Wastewater-Based Epidemiology as a Strategy for Tracking and Tracing COVID-19 at a University

Collette A. Thomas1, Matthew A. Roca1, Kristina M. Babler1, Helena M. Solo-Gabriele1, Mark E. Sharkey2, Ayaaz Amirali1, Melinda M. Boone3, Samuel Comerford2, Elena M. Cortizas3, Gabriella Cosculluela1, Benjamin Currall3, George S.
Grills3, Erin Kobetz2,3, Jennifer Laine4, Walter E. Lamar5, Christopher E. Mason6,x,y, Brian D. Reding4, Maria Robertson1, Stephan C. Schürer3,7,8, Dusica Vidovic7, Sion L. Williams3, Xue Yin3

1 Department of Chemical, Environmental, and Materials Engineering, University of Miami, Coral Gables, FL USA 6 Department of Physiology and Biophysics, Weill Cornell Medical College, New York City, NY USA
2 Department of Medicine, University of Miami Miller School of Medicine, Miami, FL USA x The HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medicine, New York, NY 10021, USA
3 Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, FL USA y The WorldQuant Initiative for Quantitative Prediction, Weill Cornell Medicine, New York, NY 10021, USA
4 Environmental Health and Safety, University of Miami, Miami, FL USA 7 Department of Molecular & Cellular Pharmacology, University of Miami Miller School of Medicines, Miami, FL USA
5 Division of Occupational Health, Safety & Compliance, University of Miami Health System, Miami, FL USA 8 Institute for Data Science & Computing, University of Miami, Coral Gables, FL USA
Sharkey et al. 2021

