 All right, well, welcome everybody and welcome back to the consortium series on bioengineering and ethics. I'm your host, Nsu Hyun. I want to first, many of you have already been through this series before, but I just wanted to go through some housekeeping issues real quickly, so everybody understands. We have a Q&A feature at the bottom, so please, as the presentation is going along and questions arise in your mind, please use that Q&A box to submit your questions to the speaker, and we will get to as many of those at the end of the talk as possible. If you have any technical issues, at that point, you might want to use the chat feature to send a message to the panelists and to the staff that could help you with that. We do have upcoming events, and you can find that under the bioethics.hms.harvard.edu subscribe address, and we're happy to have you for future events. So let me welcome our speaker. Many people have a great interest in biodiversity and environmental issues, and I thought it would be a nice change to this consortium series on research ethics to include issues such as the one that we're covering today. Back in the very beginnings of bioethics, the term bioethics used to be much more broad. It used to include not just human health and research, but also the well-being of the environment and ecology. And in that sense, we are sort of returning in this second half of our series to that broader sense of bioethics. So I'd like to welcome our guest today who's joining us from Hawaii. Mary Hagedorn is a senior research scientist at the Smithsonian Conservation Biology Institute and director of the Reef Recovery Initiative. She's worked in aquatic ecosystems around the world from the Amazon to Africa. Her innovative and interdisciplinary work uses basic science and human fertility techniques to address conservation challenges for threatened coral reefs. She's considered a leader in securing a future for marine biodiversity, and her unique team is the only group of scientists in the world developing and applying modern technology to conserve coral reefs. Mary, I welcome you to this session, and it's fantastic to have you. The floor is yours. Please take it away. Mary will speak for about an hour, and then we'll have half an hour of Q&A. Take it away, Mary. Thank you. And so it's a pleasure to be here. Can you see my slides? Yes. Excellent. Okay. It's a enormous pleasure to be here with you today. It's exciting for me as a sort of a bench scientist and a field scientist to be talking about some of the ethical ramifications of the conservation work that I'm doing. And I'm going to start out and give you kind of a heavy sort of description of everything that we do and kind of why we do it. And more towards then, we can discuss what are some of the ramifications both in terms of the work itself, but also the larger sort of social good that we can potentially see from doing this kind of work or not. And so I'm going to start by saying everyone probably knows someone who has been involved in fertility clinics. Their family has used it, friends have used it. And we've used, we're taking those same basic ideas and applying up to coral reefs. When I first started this work, nothing was known about a sort of fertility measures with coral reefs. And we had to develop our own science to be able to do this. And so we're sort of developing the science and applying it at the same time. A friend said, it's like building the plane in flight, which is really, I think, very accurate to sort of the need for this work and the need for developing new technologies and overcoming challenges and developing sound strategies for conservation for the ocean. So I'm just going to, I'm just going to broach these ethical issues right now. We'll talk about them at the end. I haven't, I repeated this slide at the end. But it's really the idea that many people do not want to see wild ecosystems managed in any way. And it's just, there's something funny about it. We manage many, many animal systems, but wild ecosystems, whether they're salmon or coral reef, they just don't want them managed or don't want to conceptualize that. And then that some of the managers of these ecosystems can be risk averse. We certainly have grappled with that during the pandemic. Also, climate change is changing the physiology of, of the organisms as we're moving. So our science has to change and respond to climate change. So it's this ever-changing moving target for us as well. And then there's a lack of understanding of really what intervention science can do for restoration of our important ecosystems. Okay. So for the, this is kind of, you and I know it's probably snowy where you guys, so I kind of, this is kind of a dig slide. This is where I work at Hawaii Institute of Marine Biology. And the nice thing about Kanioa Bay, I'm on the island of Oahu. And I work at the Smithsonian, but my lab is here. And we have all types of coral ecosystems here. We have patch reefs. We have about a 50 of them in Kanioa Bay. And we have fringing reefs and we have barrier reefs. So it's a really great natural laboratory to just be able to take a boat or a swim and get coral. It's like, on the Great Barrier Reef, it's 17 miles offshore, even to get to it. And it's very deep and rough water. So you have to have, you know, a sea-going vessel to get any coral. So coral reefs are some of the oldest and most diverse ecosystems on our planet. But