 Our speaker this evening, Peter G. Ramford, our staff at the System Engineering with Algae. Well, Peter comes to us with a long background in the environmental biochemistry. In fact, that was your PhD work that you've been working on for quite a while. But you really wanted to be an engineer. Well, I should be an engineer. Anyways, advice for the practical engineer. Well, we're glad that you're looking at the ways to put the science to work. And of course, that's what engineers do, make that scientific work, some practical work for people to use. And we're anxious to find anxious to learn what's what you've learned here in the recent work. Your recent work as technical director for the Algae Bioenergy Program at NNU. We welcome you. Thank you. Thank you for the invitation. And thank you all for coming. It's a great pleasure for me to be here. And every scientist loves to get up and talk about what they spend all their days and nights thinking about. So I've got about 30 slides for you. I'm going to kind of go through them at a semi-legally pace. If you have questions that are just really urgent then you have to ask them. Go ahead and raise your hand. I try to deal with all that slide as well as we can take questions at the end. A lot of people that contribute to this work are in the room. And I'll try to go through some of them on the slides and then maybe seem to introduce some of the other folks at the end of the talk. As Wayne said, my early interests were in environmental biochemistry. And right back when I was starting in graduate school in 1977 and 78, my thought was that biochemistry was eventually going to play a role at very large scale in terms of rebalancing systems that had become out of our own since industrialization happened on the planet. I didn't think I was going to have to wait 30 years to really have a chance of making an impact. But we've been reasonably productive along the way. We've learned quite a bit. And right now, the last five years of my career, I've spent trying to work out methods for scaling up algal biotechnology. Currently, the largest successful companies that depend on algal biotechnology are producing on less than 1,000 acres. And to get to these large scale impacts that look on paper to be very promising, we need to increase that by, one, probably, two orders of magnitude. And that's not going to be easy. So the bulk of my talk is going to be talking about cultivation barriers. But I'm going to try to put that into both the engineering and economic context for you first. And that'll explain some of the decisions that we made along the way. So in a nutshell, this first slide describes our overall thinking. The energy water environment nexus, which is what the EPSCOR NSF project is all about, is one that is both critical and imminent in terms of need for large scale new technology. And this word disruptive is an important one to define before we go too much further. A disruptive technology is one that's a little bit difficult to imagine currently. Current versions of any disruptive technology are a little bit half-faked. And yet, when the issues and barriers are overcome, they can transform an industry and put industries that have been in leadership roles for decades out of business. And an example would be the cell phone. In the early 90s, people could imagine a cell phone, but it was essentially limited by transmission towers and lack of miniaturization and software development and so on. But people kept working away on and then it's transformed our world. But if you go back even a little bit further, I was 12 years old in 1965. My best friend's dad was at the work of the highway department, hauling flasher signs around. And he had, essentially, a walkie-talkie that allowed us where he took us over to see a San Francisco Giants baseball game coming over a Donner and Pats. And he was able to haul his wife and imagine it through on a phone. So that technology actually goes back a lot further than we might imagine. So that means that by analogy, we should be able to predict what's going to be happening in 45 years in the future. And that's what I want to talk to you guys about today. What I think is going to be going on in terms of new technology at the energy, water, and environment nexus. To put this into a business context, the algal industry at a large scale does not really exist yet. And that's because the economics don't favor it. And it's really a stretch to get there. And so when I looked around at opportunities where the investment and financing might be easier, I landed on a wastewater treatment as one of those ecosystem services that represents an opportunity to get this new technology off the ground. And that's because current wastewater treatment is an energy-intensive process. Even with anaerobic digestion and co-gen recovery, we're still needing to pay, we only get about 50% of our energy costs out of that kind of a co-gen process. And yet the energy in the wastewater itself, chemical energy in wastewater is four times what you need. So we're throwing an opportunity away. And so the idea is our goal is to identify a stream of unit operations that allows to do energy-positive wastewater treatment. And if we can do that, we can take the energy savings for the operation of a wastewater treatment plant over five or 10 or 20 years, use that as collateral to get financing to go back and do the retrofit on the wastewater treatment plant. So that's the opportunity. The challenge is, how do you figure out each one of those unit operations? What are they? How do you optimize them all together, not separately, but together so that you get a process that makes sense? So the first part of my talk, I just want to introduce the team and the resources. Some of the people are here. Tanner Schaub is not. He's an analytical chemist extraordinaire. He does