 Well, hello everybody. Many thanks for joining us again today for HydroTerror's webinar series. Another great turnout today for what's a very interesting and quite complicated topic. Today we're learning all about microbial monitoring for groundwater remediation and enhanced site management. Our guest speaker is Dr Sam Rosalina, who's Vice President of Microbial Insights, which is a company based in Tennessee in the US. Before we get started, I would like to acknowledge the traditional owners and custodians of the land on which we meet today. The Panarong people of the Kulong Nation, I also pay my respects to their elders past and present. Alright, so there's a picture of our speaker, Sam Rosalina, and a little bit about Sam and microbial insights. I've had a long history of dealing with microbial insights and had the pleasure of visiting them in the US. Must be six or seven years ago now. One of the highlights of that visit was actually going up to the Smoky Mountains National Park, which is very near their office, and there's a couple of pictures there on the right-hand side of that area. It's a beautiful part of the world and definitely well worth visiting microbial insights and the areas around it. So Sam Rosalina, our speaker today. He's Vice President of Applied Innovation at Microbial Insights. He's got a Bachelor of Arts in Chemistry from Berea College and went on to complete a PhD in Analytical Chemistry at the University of Tennessee. Throughout his PhD research, Sam focused on analysis of trace-level environmental toxins, including heavy metals and toxic acids. He's an expert in the detection and quantification of trace contaminants in complex mixtures and media, including the development and optimization of innovative new analytical methods and rigorous QAQC procedures. Today Sam's going to be talking to us about analysis of microbes and in particular how those analyses can be used to inform us in our studies for remediation and site management. Before we charge into his presentation, a few details. We love to get your questions and in order for us to manage those effectively, we need you to type them into the Q&A, which is at the top of your screen as illustrated there. Why does Hydrotera run this webinar series? There's a few reasons. We're passionate about sharing knowledge. We like to facilitate education and we like to be an industry leader. Certainly our guest speaker today and this company Microbial Insights have been a leader in this area for nearly 30 years and I think we're very fortunate to have them here to help us learn a bit more about how we can use this technique in our work. So a bit about the webinar sort of completed part one. So part two, Microbial Monitoring for Groundwater Remediation and Enhanced Site Management is the topic that Sam will be going through. He'll be covering what are the tools we have available to illuminate biological potential, document petroleum and chlorinated solvent degradation, and evaluate the efficacy of amendments for treating these sorts of contaminants. Then he'll be talking in detail about natural source zone depletion, what it is and how do we assess whether the processes are occurring. And finally, he'll finish off with a case study. In part three, we will be dealing with your questions and answers. So without further ado, I will pass over to Sam. Many thanks for joining us today. Everyone and thanks for having me today. I want to especially thank Richard and everyone else at Hydrotera for allowing me to be a part of their webinar series. It's a great series and Microbial Insights is both proud to be within the webinar, but also proud to be a partner of Hydrotera. So thanks to Hydrotera for all that they do. I want to talk today about the importance of microbial monitoring, especially when it comes to groundwater remediation and enhancing conceptual site models, better understanding what's happening at a site. And so today I'll be talking about molecular biological tools or MBTs that we offer microbial insights. Again, as I was introduced, my name is Sam Rosalina and vice president of Applied Innovation here at Microbial Insights, which just means that I'm looking into the future and thinking about new innovative ways that we can kind of help all of the industries that we serve. Some of those innovative ways are thinking about some of the phenomenon that we'll be talking about today like natural source zone depletion that can be taken advantage of to really help clean up a site. So I'm excited to talk to you about some of the tools that we offer microbial insights. This is our 30th anniversary this year. And for those who are unfamiliar with microbial insights, we started as a small technology transfer. And originally focused on looking at some of the microbial lipids, which are kind of the kind of analogous to the skin cells of the microbes, and determining what what we can learn from that. Since then we've grown in both technology and innovation. And now the majority of what we offer our DNA based analysis. So the, the kind of key analyses that we offer our QPCR. What our QPCR does is it allows us to target specific genes, segments of DNA, and say who is there. So we call that taxonomic genes. So what are their name tags, which organisms are there that we we want to see organisms that we're looking for. And then what's in their toolbox we call that functional genes. So we're looking for taxonomic genes. Who's there functional genes. What tools do they have in their toolbox. To better understand the potential for biodegradation of the site. So we have hundreds of QPCR targets and that can be a little bit overwhelming if you're trying to