Abstract Methods/Materials Results Discussion

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is the Wastewater data from individual building & cluster buildings were
illness responsible for the global pandemic of COVID-19, declared by the analyzed and sent to the appropriate university personnel. If the
World Health Organization (WHO) in late 2019. This virus causes copies/L of sewage were high, the residential colleges or other
respiratory illnesses in humans; its spread is typically via direct airborne campuses were required to undergo mandatory testing, which
routes from inhaling droplets of saliva or sputum containing viral returned some positive results, depending on the location. It is
particles, or indirectly via surfaces contaminated by individuals infected Figure 2
possible that with daily sampling instead of weekly, the early warning
with the virus. Upon its outbreak, researchers focused their efforts to period could be shorter than 4 days allowing more time to identify
identify positive cases of the illness worldwide. At the University of Miami Figure 4: University of Miami sites Figure 5: University of Miami, Gables campus sewer drainage basins positive subjects and thereby possibly reducing disease transmission.
(UM), wastewater-based epidemiology (WBE) was a strategy employed Upon receipt at the laboratory, an aliquot of 10mL was removed
to detect the viral RNA shed from individuals as it can also be shed from the sample bottle into a sterile centrifuge tube for further Conclusion
within wastewater from various input sources of a building (i.e., toilet, processing for fecal indicator bacteria by culture, as a means of
• This study showed that the challenges with tracking disease
shower, sink, clothes & dish washing water). WBE, a growing strategy in confirming fecal inputs to the sampling site. Fecal coliform were outbreaks associated with the COVID-19 pandemic can be met
the epidemiological field for assessing the risk of public health against analyzed by membrane filtration using mFC agar by culturing 1 through a multi-pronged approach that integrates comprehensive
COVID-19, was instilled with methodology targeting a rapid-turnaround mL and 0.1 mL aliquots from 100:1 dilutions in sterile phosphate Figure 9: Significance of Variants at University of Miami- Pre Delta human surveillance of the disease with environmental surveillance
assessment, under 48 hours, so that University officials could inform the buffered saline. Colonies with characteristic blue color were of the virus.
community for targeted testing on a week-by-week basis to contain the quantified (Figures 6, 12, 13 ). A separate wastewater aliquot • In the case of COVID-19, the RNA of the etiologic agent of
spread amongst the University’s faculty and students. Methodology was then spiked with OC43 and then salted & acidified; once disease, SARS-CoV-2, was found to unexpectedly be excreted in
utilized included pretreatment of sewage prior to concentration with pretreated, the aliquot underwent electronegative filtration and urine and feces of both symptomatic and asymptomatic people.
electronegative filtration. Subsequent viral RNA extraction and the concentrated filter was sent to colleagues for further analysis • Although COVID-19 is a respiratory disease, and it would be
quantitative polymerase chain reaction (qPCR) assays were performed (Figure 6). expected in respiratory fluids, it also has been found in
for the nucleocapsid region of the SARS-CoV-2 genome, in addition to wastewater, allowing for an alternative approach to detecting early
other microbiological targets for inferencing phenomenon such as onset of outbreaks by measuring markers of the pathogen in
percent recovery of viral particles, molecular inhibition, and contribution Wastewater aliquot spiked with wastewater.
OC43 and was salted & acidified. Concentration for SARS-CoV-2
from human sources. In this two-year study, research was used to track • There was an observed presence of SARS CoV-2 that causes
the rate of infections, and predictions were made to supplement results Covid 19 that shows up in wastewater samples and the positivity
from clinical testing within the University’s campus and Hospital. This rates on all campuses for both faculty and students.
was novel as those who were pre- or asymptomatic rarely sought after
reported testing, skewing the effectiveness of human-reliant surveillance.
Manhole sample
1:100 raw wastewater 0.1 mL & 1 mL aliquots from 1:100 dilutions into 20
Future Directions
collection
& PBS dilution mL of PBS. Incubated at 44.5ºC for 24hrs
Results of the rapid-turnaround assessment allowed for effective Figure 6: field-to-lab overview
Figure 10: Significance of Variants at University of Miami- Delta
• Work should focus on expanding techniques and protocols for
communication from laboratory analysis to policy decision for the environment monitoring of infectious agents for the purpose of
University. From there, sub-studies to assess relationships between Results
tracking disease outbreaks
clinical cases, hospitalizations, variant analyses, methodology • Compare the significance of the different variants of public
optimization, and effective reporting of SARS-CoV-2 have branched from schools versus University of Miami cases
this work. WBE is expanding as a tool that is independent of human • Explore at post-omicron trends
testing, demographics, and can provide real-time data for the protection
of community health. References
• Sharkey, M. E.; Kumar, N.; Mantero, A. M. A.; Babler, K. M.; Boone, M. M.; Cardentey, Y.;
Cortizas, E. M.; Grills, G. S.; Herrin, J.; Kemper, J. M.; Kenney, R.; Kobetz, E.; Laine, J.; Lamar,
Methods/Materials W. E.; Mader, C. C.; Mason, C. E.; Quintero, A. Z.; Reding, B. D.; Roca, M. A.; Ryon, K.; Solle, N.
S.; Schurer, S. C.; Shukla, B.; Stevenson, M.; Stone, T.; Tallon, J. J., Jr.; Venkatapuram, S. S.;
Sampling occurred weekly on Wednesday mornings from 7:30 Vidovic, D.; Williams, S. L.; Young, B.; Solo-Gabriele, H. M., Lessons learned from SARS-CoV-2
measurements in wastewater. Sci Total Environ 2021, 798, 149177.
am to 10:30 am, starting on September 30, 2020, and continuing • Zhan, Q.; Babler, K. M.; Sharkey, M. E.; Amirali, A.; Beaver, C. C.; Boone, M. M.; Comerford, S.;
through the summer of 2022. Both clusters & individual buildings Figure 7: SARs COV-2 Positive Cases Among Faculty/Staff and Students from Fall 2020 to September 2022
Figure 11: Significance of Variants at University of Miami- Omicron
Cooper, D.; Cortizas, E. M.; Currall, B. B.; Foox, J.; Grills, G. S.; Kobetz, E.; Kumar, N.; Laine, J.;
Lamar, W. E.; Mantero, A. M. A.; Mason, C. E.; Reding, B. D.; Robertson, M.; Roca, M. A.; Ryon,
were sampled, all collected either from manholes or from lift K.; Schurer, S. C.; Shukla, B. S.; Solle, N. S.; Stevenson, M.; Tallon, J. J. J.; Thomas, C.; Thomas,
stations (Figure 1 & 2). At each site, a bottle-on chain approach T.; Vidovic, D.; Williams, S. L.; Yin, X.; Solo-Gabriele, H. M., Relationships between SARS-CoV-2
in Wastewater and COVID-19 Clinical Cases and Hospitalizations, with and without Normalization
was used to retrieve the wastewater. An ISCO peristaltic pump against Indicators of Human Waste. ACS ES&T 2022, (Water).
auto-sampler was used at two individual building scale manholes • https://coronavirus.miami.edu/dashboard/
in order to collect a weekly 24-hour composite sample (Figure 3).
Acknowledgements
Later parts of the research reported in this publication were supported by the
National Institute On Drug Abuse of the National Institutes of Health (NIH) under
Award Number U01DA053941. The content is solely the responsibility of the
authors and does not necessarily represent the official views of the National
Institutes of Health. We are very thankful to Dr. Helena Solo- Gabriele, Dr. Mark
Sharkey, Dr. Sion L. Williams, and all colleagues with whom we have interacted and
Figure 12: Figure 13: discussed best practices for wastewater sample collection, concentration, and
Top and bottom plates showing acceptable cfu counting conditions
Figure 1: Building scales Figure 2: Lift station sampling Figure 3: ISCO auto-sampler
Top plate showing ”Too many to count” (TMTC). analysis for SARS-CoV-2.
Bottom plate showing acceptable cfu counting
Figure 8: Positive Cases at the University of Miami against the 3-day moving average of Wastewater Data conditions.

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