our oil analysis. I won't be talking about his work today. It's a little bit technical, but I encourage you to invite him for a future science campaign that's some incredible stuff with his mass spectrometer. Schaub-Long-Dang is working with us every day. And some of his graduate students are here in the back on extraction and conversion of elbow biomass into bio-cold oil. Dr. Neumann Condon is here. You want to stand up? You can see you and Wayne Bedmore. He is also here. There we go. He's back up with the art. Civil Engineering, Biorecure Development, and Energy Analysis work is what goes on in Dr. Condon's lab as he is not. We've also worked with Sean and Ivy in animal science. There's another opportunity to use algae for single-cell protein for supplementing animal feed. Feeds into next month's topic of can we feed ourselves with what we can grow in New Mexico. Megan Starbuff provides us with analysis and advice and economics. And Omar Holkeen is a new professor in plant and environmental sciences who does a lot of metabolic modeling on course. This is the banner for the New Mexico Episcopal Project. The six projects are shown in the circles on the top. And the fundamental idea here is that the nexus of energy and water is defined in that first line. We need water for energy. For example, cooling towers from power plants. They use an awful lot of water. We use tremendous amounts of water for hydraulic production for the recovery of gas and oil from depleted fields. And of course, we also need energy for water, primarily for pumping and desalination. Tremendous amounts of low-quality water being produced through hydraulic production right now is just being re-injected into wells down in the ground. Two miles deep, never to see the lighted day again. And that's fresh water from ranchers and cities that are being sold to oil companies for that process. We are really mortgaging our water future for short-term gains in petroleum. That's a non-sustainable issue. And if we can come up with new sources of renewable power, we can drive, desal, plant, to recover some of the value of that water that would otherwise be lost. Another opportunity for us. And of course, everybody knows about global warming. And CO2 is a greenhouse gas. We are intent on trying to develop CO2 utilization pathways. CO2 is a nutrient, obviously, for algae. Enhancing the CO2 content increases growth rate dramatically. So we need to identify waste sources of CO2 that we can use as an input to make renewable energy. That's a system that makes an awful lot of sense. And of course, nitrogen and phosphorus fertilizer can be recycled from the hydrothermal lipofaction process. I'll talk a little bit about that as we move on. So in addition to the NSF resources that are coming through EPSCOR, we're also the recipient of a new grant from the Department of Energy called Realization of Algae Potential. I'm going to mention you as the lead institution on that. And I'm the principal investigator. And I'm lucky enough to have collaborators from Argonne, Los Alamos, and Pacific Northwest National Laboratories, Washington State University, Michigan State, a company called Alginal Biofuels out of Fort Myers, Florida, PAN Pacific Technologies out of Australia, and a refining technology company out of the Midwest called U of P Honeywell. Our approach here is to de-risk algae cultivation. And that is the long pole in the tent, so to speak, in the overall process. And it's a complex one. So part four of the topic will be on analogy cultivation. We also have some funded work to look at low energy harvesting processes. And we're going to attempt to increase the yield of our biofruits oil through genetic engineering approaches, as well as increasing the growth rate. I won't be talking about that work. It's being done by Dick Sayer at Los Alamos and is also part of the New Mexico Consortium. And then the hydrothermal liquefaction is the other technology that we can use to optimize the recovery of biofruits oil from algae and get the yields to the point where the economics start to make sense. So the second part of the talk is basically to give you a status of where we are with all the biotechnology and why we're interested in doing it in the first place. So this is a slide that came out in the publication by a fellow by New Zealand, everybody named Manchester in 2007. It's been shown in a zillion times in conferences around the world ever since. And it just points out what sort of yield you can get in terms of leaders per hectare from different crops. And we can see here from corn oil, we're around 172. Two different yields of algae were way, way, way above that. The best one that's actually in business right now is the oil palm plant, where that can only grow in the tropics. And the oil palm productivity is pretty good. Downsides on that are that you have to clear rainforest to grow it. And the carbon footprint associated with clearing the rainforest is estimated to take maybe 75 or 80 years per question to grow there. Presuming on that slide that all the available forms of transportation are converted in 70s and burn and oil as opposed to gasoline or is it based on the present division between most of the gas engines and some oil there. So this is only the yield of vegetable oil. There's no downstream processing. But you said 50% of transportation fuel needs. Most vehicles, as far as I know, can't burn oil. That's correct. So are you presuming that everything would be converted to an oil burning engine? Vegetable oil is very easy to convert into a variety of different fuels with technology that exists today. The feed stock is the real issue. So for example, global alternative energy is a small company out of El Paso, LinkedIn, Sea of Alley Transport. They have a biodiesel facility. They use vegetable oil and cooking grease. And they produce about 14 million gallons of fuel, a lot of diesel fuel a year out of that. 80% of their costs are feed stock. 