choose a la carte which targets to look for at your site. So instead, what we offer are something called quanta rays. And these are arrays of QPCR targets that we can look for simultaneously. So you, instead of sampling for groundwater and then selecting QPCR targets you can instead sample that groundwater and send it to us to look for specific suite of targets related to chlorinated solvent degradation, for example, or petroleum hydrocarbon degradation. So I'll be talking about a few of those different quantity platforms that we offer here within this webinar. We also offer a couple different isotopic analyses stable isotope probing is a very powerful tool for conclusive proof of petroleum hydrocarbon biodegradation. So if you have a hydrocarbon site highly recommend looking at that compound specific isotope analysis similarly is very useful for chlorinated solvent sites. It also can provide conclusive evidence of degradation. But it can be any type of degradation it can be biotic. It can be a biotic. And so CSA is a very powerful tool for for chlorinated solvent sites. We also have a number of passive sampling devices. Probably best known is our bio trap it's unique to microbial insights. It's a passive sampling device that you can deploy in a pre existing monitoring well. It allows the native microbes to colonize that trap. So you get an understanding of which microbes are actively colonizing within within your groundwater at your site so it's a powerful way to look at community there. And then we can run any of our molecular biological tools or MBT is off of that bio trap so you can run stable isotope probing you can run any of our QPCR or quantary analyses or you can look at the compound specific isotope analysis. We also have something called in situ microcosms in situ microcosms are like a beautiful balance between full scale pilot test and in a bench top lab study. Instead of a bench top microcosm where you're biasing what works well in a lab or you're taking samples from the environment moving them to a pristine laboratory. In situ microcosm also can be deployed in a pre existing monitoring well. It's an advanced version of our bio traps but it allows you to compare different amendments within your groundwater at your site to make sure that amendment a works better over any or for example if you wanted to to bio augment you could test different bio augment and see the survivability. Now one important thing about microbial insights is that we're strictly a lab we don't sell any kind of products. So that allows us to be unbiased in our determination and in our data analysis. We're trying to sell you anything on the back end. What we want to do is provide actionable data to help you decide what steps to take next at a site. So if there's any takeaway that I hope you bring from this talk it's it's when and how to use molecular biological tools so often I think people make the mistake of only thinking of strategies like monitor natural radiation or enhanced bioremediation when it comes to monitoring for microbes in in the groundwater and the soil at your site. And I think that's fair I think that monitoring for the microbiology for strategies like M&A and enhanced bio. That's really important it's important to know what microbes are there because a lot of your strategy revolves around that. But but I think it's also important to monitor for microbes, even if you're taking a more aggressive strategy so for example if you're using in situ chemical oxidation. If you're injecting permanganate or persulfate monitoring for microbes is surprisingly important and here's why. When you do an injection of in situ chemical oxidation. You will you destroy a lot of the contaminant mass what's left over that's maybe still stuck on the soil or it's small fractions that the oxidation didn't reach the injection didn't reach. That can back to fuse or it can re dissolve back into the groundwater so now you have a lot less contaminant but it's very dilute within the plume and this is when bio the microbes that are natively present can move back into town and and polish that off we call it bio polishing so destroying that last fragment of contaminant that's left. Often people think that when you do in situ chemical oxidation or aggressive treatments like thermal treatment that it wipes out the microbial population. And though it does certainly decrease the population what it does is it provides competitive advantage to the microbes of interest that can degrade the contaminants that are left over. Eventually you're bringing down the population of the microbes, but then after the isco moves through the right microbes move back into town and grow rapidly to degrade the contaminants that are left over. Similarly with thermal and we've seen this through a number of studies with organizations like cascade environmental thermal treatment does a really good job of destroying and removing. High concentrations of contaminant mass at a source area especially, and again that high temperature does destroy a lot of the microbes that are present really decreases their population. But what we see with thermal is that you get kind of this halo effect of slightly elevated temperatures around where the thermal treatment is happening. The ground water and soil are warm instead of hot and what that does is it incubates the microbes of interest and so while you're getting really aggressive mass removal and destruction at the source. You're also getting enhanced bio remediation down gradient. And so the key here is that there's always microbes present at your site in your groundwater in your soil. So often that includes indigenous microbes that