emerging diseases or antibiotic resistance (Wyoming, 2021).
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West
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Figure 19. Wyoming State SARS-CoV-2 Wastewater Surveillance Dashboard (Wyoming PHL, 2021).
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59
6.2.6 Hampton Roads Sanitation District in Hampton Roads,
Virginia
The HRSD is a wastewater utility serving 20 cities and counties in southeastern Virginia. HRSD treats
wastewater for more than 1.7 million people over a service area of nearly 5,000 square miles by
operating 17 WWTPs of varying sizes. HRSD began a pilot microbial source tracking program to detect
surface waters microbes in 2014. This provided HRSD with molecular experience, as well as the
sampling and analytical equipment necessary when the COVID-19 pandemic started. As soon as CDC
developed the clinical SARS-CoV-2 RT-qPCR diagnostic panel, HRSD shifted their focus to quantifying
SARS-CoV-2 RNA in local raw influent wastewater. In early March 2020, HRSD began a SARS-CoV-2
wastewater surveillance pilot study using existing utility funding (HRSD, 2021 b). In collaboration with
experts from the University of Notre Dame and Ohio State University, HRSD then published a research
paper on the results of their 21 -week pilot program that analyzed WWTP influent from the nine large
WWTPs in the region. The manuscript includes details on the program's analytical methods and data
analyses (Gonzalez et al„ 2020).
HRSD has continued weekly sampling of WWTP influent for SARS-CoV-2 beyond the pilot program for
the large WWTPs in the region. In addition, HRSD participated in the WRF's interlaboratory methods
evaluation (Pecson et al„ 2021) and continues to refine their analytical methods over time as more
information on SARS-CoV-2 becomes available (HRSD, 2021b).
HRSD displays up-to-date surveillance results on their HRSD COVID-19 Surveillance Dashboard as a
graph of the total Hampton Roads viral SARS-CoV-2 load in local wastewater over time compared to
clinical cases (see Figure 20). The figure includes the timing of state and local restrictions. HRSD's
dashboard also presents the SARS-CoV-2 concentration spatially over time in a dynamic image. Figure
21 shows a snapshot in time of the spatial variation in normalized SARS-CoV-2 loading across the
region. HRSD also provides wastewater results to the Virginia Department of Health and uploads the
results to CDC's NWSS (HRSD, 2021 b).
HRSD also analyzes wastewater samples for other Virginia government surveillance programs at-cost.
As of April 2021, HRSD was analyzing wastewater samples from all 40 Virginia Department of
Corrections facilities, as well as wastewater samples from some military barracks located in Virginia. In
addition, HRSD has analyzed samples from the Western Virginia Water Authority in Roanoke, Virginia,
since May 2020 (HRSD, 2021 b).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Daily New Clinical Cases and Viral Load in HRSD Treatment Facilities
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overtime (HRSD, 2021 a).
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Log Viral Load
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Figure 21. HRSD's dashboard presents the SARS-CoV-2 wastewater concentration spatially throughout the collection system
overtime (HRSD, 2021a)
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62
6.2.7 Houston, Texas
The Houston wastewater surveillance program began in March 2020, led by HHD and in collaboration
with Rice University for technical expertise and laboratory analyses. Baylor College of Medicine also
provides laboratory support (HHD, 2020; Rice, 2020). The program was initially funded by Rice
University an $200,000 ($35,000 subaward to Rice) NSF RAPID grant (Award #2029025), funding from
CDC Foundation of $65,000, and received additional funding to expand through the CARES Act
(Houston, 2021; NSF, 2020o). Houston also reprogrammed some of their funding received from CDC's
ELC cooperative agreement to support the wastewater surveillance program (Houston, 2021).
Houston Water collects weekly wastewater samples at the influent of all 39 WWTPs in Houston, while
HHD collects samples at other manhole and lift station locations throughout the city. Figure 22 presents
a map of the areas the Houston surveillance program covers. As of March 2021, the program includes
monitoring at 25 nursing homes, 51 K-12 schools, a correctional facility, and two homeless shelters, as
well as at 30 lift stations to capture smaller segments of the community. Houston collects 24-hour
composite samples at all locations, with the exception of schools, where the city collects six- to eight-
hour composite samples to represent typical school hours (Houston, 2021).
HHD initially sent samples to a commercial laboratory but subsequently switched to local universities,
Rice and Baylor, to reduce costs and minimize sampling result turnaround time. HHD worked closely
with researchers at Rice University, who have provided technical expertise and laboratory analysis
throughout the duration of the program, but also sent samples to Baylor for validation and
standardization of the analytical methods (Houston, 2021). Although the universities used different
methods (i.e., Rice used RT-ddPCR and Baylor used RT-qPCR), the universities worked together to
optimize their approaches and analyzed their samples in triplicate to further verify results (Houston,
2021). For example, in order to choose the best method for standardization, the researchers compared
five different RNA concentration and extraction methods. As the program grew, Rice also supported
the transition of some of the wastewater sample analyses over to HHD's laboratory. HHD identified the
flexible and adaptive nature of these academic partners as a key contributor to the success of their
program, noting the rapid evolution in how data had to be analyzed, interpreted, and communicated to
the public as more information became available (Houston, 2021).
HHD uses the wastewater data, along with other public health information, to identify "hot spot zones"
for targeted public health interventions. HHD develops weekly reports that summarize available data for
each of the 105 zip codes in Houston. HHD then identifies "hot spot zones" based on a detailed review
of individual COVID-19 testing results, comparisons of wastewater results to the prior week and to
peak levels observed in the summer of 2020 (see Figure 24), vaccination rates, and known presence of
COVID-19 variants. HHD meets every other week to discuss the zip code summary reports, and in
general, considers the wastewater data to be the most important metric (Houston, 2021).
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63
CEDAR BAYOU
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Figure 22. Areas of Houston covered by the wastewater surveillance program (Houston, 2021).
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Pattern of One Week by Percent of July 6, 2020 by End Date
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Figure 23. Percent change in wastewater results from July 6, 2020 (Peak) for areas of Houston covered by the wastewater
surveillance program (Houston, 2021).
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65
When HHD identifies new zip codes for public health intervention, they set up additional temporary
vaccination and individual testing sites. HHD also identifies support in the local community (e.g., local
churches or faith groups) to help connect with residents and determine barriers to testing and
vaccination, as well as to schedule appointments and secure transportation to testing and vaccination
sites. HHD provides funds to the local community groups for their involvement. Where wastewater
samples are collected at individual facilities, HHD provides those facilities with the wastewater results
weekly (Houston, 2021).
HHD attributes much of the success of their program to collaborations with researchers at Rice and
Baylor, who supported the laboratory methods research while understanding the critical need for
wastewater data to be actionable and to Rice for statistical interpretation and forecasting (Houston,
2021). Rice has recently begun quantifying two specific mutations in wastewater samples that are
characteristic of the United Kingdom variant (B. 1. 1.7). Based on initial data, Rice is now working with
HHD on sequencing the whole SARS-CoV-2 genome. While this approach does not definitively
determine whether the variant is present, it provides information on the likelihood of variants being
present among the population contributing to local wastewater. HHD uses the wastewater mutation
sample data as a screening mechanism to identify communities where the mutations might be present
and to help HHD determine where to allocate resources for genome sequencing of positive personal
tests (Houston, 2021).