20% is the plant and the operation. So it really is the feed stock for these operations that are driving the economy. So if we translate those numbers into a map of the land required to display 15% of the US transportation fuel usage, and this is a slide that we borrowed from Sapphire. I'm going to bring Davis in the back of the room from Sapphire. If you want to talk to Bryn about the Sapphire operation, you can grab them after the seminar. The bottom line is that forest waste takes this much space. This is projected on the map of the US, obviously. Corn star, talking about 50 million acres. Corn ethanol, 90 million. Tree farming, 70 million. Switchgrass is a tall, rapid-drawing C4 plant. 90 million acres in here. So what you need for algae. Nevertheless, it looks small on that map. It is gigantic. And the amount of water involved is gigantic. And it is not a simple matter whatsoever. The New Mexico advantage is that algae growth is obviously driven by solar radiation. And in actual fact, we have too much light flux here. It's difficult for the algae to use at all. As a matter of fact, they evolved to take up more than they need and be wasteful as a competitive strategy to prevent competitors from moving in on them. And so one of the opportunities for genetic engineering is to re-equilibrate their light harvesting system. So they only take up as much as they need to grow. That's the good news. You can do that. It's been demonstrated. And the benefits have been measured and documented. The advantage is it's an anti-competitive strategy. If you do that, the genetically modified algae are going to be less competitive in an open environment. And the problem is going to be taken over rapidly by wild type strains. So you've got to figure out a way to protect your new strain from wild type competitors that are going to come in and remove that advantage. The big New Mexico disadvantage is water. And evaporative water losses in New Mexico are around 8 to 9 feet per year. And so that means that your water is going to become more and more salty. Your makeup water, if you're using on brackets or underground water source that's high in total dissolved solids, your salt is going to creep up enough. And pretty soon you're going to be growing only strains that can grow in the Great Salt Lake or other hyper-sailing bodies of water. Or you're going to have to come up with a new source of fresh water. In New Mexico, that's a non-starter. Everything is spoken for already. And there's really no new water. So water conservation was my first and primary design principle when I just got back into this game about six years ago. How do we figure out a way to grow algae in New Mexico without any evaporative water loss? And this next slide kind of demonstrates why it's important. It's from a paper by a friend of mine by the name Jason Klan, who works at Utah State University now. He did an analysis of the water usage and evaporative makeup water here in their miscolor. And he just looked at four different types of fuel products that you could make from algal starting material. And the bottom line is no matter how you slice it, no matter how you process it in your downstream, the water usage is dominated by your evaporative makeup water. And again, this is a non-starter for New Mexico. Good news for the Southeast and Gulf Coast region where they have plenty of water. Bad news if you're interested in economic development here in the Southwest. So I think I've covered a lot of this. Let me just jump in here and reiterate that what we're interested in are alternative water sources that aren't being utilized productively right now. They include both municipal and agricultural wastewater. And I'm thinking primarily about feed logs, dairy waste, and so on, and where there's both nutrients and water that's otherwise evaporating and being lost. We also have produce water from oil and gas extraction. For oil wells, the amount of water that comes up with the oil goes up as the age of the field increases. For gas extraction, it's just the opposite. Nevertheless, there's an awful lot of oil that has to be disposed of. And right now, producers pay to have it disposed of as well. There are brackish and saline sources that didn't need to be cleaned up. And there's a small company in New Belen, in Mexico, called Algorado Biotechnology. They have a business for doing that with algae. So those are some of the opportunities that we're looking at. And that's kind of the state of where we are with large-scale algae biotechnology. So part three I want to give you just a short introduction to the notion of industrial ecology. And I'm going to go into too much detail on the slide. The basic notion is that the industrial revolution was driven by being able to take a source material, do some manufacturing and clean up on it, and make a product that had value, and then throw the waste downstream and down the road and let somebody else worry about it. And while there were not so many humans, that process worked OK as the population grows and the waste products become more troublesome and we end up with environmental issues, basically industrialization takes us away from the chemical steady stakes that the earth established in the pre-industrial era. And as population grows, we have to begin to actively manage those. And on an industrial scale, we're talking about finding uses for every waste stream from any industrial process. And they'll turn those waste streams into an input for another industry, such that the sum total of all those transactions ends up being a net zero. And that's where humanity is going to have to go if population prediction is for the 21st century culture. There is no choice about this, ladies and gentlemen. We