are able to degrade the contaminants that you're struggling with at your site or dealing with in your strategies. And so, you may as well run the analysis incorporated into your conceptual side model and show that not only are you doing strategy a, but simultaneously, the microbes are working for you as well. So it's data that you don't want to miss out on when it comes to proving that contaminants are being degraded at your site. So the quandary which I mentioned previously is really key for doing this. It's really useful for monitoring microbes of interest. And the way that it works, and I won't go into too much detail in the science through this talk, but the way that it works is you grab a groundwater sample or a soil sample in the same way that you would grab for a lot of different analyses, low flow pump with groundwater. And from there, what will happen is you'll within that groundwater or soil sample you'll also be pulling out the microbes that are present in those. And we will then extract the DNA from those microbes that DNA is what we're looking for to say who is there, and what functional capabilities do they have what tools in the toolbox. So we can extract that DNA. It goes on to a chip or an array and within that array are many sub arrays and what that allows us to do is run QPCR analysis on lots of targets simultaneously. And so we're able to quantify these genes, a number of different genes from a single target at the same time. So we can then look as I said we can look at functional genes what tools are in the toolbox we can look at taxonomic genes, what name tags are present, which microbes are we looking for. And that depends on which quandary that we've selected we have a number of different quantities. We began many years ago with quandary clore, which is specific to sites with chlorinated solvent impacts. We have the quandary Petro for petroleum hydrocarbon sites. Shortly after we introduced the quandary mic which is useful for for showing that the potential for microbiologically influenced corrosion. In this case, you know, we're looking for bad microbes that can cause problems, whereas for all of our other quantities we're looking for good microbes that can help us remediate. And then now we have two newer versions of the quandary. We have the quandary in SCD for natural source zone depletion and I'll be talking about this in in depth today. And we have the quandary BGC which is for biogeochemical analysis so a strong way of enhancing the biogeochemical data that you might be already grabbing at your site. So briefly, what does this look like I'm going to show you a couple quick examples. The quandary Petro is one of our quantities and it includes over 20 different gene targets related to petroleum hydrocarbon biodegradation. So when we look at the results of just a subsection of those gene targets, we can see some of the gene targets here along the X axis, and you don't necessarily have to know what they are. But our reports will describe in depth with these gene targets are what they're related to along the Y axis we have our quantification and this is really key. So the quandary is important is it gives you these values that you can use as key performance indicators, or to better understand how to enhance the bio at a site. And so on the, on the left, again, our concentrations are cells per milliliter the concentrations and the quantification is key to apply because it provides things like percentile ranking. So up here you can see that the potential for anaerobic degradation of poly aromatic hydrocarbons is in the top 90th percentile at one of these wells. And so the quantification part is really key to understanding what's happening at the site. So this is just a subsection of our quandary Petro. So these targets related to these pH bio degradation. On the left hand side, we've grouped together aerobic pH degradation functional genes related to aerobic pH degradation. And on the right hand side, we've incorporated functional genes related to anaerobic pH degradation. And so you look at the data, we can look at the light gray well 17d is a background well it's unimpacted by any of the hydrocarbons at this site so no BTECs no pHs. And we do see that there are no functional gene targets related to pH degradation that were detected at this well, which makes sense because there's no impact. Our up gradient well near the source is this darker blue 7c. For upgrading it while we see a good potential for aerobic pH degradation. While our down gradient well 44 this light blue has a really high potential for anaerobic pH degradation so down gradient, much better chance of moving anaerobic allowing these pH degraders to take effect. Another subsection of this data and this is from the same site are BTECs degraders so aerobic BTECs degradation on the left hand side anaerobic on the right hand side. Again, you can see anaerobic potential down gradient is pretty high for BTECs, while upgrading aerobic is is the better potential. Similarly, we have the quandary chlor which is related to chlorinated solvent sites this includes over 29 different gene targets, specific to chlorinated solvent biodegradation, and it looks pretty similar. Down here on the bottom, we have a number of different gene targets, functional genes, the tools in the toolbox, like final chloride reductase. We also have a number of taxonomic genes so the name tags of the organisms we're looking for. One you may recognize is dehalic equities or DHC. This organism is really good at degrading PCE all the way down to harmless ethene and it can do that on its own. But there's a number of different degraders present and again