Beyond COVID-19, HHD and Rice are exploring future uses of wastewater surveillance (e.g., influenza
and other viral pathogens) with hopes of developing a long-term robust monitoring program for the city
(Houston, 2021).
6.2.8 Tempe, Arizona
In 2018, Tempe and Arizona State University's (ASU's) Biodesign Institute initiated a program to study
opioids in wastewater with funding from the city council's Innovation Fund (Tempe, 2021 d). For this
effort, the city analyzed samples from five WWTPs for various parent drug compounds (e.g., fentanyl)
and their metabolites (e.g., norfentanyl) (Tempe, 2021c). Tempe used the results to provide insight on
illicit drug use in the city and to support public health decisions such as where to allocate resources and
strategies for reducing opioid use. To foster community buy-in for this project, Tempe held a town hall
meeting where a diverse panel of experts, including public health officials and the fire chief, described the
project and then presented the data in the form of an online data dashboard. Tempe credits this meeting
and the development of a publicly available dashboard as establishing the necessary foundation of
transparency and trust with community members for the opioid wastewater surveillance program
(Tempe, 202Id).
When the idea of testing wastewater for SARS-CoV-2 came about in February 2020, the city was able to
quickly establish a program by leveraging community support and lessons learned from the city's existing
opioids program, as well as systems like the partnership with ASU for laboratory analyses and the online
data dashboard for disseminating results. Tempe began wastewater surveillance for SARS-CoV-2 in late
March 2020 from one sewershed and expanded throughout the city shortly after (Tempe, 2021 d). As of
February 2021, Tempe collects 24-hour composite samples two to seven times per week in seven
sewersheds (Tempe, 2021 b). ASU then performs the laboratory analysis of SARS-CoV-2 via RT-qPCR
on the wastewater samples using funding from NIH and NSF grants. Tempe presents the results in real
time on an online data dashboard for use, along with other public health data, to drive decisions (e.g.,
when to close or reopen certain establishments) (Tempe, 202Id).
Tempe's success piqued the interest of, and partnership offer from the neighboring town of Guadalupe,
which has a large population of essential workers and low-income and minority families. Guadalupe has
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
66
since partnered with Tempe and ASU to monitor wastewater for SARS-CoV-2. Tempe posts the
Guadalupe results on Tempe's online dashboard. Guadalupe's Mayor and Council use the results to
support its public health messaging (e.g., the importance of social distancing) and decision-making
(Tempe, 202Id).
Tempe's online data dashboard presents an interactive bar chart with average weekly SARS-CoV-2
wastewater results for each geographic collection area (see Figure 24) (Tempe, 2021a). The dashboard
also includes an "event timeline" that indicates when the city implemented certain public health
measures that may be tied to trends observed in the wastewater data (e.g., when dine-in restaurants
were closed, when schools were closed, when the stay-at-home order was lifted) (Tempe, 2021a).
Tempe also developed a timeline with user-friendly background information on local and state events
related to COVID-19 that include events like (Tempe, 2021a):
¦ 3/14/2020 - Arizona closed schools statewide
¦ 3/29/2020 - Arizona issued a stay-at-home order
¦ 5/14/2020 - Arizona's stay at home order ended
¦ 6/10/2020 - COVID-19 testing became available in the Town of Guadalupe
¦ 6/19/2020 - Maricopa County required masks in public spaces
¦ 12/15/2020 - Tempe began phase IA of the vaccine roll out
¦ I /28/2021 - Arizona detected the first case of the U.K. COVID-19 variant
¦ 3/14/2021 - Tempe Elementary School District and Tempe Union High School District resumed
in-person learning
Tempe's wastewater program for SARS-CoV-2 has offered an affordable way for the city to track
COVID-19 in the community and has, in some cases, served as an early warning of increased COVID-19
spread (Tempe, 202Id). Tempe has also used the data to understand the distribution of COVID-19
cases throughout the city and to determine where to deploy additional services to educate the public
and increase awareness on implemented public health measures. A key driver of this effort was to
address equity issues within the community and determine if there were disproportionate
concentrations of cases in certain locations. As an example, early in the program, Tempe observed
elevated concentrations of SARS-CoV-2 in one area. Based on the wastewater data and the community
demographics, Tempe recognized the need to provide additional resources and launched an educational
campaign for the residents focused on reducing COVID-19 risk. City workers campaigned door to door
throughout the community and distributed educational materials in Spanish and English, face masks,
stickers, and care packages. After the campaign, the city observed slight decreases in SARS-CoV-2
wastewater measurements (Tempe, 202Id).
Beyond COVID-19, the city is evaluating options for future wastewater surveillance for other
biomarkers that can indicate the presence of other viruses, drugs and alcohol, and other health concerns
(e.g., asthma medicine) to aid in city planning (e.g., public education and outreach, allergy-friendly tree
canopy selection) (Tempe, 202Id).
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rif COVID-19 Wastewater Results
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Figure 24. The City of Tempe's wastewater dashboard for SARS-CoV-2 with a map of the areas sampled (Tempe, 2021 a).
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68
6.2.9 Clemson University in Clemson, South Carolina
The Clemson University wastewater surveillance program, led by Dr. David Freedman, a professor in
the Department of Environmental Engineering and Earth Sciences, began in late May 2020. Backed by
university funding as part of Clemson's initiative to return students to campus amidst the COVID-19
pandemic (Clemson, 2021 b), the program tracks both student transmission of the SARS-CoV-2 virus on
campus, as well as student and public transmission of the virus in the city of Clemson and town of
Pendleton, South Carolina (Clemson, 2021a).
Once or twice a week, 24-hour composite samples of influent wastewater are collected from an on-
campus WWTP and two off-campus WWTPs (Clemson, 2021a; Clemson, 2021 b). The samples are then
sent to SiREM, a commercial lab in Knoxville, Tennessee, which analyzes for the virus using RT-qPCR for
the Nl and N2 genes of SARS-CoV-2 (Clemson, 2021b; Clemson, 2020). Beginning in early March of
2021, Clemson expanded their analysis of the SARS-CoV-2 virus in wastewater to include the B. 1. 1.7
variant (i.e., the United Kingdom [U.K.] variant) and the BI.35I variant (i.e., the South African [S.A]
variant). The laboratory uses the same molecular test used to quantify virus, except that the test is
highly specific for the RNA that is uniquely associated with the variants.8 Wastewater results for each
variant are reported as percentages, where the variant RNA copies/L are divided by the total RNA virus
copies/L (Clemson, 2021 b). When none of the variant RNA is detected, the percentage is zero. When
all of the total RNA virus detected consists of a variant, the percentage is 100.