in every ocean, they are under siege. So they're all in trouble. There's not a single place in the world where they are not. And just to give you some perspective as to why corals are important to us, we live on an ocean planet and the oceans produce about 50% of our oxygen for us. Corals are involved in that oxygen production pathway. So they help us breathe briefly. Corals are ecosystems engineers like trees. And they are both animals and habitats, just like trees are both plants and habitat. And so they're a critical habitat on earth because they do change our ecosystems and they actually change our weather. Coral reefs provide homes for one quarter of all marine life on the planet. And that's, you know, at some point in a marine organisms life, they will live on a coral reef. And so they're critical nursery habitats all over the world. They are also a source of new medicines such as antibiotics, cancer, HIV, AIDS drugs, NIH has been collecting corals because they're so they're such old animals on the planet. They have chemical defenses that are very unique just like trees. And so there's some very unique chemicals that we will use from corals in the future. We also form natural storm barriers for our coastlines and cities. I just had, you know, we just had a tsunami here 10 days ago or so. And it wasn't a big tsunami that for Katanga it was, but for Hawaii was it only threw boats up on the shores. But you know, they the coral reefs here do protect our homes and I'm very grateful for them. And they also provide livelihood and food and fishing, such as fishing and tourism for people and for about 500 million people. So they're critical to feed us and to, you know, keep us moving, you know, in terms of our livelihoods. Reefs also contribute about $350 billion annually to the global economy. So they're a major financial driver as well, you know, in terms of, you know, our planet. So I know you've heard a lot probably about climate change, but coral reefs were in trouble long before climate change started happening. And part of it was the local stressors. And in the upper right hand corner you see a sort of aerial, you know, a space shot of a river that's spewing sediment, that brown sort of stuff that's coming into the water. And then a boat which is overfishing. And the damage to coral reefs prior to climate change was extensive. Much of the Florida reef track was damaged by local causes. Think of the farms and things like that that are close to Florida or upstream of Florida, whose rivers spew nutrients, fertilizers, you know, all sorts of petrochemicals, et cetera, into the water. And so I think it was an unobserved or perhaps unpaid attention to fact that the corals were struggling and dying, you know, long before climate change started. But and so it's true not only in Florida, but on the Great Barrier Reef as well. So it hasn't just been climate change. It's been our inattention to local conditions. But of course, climate change is exacerbating things now. And for those of you who this is a new concept to, we are overusing fossil fuels. And when we burn gasoline or coal or whatever it may be, we put in greenhouse gases into the atmosphere. And those are particles that surround our earth and create a blanket as we're around the earth. And when sunlight penetrates our atmosphere, many much of that radiation goes back out into space, it gets bounced back into space. But infrared, which is warm, you think of those those infrared lamps you see at restaurants that warm your dinner or keep it warm. Those those get reflected back by the particles, so more particles, more reflection, more warmth. And so it as our atmosphere warms, it's warming our oceans. In addition, oceans act as a sink for carbon dioxide. So think of soda, if you will, or pop, since you're in Massachusetts. And, you know, maybe your mom or your dad said, don't drink soda, it's not good for your teeth when you're a kid. And isn't the sugar that's bad for your teeth? It's the acid that forms the carbonic acid when CO2 dissolves in water, forms carbonic acid, which can etch and destroy your teeth. And coral, corals are an animal, and they have tissue, but they also have skeletons. And that skeleton is much like our teeth. And so it can be eroded. So we have a, we really have a double edged sword that's going on with the global causes in terms of ocean warming and acidification, they both lead to coral stress, which can lead to bleaching, which I'll describe in a second here. Disease, corals get disease, and reproductive loss and potentially death. So in terms of coral bleaching, it has always happened on coral reefs. And if you look back in the fossil record, you can see it. And perhaps, you know, I'm just going to throw out a number there, but it's probably fairly accurate. Maybe once every 100 years, you might see a bleaching event on a coral reef, historically. And, but since 1980s, NOAA has been keeping track of them. And mostly they were local. If you look at the sort of X-axis here, you see 1980 down at the bottom here. And until about 1998, we had about 370 local bleaching events around the world. But then in 1998, there was a global one, which is this huge peak. And during that time, 15% of the world's corals died. And since that time, it's only gotten worse in terms of the amount of the number of bleaching events that are happening. We now have close to 3,700 happening, and it's just getting worse and