there's percentile rankings come into play. What percentile compared to sites around the world are these numbers are they high or are they low. So this is a two time point analysis at the same location at a site. The blue is the baseline, so pre injection and the red is a post injection. And just from these two time points, a lot of questions were answered at the site including did the injection work. There was an electron donor injection needed with the pre injection, and then after the injection showing that not only did it work but it increased some of these targets to the top 98th percentile compared to sites around the world. Both of these examples were actually utilized to to obtain an M&A ruling by a regulatory body so both of these move towards monitor natural attenuation. Again that may not always be the strategy, but it is important to note that this data is really key for showing the potential for complete biodegradation at a site. So I'm going to switch gears a little bit and talk briefly about natural source zone depletion. And for those of you who are unfamiliar with the term, natural source zone depletion is a relatively new concept. It's a phenomenon that scientists have noted results in pretty rapid decrease of source zone L napple. So liquid light non aqueous phase liquids. And so for a long time we knew that anaerobic biodegradation of constituents of petroleum hydrocarbons could be degraded biologically. And so we knew that that was occurring. But it wasn't until more recently that this natural source zone depletion came into light. It's a very complex phenomenon and just to kind of highlight how complex it is. This is a figure from a lorry paper from 2019 and it includes a lot of different components of what could and should be modeled if if there were models for natural source zone depletion. So a lot of those are physical processes, like volatilization and dilution dissolution. A lot of them are microbial processes. Some of them are things like microbial byproducts like biofilms or bio surfactants. And then there's a lot of chemical chemical portion to this so there's a lot of play between the chemistry the geochemistry the biochemistry the biology. The chemical properties. So it's a very complex phenomenon that we're still learning a lot about. So the easiest way to kind of break it down in terms of the biological portion is what we found is this garg paper which gives an overview of natural source and depletion from 2017. So it breaks natural source zone depletion into segments in terms of depth within the soil. And so you'll see that where the L napple exists there's saturated L napple and there's unsaturated L napple zone, the unsaturated zone. Within that we can expect to see things like methanogens fermenters these fermenters break down the L napple and produce products that the methanogens can then convert to methane. We can look for things like bio surfactant production. This is a functional capability that a number of organisms have to produce essentially a detergent and that detergent can pull some of the petroleum hydrocarbons out of the soil pores and into the water. So it makes the degradation more rapid. And it also incorporates some of the physical process. We also have looking into of course like the redox situation of the site, what kind of reducing conditions are present. And then we have kind of up near the surface methanotrophs in the aerobic portion of the site. The methanotrophs are able to take the methane that the methanogens produce, turn it into CO2. So the presence of methanotrophs. It's a pretty clear indicator, along with the combination of methanogens and fermenters that NSED is likely taking place at a site. So breaking it down in this way is helpful. But then the question is, why is this important? Why do we necessarily care about NSED or monitoring NSED? And the reason is this, is that through a large overview or analysis of lots of different sites undergoing NSED, Garg was able to see relatively high rates of source removal under natural conditions. So for example, over, you know, 2100 to 7700 gallons per acre per year, or that comes through about 20,000 to 72,000 liters per hectare per year. So it's not a number that's worth turning your back on. It's worth investigating more. If there's a potential that this is happening at your site, these are rates that are seriously helpful in terms of remediation timelines cleaning up the site. So then the next question, of course, how do we get there? How do we move towards natural source on depletion or see if it's present at our site? And just like anything, if we're trying to get from point A to point B, we want to draw a map. If you were to ask me to draw you a map to get from Melbourne to Adelaide, and I drew this map, this would not be a helpful map. It doesn't give much context. You don't know even which direction is north versus south. You don't know if there are any barriers in the way. And most importantly, it provides no context for how to get from point A to point B. There's no bus routes, there's no train schedules, there's no highways, or even, you know, city blocks. And so this is not a very helpful map. Even if I were to throw in more context, like where is that in relation to Sydney? This is not a useful map. And similarly, this is analogous to just looking at the chemical and geochemical parameters. So right now, for natural source on depletion, there are some in-depth and very technical ways to measure natural source on depletion once you know it's occurring already. But the easiest way to identify if it's occurring currently is looking at the chemical and geochemical parameters. While these