have to do this. And there are fortunes to be made in doing it. Fortunes to be lost as well, giving their own bed. So let's be clear about that. There's lots of folks that have already had great ideas that turned out to not work. So we need good science. We need good engineers. And we need those folks and disciplines talking to one another to sort this out. So let's kind of take a look and step back a little bit. How many of you are watching the new Cosmo series on TV? It's pretty good, isn't it? I'm sorry my tongue isn't that good. But I don't have their question. So I went to the internet. I found just a couple of pictures that I thought would be useful in describing this sort of pre-industrial ecological boundary. And in this kind of a scenario, you have a land environment adjacent to a body of water. And there will be waste coming from the land from runoff and the activities of plants and microorganisms that degrade the plant material and animals and so on. And that will runoff into the water. And it will carry with it nitrogen and phosphorus. That's what the N and P stands for. And what happens in the presence of water, N and P, and sunlight in CO2 is that you get algae to grow. And those algae can, in turn, set up an aqueous-based ecosystem in which the algae are eaten by invertebrate rotifers and ciliates and those are eaten by little fish and little fishery and by bigger fish and so on and so forth. And the whole thing is more or less in balance. And you never end up with a situation where there's too much algae for the ecosystem to keep it in balance. When population goes up, you end up having much higher productivities from the use of artificially derived nitrogen and phosphorus. We use petroleum and a process called a hover process to make ammonia in huge quantities. And that was responsible for the green revolution of the 20th century. Unfortunately, we put too much of that nitrogen into our fields. And it runs off into our aqueous system here. And now we have way more nitrogen and phosphorus. We end up with more algae than we can utilize. The algae, instead of just being eaten by the invertebrate organisms and them being eaten by the fish, the algae settle down into the water column. And they start to be degraded by bacteria. And the bacteria are consuming oxygen in the process of doing that. And then the whole system becomes anoxic. There isn't enough dissolved oxygen in the water column to support the life of the fish. You get dead fish. The whole ecosystem collapses. That's our dead zones. Sorry? That's our dead zones in the ocean. Is it dead zone just like this? At the end of every major river in the world is the result of this overuse of rivers as a pipeline for pollution. So this is also an opportunity. We can intercept these streams of nitrogen and phosphorus rich material, use sunlight and carbon dioxide from the atmosphere, and do this in a controlled fashion. And then work that biomass into usable energy products. We now are making laminate for rent. And that's the whole idea. But how do you do it at industrial scales? That's the key. And so just put one slide of chemistry in for you. This is a chemical equation for photosynthesis in which we have water and carbon dioxide and the nutrients that we've been talking about, plus sunlight, producing products over here. And the form of oxygen is just a chemist's notation for carbohydrate. Lots of other things are made from that carbohydrate and oxygen. And this is shown as one carbon because it's a balanced chemical equation. We have one carbon on the left and one carbon on the right. Look at, you can count up all the oxygens and hydrogens. They're balanced, too. This equation goes to the left as well. That's how animals make their living. Everything that we take in, eat, and respire into CO2 and water ultimately comes from photosynthesis. And the free industrial earth, these arrows that flow through those in both directions was roughly equal. And what's happened over the industrial era is that we're using more and more of the photosynthesis reaction as being overwhelmed by the respiration reaction. And it turns out it's not us respiring. It's causing the CO2 accumulation. It's that this reaction going from right to left is the same as it is for combustion. Respiration and combustion are essentially the same reaction. It's the respiration is done in a controlled fashion inside cells. And combustion is done in an engine. So combustion and petroleum is overloading this equilibrium going back to the left. And we have to balance that out. That's biochemistry and a very large scale solving the environmental problem. So the solution, or a solution, if we're going to tackle wastewater treatment first, because we think we can finance the plant retrofit based on energy savings, we have a series of needed operations in which we start with an area in which we're going to grow all the biomass. We need a harvesting system. We can fractionate that harvest if we want to. Part of what we can do is pull out the nutrients at this stage and put them back into production of more biomass. There's technology that the guys in the bathroom here are working on today to realize that return. And then from there, we can go to this process. There's a hydrothermal lipofaction I'll explain in the next couple of slides. It's basically like a pressure cooker. Heat up a liquid solution under with containment so that the pressure builds at the same time. If they were cooked with a pressure cooker, it rapidly cooks stuff much faster than it would otherwise because of the combination of the heat and the pressure. So if we tune our system right, we're going to get a CO2-rich gas. That CO2 can go back to our biomass production system to increase the growth rate. We're going to get a solid residue, that solid residue or biochar of several different uses that we