Clemson's surveillance program reports each sampling location's SARS-CoV-2 results in copies/L on its
online COVID-19 Wastewater Dashboard (see Figure 25) (Clemson, 2021a). The dashboard ranks the
severity of the SARS-CoV-2 copies/L in wastewater by impact level. The impact levels allow the public
to interpret the potential risk of COVID-19 from the wastewater more clearly. Dr. Freedman created
the ranking system by comparing cases and surveillance results from European cities (e.g., Paris,
Barcelona) to Clemson's results. The impact levels are as follows (Clemson, 2021b; Clemson, 2020):
¦ less than 4,000 virus copies/L = no impact
¦ 4,000 to 9,999 virus copies/L = minimal impact
¦ 10,000 to 99,999 virus copies/L = potential increasing impact
¦ 100,000 to 999,999 virus copies/L = potential moderate impact
¦ greater than 1,000,000 virus copies/L = potential significant impact
As of April 2021, the dashboard also includes results for the B. 1. 1.7 variant (see Figure 28) (Clemson,
2021a).
8 SiREM quantifies variants in wastewater samples for Clemson University using the same RNA extracted to
quantify the total SARS-CoV-2 virus. SiREM first uses the TaqPath COVD-19 PCR assay kit to quantify the total
virus. This kit detects the S gene, N gene, and ORFlab gene sequence. SiREM then uses a second PCR kit (GSD
NovaType SARS-CoV-2 ID PCR) to detect the U.K. variant and the S.A. variant. Based on differences in Ct levels
during the PCR reaction for RNA that is subjected to both kits, SiREM delineates what percentage of the total
virus counts are attributable to the U.K. and S.A variants. Differences in Ct levels are a result of mutations to the
S gene that are present in the variants (Clemson, 2021 b).
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69
CLEMS5&N COVID-19 WASTEWATER TESTING
UNIVERSITY
Wastewater Samples as of 7/1/2021
Legend (hover for description)
© 2021 Mapbox © OpenStreetMap
Figure 25. The Clemson University COVID-19 Wastewater Dashboard color codes the impact level on a map of each
WWTP's collection system area (Clemson, 2021a).
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70
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Figure 26. The Clemson University COVID-19 Wastewater Dashboard demonstrates the impact level and virus copies/L in
each WWTP influent sampling point over time. The dashboard also includes variant tracking results (Clemson, 2021 a).
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71
Both the city of Clemson and Clemson University are using the wastewater surveillance program for
their COVID-19 response (Clemson, 2021 a; City of Clemson, 2021). In June 2020, when the city council
began considering a mask ordinance, the city manager invited Dr. Freedman to present the elevated
SARS-CoV-2 wastewater results from the city's Cochran Road WWTP (City of Clemson, 2020a).
Shortly after hearing about the health concerns surrounding COVID-19 levels in the community, the city
instituted a face mask ordinance citing the wastewater surveillance data and continues to do so with
each subsequent extension of the ordinance. In October 2020, the city council extended the face mask
ordinance until Christmas based on new elevated levels of SARS-CoV-2 in wastewater (City of Clemson,
2020b; Clemson, 2021b).
Clemson University increased the frequency of testing for SARS-CoV-2 in the wastewater after students
returned to on-campus dormitories in Fall 2020. The wastewater results had detectable SARS-CoV-2,
which did not align with the university's expectation that COVID-19 would be minimal on campus since
students were required to have a negative COVID-19 test before and after arriving. Soon thereafter,
Clemson University ramped up their individual testing plan for the fall semester to incorporate weekly
testing for all students who live on campus. This was extended in the Spring 2021 semester to include
weekly testing of any person who accessed a university building (Clemson, 2021 b).
Clemson plans to continue monitoring the wastewater into the Summer of 2021, with declining
concentrations anticipated in response to increasing levels of vaccination (Clemson, 2021b).
6.2.10 University of Arizona in Tucson, Arizona
The University of Arizona is leading and participating in multiple wastewater surveillance programs for
SARS-CoV-2, much of which is spearheaded by Dr. Ian Pepper, Dr. Charles Gerba, and their team at the
Water and Energy Sustainable Technology (WEST) Center (University of Arizona, 2021; University of
Arizona, 2020b). The university's work in this field began in March 2020 when Dr. Pepper posted an
advertisement on the WEST Center's website for SARS-CoV-2 analyses of the Nl and N2 gene variant
in wastewater that utilities could receive for a fee. The WEST Center was able to quickly set up a
laboratory for this work given their prior experience testing wastewater for other viruses, which also
meant they already had the necessary equipment. Between March and the end of 2020, they analyzed
approximately 350 samples from utilities throughout the U.S. and in Canada. In addition to providing
SARS-CoV-2 wastewater results, Dr. Pepper and his team prepared summary reports with each data
package that included a brief interpretation of the results and comparisons to the number of clinical
cases in the local area at the time (University of Arizona, 2021; University of Arizona, 2020b).
In June 2020, the university created a task force to develop a plan and coordinate logistics for
monitoring COVID-19 on campus with both personal testing and wastewater surveillance. The
University President was supportive of the wastewater surveillance program and provided funding from
the university (University of Arizona, 2021). Shortly after the wastewater program began and students
returned to campus in late August, the university detected SARS-CoV-2 in the wastewater from one of
the dormitories. Based on the wastewater results, the university tested all students in the dormitory,
ultimately identifying two asymptomatic students who tested positive for COVID-19. The students were
immediately isolated and SARS-CoV-2 was not detected in wastewater collected the next day—a
success story that has garnered national attention (University of Arizona, 2021; University of Arizona,
2020a). The university has repeated this success and potentially prevented outbreaks nearly one
hundred other times, where SARS-CoV-2 detected in wastewater triggered personal testing that
identified students with COVID-19 for isolation. In general, wastewater testing was found to be a good
predictor of COVID-19 cases. When SARS-CoV-2 was detected in wastewater, the university often
found students who tested positive for COVID-19 in the same dormitory. Conversely, when SARS-
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72
CoV-2 was not detected in wastewater, students often did not test positive for COVID-19 (Betancourt
et al„ 2021).
The university currently collects grab samples three times per week at 18 dormitories on campus and
analyzes the samples on the same day they are collected (University of Arizona, 2021). To ensure that
the wastewater data reflect a distinct population and are actionable, samples are collected at
dormitories with a single sewer line. Tied to this, the university has developed action levels for their
campus wastewater surveillance program (see Table 6). For example, non-detect wastewater results
require no action, while concentrations between 1,000 and 10,000 copies/L trigger 20 percent random
personal testing of dormitory residents. This program is a strong example for how rapid wastewater
results and an efficient alert system can help minimize the spread of COVID-19 (Gerba, 2021).
Table 6. University of Arizona's levels of concern and associated actions for
ranges of SARS-CoV-2 wastewater concentrations.