are very important, they don't give you much context. So we can look at things like we know that fermenters produce some of these targets, and we know that methanogens produce this, but we don't know how they all interact. And so this is why incorporating molecular biological tools like the Quantra NSCD is really important because it provides much more context for how these chemical and geochemical parameters interact. And so now we have, you know, we have those streets. We have one-way streets, and we have highways. We know how all of these targets interact. And so these gold boxes are gene targets that are incorporated in the Quantra NSCD. And by utilizing the Quantra NSCD alongside our other lines of evidence, like our chemical and our geochemical, we're able to much better understand what's happening at the site. So here we can see things like fermenters, which can degrade the LNAPL directly and produce all of these different byproducts like alcohol, formate, hydrogen, volatile fatty acids. You don't necessarily need to know all of the biochemistry, but as long as you understand how these things connect, it's very useful because now we can see if there's fermenters and they're doing their job well, then we have methanogens which are able to convert a lot of these products into methane. If we have methane, we likely have methanotrophs, which are able to use lots of different electron acceptors to turn methane into alcohols, which can then be converted into CO2. And so all of these play important roles within the NSCD microbiome. We also have, as I mentioned, the biosurfactant production. This would be a functional gene, a tool in a toolbox. The biosurfactant production, like I mentioned, is able to pull these petroleum hydrocarbons out of the soil pores and dissolve them into the groundwater. And so also utilizing the Cointre Petro in conjunction with Cointre NSCD is very useful because now there are gene targets that can go after the dissolved petroleum hydrocarbons that are in the groundwater thanks to the biosurfactant production of some of these other microorganisms. So, streaming these analyses together and monitoring the microbes at a site can really add to your overall understanding of what's happening at the site and can add to whatever strategy that you're using. So as an example of that, this is a site where there was an Elnapolis source area, quite a bit left as you'll see. And then the utilization of Cointre Petro and Cointre NSCD were incorporated into this site management early on, which ended up saving quite a bit of money over time. So looking at some of the basic parameters, the chemistry and some of the geochemical parameters, we're looking at B-tex concentrations in the source area very high, so as high as 60 milligrams per liter. As we move down gradient that decreases below detection limit in some locations or around one milligram per liter, but even in the mid plume we're seeing relatively high concentration to B-tex, 5 to 10 milligrams per liter. If we look at the methane, we see that methane is being produced somewhere around the source area. We see no methane in our background well as can be expected. And we see a little bit produced kind of on the outskirts of the plume as well. So without the Cointre data, this would be pretty much all that we had to go off of. But instead we can turn to some of the Cointre data and understand what are the different potentials for biodegradation at this site. So let's start with some of the Cointre Petro targets. These are a number of different gene targets related to aerobic B-tex degradation. We see the degradation of benzene toluene, ethyl benzene, xylenes under aerobic conditions. And we see within the source area less potential for aerobic degradation. But down gradient where we would expect it to be more aerobic, we see quite a bit of potential for aerobic degradation of B-tex compounds at this site. This is good to know. It means that likely degraders are already degrading some of these B-tex compounds. It may also be why our concentrations dip to below detection limit in some locations near the outskirts of the plume. Now here's where a background well comes into play and is really important for microbial monitoring. Some genes are a little bit more ubiquitous than others. And so we do see aerobic B-tex degrading genes present outside of the plume. And that gives us context for the ones that are detected inside of the plume. So we can say that these locations have aerobic B-tex degradation potential similar to background. And so having that added context is key. You might also ask why do we see high concentrations of aerobic B-tex degradation in the source area where we would expect it to be anaerobic. Oftentimes what we'll see are micro-araphilic heterogeneities. In other words, our soil and our groundwater is not as homogeneous as we'd like to portray in some of our pictures like this, right? So there can be aerobic pockets within that soil. And so we might see the potential for aerobic degradation. Or it could also be that there are organisms that are present that have these tools in their toolbox and are ready to use them should the site switch to aerobic. So important to keep in mind some of that context as well. Now let's look at anaerobic-tex degradation. This is the degradation of toluene, ethylbenzene, and xylenes under anaerobic conditions. This is a gene BSSA specific to this function, it's a tool in a toolbox. In the background, well, we don't see any of this gene target. But we do see relatively high concentrations in the source area, medium concentrations in some of the down gradient source area, and in the middle of the