Collection dates, estimated cases and the estimated prevalence rate for the Angola sewershed are shown in this graph. Numbers came from testing through BioBot Analytics using samples taken weekly from the Angola Wastewater Treatment plant.

ANGOLA — As the school year begins, the Angola Wastewater Treatment staff have a plan in place to help potentially identify COVID-19 in wastewater by testing samples from the collection system serving Angola High School, Angola Middle School and Ryan Park Elementary School as well as a system on the Trine University campus.

Treatment plant staff and BioBot Analytics of Cambridge, Massachusetts, have been working to identify and quantify COVID-19 in wastewater since late March using a program that is continuing to develop even now.

In March, Wastewater Superintendent Craig Williams read an article about BioBot and how the founders took their model — originally testing wastewater to track opioids — and looked at how their sampling and testing knowledge could be applied to help identify COVID-19 in the wastewater as well.

Angola is one of the many places doing similar testing, according to Williams.

“BioBot developed the method, and we send a sample once a week,” Williams said.

Since March, samples from the Angola city sewershed served by the plant have been sent in. Within three to five days, the analysis is completed and BioBot sends results back.

“I forward the reports to the Steuben County Health Department, Cameron Hospital, the Angola COVID-19 Task Force and other city leaders,” Williams said. “By testing for COVID-19 in wastewater, we are able to get a snapshot of the infection rate of the local population and by observing trends, may be able to provide advanced warning of an increase in cases to public health officials.”

Testing protocols

BioBot’s testing protocols include sample pasteurization to inactivate coronaviruses, removing bacterial cells and using a polyethylene glycol based precipitation method to concentrate viruses in the sample, extracting the RNA genome and using a quantitative polymerase chain reaction-based method to detect and quantify the SARS-CoV-2 virus and an associated fecal normalization control.

A virus genome is used to create a standard curve that gives the answer of copies per milliliter of sewage.

To measure virus concentration in sewage, measurements are converted into a case estimate using the following equation:

Number of infected people = (total amount of virus per day) / (virus shed per infected person per day)

Estimates from BioBot may not match confirmed community case numbers for a number of reasons such as that clinical testing may not represent the total infected population and that the virus may start being shed before patients develop symptoms and seek out testing.

Methods are adapted from Centers for Disease Control and Prevention protocols with the approach relying on detecting genetic fragments of the virus in stool. It does not determine if the virus is dead or active.

Case estimates

Case estimates, according to the July 27 data set report, should be evaluated as beta results when comparing to reported COVID-19 cases.

In March, BioBot published the first study — not yet peer reviewed — to estimate numbers of people infected based on sewage levels collected from a Massachusetts metropolitan area. On March 25, that area had approximately 446 confirmed cases of the virus.

Sewage analysis from the area estimated up to 115,000 people infected and shedding the SARS-CoV-2 virus.

The study can be found at

The first data set for Angola, dated March 25, estimated 60 cases could be in Angola. Since that date, numbers have ranged from a low end of 20 to approximately 740.

Sampling expands

Starting on Aug. 16, sampling will expand because of a program coordinated by the Indiana Finance Authority.

Sampling will be done four times a week at the wastewater plant, three times a week on the Trine University campus, and at a point in the collection system serving Angola High School, Angola Middle School and Ryan Park Elementary School once a week.

“All of this information will be provided to the health department, the task force, Cameron Hospital, Trine University and the Metropolitan School District of Steuben County,” Williams said. “Our hope is to provide data that helps public health officials, management at Trine University, MSD and the city to identify and respond to apparent case increases before they become a public health emergency.”

While testing at, say Trine, won’t be exact as far as what residence hall numbers are from, Williams said the information should be enough to get a good risk analysis feedback loop going.

Sampling at Trine will include residence halls, the MTI Center and the Athletic and Recreation Center.


Initially, Williams said his department re-prioritized a few things to make sure funds were available to do the testing. As things progressed a bit, he said conversations with Deputy Clerk Ryan Herbert led to Herbert calling the finance authority to see if testing cost would be reimbursed.

As it turns out, yes, so that has enabled the testing to go on much longer than initially thought possible.

Williams said as of now, it looks like the testing program will be able to continue into October.

“The finance authority has been just fantastic to work with,” he said.

A large undertaking

As testing increases, Williams said it will be a bigger task for his staff, probably using one or two more man hours daily to get it all done, but its something he feels will be worth it.

“It will be challenging, but worth it,” he said. “We feel the data will get the feedback loops, to help to get some level of normalcy back.”

Aug. 3, when Williams told the Angola Common Council what he was doing with the testing, Councilman Dave Olson was quick to give accolades.

“Congratulations to all of you on that,” Olson said. “This is a fantastic plan that could potentially stop a huge crisis.”

Williams said the hope is to continue to foster an environment that allows the community to continue recovering from the pandemic.

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