Level of
Concern
Wastewater SARS-CoV-2
Concentration Order of Magnitude
(gene copies/L)
Action Item
0
Non-detect
No action
1
10 to less than 1,000
Enhanced awareness and disinfection
2
1,000 to less than 100,000
20% random testing of residents
3
100,000 to less than 10,000,000
40% random testing of residents
4
10,000,000 or higher
All residents tested
Based on the success of the on-campus wastewater surveillance program, the university's Yuma Center
of Excellence for Desert Agriculture received funding from the Arizona Department of Health in January
of 2021 to monitor wastewater in Yuma County. Dr. Pepper's team helped colleagues establish an off-
campus laboratory and develop a sampling plan. This plan involved dividing the county into I 3 distinct
regions and then identifying "high-risk" facilities (e.g., schools, nursing homes) for personal testing when
community wastewater samples show increased levels of SARS-CoV-2 (University of Arizona, 2021;
Ducey, 2021; Gerba, 2021).
In addition to these implementation programs, Dr. Pepper and the WEST Center team are actively
researching methods for predicting numbers of infected individuals with wastewater data. For example,
they are currently using personal testing and wastewater data collected on campus to predict the
shedding rate of the virus in feces. The university is also testing the wastewater for mutations of the
SARS-CoV-2 virus to evaluate the likelihood of variants present (University of Arizona, 2021).
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73
7 Wastewater Surveillance Lessons
Learned
EPA identified 14 states with large-scale SARS-CoV-2 wastewater surveillance programs, along with 160
local communities or academic institutions conducting wastewater surveillance. Each of these groups
relied on different approaches for their program, from analytical methods to funding mechanisms to
communication of the results. All the programs showed that wastewater surveillance can be an
important and effective tool for early detection of SARS-CoV-2, giving the programs the ability to take
action to prevent the continued spread of COVID-19. The programs also noted that adequate and
sustained funding was critical to their success and that without it, a surveillance program would be hard
to maintain.
EPA selected ten of these programs to highlight in the case studies presented in Section 6.2. These case
studies present the different approaches for implementing programs and analyzing wastewater data to
track the presence of SARS-CoV-2. The technical leads for the case study programs noted that much of
their success was dependent on four key aspects of their wastewater surveillance programs:
collaboration, flexibility, transparent communication, and adequate funding. In terms of collaboration, the
wastewater surveillance programs leveraged resources through establishing partnerships and
relationships with entities within their programs, as well as with external groups. For example, the
Wyoming wastewater surveillance program initiated its own outreach efforts, but also relied on
outreach support from the Wyoming Association of Rural Water Systems. These collaborative efforts
helped educate utilities on the benefits of wastewater surveillance, ultimately leading to their voluntary
participation in the state's program (Wyoming, 2021).
The success of these programs was also tied to their ability to be flexible and open to the rapidly
developing science around wastewater surveillance for SARS-CoV-2. This flexibility included adapting to
ongoing developments in analytical methods and data use by public health officials, as well as availability
of necessary materials for sample collection and data analysis. For example, the Michigan wastewater
surveillance program originally used RT-qPCR for laboratory analysis, but later shifted to RT-ddPCR due
to supply chain issues with some of the materials needed for RT-qPCR methods (Michigan, 2021a). As
another example, the Ohio wastewater surveillance program initially launched their program with
weekly wastewater sampling, but then changed to twice per week as it provided more meaningful and
actionable data for the local health departments (Ohio, 2021).
Program leads also highlighted the necessity of transparent communication within their organizations
and with stakeholders in order to gain community support. For example, the Ohio wastewater
surveillance program began after Ohio Governor DeWine became aware of wastewater surveillance.
This led to a successful program because of the support from the highest levels within the state,
including leadership at the ODH and Ohio EPA (Ohio, 2021). As another example, the opioid
wastewater surveillance program in Tempe, Arizona, established strong community buy-in by having
public health officials and the local fire chief share information during town hall meetings. Tempe was
able to build on these relationships and the confidence established within the local community to quickly
implement a wastewater surveillance program for SARS-CoV-2 (Tempe, 202Id).
The programs also identified that adequate and sustained funding was critical to their success in
providing consistent support for public health. For example, Michigan's wastewater surveillance pilot
project ended in February 2021 after spending all their CARES Act funding. Some of the participating
sites continued wastewater surveillance using other funding sources, but some sites had to stop
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74
sampling. MDHHS was able to reestablish the wastewater surveillance program using funding from
CDC's ELC Emerging Detection Enhancement award with tentative plans to start in June 2021 (Peters,
2021). On the other hand, Wyoming had sufficient funding to incentivize community participation by
paying the communities $300 per sample, providing bonuses to communities that consistently sampled,
and paying technical assistance providers for onboarding communities into the program. Wyoming used
CARES Act and CDC ELC funding, originally budgeting for 50 to 100 communities participating. With
the 30 communities that Wyoming included in early 2021, they estimated there were about 30 months
of funding remaining (Wyoming, 2021).
Wastewater-based surveillance is a new and changing field. This report highlights the unique aspects of
some of the wastewater surveillance implementation work and lessons learned. This rapidly evolving
field can be an effective tool for early detection of SARS-CoV-2 and other pathogens in the future,
especially among disadvantaged or vulnerable populations where clinical testing may not be widely
available.
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Appendix A.
Summary of Wastewater Surveillance Programs
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-1. State wastewater surveillance programs.
State
Lead Agency
Collaborators/
Partners
Sampling Start
Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard
Dashboard Description (Link
Included in Reference)
Colorado
Colorado
Department of
Public Health
and Environment
Colorado State
University,
Metropolitan State
University, GT
Molecular
August 2020
22 WWTPs
Unknown
Yes
Map of sewersheds with a
timeseries of wastewater results
in copies/L. Also includes a bar
chart of personal testing data
(Colorado DPH, 2021).
Connecticut
Connecticut
Department of
Health
Yale University,
Connecticut
Agricultural
Experimental Station
August 2020
7 WWTPs
RT-qPCR
Yes
Map of WWTP locations, chart
of wastewater N1 and N2 data in
copies/L with a seven-day
trendline, and a chart of personal
testing data (Yale University,
2021a).
Indiana
Indiana Finance
Authority
120Water,
University of Notre
Dame, Indiana
University, Microbac
August 2020
15 WWTPs, 44
other locations
RT-ddPCR
and RT-
qPCR
No
Not applicable. Indiana published
a report with the results (Indiana,
2020).
Kansas
Kansas
Department of
Health and
Environment
University of Kansas
May 2020
12 WWTPs
Unknown
No
Not applicable. No centralized
dashboard, but some cities have
posted results (University of
Kansas, 2020; Lawrence, 2021).
Maryland
Maryland
Department of
the Environment
Maryland
Department of
Health, St. Mary's
College, MetCom
Pilot phase:
Summer 2020
Follow-on
phase: Winter
2021
2020 pilot phase:
27 low-income
housing areas and
10 correctional
facilities
Follow-on phase:
As of May 2021, 22
sites
RT-qPCR
Yes
Map of sampling locations with
color-coded map markers based
on SARS-CoV-2 trend with plots
of copies/L over time (Maryland,
2021).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-1. State wastewater surveillance programs.