can utilize, one of which is simply to burn it to produce the heat for the hydrothermal lipofaction. It's also good as a soil amendment. There's a variety of different things you can do with this solid residue and a biofuel. So those are the fundamental unit operations that we're trying to figure out how to make this work at scale. So let me just take a sidebar here and say, if you're driving to the airport, E&L pass out and you don't take the Anthony Gap, but you go down through town, after you pass downtown, if you look up to your right, there's a refinery down there. Old Western refinery. And if we were to supply Western refining with all of their feedstock needs, based on current cultivation yields, you would take somewhere between 25,000 to 30,000 acres of ponds to do that. And right now, the cost per acre upon is a little bit difficult to nail down. You hear estimates as low as $40,000 an acre and as high as $150,000 an acre or a whole lot. So we're talking about 100, that's a lot of money. You have to fly 30,000 times either one of those numbers. It's a big number. So we need to be able to drive down the cost of production. We need to be able to drive up yields so that we don't need to spend that much money. It's a big problem. So harmonize all these different unit operations so they work well in concert. It is not a simple task. Now if you take a look at the current risk assessment for each of these unit operations, this is where we have the greatest risk. And the risk is based on the ecosystem of the open ponds or PVRs at large scale are sensitive to invasion by those little invertebrate grazers that I was talking about earlier. They love algae. They'll find it if it's there and they can kill a crop overnight. There's viruses that can attack algae. There are pathogenic fungi that will come in and wipe out the crop. There's a lot of issues around choosing the right organism that can survive at very, very large scale. Crop protection for algae is a fundamental agronomic need like it just like it is for weed or corn or tomatoes or anything else. And that crop protection science is in its infancy. A lot of the work has been done at Sapphire. Unfortunately the company keeps that technology under wraps and so they know a lot more than the rest of us do. And hopefully they're gonna make a smashing success out of applying the crop protection knowledge to open ponds to show that that can work and that would be a huge boost to everybody in this field. But it has yet to be demonstrated. And so the financial analysts, the engineers and the scientists all agree this is the biggest problem. Our thing is not a problem but it's a risk because the energy of harvesting is so hot. Algae don't get very dense in water and so you're gonna have to remove 99% of the weight of your harvest is water. You gotta remove it all before you can do anything with it. So that's why it's a high risk. The rest of the stuff is technology to deal with it. It's a matter of whether or not you can clench together your supply chain and the expense associated with doing it. So I mentioned several times now so we view wastewater treatment as a bridge. The design criteria for an energy positive system I've listed here. We need a, essentially this is how we're approaching it. We decided right off the bat we're only gonna try to do something for arid climates where sunshine is high and you have to deal with evaporative water loss system. So everything is tailored for desert or warm environment. So this would work in the southwestern US. It might work okay in the southeastern US probably any subtropical or tropical area of the world. I'm gonna remind you that there are two billion people in the world that have no access to wastewater treatment whatsoever with huge public health consequences. And so this is a part of the world that needs this technology. We're not talking about upgrading or replacing we're talking about new technology. And so even though I didn't write it here one of the design criteria in the back of my head is that you would want this to work at a village scale a city scale and a big city scale. And if you think about arid places oftentimes there's a lot of land available not too far away that isn't very expensive. So those are good aspects of focusing your design criteria on regions. So we're gonna use a closed plastic bag system that'll prevent evaporation, odors and will experience presence over heating. So then we're gonna need an algae that likes it hot. So where do you go? You go to a hot springs where algae evolve to deal with warm temperatures. And so I'll be telling you about a wonderful organism called the Goldy area sulfur area that fits that bill. There are at least three different energy products that we think we can tailor to a specific site. You can either get bio crude oil, biogas for distribution or you can turn that biogas into electricity. The technology for doing all of that is available. You need to focus down as the beach stock, the hardest thing and so on. We need efficient CO2 utilization inside our closed system and we need a CO2 supply. And so co-citing of this kind of a system with the CO2 source becomes a very important criteria. It's not necessarily something that we're designing for but it's something that will drive where these plants will arise. And of course the nitrogen and phosphorus are the key nutrients for algae growth and we need a source of those. It needs to be inexpensive enough to match your needs. So our current technology, and I won't go through this in too much detail, but the basic idea is you have a solid liquid separation. So liquids go into a tank and you bubble that tank with air or pure oxygen and microorganisms start to grow on the carbon in there. There's a fundamental imbalance of carbon, nitrogen and phosphorus in wastewater. There isn't enough carbon in