Wastewater-Based Epidemiology Initiative’s interdisciplinary team tracks COVID-19-triggering virus

So far, many tests for the coronavirus have involved air, its main transmission medium. (Hence, face masks’ sudden emergence as couture). But an enterprising, interdisciplinary group of Norwich students and faculty has started looking for the virus in another “essential element,” water.

Since September, the group has been developing a plan to test wastewater from the Northfield Wastewater Treatment Facility and campus manholes to look for viral RNA, a genetic signal, to detect COVID-19. The testing works as an early screening tool because the virus can be shed in feces even when infected people are asymptomatic.

Dr. Tara Kulkarni, an associate professor of civil and environmental engineering, is co-directing the university’s project, dubbed the Norwich University Wastewater-Based Epidemiology Initiative. The project took its first sample from the Northfield Wastewater Treatment Facility at 242 Dog River Road on Sept. 30; additional samples were collected there Nov. 4, Nov. 11 and Nov. 18. Results were COVID-19-positive on samples from the latter two dates, unsurprising, Kulkarni said, given COVID-19’s documented local presence.

Norwich’s project involves civil engineering, construction management students (to build equipment and conduct the sampling); chemistry and biochemistry students (to analyze the samples) and humanities students (to record podcasts and promote the project).

The federal Centers for Disease Control and Prevention has said wastewater testing can portend COVID-19 outbreaks by up to seven days before confirmed cases appear in public health data. The CDC created a website to which state, tribal, local and territorial health departments can submit wastewater testing data for a national database. In Vermont, Burlington has been testing sewers for COVID-19 since August.

Norwich’s team collected samples from three campus locations Nov. 5 and Nov. 14 and analyzed them Nov. 19 in university labs. Goodyear and Gerard hall samples yielded positive results, triggering a move to dorm quarantine from campus quarantine and requiring resident students to stay in their rooms, wear masks and wash their hands.

In a Nov. 20 Update from the Hill memo, Norwich President Mark C. Anarumo said several hundred students from these two dorms completed targeted testing for COVID-19 at Plumley Armory that day.

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Dr. Tara Kulkarni, an associate professor of civil and environmental engineering, works with students from the Norwich University Wastewater-Based Epidemiology Initiative team on Oct. 14, 2020. (Photo by Mark Collier.)

Senior construction management majors Conner Ruland and Looknauth Mahadeo of the initiative’s technical team, directed by lab technician Eric Wood, assembled an autosampler to test water from campus manholes. The autosampler lets the team collect a 24-hour composite sample, meeting CDC best practices.

Pilot testing continued through three other major manholes on campus before students began leaving campus over the Nov. 20 weekend for Thanksgiving. Kulkarni said sampling will likely occur at least once weekly when students return to campus in mid-January for the spring semester.

Norwich’s project involves civil engineering, construction management students (to build equipment and conduct the sampling); chemistry and biochemistry (to analyze the samples) and humanities students (to record podcasts and promote the project). Professors include Kulkarni, Communications Lecturer Dr. Stephen Pite and Chemistry Lab Coordinator and Lecturer Marie Agan.

Kulkarni said work will continue through the academic year. Students and faculty will then assess whether the work should keep focusing on COVID-19 or transition to other wastewater-based epidemiology aspects for long-term research.

Origin story

Netherlands researchers in July published the first report of analyzing wastewater — from home and building use (i.e., feces-containing water from toilets, and water from showers, sinks) and nonhousehold sources (e.g., rainwater and industrial use) — to test for RNA signals of SARS-CoV-2, aka the novel coronavirus.

The CDC has said wastewater testing has previously helped to detect diseases, including polio, early. (Wastewater testing in Israel for poliovirus in 2013 and 2014 prompted the immunization of 1 million children, The New York Times reported, halting the outbreak before any cases of paralysis were reported). Wastewater testing has also been used to detect opioids.

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Norwich University Lecturer and Chemistry Laboratory Coordinator Marie Agan observes a wastewater tank at the Northfield Wastewater Treatment Facility in September 2020.  (Photo by Norwich University Wastewater-Based Epidemiology Initiative.)

Norwich’s project will eventually merge with efforts by the University of Vermont, and St. Michael’s College. Efforts have begun to seek collaborative funding to research broader impacts of using wastewater in epidemiological analyses to bolster community health. 

Kulkarni said careful collaboration can ensure findings are accurate and reliable and wide faculty involvement might help attract more research funding to diversify the project. She and Agan agreed that myriad viewpoints enriched the experience.

“Using these multidisciplinary teams to fix these problems is what we do here at Norwich,” Agan said in an interview with the communications students involved in the project. “I read a quote … ‘Build a team that’s so good that you can’t pick out its leader.’ I think that’s what we’re doing here.

“Find people who know things that you don’t, or find people who know things that you do know, it’s good to have diversity of thought in your research team.”

National effort

With its project, Norwich joined a collection of universities and labs nationwide in using wastewater to test for COVID-19. NPR reported Oct. 26 that more than 65 U.S. colleges are testing wastewater to monitor coronavirus spread; others include Syracuse University, Colorado College, Louisiana State University and the University of California, San Diego. Researchers have said dorm wastewater testing works for colleges because researchers can pinpoint groups who might be infected because they can tie wastewater to specific dorms. 