plume. We don't really see any potential for anaerobic-tex degradation along the outskirts of the plume, which makes sense because we know that they're relatively aerobic. So then let's start looking at some of the NSED targets, the natural source zone depletion targets. We know that at the base of that structure were fermenters. The fermenters are really key for producing the byproducts that are then utilized by the methanogens and then the methanotrophs. They're the ones that kickstart the whole process of natural source zone depletion by fermenting the LNAPL present at the site. And sure enough, at the source zone, what we see are high concentrations of fermenters, which is what we would hope to see for NSED. As we move down gradient, we see mid-range concentrations of fermenters, but again, they're about the same as our background concentrations. Now what about methanogens? Methanogens are also relatively high near the source area. Down gradient we see on the outskirts of the plume, lower concentrations, but there's still methanogens present. And this actually matches up well with our methane map. Where did we see methane concentrations high, medium, and low? Last, we want to look at things like our biosurfactant genes. These are gene targets related to that detergent production that some microbes can do. And this is again useful for pulling the petroleum hydrocarbons out of the soil into the groundwater for more rapid degradation. Here we also see some concentrations, mid-range concentrations of biosurfactant producing genes here at the site, both in the source area and some locations down gradient. But mostly seen at the source area, where we would expect an NSED to be taking place. So overall at the site, between the two different analyses, single time point, but trended down gradient. What we saw was a good genetic potential for aerobic V-tex degradation throughout the plume, anaerobic text degradation in the source area of the plume. But we also see a good potential for natural source zone depletion based on some of just a few number of these NSED targets. Again, the NSED, the Quantre NSED incorporates over 20 different gene targets related to natural source zone depletion. But just due to time, we didn't have a chance to show you all of those targets. But the key here is that it's likely occurring at the site. So the site manager was then able to move to a more technical strategy to monitor the natural source zone depletion and actually monitor some of those rates. Now I want to say that we have one client that has a very large portfolio of petroleum hydrocarbon sites. And what they found by using our MBTs is that by performing about, we'll put it in Australian dollars, but by performing about 12,000 Australian dollars worth of microbial monitoring upfront were throughout the process that they were able to save overall about 500,000 to 800,000 Australian dollars over the course of the project. And that's because they're able to understand what's happening at the site better, incorporate that biodegradation alongside their other strategies and avoid switching strategies when they don't need to based on the data that we provide. So what does the Quantre report look like? This is a portion of that report. Some of the gene targets on the left, the sample names in the concentration. This was a soil sample. So this is cells per gram instead of cells per milliliter. But then the question is what does this mean? How do I know what this data actually tells me? Is this high or is this low? So also within those reports we provide heat maps to give you a better context of what those concentrations are in reference to other sites around the world. Are these high concentrations or are they low? Now the way that we're able to provide those heat maps and also those percentile rankings that I mentioned earlier is our microbial insights database. So this is a database that's over 25 years old. It incorporates over 100,000 field samples from around the world, literally from all seven continents. We just recently got samples from Antarctica and so we were very much looking forward to being able to say all seven continents. And so through our QPCR database, we're able to provide much more context for that data. It allows you to look at your data and say, compared to sites around the world, is this high? Is this medium low? Do I need to do something to enhance these numbers? Or should we leave it as is? Is this good for M&A? Is this good for post-insitu chemical oxidation? And so by having this database, we're able to give you much more than just a number on a piece of paper. So some key things that we recommend in terms of using microbial monitoring and data. We don't expect you to be a microbiologist or to have microbiologists on your team. And so we always recommend to treat MI microbial insights as your microbiologist. You can ask us questions, talk through a site with us. We are very customer service oriented as is Hydrotera, our Australian partners. We also recommend generating actionable data, so thinking about what questions you want answered at the site. And then working with us to figure out the tools that are best for answering those questions. And using the database to add context is really key. So using that database, incorporating your own, we also allow clients to upload their own information, like chemistry, geochemistry at a site to kind of better hone in usefulness within that database. But even just the microbial data alone, using the database actually provides quite a bit more context for you. And then the last but not least that you saw throughout this presentation, trending the data is really