that wastewater to support the utilization of all of the nitrogen and phosphorus. So when the carbon is depleted, the microorganisms stop growing and there's still too much nitrogen and phosphorus in there. So you put that out into the environment and you end up with eutrophication and dead salons and salons. So we end up having to go to a denitrification, which we take that valuable nitrogen resource and convert it into M2, which is 80% of what we're breathing in and out right now is nitrogen gas. It's inert, doesn't react well, has no value. So we take a valuable commodity to prevent eutrophication and we turn it into something that's useless. That's how we do wastewater treatment these days. You can do some energy recovery through anaerobic digestion, but you only get about 50% of what you need. In our post-technology, photosynthesis in CO2 provides the balance between carbon, nitrogen and phosphorus. So you get in one step removal of all of the carbon, all of the nitrogen and all of the phosphorus. This word mixotropic simply means that you want to know you that can both do respiration and photosynthesis. Remember, respiration was going from right to left on my chemical equations slide and photosynthesis was going from left to right. There are plenty of algae out there that will take that thing in both directions. So then that will remove the biological oxygen and instead of your activated sludge microorganisms. And then you're gonna end up with a lot more biomass over here than you do over there. If you're trying to treat, as we treat sludge now, that's bad. But if you have a good process like hydrothermal lipofaction for turning that into one of those three different fuel products, that's good because it means you need to get to an energy-positive plant. And then the hydrothermal lipofaction allows us to recycle our nitrogen and phosphorus, sell it for agricultural production or make more algae. We're interested in ramping up the amount of biofuel that we produce. So maximizing biomass is the key to making the energy-positive wastewater treatment work. So here's a diagram about what hydrothermal lipofaction looks like. You basically have a biomass source and a sorority pump and most of this is involved in heat exchange. This is your lipofaction reactor and what you repressurize and heat. And the products end up going to a separator this whole bit here is about energy recovery and water recovery and this is kind of what the bio-cruise looks like. This is from a group run by a fellow by the name of Savage at the University of Michigan. He's done a lot of pioneering work on hydrothermal lipofaction. You can see it's pretty VISTA stuff. Nevertheless, technology exists today to turn this into jet fuel, green diesel, gasoline, whatever kind of fuel you want. Dealing with this stuff is not the right limiting step. Okay, last part of the talk. How do we get around the barriers to large-scale electrical division? And I've chosen two pictures on here. They are pictures of systems that were horrible failures. On the left there is a company out of Arizona that was working with the Arizona Public Utilities and they had a fair amount of money and a pretty illustrious group of founders and it failed utterly in terms of its promise. It's a picture of their bioreactors and they lost their shirts and went out of business in 2008 or 2009, I think. This one on the right is much closer to home. This is a company out of, and it's right across the river in Texas along, I don't know if you can remember, but the first one, not Antony, but immediately West of Antony. The company is named Valsent and that was a parent company and Vertigro was the local outfit. And the idea here was that he had a tangy bag so you just put him in a greenhouse and the idea was a great one in that you want to distribute the light in a way that is maximally tuned to the algae's needs and increasing the surface area. It's a little bit of a no-brainer. I think, well, you can go up and dilute the light. And so, conceptually, it was a beautiful idea. In practice, they couldn't find an algae that didn't form a biofilm on these surfaces that they couldn't clean up. And so, this idea also failed. It's being tried again by a company called Algenon out of Fort Myers that I'm collaborating with and hoping that they solve that biofilm problem. And I question them about it and they don't tell me about it. But they seem to be betting the bank on it. And so, we'll see. New technology does come along. We know a lot more about biofilm information than we used to, but it is not a simple process. And I am and remain skeptical. So, this is what we have at New Mexico State. There's three different types of bioreactors. One is from a company called Solix Biofuels. I was associated with Solix for a little bit more than a year as their vice president for biotechnology. Their main product was a system for growing algae in a water basin that allows for temperature control, optimum light distribution, and CO2 distribution, and algae grow like bunnies in this thing. They love it. They grow about 10 times faster than they would in an open pond. That's the good news. The bad news is this thing is ridiculously expensive. And the panels are highly engineered to make them work well, but they have to be replaced probably at least twice a year. And the labor and cost associated with building and switching them out didn't make sense at scale. Solix is still in business and what they've decided to do is grow these algae for high-value products, products that you can sell for thousands of dollars a kilogram. And then you don't need a huge footprint. You can afford to spend a lot of money on your system and make a go of it economically. And they're competing directly with a company called Solix on it. Their approach to getting around the scale of problem was to do