Kentucky.

Health is funded in part by a grant from the foundation for a healthy Kentucky.

>> Infectious diseases can be tracked using wastewater.

Now, I don't know how, but I know a guy who does stay with us.

As we talk about wastewater epidemiology with Doctor Eric Rhodes Co next Kentucky Health.

♪ ♪ ♪ To appropriately treat someone with an infectious disease.

The offending agent, whether bacteria fungus or virus must be identified.

>> Identification of the pathogen also allows for when necessary the institution of measures to protect the patient's family.

Co-workers and the community from potential spread.

Techniques for painting material for now says familiar to all of us and include phlebotomist for blood swabbing about nose or throat.

The cough and sputum and yes, providing a stool sample.

Not this kind of stool sample, but this one.

These techniques are good for evaluating individuals, but there's an alternative and less intrusive way to detect the presence of a pathogen in our neighborhoods and communities.

And that is through the sewer system.

The talk to us about wastewater epidemiology we have is our guest today.

Doctor Eric Rhodes Co but the road to graduate from Rockhurst University with a BS and math computer science and biology.

And then attendant Rensselaer Polytechnic Institute for MS in computer Science.

And later obtained a doctor of science in computer science at Washington University in St. Louis, Missouri.

He's currently professor of biochemistry and molecular genetics at University of Louisville.

And so should director of can Kentucky about medical research infrastructure network and Biles Informatics Corps director, Kentucky by medical Research Infrastructure Network, Doctor Rhodes.

Eric, thanks for being with us today.

Thank you for having me.

I use the term wastewater epidemiology so you surveillance or the 2, can they be interchange?

>> Yes, so we we think about the So we've been doing starting with the sars-cov 2 looking for within the wastewater in Jefferson County, Kentucky.

>> So what is the premise of using our sewer system.

Which I assume wastewater and that sewer system as a means of detecting following the pathogen like SARS or anything else.

>> So if we think everyone hoops, right?

So everyone is going to be contributing to the wastewater system.

And so we'll be able to measure on a more accurate basis.

What's actually in the wastewater as opposed to relying on diagnostic tests.

So we take Sars-cov 2, for instance, early on, people may have gone and gotten a PCR test and we would know those results.

But later on, as we got the at-home where people could contest themselves.

We believe having that information and knowing positive rates was an is as easy to get So instead of doing we thought of using a wastewater measuring what's in the actual area.

So we've been looking at 17 different catchment facilities throughout Jefferson County, Kentucky.

So we have a coverage of the Hole County as a whole and be able to measure what's in the population more accurately.

>> Welcome back to those catchment communities in a minute.

But is the premise in so if someone's at home or they're at work, as you said, there's a book by that title.

By the way, everyone P****.

So that's how the storm gets into the sewer system in you evaluate it.

>> Right.

So that's what we're looking at is is the wastewater and looking at it several different locations.

But yes, the people are contributing to that and then the amount of bacteria, the amount of viruses that are in the way SWAT.

>> Clearly there's not one guy that takes his bucket and goes and then down into the sources of who are the people that are your partners that helped in this endeavor?

Right.

So it starts with Metropolitan sewer district and nasty.

And so they.

>> What they're doing at these locations that we have for taking what's Composites sampler, which is step down, basically into the wastewater over 24 hour period.

And it does a slow it's low capture of wastewater over that time.

So it's not just at whatever is in the wastewater that point in time.

But over 24 hour period, theoretically everybody will if use the restroom and facility and we'll have contributed to that will be able to measure more accurately from and one day period.

>> Given the current technology.

Are you measuring you're looking at bacteria or is the potential for looking at bacteria, viruses, fungi, other infectious pathogens or what is it and what are some of the things that you can see?

>> Right.

So when we're looking at the wastewater, these are going to be any pathogens that are frequently so in this case are going to be a lot of bacteria that we look at.

We can look at viruses as well.