worse. So the state of the reefs are not good. As I told you, local and global stressors are impacting reefs. Bleaching is becoming more frequent across the globe. Now, in this image, you see a good example of bleaching. So coral live in close symbiosis with algal, algal cells called symbiotase, symbiotase, symbionts. Anyway, they changed the name just recently. And so these are algal cells that produce that are photosynthetic and live inside their cells, much like we have mitochondria that give us energy. Symbionts give coral energy. They take the sunlight and through photosynthesis produce sugars that power the cells of the coral by feeding them these sugars that can produce proteins and fats. And during a warming event, like it usually is an El Nino year, you'll get warming events. And during El Nino years, if you're on a coral reef, if you should ever spend any time in the tropics, you know, it's sun and shade constantly. There's always the, because we make cumulus clouds in the tropics. And so you have these puffy clouds that are always going by and blocking the sun. And so there's not this and just really, you know, sort of the ancient mariner kind of scene where it's just, you know, just sunlight constantly. It's, it's, but during a during an El Nino year, it is, and so you get a lot of much more intense sunlight and the waters are warming and this stresses and over the stresses, the symbionts, their photosynthetics apparatus gets damaged, they stress, they produce Ross, which is a reactive oxygen species and the corals kick them out. And so this can, they can turn from their usual brown or green and the symbionts produce the color for corals. And so you can see them go from a normal color to a bone white color in just minutes, like a half hour, that they can just release all their symbionts and they release them into the water. If they do not get them, now the coral is not dead at this point. It is, it is, it's kind of like the invisible man, you're, you're looking through its tissue, which is a thin veneer on the skeleton and that you're seeing the bone white skeleton. But if they do not get their symbionts back in about 10 weeks, they will die. So as I said, many, many species ecosystems around the world have lost their, their, their coral. So what we are proposing is to use cryopreservation and what we're doing is using cryopreservation to try and help coral reefs. And we believe that through cryopreservation, we can save the genetic and species diversities of coral reefs. Now, just for non science majors, genetic diversity is the number of animals in a population. You want to have as many as possible. So you don't get inbreeding or inbreeding depression and biodiversity is just the number of species you want to have as many ecosystems do better the more diverse they are. They can respond to, to natural disasters if you have more species. So cryopreservation is the marriage of animal husbandry, physiology, development, engineering and quality, quality control and biophysics at cold temperatures. So it's this amalgam of different sciences that are never taught pretty much in, in most universities classes. If you're an engineer, you may get some biophysics and heat, heat transfer and things like that, that deal with cryopreservation, but it's not often taught in, in schools. So the goal with cryopreservation is a sort of a triangle where you have a cell in the center, which is the blue triangle. And you have water permeability, which is water must come out of the cell and it dissolves across the membrane. Some cells have water channels, but most cells, it dissolves across the memory. So you want to get water out, because you don't, the main thing about cryopreservation is you don't want ice crystals to form inside the cells. They can form outside the cells, but not inside it. If they form inside, it kills the cell immediate. It forms, it forms spears that basically, you know, ice, ice spears that pierce the cells. So you want to get an antifreeze or cryoprotection into the cells and that, that dissolves across the membrane as well. And then you need to know the freezing rate, how fast this is all happening. So, because you don't want ice crystals to form in the cell, the freezing rate is critical. And so for many cells, you have to figure this out ahead of time. And there's other parameters that you have to look at, whether the membranes themselves are sensitive to permeability. Some, some cells, especially coral, unfortunately, things that have a lot of fat in them, the membranes destabilize as they move down towards zero degrees. And it has to do with the composition of the membrane. And as they move from, say, a room temperature, or in the case of coral, 26 degrees or so, down towards zero degrees, you get destabilization of the membranes and holes open up the membrane. And the cells just basically fall apart. So if you're using standard cryopreservation protocols like sperm freezing, which is a slow freezing, and it's done many, many animals around the world, humans do that. We can freeze sperm of almost anything. And because they're very small and relatively easy to freeze. And so basically, you're going to get water out and you're going to put a cryoprotectant in. And the process is relatively simple. Here you are, you know, approaching zero with your, with your cell, and you're actually going to use ice crystals in the slow process. You're going to bring water