key. You want to look at background concentrations. You want to look at concentrations either over time or moving down gradient, different portions of the plume. Trending the data is really necessary because it gives you much more information and relative comparisons to better understand what's happening at the site. Again, I want to stress that, as I mentioned before, we don't sell any products. We're strictly a laboratory, so that allows us to be unbiased in our interpretation of the data. And so there's no fear of us trying to sell you a culture or a product in order to keep you. We have locations all around the world. As you probably know, we have a location in Adelaide, Australia, and so receiving samples is very easy for us. And then last but not least, of course, our partners Hydratera, which we're very thankful for both again in allowing us to be part of their great webinar series, but also for being fantastic partners in Australia or boots on the ground in Australia. If you have questions about what we offer, I recommend that you reach out to Richard or anyone else at Hydratera. And if you want to reach out to us directly, you can email us at infoatmicrobe.com. But our Hydratera partners are really key for our Australia customers, and they can direct you to us if they need to, but they're always there to answer your questions. So again, I want to thank all of you for being here today. I want you to take away the fact that microbial monitoring is really useful no matter what your strategy is. Truly, there's always microbes present, and if they're degrading the contaminants alongside your strategy, it's worth knowing that. And again, want to thank you Richard and everyone else at Hydratera for setting this up. I'm very happy to be able to do this. Thank goodness for technology that allows me to present in Australia while being asleep in my bed. So thanks for allowing me to pre-record this and present to all of you today. Again, reach out if you have any questions and thanks again. Well, I'm sure you'll all agree that that was a very comprehensive presentation. And it's truly amazing how advanced we've become in terms of being able to assess and predict remediation. I had a few sort of personal takeaways that I've taken out of this presentation. Secondly, there's always microbes present. It's kind of nice to know that they are there and they seem to be able to, in many cases, lead to action and sometimes we need to stimulate them, but typically they are present. Secondly, the tools we've seen today provide valuable insights into the efficacy of those microbial populations for assisting with remediation. So not only these days do we know that they're there, but with the ability to sort of characterize those genes that actually do the work. We know a lot more about a lot more certainty that they can actually do the job we need them to do. It's really important to understand I think that in situ remediation does occur and sometimes the rates of that remediation are significant. It's not about us doing much at all. So getting these sort of metrics early in the process, like actually in the assessment process, allows you to be better informed about that before you commit to large scale remediation works. Because in some instances it may not be worth doing much remediation at all if you know that you're getting significant degradation anyway. So very important to think about that. The assessment technologies that Sam's gone through today do provide predictive indicators of degradation of chlorinated solvents, petroleum hydrocarbons and microbial corrosion. And so they're definitely worth keeping in the back of your mind for assisting your clients with their sites. If you're a site owner, something that you would be hoping that you're being advised on to help you assess what level of cleanup that you need to do. Finally, monitored zone, source zone depletion is becoming more commonly discussed. And these sort of indicators really do provide us with a lot more understanding of all the factors affecting the rate of that depletion. And whether or not we can have certainty that it is actually occurring. So they were my take aims without further ado, I think let's move over to Q&A and we've got quite a few early bird questions, which we will go through now. Thank you very much. Well, we've had several early bird questions come in today. The last week and many things for those. I'm going to move to looking at those now and read those out. I've spoken with Sam earlier today to get his input into these questions and I will do my best to answer your questions now. So question number one, are there any tools for real time dissolved oxygen monitoring in groundwater bores? Well, there are many and certainly that's a core part of HydroTerror's business and we offer them both for sale and rental and you can have them connected up for telemetry purposes. Or you can have them with data logging devices. There's a range of different types. Often we use optical DO sensors these days because they are more reliable in the long term. Certainly, but feel free to give us a call so we can help you out there. Next question. Environmental health risk assessment brackets microbial of leachate, microbes in leachate. So we had a little bit of trouble understanding the emphasis here and then we concluded that the question related to is it possible to track the or use the microbial analysis that microbial insights have to evaluate the health risks of microbes? And the short answer is yes. They certainly do that for water quality where you're looking at drinking water quality, for example, and trying to work out what the source of the