State
Lead Agency
Collaborators/
Partners
Sampling Start
Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard
Dashboard Description (Link
Included in Reference)
Michigan
Michigan
Department of
Health and
Human Services
Michigan
Environment, Great
Lakes, and Energy;
universities
September 2020
270 WWTPs and
facilities
RT-ddPCR
Yes
Map of testing sites and relevant
detail such as lab name, grant
award and amount, start date,
sample type, and partner name
(Michigan EGLE, 2021).
Missouri
Missouri
Department of
Health and
Senior Services
Missouri
Department of
Natural Resources,
University of
Missouri
July 2020
50 WWTPs
Unknown
Yes
Map of WWTPs with color-
coded points based on
wastewater trend, and a
timeseries plot of viral copies/day
(Missouri DHSS, 2021).
New Mexico
New Mexico
Environment
Department
None
December 2020
22 congregate
facilities
Unknown
Yes
Timeseries plots of copies/L
sampling results over time for
each facility, along with current
level of concern and current
trend (NMED, 2021a).
North
Dakota
North Dakota
Department of
Environmental
Quality
North Dakota State
University
July 2020
21 WWTPs
Unknown
No
Not applicable. No public
dashboard (Dura, 2020; Cooper,
2021).
Ohio
Ohio
Department of
Health
Ohio EPA, Ohio
WRC, U.S. EPA, and
other participating
universities
July 2020
66 WWTPs
Unknown
Yes
A map of WWTPs color-coded
to show the status of SARS-CoV-
2 in wastewater (e.g., increasing).
Also includes a timeseries plot
for each WWTP with showing
million gene copies/day and
average N2 gene copies/L over
time (Ohio DOH, 2020).
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-1. State wastewater surveillance programs.
State
Lead Agency
Collaborators/
Partners
Sampling Start
Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard
Dashboard Description (Link
Included in Reference)
Oregon
Oregon Health
Authority
Oregon State
University
August 2020
42 WWTPs
Unknown
Yes
Map of WWTPs that indicates
whether SARS-CoV-2 was
detected or not detected in the
week of monitoring; can adjust
the date to see over time
(Oregon Health Authority, 2021).
Utah
Utah
Department of
Environmental
Quality
Utah Department of
Health. University of
Utah, Utah State
University, and
Brigham Young
University
April 2020
42 WWTPs
Unknown
Yes
Map of WWTP sewersheds with
timeseries plot of wastewater
data in million gene
copies/person/day and daily new
individual testing cases (Utah
DEQ, 2021).
Wisconsin
Wisconsin
Department of
Health Services
Wisconsin State Lab
of Hygiene,
University of
Wisconsin-
Milwaukee
October 2020
65 WWTPs,
including 4 tribal
WWTPs
Unknown
Yes
Map of WWTP sewersheds with
a timeseries plot of SARS-CoV-2
in million gene copes/person/day
and the seven-day average
individual case rate of COVID-19
(Wisconsin DOHS, 2020).
Wyoming
Wyoming Public
Health
Laboratory
Wyoming
Department of
Health, University of
Wyoming, Wyoming
Association of Rural
Water Systems
July 2020
34 WWTPs and 13
facilities
RT-qPCR
Yes
Map of WWTPs color coded by
percent of modeled prevalence of
COVID-19 infections based on
wastewater results. Also includes
timeseries plots of modeled
prevalence, Ct count, and
copies/mL for WWTPs and
facilities (Wyoming PHL, 2021)
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
University of
Alaska-Anchorage
AK
Unknown—program details were not researched (see Section 3)
University of
Alaska-Fairbanks
AK
Unknown—program details were not researched (see Section 3)
Birmingham
Southern College
AL
Unknown—program details were not researched (see Section 3)
University of
Arkansas
AR
Unknown—program details were not researched (see Section 3)
Gilbert
AZ
Arizona State
University
May 2020
3 areas in
collection system
RT-qPCR
Yes
Map of sewershed areas for each
sample location with a timeseries
bar chart with average weekly
results in copies/L (Gilbert,
2021).
Tempe and
Guadalupe
AZ
Arizona State
University
March 2020
8 WWTPs and
sewersheds
RT-qPCR
Yes
Map of sewershed areas for each
sample location with a timeseries
bar charts with average weekly
results in copies/L (Tempe,
2021a).
University of
Arizona
AZ
University of
Nevada- Las Vegas
March 2020
18 dorms
RT-qPCR
No
Not Applicable. No public
dashboard (University of
Arizona, 2020a; University of
Arizona, 2020b; University of
Arizona, 2021).
University of
Northern Arizona
AZ
Unknown—program details were not researched (see Section 3)
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Navajo Nation
AZ
Unknown—program details were not researched (see Section 3)
Alameda County
CA
Unknown—program details were not researched (see Section 3)
City of Davis
CA
University of
California - Davis
November
2020
1 1 sewersheds
Unknown
Yes
Timeseries plot for each
sewershed with virus
concentration normalized to
fecal strength, city-wide
wastewater results, and
detection limit (Healthy Davis
Together, 2021).
City of Palm Springs
CA
GT Molecular
August 2020
1 WWTP
Unknown
Yes
Weekly reports with timeseries
plots of copies/L and trendline;
includes significant events. Also
evaluating for variants (Palm
Springs, 2021).
Contra Costa
County
CA
Unknown—program details were not researched (see Section 3)
Loma Linda
University
CA
Unknown—program details were not researched (see Section 3)
Los Angeles County
Sanitation Districts
CA
University of
Arizona,
California State
Water Resources
Control Board
August 2020
2 WWTPs
Unknown
Yes
Timeseries plots with weekly
average copies/L, seven-day
average new COVID-19 cases,
and three-day average LA
County hospitalizations (LA
County, 2021).
Marin County
CA
Unknown—program details were not researched (see Section 3)
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Mariposa County
CA
Biobot Analytics
May 2020
3 locations
Unknown
Yes
Timeseries plots with data from
each of the three sampling
locations as virus
concentration/L (Mariposa
County, 2021).
San Francisco
County
CA
Unknown—program details were not researched (see Section 3)
Santa Clara County
CA
Santa Clara
County
Department of
Environmental
Health, City of
Palo Alto, City of
San Jose,
Sunnyvale, Gilroy,
Stanford
University
October 2020
4 WWTPs
Unknown
Yes
Timeseries plot for each WWTP
with the S and N genes
normalized to pepper mild
mottle virus (Santa Clara
County, 2021).
Stanford University
CA
Unknown—program details were not researched (see Section 3)
University of
California-Berkeley
CA
None
August 2020
5 locations on
campus
RT-qPCR
Yes
Map of sampling locations with
icons indicating detected or not
detected SARS-CoV-2 or no
sample collected (UC Berkeley,
2021).