it in fermenters. Forget about photosynthesis, just feed them sugars and gigantic industrial fermenters and get them to make the high-value products that you want. And so it gets a bit of a game between Sapphire and Solix. I'm just seeing if you can survive. These are the first generation algae and olfactory actors on the lower right here. This is their Fort Myers facility. Each one of these things is about 50 feet long. They have about 18 centimeters of depth in the bottom. They have a drive that provides mixing and just kind of a water boil that goes back and forth and creates eddies that provide good mixing. And we're in the process of setting up a whole field of these things that are just for the Fabian Garcia egg science center. These are the ones that they abandoned. I like them because algae never get on this surface. There's no fouling that happens here. The major risk is that they're over-engineered and they may end up having a biofilm in the bottom that could end up attracting critters that are gonna potentially make continuous operation difficult. We have no data on how long these things will last. For the amount of money going into them is probably they're gonna need to last a year to three years in order to make the economics work. So I think the jury is out on this one for large-scale use. It might work, it might not. Definitely not here. The one up here on the top is actually Bryn Davis' design from Sapphire. And it's the simplest of the lot. It has, basically, it utilizes plastic tubing that you can buy from moving air around in very large bean houses. It's already treated for UV resistance. You use sandbags to create a berm around the outside, an essential berm down the middle, and you stick a paddle wheel inside that bag to chase the water around in a circle. The enriched air inside this bag and this bag was CO2. And we're gonna basically have a bake-off between this design and this design out of David and Garcia. My money's on this one because it's simple and it's cheap. These things have some cool features too, but we'll find out. And I can't tell you the answer yet. So what sort of temperatures do you reach in those systems? This is data from Alginol, collected in Sonora, Mexico, which will be a little warmer than what we have here. And it shows the seasonal variation. This is August of 2009. This is winter 2010, summer 2010, winter 2009. And you can see these are the diurnal variations and the diurnal variations have the sun responding on nature to it. So the red lines define the temperature range in which the hot spring is algae and what I'm gonna tell you about next, light to grow. And between the green bars, the temperature range where an organism called chlorella, that we're also working with, likes to grow. Both of those organisms can be grown in fermenters at very high density. So the notion is that you could do a seasonal crop rotation between chlorella and galliareal and keep your production going year-round. And we'll be testing that out with our diurnal. So, goldieres, sulfur areas are red, unicellular, eukaryotic algae. It grows between pH zero and four to give you a marker. Coca-Cola is about pH two and a half. Lemonade is about the same. It's pretty acidic. Things don't like to grow with that pH. If you leave your coke out on the table top for a week and look at it, there's still a whole lot in there, right? It's not like lots of milk or something. That's good for us. They grow up to 56 degrees centigrade. They're both photoautotropic and metotropic. That means they can take that chemical reaction in both directions. They grow on a very large number of carbon sources. I'll tell you a little bit about that. In the next slide, in this Robisco discrimination, it just means that there's an enzyme inside of all plants and algae that fix CO2 into carbohydrate. And these red algae are really good at that. Is there a must to attack them because it might spend a sitting? Is there a must a virus or a part of an eye or whatever that would attack them? Yeah. So if you didn't hear that in the back, she's making the observation that acidic pH is, because there's so many less organisms that can grow, that whole ecology in that system is really simple. And so there's fewer viruses, or at least we presume, but there are fewer viruses, fungi, and grazers that will cause problems. So far, that's proven to be so. But we need bigger scale operations, run over long periods of time before we're gonna discover what the problems might arise. I'm not naive enough to think that there aren't issues that have arisen. And I think we need to know and understand more about the ecology of those Yellowstone hot springs than we know right now to be able to predict what might go wrong and head it off before it happens. But that's another grant and another agency. Okay, so this is just the temperature and pH limits of life. And so if you look at those two different dimensions, if you're way down here on the pH scale, and you're relatively high on the temperature scale, they've eliminated most of everything. That's the whole idea here. And that's what this slide is just to tell you. Most of the algae that's grown in the world today is just spirulina, and it can grow up to pH 11, 11.4. And that's part of the reason why it survives. It also has this curlicume ornphology that makes it difficult for the grazers to consume it. It's just little green balls, they can just sweep them in. But if it's a corkscrew, it's a little bit different. So there's a couple of different reasons why these guys might be the most robust of the algae that are grown currently for commercial use. So here's a slide on the different carbohydrates and amino acids and stuff that the grow the area you can use for growth in the dark. Here's a reference to the genome project. So we know the full genome of the area. And we know