And we could potentially a look at other fungal as well.

So well, we've been focusing on and we have data now for the past month or so on a on a weekly basis, looking starting with a lot of respiratory viruses that are interesting, including Sars-cov 2 looking at monkeypox.

I'm looking at polio.

So we've had those.

And so it's come and then obviously is importance worldwide.

But then looking at other things, such as at no viruses or boca viruses, things that or more common looking at influence.

And so we're working on figuring out a way to be able to to detect those on more an easier bases we're also looking at bacteria, but specifically since bacteria, genomes are much longer and would swamp out most of what's in the wastewater in terms of the pathogens.

We're focusing on cassettes are genes that related to antibiotic resistance.

So looking at bank, my son resistant genes, Memphis on resistant genes.

And so looking at specific regions.

But then those bacteria.

So >> when you talk that's going too because, you know, we all are concerned about South Sudan about of resistant bacteria.

Thing so you might be able to pick up an area where there's a give a greater concentration.

A virus or again, one of And it's about it was just better.

Can you then go back and say, OK, we've narrowed it down to this area and we need to do something at that site.

>> Right.

And that's that's the hope is that by using these these pan pathogen models that once we see something that that is spiking up or is coming up, that that can be used to make some sort of public policy decision in that area that at that given a point in time.

So as one example of Sars-cov 2 looking will be able to measure win 1, 1, areas more highly concentrated.

We're also looking at the whole genome sequencing, the whole 30,000 base genome and being able to look at individual mutations along those 30,000 bases and being able to detect, then what are the variants of concern?

So obviously now it's Omicron.

But are we are we seeing those variants?

Are we seeing particular variance in certain locations?

And now that now what we know in terms of treatment, certain variants are more treatable by a particular targets.

And so that that's also knowing having that information may give physicians a more direct a treatment more immediately to to try first.

>> Tell me about this map of the catchment area that so how do you divide out the county or it is just to think how you look at the metropolitan area.

>> Right.

So so right now we're looking at a Jefferson County.

We have pretty good coverage across the whole We have 17 different catchment sites.

5, those our water quality treatment facilities.

We've been looking at all 17 sites for Sars-cov 2, at least for the for a little bit over a year.

We're now focusing on the 5 water quality treatment sites which are basically cover the whole county as well as 3 problematic areas where we've seen a lot more diversity showing up with Sars-cov 2.

And so we're looking at that.

And so it's a smaller population, contributing to those.

So >> I kind of think back when I played in the band, a member band director told everybody that play in particular, he's and then he told the right half where I was sitting in the right half that he said the back half.

I was in the back end then said the last line I was in the blast line that he stood in front of me and told me the play.

That's what I quit ban.

But some wondered.

>> Can you isolate out too?

And individual businesses community center home nursing resulting hospital.

>> All right.

So you could look at the wastewater, this coming out of certain facility.

And so the environments to get the University of Louisville has done some Sam plane for rent, for instance, within correctional facilities throughout Kentucky.

We've also been working some some researchers at the University of Kentucky that have access to nursing home facilities throughout the eastern part of the state as well some other areas such as looking at rest looking at it.

The transient as people are moving through the state.

And now those have been used a lot for for illicit drugs.

Throughout these regions, but but they could be used for for measuring the Pan pathogens as well.

Anything where we're sampling wastewater will be able to do.

And as long as you have a control of where that's coming from, you can do a more localized level.

All right.

So you have this voluminous.

Amount of material.

That is coming down.

>> How can you possibly tell me that in all this Pu you can pick out viral particles and with some degree of accuracy say yes, this concentrations going up.

Why isn't it obscured by all the other stuff there?

How do you do these tests?

>> Yes, so if we think about it, when you when you go, when you've got get your nose swabbed for for getting a test for sars-cov 2, what are they doing?