University of
California-Irvine
CA
Unknown—program details were not researched (see Section 3)
University of
California-Merced
CA
Unknown—program details were not researched (see Section 3)
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A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
University of
California-San
Diego
CA
None
Summer 2020
640 campus
buildings
RT-qPCR
Yes
A campus map that highlights
buildings different colors based
on if they are monitored/not
monitored and if SARS-CoV-2
was detected/not detected in the
wastewater (UC San Diego,
2021).
University of
California-Santa
Barbara
CA
Unknown—program details were not researched (see Section 3)
University of
Southern California
CA
None
October 2020
Dorms, athletic
facilities, and office
buildings
RT-ddPCR
No
Not applicable. No public
dashboard (USC, 2021).
Colorado College
CO
Unknown—program details were not researched (see Section 3)
Colorado State
University
CO
None
Beginning of fall
semester 2020
17 locations on
campus, focused
on dorms
RT-ddPCR
No
Not applicable. No public
dashboard (CO State, 2020).
University of
Colorado-Boulder
CO
None
August 2020
23 sites on-
campus (residence
halls, other
buildings)
RT-qPCR
No
Not applicable. No public
dashboard (CU Boulder, 2020).
University of
Denver
CO
Unknown—program details were not researched (see Section 3)
Metro State
University
CO
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
University of
Connecticut
CT
None
June 2020
1 WWTP and 14
buildings
RT-qPCR
Yes
Timeseries plot of the ratio of
the E and N1 genes to a fecal
indicator virus on collection
dates for each of the sampling
locations (UConn, 2021).
University of
Hartford
CT
Unknown—program details were not researched (see Section 3)
Howard University
DC
Unknown—program details were not researched (see Section 3)
New Castle County
DE
Biobot Analytics,
University of
Delaware
April 2020
3 WWTPs and
sewersheds
PCR
Yes
Timeseries plot of the aggregate
sewer system viral copies/L and
rolling seven-day average
confirmed COVID-19 cases. Also
includes latest viral copes/L for
each sampling location and
timeseries plots of viral copies/L
with 95% confidence interval
shading for each sampling result.
Also includes sewershed maps
for each sampling location
(NCCo, 2021).
Broward County
FL
Unknown—program details were not researched (see Section 3)
Florida Atlantic
University
FL
None
March 2020
Campus lift
stations
PCR
No
Not applicable. No public
dashboard (Randall, 2020).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Loxahatchee River
District
FL
Biobot Analytics,
Florida
Department of
Health
May 2020
1 WWTP
qPCR
Yes
Timeseries plots with copies/L in
log scale and linear scale are
compared to new clinical cases
and seven-day rolling average
clinical cases (LRD, 2021).
Miami-Dade County
FL
Biobot Analytics
March 2020
3 WWTPs
Unknown
No
Not applicable. No public
dashboard (WaterWorld, 2020).
Ringling College of
Art and Design
FL
Unknown—program details were not researched (see Section 3)
Rollins College
FL
Unknown—program details were not researched (see Section 3)
University of Florida
FL
Unknown—program details were not researched (see Section 3)
Athens-Clarke
County
GA
University of
Georgia
June 2020
3 WWTPs
RT-PCR
Yes
Timeseries plot of total viral
copies with a 95% confidence
interval, predicted total viral
copies, along with daily and
seven-day rolling average
COVID-19 reported cases
(UGA, 2021).
Emory University
GA
Unknown—program details were not researched (see Section 3)
University of
Hawaii-Manoa
HI
Unknown—program details were not researched (see Section 3)
City of Boise
ID
University of
Missouri, Boise
State University
May 2020
2 WWTPs
Unknown
Yes
Timeseries plot with wastewater
results in copies/L and a plot of
confirmed and probable COVID-
19 cases (Boise, 2021).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
City of Moscow
ID
Biobot Analytics,
University of
Idaho
May 2020
1 WWTP
RT-PCR
No
Not applicable. No public
dashboard (Argonaut, 2020).
Twin Falls
ID
Unknown—program details were not researched (see Section 3)
University of Idaho
ID
None
May 2020
9 campus
locations
RT-PCR
No
Not applicable. No public
dashboard (Argonaut, 2020).
Chicago
IL
University of
Illinois - Chicago,
Northwestern
University,
Argonne National
Laboratory,
Metropolitan
Water
Reclamation
District of
Greater Chicago,
Chicago
Department of
Public Health
Unknown
3 WWTPs and
sewersheds
Unknown
No
Not applicable. No public
dashboard available (UIC, 2020).
Kendell County
IL
Yorkville-Bristol
Sanitary District,
RJN Group, GT
Molecular
November
2020
1 WWTP
Unknown
Yes
Map with WWTP sewershed,
timeseries plot of virus
concentration and number of
positive COVID-19 tests and
average COVID-19 positivity
percent for Kendall County. Also
includes wastewater result trend
(YBSD, 2021).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Northern Illinois
University
IL
Unknown—program details were not researched (see Section 3)
University of Illinois
at Chicago-School
of Public Health
IL
Unknown—program details were not researched (see Section 3)
City of Carmel
IN
Biobot Analytics,
University of
Notre Dame,
Pace Analytical
May 2020
1 WWTP
Unknown
No
Not applicable. No public
dashboard available (Carmel,
2020).
University of Notre
Dame
IN
None
July 2020
Throughout
campus
RT-ddPCR
No
Not applicable. No public
dashboard available (Zacharias,
2020).
City of Lawrence
KS
University of
Kansas
June 2020
2 WWTPs
Unknown
Yes
Map of the sewersheds for each
WWTP along with interactive
timeseries plots with the N1 and
N2 copies/L and average in
copies/day, along with the 14-day
rolling average new COVID-19
cases (Lawrence, 2021).
University of
Kentucky
KY
None
September
2020
Campus dorms
PCR
No
Not applicable. No public
dashboard available (Chapin,
2020).
University of
Louisville
KY
Unknown—program details were not researched (see Section 3)
Murray State
University
KY
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Louisiana State
University
LA
Baton Rouge
June 2020
Throughout
campus and
surrounding
community
qPCR
No
Not applicable. No public
dashboard available (Rddad,
2020).
Tulane University
LA
New Orleans
January 2020
Throughout
campus and
surrounding
community
Unknown
No
Not applicable. No public
dashboard available (Zobel,
2020).
Massachusetts
Institute of
Technology
MA
None
October 2020
7 campus buildings
RT-qPCR
No
Not applicable. No public
dashboard available (Winn,
2020).
Northeastern
University
MA
Unknown—program details were not researched (see Section 3)
Town and County
of Nantucket
MA
Biobot Analytics
April 2020
1 WWTP
RT-qPCR
Yes
PDF with weekly results that
includes a timeseries plot of viral
copies/L sewage with the daily
and seven-day rolling average of
new COVID-19 cases. Also
includes comparisons of
wastewater to samples
nationwide. Includes Biobot's
COVID-19 incidence estimate of
new cases/day based on the
wastewater results (Nantucket,
2021).