from the analysis of that genome that it has transporters for all of these different things from different monosaccharides, both hexoses and pentoses, sugar alcohols, glycerol, disaccharides, acetate, amino acids, polysaccharides. It is quite versatile in terms of what it can utilize. It's a little bit of a mystery as to why it's so good at it. If you look at the number of carbon transporters in that genome, it's about the same as what you see in wood rot fungi or other filamentous fungi that have enormous heterotrophic cannibalic capabilities. And why this caldiary is essentially equivalent to them isn't quite understood yet. Is that growing a chocolate cake there? I'm sorry? Is that growing a chocolate cake there? No, it just meant there was a chocolate cake is a source of sugars. Put the chocolate cake in some water, add the caldiary, put it in a dark, ittle girl. Oh, great. All right, these are electron micrographs of caldiary taken by a colleague of mine, Ursula Goodenoff at Washington University in St. Louis. This is what they look like when they're growing rapidly in a photosynthetic mode. And when they become depleted of nitrogen, they de-differentiate that chloroplast that shrinks mildly. And you see all these granules in the cytoplasm, that starch. That's just polymerized glucose. And you see these liquid bodies form as well. These are the sources of oil for conversion to biofuel. The great thing about hydrothermal liquid is that it could produce high yields of bio crude oil from either this material or this material. And that's important because it takes time for these things to convert into these things. And if you don't have to wait for that, then your land can be more productive. Your yields per unit time are going to go up. But nevertheless, we know that the genetics in biochemistry for synthesizing vegetable oil, which is essentially what these liquid bodies are, exist in this organism and it can be exploited by genetic means. All right, coming into the home stretch here. This is data from last year at the Fabian Garcia Science Center with Goldie area growing in the sapphire style photobioreactors, starting on May 18 and going through June 6. And they go through a typical lag phase or they're adapting to their new conditions. The red line represents the daily temperature variation. And then they hit a region of growth where it is more or less linear all the way of time. And they achieve about 2 and 1 half grams per liter in that period of time. If we kept adding nutrients and let it go, this would continue to go out. We haven't yet identified what the upper limit in the sapphire bag system might be. But 2 to 3 grams per liter is our target. So we were happy that we achieved it. This is the first experiment that we did, actually. And our yields were about 16 and 1 half grams per meter squared per day. Our target for our DOE grant is about 20. So we're close, even with the wild type. So we were really delighted with these results. This is a result from the Nash-Silver Unhub who is in the audience in the back of the room here. This is his first paper on measurement of the rate of removal of nitrogen and phosphate from a wastewater system. And in showing that in systems that are very similar to one that I just showed from the last data, we get removal of both phosphate and anionic nitrogen within several days, achieving discharge limits. And so this is our proof of principle data on the nitrogen and phosphate removal rates for Goldy area. We're working now on the biological oxygen demand. That's the removal of the carbon sources and that work is ongoing. And I'll invite you back to see that. OK, last data slide. We'll work with Professor Adrian Ock in the Plant Environmental Science to look at survival of bacteria that might be deleterious that are in our wastewater treatment system now. And so what they did is they isolated an E. coli strain 15v6 that was highly resistant to multiple antibiotics. And there's good data in the literature that resistance to multiple antibiotics correlates with survival. And basically what we did is we looked at two different temperatures, 40 and 48 degrees and four different pH values. And the bottom line is that the lower pH is particularly 1 and 2, survival at either temperature was measured in minutes. If we go to a little bit higher pH, but higher temperature, then they can last for a day. The bottom line is that the conditions under which we're growing Goldy area are not conducive to the growth of human bacterial pathogens. And in point of fact, we did an eight-day selection under those Goldy area growth conditions with primary untreated sewage water would have had all kinds of viruses and bacteria in it. At the end of eight days, we plated that material on very rich bacterial auger. We had a single survivor. It was a bacillus, like an informal strain. There were no other bacteria that were viable at the end of that selection. And one of Dr. Condon's students is studying the physiology and biochemistry of that bacillus strain in case we should need it to speed up the removal of biological oxygen demand. We don't think that's going to be necessary, but that could be an important resource. So to summarize, the last part of the talk, the Goldy area growth as well, with little risk of culture failure and observed any so far, most microorganisms die into the same conditions. So that's good. We think this is going to be a stable, robust system. And it's inexpensive. We're going to get more biomass since we're going to get more energy from the photosynthetic wastewater treatment process. And we hope that this will translate to underserved global communities in tropical and semi-tropical areas. So last slide just to acknowledge is, again, my collaborators and funding sources, thank you for your attention and patience. I'm happy to take questions.