So they're looking at the actual presence of that virus in your nasal swab and really disregarding everything else and the way that that that's done with Sars-cov 2 with those tests, the PCR test is really to signing a primer, which is a series of a base, is about 20 basis long that the flank hit one region of the 30,000 base genome.

Another set that that flank another 20 bases and you sequence across as sincere, actually sequencing the genome.

And it's only going to be the genome of sars-cov 2.

So if you're infected with influenza, it's not going to check the influence and less you detect primers specifically for those.

And so what we've done with the Sars-cov 2 is we have a a we're that we're using from company that has these flanking.

So when when you go to get tested for Sars-cov 2, just see if you have it.

It's basically only looking at 3 regions of the genome that they pay.

But what we do is we're actually using a kit that has frank these flanking regions to cover the whole genome wide.

And they're also overlapping.

So if there are any mutations in here that don't allow it to actually amplify, we can actually still sequence those well.

And so what we're doing with the Pan Pathogen model us a little bit different.

So instead of amplifying these regions like we do a sars-cov 2, what we do is we've designed a set of of although Nucleotides which are complementary to the that the DNA or RNA, depending upon but type of virus, it is and it gets all the material gets passed through saw than that nucleic acids gets passed through and it's only captured if we have probes that are complementary to.

And so if we have a particular bacteria that's found very high levels were not interested in is not to buy into any of these fall through.

And so we'll have these capture beads that have the sequences that were interested in sequencing.

And then those are then taken run a DNA sequencer at the sequencing Technology Center and University of Louisville and then we 100 million reads of whatever's in the wastewater at the sites.

Well, and then what we do is then we take those rates that we map them against a database of what we know.

We're trying to collect and then we can see across the whole genome of these.

How much of every single base is covered?

Are there any mutations in these if we get enough reads and then we can all I also normalized that.

So with the sars-cov 2, the way that the normal station has done is by looking at a it's a the virus associated peppers and peppers, the senators that one made so that you eat.

Okay.

So this fire says associate with peppers, many peppers.

You also eat the virus that passes through your system and then we can measure what level is that particular virus in this location?

And then we can normalize that across locations because theoretically it should be found at a basic and all the sites and with the frequency of.

>> But he next restaurants around with all the peppers.

I'm sure you have a widespread base.

I just got to get this feeling, though.

Number one, there's got to be a tremendous amount of noise because of all the other stuff that's happening.

So I get to envision that there's a conveyor belt that's going by and you're sitting there going.

Yes, is to just get pulled out and for the value in the rest of the stuff you let pass on by.

>> Right.

And then so that's really what was happening.

So we are pulling out specific things that we're interested in and recognize recognizing that this is what I want.

This is what I'm in a sequence.

All this other stuff.

I'm not interested in.

And so as a what currently doing there are 29 respiratory viruses that we're looking at.

And we're actually looking at 50 different strains within within those.

And that's part of a respiratory virus package where they have these all the nucleotides I created.

But then we have an additional 40 or so bacteria, bacterial genes as well as other viruses that we're pulling off as well that we've designed those all going to play time.

>> They used the term How does that actually work?

So the what is that?

Yes, the way that the PCR work with the season was in.

So it stands for Polymerase chain reaction.

And so the way that it works is you start with one strand of DNA.

Okay.

Okay.

So we think about this with Sars-cov 2.

You're going to have many copies of that and your and your nose.

>> And and so what happens is one piece is complementary to one region heading in one direction of the ACS.

Jason tease for that region.

You have another that's complementary in the other direction.

And so DNA is a double strand molecule.

Yeah.

You're using a single strand of molecules.

A 10 play adding on these primers and then it sequences from the beginning of this tree to the end of this street.

And so you have now, instead of having one copy of that region, you have 2 copies.

We continue this process to goes to 4 for ghost coast to 16 16 goes.

32 a doubling process every single time.

And so what what you can do to measure the amount of fires that you have you may have seen if you've had a test.