University of
Massachusetts
Lowell
MA
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Williams College
MA
Unknown—program details were not researched (see Section 3)
Frederick County
MD
CosmosID
May 2020
1 WWTP
RT-qPCR
No
Not applicable. No public
dashboard available (Frederick
County, 2020).
Mount St. Mary's
University
MD
Unknown—program details were not researched (see Section 3)
St. Mary's College of
Maryland
MD
Unknown—program details were not researched (see Section 3)
St. Mary's County
MD
St. Mary's County
Health
Department, St.
Mary's College of
Maryland, St.
Mary's County
Metropolitan
Commission
July 2020
9 WWTPs
Unknown
Yes
Map of the WWTP service areas
along with timeseries plots for
each of the WWTPs with the
virus copies/L (St. Mary's, 2021).
University of
Maryland
MD
None
September
2020
Throughout
campus
Unknown
No
Not applicable. No public
dashboard available
(Neugeboren, 2020).
Washington
Suburban Sanitary
Commission
MD
University of
Maryland,
Montgomery
County, Prince
George's County
Unknown
6 locations
Unknown
No
Not applicable. No public
dashboard available (UMD,
2020).
Saint Joseph's
College of Maine
ME
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
University of Maine-
Orono, Fort Kent,
and Presque Isle and
University of
Southern Maine-
Gorham
ME
CES, Inc., Orono,
Farmington
July 2020
9 campus
locations and 2
WWTPs
Unknown
Yes
Tables for each sample location
with positive/negative/
indeterminate categorization and
viral equivalence/L for positive
results (Maine, 2021).
University of
Southern Maine
ME
Unknown—program details were not researched (see Section 3)
Albion College
Ml
Unknown—program details were not researched (see Section 3)
Alma College
Ml
Unknown—program details were not researched (see Section 3)
Central Michigan
University
Ml
Unknown—program details were not researched (see Section 3)
Eastern Michigan
University
Ml
Unknown—program details were not researched (see Section 3)
Ferris State
University
Ml
Unknown—program details were not researched (see Section 3)
Grand Valley State
University-AWRI
Ml
Unknown—program details were not researched (see Section 3)
Hope College
Ml
Trident
Laboratories,
Holland BPW
Unknown
9 campus
locations and 1
WWTP
qPCR
No
Not applicable. No public
dashboard available (Hope
College, 2021)
Michigan State
University
Ml
None
May 2020
Campus manholes
Unknown
No
Not applicable. No public
dashboard available (Lavery,
2020).
Oakland University
Ml
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Saginaw Valley State
University
Ml
Unknown—program details were not researched (see Section 3)
University of
Michigan
Ml
None
Unknown
Throughout
campus
Unknown
No
Not applicable. No public
dashboard available (UMich,
2020).
University of
Minnesota
MN
Unknown—program details were not researched (see Section 3)
University of
Minnesota Duluth
MN
Unknown—program details were not researched (see Section 3)
University of
Minnesota
Rochester
MN
Unknown—program details were not researched (see Section 3)
Missouri State
University
MO
Unknown—program details were not researched (see Section 3)
University of
Mississippi
MS
Unknown—program details were not researched (see Section 3)
Carroll College
MT
Unknown—program details were not researched (see Section 3)
Montana State
University
MT
Unknown—program details were not researched (see Section 3)
Appalachian State
University
NC
University of
North Carolina
Fall 2020
8 dorms
RT-ddPCR
No
Not applicable. No public
dashboard available (Buffy, 2021).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Jackson County
NC
Tuckaseigee
Water and Sewer
Authority, Jackson
County
Department of
Public Health,
University of
Wisconsin-
Milwaukee,
Dogwood Health
T rust,
Mathematica
July 2020
1 WWTP
Unknown
No
Not applicable. No public
dashboard available
(Mathematica, 2020).
Raleigh
NC
North Carolina
State University
March 2020
1 WWTP, 2
residential
sewersheds, and
hospital
Unknown
No
Not applicable. No public
dashboard available (Sherman,
2020).
North Carolina
State University
NC
Unknown—program details were not researched (see
Section 3)
University of North
Carolina-Chapel Hill
NC
Unknown—program details were not researched (see
Section 3)
University of North
Carolina-Charlotte
NC
None
Unknown
20 locations
throughout
campus
Unknown
No
Not applicable. No public
dashboard available (UNC
Charlotte, 2020).
University of North
Carolina-
Wilmington
NC
Unknown—program details were not researched (see
Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
North Dakota State
University
ND
Unknown—program details were not researched (see Section 3)
University of
Nebraska
NE
Unknown—program details were not researched (see Section 3)
Darmouth-
Hitchcock Medical
Center
NH
Unknown—program details were not researched (see Section 3)
Keene State College
NH
City of Keene,
Cheshire Medical
Center
August 2020
Campus and off-
campus WWTPs
Unknown
Yes
Bar chart with sampling results
for both sampling locations along
with two trendlines in viral
copies/L. Chart also includes
significant events. Note that the
dashboard does not include the
entire timeframe (Keene, 2021).
University of New
Hampshire
NH
None
August 2020
Throughout
campus
RT-ddPCR
No
Not Applicable. No public
dashboard available (UNH,
2020).
Bergen County
NJ
Columbia
University, Bergen
County Utilities
Authority,
AECOM
March 2020
1 WWTP and
throughout
sewershed
RT-qPCR
No
Not applicable. No public
dashboard available (AECOM,
2020; Chen, 2020).
New Jersey Institute
of Technology
NJ
None
September
2020
Each occupied
dorm
PCR
No
Not applicable. No public
dashboard available (NJIT, 2021).
New Mexico State
University
NM
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
University of New
Mexico
NM
Unknown—program details were not researched (see Section 3)
University of
Nevada-Reno
NV
Unknown—program details were not researched (see Section 3)
University of
Nevada-Las Vegas
NV
Unknown—program details were not researched (see Section 3)
Clarkson
U n ive rs ity-Potsdam
Hill Campus
NY
Unknown—program details were not researched (see Section 3)
Colgate University
NY
Unknown—program details were not researched (see Section 3)
Columbia University
NY
Unknown—program details were not researched (see Section 3)
New York City
NY
New York City
Department of
Environmental
Protection, New
York City
Department of
Health and Mental
Hygiene
April 2020
14 WWTPs
Unknown
No
Not applicable. No public
dashboard available (Kilgannon,
2020).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Onondaga County
NY
Syracuse
University, State
University of New
York College of
Environmental
Science and
Forestry, State
University of New
York Upstate
Medical University
August 2020
6 WWTPs
Unknown
No
Not applicable. No public
dashboard available (Cox, 2020).
Queens College,
City University of
New York
NY
Unknown—program details were not researched (see Section 3)
Rochester Institute
of Technology
NY
None
August 2020
15 locations
throughout
campus
Unknown
No
Not applicable. No public
dashboard available (Auburn,
2020).