microbial impact that's been identified in that drinking water, for example. So in the context where you're looking for, has there been an impact to say beneficial use of drinking water from leachate? You can certainly use the analysis, the analytical techniques that microbial insights have to help you trace what the source of that microbial impact is. That's covered everything that Sam said on that. Question three. Any suggestions where this data or other microbial site data would be indicative of increased soil carbon? Well, this was a really interesting question and the answer to that is yes. There's a lot that can be done with this sort of data. So the first part of it is to say that research has shown that if you look at the ratio of microbes to fungi, that's an important thing to know about. So if that ratio gets out of kilter, that affects how much soil carbon is actually there. So there's a direct relationship between soil microbial activity and soil carbon and the microbes and fungi have an incredibly important part to play in that. So understanding that ratio is an important indicator of whether or not you're actually getting sequestration processes occurring. The second part of this is that microbial insights do have really good ways of identifying whether certain specific activities and microbial reactions actually happening in the soil. So they can also map those processes and they have a name for that. They call that next generation sequencing. But I guess the short answer is yes and it would be a good thing to reach out to microbial insights for further information on that. Question number four, how can computer vision or technology be helpful in microbial monitoring? So we took this question to be focused on what an additional data analysis can be undertaken on the primary data that's collected by microbial insights. So keep in mind the data that microbial insights collects is all kept on a database that they retain and they sort of quarantine your specific data in there. And that data is largely about those sorts of parameters that we've looked at today in the webinar. What they have indicated that and what is very useful is to get additional site specific data from the consultants doing the work. Some of those site specific contaminant concentrations and other indicators of those sort of secondary indicators of whether or not biological activity are occurring. So bringing those two data sets together and doing some analysis in the database that microbial insights have does sound really valuable. That's probably one of the main ones. They've also been very effective at combining some of these data sets together to create spatial plots, which allow you to identify more clearly by combining more site specific data with theirs. To really identify what areas of the site those pockets that would benefit most from some specific augmentation with some kind of additive. I think that's really important to know. All right. Next question. Any tools to measure cell activity, therefore 10 cells running at 100% or 100 cells running at 10%. This might help formulate what nutrients are required. The analysis that microbial insights do, they have a very specific way of achieving what I think this question is getting at, which is will the site benefit from additives. And the key analysis that is most official, I have written down here. In situ microcosms, these can be deployed in pre existing monitoring wells, and they allow comparison between different wells to allow us to assess, even within the single site, which parts of the site would benefit most from that formulation. So getting back to the emphasis of the question. Are the cells running at 100% or running at 10% that's exactly how you would do it using their technology. So it sounds like a really practical approach to put to it before you start making recommendations on adding amendments is to actually do those studies. Microbes that really do the work. Final question. How does the transmission of data or information? How is that managed from the remote location? Well, the short answer is microbial insights collect samples, whether they're on their customized sampling devices or whether they're just traditional samples of water and soil. They take them back to in the case of Australia back to Adelaide for analysis. And they don't specifically have any remote monitoring technology. However, what hydratera does is a lot of that sort of work, collecting remote data and transmitting that data back. And that data can be combined with the outputs from microbial insights. And I think that's where the key here really is is if you can have that site specific sort of time series data. So I think that being indicators of that biological activity combined with the actual hard data from microbial insights on what genes are present and how they can be stimulated. I think that leads to a really powerful way to be able to advise your clients on how to improve their in situ remediation. That's the end of the early bird questions today. Unfortunately, due to some logistical challenges today, I'm not going to be able to field the other questions that have been raised during the Q&A during the webinar. Apologies for that, but I've had a flight change which has disrupted my availability for that. So I would really like to thank you all for attending today and any of the questions that you have raised today in the webinar we will send emails out to the recipients of this webinar. So you will get some answers to those in writing. But many thanks for attending the webinar today and really appreciate Sam's involvement in this one and also your own. Many thanks. Cheers for now.