And you've tested positive what's called a CT value and that CT value is going to be the cycling value and the lower that value is tells you that it was that many cycles that you had the form of this PCR before it was detected.

Okay.

So if it was detected after the second cycle, that's really bad.

I mean, there's a light.

So there's a lot there that says you got a notice that says we rarely rarely see anything under 10.

But you see something 10 to 15.

That means that you have a lot of virus hanging out.

You see something to the level of 40.

You don't have any virus and they're so what we see typically in the wastewater now with levels going down trending downward CT values that maybe 30 34 35 still detectable, but not as high.

And each one of those, there's a 10th.

There's a a 2 fold difference right between 34 and 35.

>> So if I'm out walking and I have my binoculars, not see a cooler way away in 100 times.

Magnification don't have to sweat.

But if he is that 2 times magnification, I got a problem.

I think I think I guess, right?

So so by looking at that you're able to to detect so what we're doing, the sequencing and doing the normal station, you can see the coverage and then we can.

>> And part of it up to is figuring out what's there is as a baseline.

So there's always going to be something in the wastewater, Your your body has to function with bacteria to break down foods, for instance, in in your gut.

other areas and called this culture.

Micro includes all of these.

So they're all part of our everyday process.

They're going to be part of the foods that we eat and most of them are not going to be harmful and are in fact, are going to be beneficial to us.

But it's when we get those things that now there's a level at which is is not normally found that that's what we need to KET an eye on it and figure out what might be both the cause and a consequence.

I see a tremendous benefit to us all for this.

But there's got to be a cost who's picking this up?

So with the sars-cov 2.

It's we've been funded by a couple of different places.

One has been the Rockefeller Foundation and their pandemic Prevention institute.

And and really what they're interested in, it's not as we're looking at it in terms of what's happening locally for us in terms of Jefferson County.

But they're also looking at that in terms of what's happening worldwide.

And so we've worked with a number of researchers, for instance, in Chicago that that had a lot more catchment areas and looking at what's in their waste water, including Airport some researchers and Oklahoma kind of doing this.

The same thing Kiwanis is actually done this.

Look at as airplanes would arrive and to Australia and kind of quarantine until they KET.

Was this flight a claim flight or not in terms of star see could be done that quickly.

It could be done that quickly.

Looking at the by the time you go through customs and get down the down the road that that within a short amount of time that that you could have an answer.

And then we've also had support the metro Louisville Department Metro Public Health and Wellness is keenly interested in this and figuring out what's in the wastewater.

We've had some collaborators with a Jefferson County Public that had some interest in more.

The clinical samples which we've been doing sequencing of as well.

But they have an interest in what we're doing and what we're seeing in certain areas because that can also affect, you know, micro changes that they could bake.

And, you know, if we've seen a certain virus or were seen an uptick in something, a certain area where there might be schools and maybe they can make decisions on how intermix students might be and in the classroom during this time.

So the data that you're getting now can lead to actionable public policy.

Within days of cells, right?

So our typical from one week we get the typically rise on a Monday.

And the part of that goes your offense, you know, makes it does some of the sequencing work with PCR analysis just to to detect levels that were seen and they have that within a day.

And then when we do that, the whole genome sequencing, it takes a couple of days to process.

That takes about 19 hours to sequence it and takes about half a day to to do the analysis of that variant detection.

And so generally by Friday, we have an answer for data that came in on on Monday.

>> I am sure that there will be some who will fear big brother and say that you're a truce of and intruding I'm intruding a live and I don't want you to and be able to sample me.

How specific can you get back to a point where you say this block is where something this house?

>> Yes.

So with what we're doing, we're not at that level of granularity.

So we're looking catchment sites that the larger catchment sites, maybe 100,000 individuals that are contributing to that the smaller catchment sites may be, you know, on the order of a couple of 1000 individuals.

Now, theoretically, you could do that.

You could map back to an individual house that becomes very expensive.

because now you're having to to track back and how do you know to track back to that particular house and not all the houses and in particular neighborhood?