Siena College
NY
None
August 2020
9 areas of campus
Unknown
No
Not applicable. No public
dashboard available (Siena, 2020).
St. John Fisher
College
NY
None
August 2020
All dorms
Unknown
No
Not applicable. No public
dashboard available (St. John
Fisher, 2020).
St. Lawrence
University
NY
Unknown—program details were not researched (see Section 3)
State University of
New York-Albany
NY
Unknown—program details were not researched (see Section 3)
State University of
New York-Canton
NY
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
State University of
New York-College
of Environmental
Science and Forestry
NY
Unknown—program details were not researched (see Section 3)
State University of
New York-
Morrisville
NY
Unknown—program details were not researched (see Section 3)
State University of
New York-Oneonta
NY
Cornell, Syracuse
University,
Upstate Medical,
Quadrant
Biosciences
August 2020
3 points
throughout
campus
Unknown
No
Not applicable. No public
dashboard available (Kelvin,
2020).
State University of
New York-Oswego
NY
Unknown—program details were not researched (see Section 3)
Stony Brook
University
NY
Unknown—program details were not researched (see Section 3)
Kent State
University
OH
Unknown—program details were not researched (see Section 3)
Kenyon College
OH
Unknown—program details were not researched (see Section 3)
Ohio University
OH
Unknown—program details were not researched (see Section 3)
University of
Oklahoma
OK
Unknown—program details were not researched (see Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
City of Corvallis
OR
Oregon State
University,
Oregon Health
Authority
July 2020
5 sewershed
locations and 1
WWTP
RT-qPCR
Yes
Map of sampling locations with a
timeseries plot of copies/L for
each sampling location and for all
sampling locations overlaid
(Corvallis, 2021).
Borough of Indiana
PA
Biobot Analytics
April 2020
1 WWTP
Unknown
Yes
Timeseries plot of average daily
flow, viral copies, cumulative
COVID-19 cases, and weekly
COVID-19 case change. Also
includes a trend indicator with
decreasing, no change, or
increasing trends (Borough of
Indiana, 2021).
Erie County
PA
Erie County
Department of
Health, Biobot
Analytics
May 2020
1 WWTP
Unknown
No
Not applicable. No public
dashboard available (Erie News
Now, 2021).
Franklin and
Marshall College
PA
Unknown—program details were not researched (see
Section 3)
Lehigh University
PA
Unknown—program details were not researched (see
Section 3)
Penn State
University
PA
None
September
2020
Campus WWTP
and sewersheds,
off-campus
WWTP
Unknown
No
Not applicable. No public
dashboard available (Lajeunesse,
2020).
Susquehanna
University
PA
Unknown—program details were not researched (see
Section 3)
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Clemson University
SC
City of Clemson
May 2020
3 WWTPs
RT-qPCR
Yes
A map of collection areas color
coded by the current impact
based on the wastewater results.
Also includes timeseries plots of
results in viral copies/L for each
WWTP and timeseries plots of
variant ratios in percentages over
the same time (Clemson, 2021 a).
University of South
Carolina
sc
Unknown—program details were not researched (see Section 3)
City of Chattanooga
TN
Biobot Analytics
May 2020
1 WWTP
RT-qPCR
Yes
Bar chart with SARS-CoV-2
concentrations. Also includes
Biobot weekly reports with
normalized viral copies/L sewage
compared to daily and seven-day
rolling average COVID-19 cases;
includes comparisons of
wastewater to samples
nationwide and Biobot's COVID-
19 incidence estimate of new
cases/day based on the
wastewater results
(Chattanooga, 2021).
Gallatin City-
County Health
Department
MT
Archer
Biologicals, LLC;
Montana State
University
March 2020
5 WWTP
qPCR
Yes
Timeseries plot of N1 and N2 in
genome copies/L compared to
the number of COVID-19 cases
for Bozeman. Remaining
WWTPs present table with
results (Healthy Gallatin, 2021).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
University of
Tennessee
TN
None
Unknown
At least 10
campus buildings
Unknown
No
Not applicable. No public
dashboard available (Crawford,
2020).
Baylor College of
Medicine
TX
Unknown—program details were not researched (see Section 3)
Houston
TX
Houston Health
Department, Rice
University, Baylor
College of
Medicine
May 2020
39 WWTPs
RT-ddPCR
and RT-
qPCR
No
Not applicable. No public
dashboard available (Houston,
2021).
Texas A&M
University
TX
Unknown—program details were not researched (see Section 3)
University of Texas
TX
Unknown—program details were not researched (see Section 3)
University of Texas-
San Antonio
TX
Unknown—program details were not researched (see Section 3)
Utah State
University
UT
Utah Department
of Environmental
Quality
July 2020
6 communities at
Logan campus, 4
dorms at Eastern
campus, and 2
dorms at Blanding
campus
Unknown
No
Not applicable. No public
dashboard available (USU, 2021).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
Hampton Roads
Sanitation District
VA
University of
Notre Dame,
Ohio State
University
March 2020
9 WWTPs
RT-ddPCR
Yes
Timeseries plot of viral load for
all WWTPs normalized by
WWTP flow rate with new
clinical cases. Also includes a
mapping of results normalized to
population over time for each of
the service areas (HRSD, 2021a).
Stafford County
VA
Biobot Analytics,
Harvard
University,
Massachusetts
Institute of
Technology, and
Brigham and
Women's
Hospital
April 2020
2 WWTPs
Unknown
No
Not applicable. No public
dashboard available (Stafford
County, 2020).
University of
Virginia
VA
None
September
2020
15 dorms and
university hospital
Unknown
No
Not applicable. No public
dashboard available (Whitman,
2020).
Virginia Tech
VA
None
September
2020
15 dorms
Unknown
No
Not applicable. No public
dashboard available (Adams,
2020).
-------
A Compendium of U.S. Wastewater Surveillance for COVID-19 Public Health Efforts
Table A-2. Municipality and university wastewater surveillance programs.
Locality Name or
Academic
Institution
State
Collaborators/
Partners
Sampling
Start Date
Types and
Number of
Sampling
Locations
Analytical
Method
Dashboard?
Dashboard Description
City of Burlington
VT
Vermont
Department of
Health,
Dartmouth
Hitchcock Medical
Center, University
of Vermont,
GoAigua
August 2020
3 WWTPs
Unknown
Yes
Timeseries plot of copies/L for
each of the WWTPs; timeseries
plot of percent of variant
(Del6970 [Bl 17], N50IY[BI 17],
and E484K [Unknown]) for each
WWTP starting February 2021
(Burlington, 2021).
Norwich University
VT
Unknown—program details were not researched (see Section 3)
University of
Vermont
VT
Unknown—program details were not researched (see Section 3)
University of
Washington
WA
Unknown—program details were not researched (see Section 3)
University of
Wisconsin-Oshkosh
Wl
Unknown—program details were not researched (see Section 3)
West Virginia
University
WV
Unknown—program details were not researched (see Section 3)
University of
Wyoming
WY
Unknown—program details were not researched (see Section 3)
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