In August, the University of Arizona said wastewater tests might have prevented an outbreak. After a wastewater test from a dorm came back coronavirus-positive, the university tested the 311 people living and working there and found two asymptomatic but COVID-19-positive students, The Washington Post reported. The infected students were immediately quarantined. On Nov. 11, the NNY360 news website reported that St. Lawrence University called for a precautionary quarantine at a dorm following a COVID-19-positive wastewater test.

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Norwich University Wastewater Based Epidemiology Initiative team members from left, Conner Ruland, Looknauth Mahadeo, Shi Miao Yuan and Jack Schultz, all members of the Corps of Cadets, stand by a campus manhole in September 2020. (Norwich University Wastewater-Based Epidemiology Initiative.)

In New England, the Massachusetts Water Resource Authority awarded Biobot Analytics a $200,000 contract for a six-month pilot study of wastewater at the Deer Island Treatment Plant in Winthrop, Massachusetts. The authority has given Biobot Analytics water for sampling since March.

“Analysis of wastewater for the genetic signal (viral RNA) of the SARS-CoV-2 virus that causes COVID-19 is proving to be a cost-effective approach to providing population-level screening for outbreaks of COVID-19,” the authority said in a statement.

Agan, who’s working on the project as part of her graduate studies in analytical chemistry, medical lab science and public health at the University of Vermont, said Norwich’s wastewater testing methods follow CDC protocols. It’s using a real-time reverse transcription polymerase chain reaction test that amplifies the RNA to test for the coronavirus’s presence. The approach relies on detecting genetic fragments of the virus that are excreted in stool by qPCR analysis, which does not determine whether the virus is dead or active. 

To validate the polymerase chain reaction samples, Agan said she’ll run loop-mediated isothermal amplification reaction tests, which also amplify RNA. Norwich’s first samples, collected from the Northfield Wastewater Treatment Plant weekly, were first sent to outside labs for testing, including Biobot, of Cambridge, Massachusetts, and Quadrant Analytics in Syracuse, New York.

In-house aspirations

Agan and Kulkarni have said Norwich hopes to move testing in-house to save costs and give students hands-on analysis experience. Seniors Gracie Dominguez and Samantha Gonzalez and graduate student Mallory Dutil, a 2020 Norwich graduate who’s pursuing a master’s degree in civil engineering, will work with Agan in Norwich’s lab.

The project’s students meet in weekly GoToMeeting gatherings to compare notes and debrief. In an October meeting, many said they liked the project’s altruism. By finding early COVID-19 warnings, they said, they could meet an immediate need — keeping the university, and the Northfield community healthier and safer. By developing equipment, data sets and methodology, they said, they could meet a future need — knowledge and systems that can apply to other projects.

“We’re setting up a plan that the school can potentially adapt,” Mahadeo, the Norwich technical team member, said. “They can use this for a situation like this or another one that involves like, a natural disaster. (It’s) flexible.”

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From left, Norwich University Lecturer and Chemistry Laboratory Coordinator Marie Agan and construction management major Shi Miao Yuan, teammates on the Norwich University Wastewater-Based Epidemiology Initiative, collect a wastewater sample from a campus manhole in October 2020. (Photo courtesy Norwich University Wastewater-Based Epidemiology Initiative.)

Dutil, who graduated in August with degrees in chemistry, engineering and environmental science, and whose master’s focus is environmental/water resources engineering, said she’s looking forward to returning to the lab and helping people in Northfield, where she’s long lived. Ruland also welcomed the chance to help the community.

“This is something we can do to help not only Norwich, but you know the Northfield community,” Ruland said. “(We’re) able to be involved and be part of the solution compared to just talking about it.”

Jack Schultz, a senior education communications major who’s been recording podcasts and interviewing his student peers for the project, said, “This is kind of my way to … be able to help out what’s going on around the world.”

Jamie Heath, a sophomore business management major and Schultz’s communications partner on the initiative, said she was enjoying learning new viewpoints.

“It’s different than anything I’ve ever done before,” she said during her interview with Kulkarni. “My brain doesn’t really work the way a scientist’s brain does. So, it’s cool to hear different perspectives and all the science talk.”

Kulkarni said the initiative may add students and faculty as it evolves. In an interview with the initiative’s humanities students, she said mathematics majors, for example, might introduce models for data analysis or business students might examine the pandemic’s effect on community economics.

Agan said she hoped the project might inspire Norwich to expand its science, technology, engineering and mathematics curriculum, perhaps adding courses in clinical science, clinical diagnostics or medical laboratory science.

“With the need for clinical lab scientists, especially in the military and in the civilian sector, we as a college are uniquely poised to respond to this need,” Agan said. “Over the next five years … clinical science is poised to grow significantly, especially with emerging infectious diseases like COVID-19. There’s an opportunity to go beyond this testing.”

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Wastewater-Based Epidemiology Initiative personnel:

Technical team: Students, Elias Bou-Chahine, Looknauth Mahadeo, Conner Ruland, Nirmal Tamang and Shi Miao Yuan. Faculty lead, Tara Kulkarni.

Scientific team: Students, Gracie Dominguez, Samantha Gonzalez, Mallory Dutil. Faculty lead, Marie Agan.

Humanities team: Students, Jack Schultz and Jamie Heath. Faculty lead, Stephen Pite.

(Slideshow photos by Mark Collier and Norwich University Wastewater-Based Epidemiology Initiative.)

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