So we are cognizant that that there are our concerns.

I'm with, you know, what we find and where we find it and who we share.

That data I'm really And so what we do is once we've we've gotten our our answers, we share that with public health and wellness department and then they make a decision on where it goes from there.

>> I would imagine that hospital systems and insurers would be very much interested in knowing what the levels of exposure or potential exposure are in the community.

The >> yes.

So I do think hospitals are are interested in this.

And that's one thing that could be done with this is actually looking at what's in the hospitals themselves and what might be the circulating that could could help them make some decisions as well.

So, you know, that gets back into having a little bit too much information and that might not be.

>> Which I think in terms of staffing, if I know that we're early on, there's the potential spike, what we need to shift and icus, respirators and all.

That's right.

So the and that's one thing that we've throughout.

And I think we had 87 weeks where we're doing the sars-cov 2.

>> So one of the things we're looking at as well as hospitalizations.

So, you know, some variance are are worse than others to short session.

Cern show.

And so looking at the trends with that from when we detected 2, when the hospitals are going, I'm going to look at worldwide too.

And so, yeah, that information could kind of effect.

It's it with Sars-cov 2.

There's going to be a delay right from when you get it detected, too.

When you start seeing people showing up in the hospital.

>> Is this technique of using wastewater surveillance?

>> Is it complimentary to contact tracing or does it's a plan that now?

So I you know, I think that the contact tracing is is is really looking at once a person becomes infected and looking at the other people that could be infected and and trying to help control the spread of a virus.

And we've seen some things in the wastewater where particular variant pops up in a location and then we never see it again.

And so this is an indication that somebody has gotten a variant that we just don't see.

Typically here, they've done the right things.

Yeah, that they've quarantined and quarantines work.

They go and we actually saw with with Delta.

We saw Delta appear about 2 to 3 I guess, was about 2 months before her.

It was really popped up in one location in the wastewater and was gone.

And then once it came in, we found at multiple locations.

It grew pretty quickly.

>> Thankfully, we've got about a little over a minute.

I want one of the 3 things you would want us all to know about wastewater.

So that is good about this.

>> So with the with the wastewater surveillance, I think it's it's good way to to help measure the pathogens that are in the environment.

I guess I don't use it as a scare tactic because we're seeing something doesn't mean that it's necessarily a bad thing.

We're just keeping an eye on what's going on there are also some viruses, you know, for instance, respiratory viruses that were interested in that aren't weekly shed at the same level.

And so we're thinking about ways in which we can measure other sort of input such as air ventilators or or things like to maybe pull out some of those.

And so it's a way in which we can also very easily pivot to what other whatever may be coming down the road and to be able to detect going forward.

So you're kind of excited about the sort of stuff.

Yes, I'm excited about So my area is fire from attics.

And so I'm on the analysis end of the things.

And so I'm excited because when I'm looking at the data, I'm really the first one that knows what's what's going on?

Because on the one that's that's looking at the data and it's it's interesting because it's changing all the time.

>> Well, when I see you run into the store and getting all the pack to that and all the other stuff I know I need to start worrying about what's going on.

And this is just very interesting.

I want to thank you for taking time out of your busy day to come down and talk and try to explain to someone like me, find the festive but knows nothing about it.

I want to thank you for being with us today.

Also, I hope that you have a better understanding of what wastewater surveillance is and not understand how it functions as part of a armament area use of fighting infectious disease.

>> If you want to watch this show watch an archived version of past shows.

Please go to www DOT KET Dot or if you have a question or comment about this on the shows, we can reach that.

Okay.

Why health at KET Dot Org.

I look forward to seeing you on the next Kentucky Health and I'm sure you're going to take a look at that, too, that you're flushing down the toilet and wonder what information are you passing along?

Thank you.

See you again >> Kentucky Health is funded in part by a grant from the foundation for a healthy Kentucky.