out as you're seeing here. And your cryoprotectants are entering the cell at about minus five degrees. Ice crystals are forming in the solution. And then if you go too fast with the freezing rate, because it's a rate, water has to come out and cryoprotectants have to go in. If you go too fast and ice crystals start forming in the cells, and that's bad. But if you get it right, then what happens is you get this dehydration, the cells shrink down, and you concentrate your solutes in your cells, and you have concentrated cryoprotectant in there, and they can freeze and then everything's fine. But there's another way to do that. And this is the most common way with human embryos is to use a process called vitrification. And in vitrification, you want to form a glass of your solute. And basically, this is how it happens. Use your cell at 22 degrees. And instead of waiting and going down to zero degrees, you do most everything on the bench. So your cell at 22 degrees, you have a much more concentrated vitrification medium. So for many, many sperm cryopreservation protocols, you might use a 10% DMSO, which is what we use for coral, or a 1.25 molar solution, however you want to think about it. And so it's not very toxic. And it's not very, you go from something like, in the case of a coral cell, which is homeostasis is at 1,000 milli-osmol instead of 300 milli-osmol molar for a vertebrate cell, we go from 1,000 to say 2,200 milli-osmol during the process, which is not too osmatically stressful. But in the case of vitrification, we are going from, say in a vertebrate cell, 300 milli-osmolar to five molar or six molar. So it can be very stressful. And this is what some of the hurdles that we're having for organ cryopreservation as well as the toxicity and getting these very sort of viscous talk, relatively toxic solutions throughout organs. But for individual cells like embryos can work really, really well because they're tiny and you can get the vitrification medium and the water out fairly quickly. And so then what happens is that once you've gotten the cells to the right shrinkage, the amount of concentrated cryoprotectin inside the cells, then you just plunge them directly into liquid nitrogen and it forms a glass. So instead of forming ice, which you can see with your eye when you do it, you have this like if it's an embryo, you have this beautiful embryo that's clear as a bell, and it looks like a crystal. Okay, so what are some of the solutions and challenges for coral cryopreservation? First of all, coral are some of the most reproductively limited animals on the planet. And I put up the panda here because I'm from the Smithsonian and we have pandas. And it's always this amazing time of year when the pandas are going to spawn and everyone looks forward to it and with a panda cam and it's all very good. But the pandas only spawn like the males are reproductive for about six weeks out of the year and the females for about two or three days out of the year. So it's very, very limited and they use hormones to determine estrous cycles, etc, etc. But coral only reproduced for an hour, 45 minutes for two nights a year. So an hour and a half total a year. And they're in very remote parts of the world with that which are hard to get to like Tahiti and Maria and Hawaii and that are hurricane prone. And so sometimes we can be forced off our, you know, our field sites by bad weather, which has happened. They're also stuck to the bottom, they're immobile and so they have to synchronize their breeding. So you can imagine if you have one coral here and one coral here and if one spawns on one night and one spawns two nights later, nothing's going to happen because the eggs in the sperm have to cross within your neighbor. They don't do well with self fertilization. Corals are hermaphrodites. They produce both eggs and sperm, which is very unique for an animal species and makes things more complicated. But so there's some challenges and interesting things about corals. And so I'm going to show you what they look like when they reproduce. So here are really important concepts and I hope you take this away. Corals are animals. They're not plants and they're not rocks. They have some of those features because they have a skeleton, which is rock like we do too. And they have algae that live inside of them, which are plant like, and you can fragment them. Corals can reproduce sexually this way with eggs and sperm, but asexually like a plant can with a fragment and we'll talk more about that later. So here is an individual coral polyp in this blue circle and corals from colonies. So one coral polyp settles and then it breaks into two and then into four and then on from there. So they're colonial animals that are mostly genetically identical in the colony. When they breed, many species produce produce egg sperm bundles. And what this is at least large spheres filled with eggs. And you can imagine, think of a cluster of grapes with a packet of sperm in the center. And those are filled with lipids and those rise the surface. And you'll see that in a little bit in the video I show you. And when they break apart on the surface, they form the slick of beautiful pink eggs with sperm and you can actually when the great barrier responds, you can see the reproduction from space. That's how amazingly extensive it is. It's amazing to see. So here's the video and here's a coral reproducing. There's an egg sperm bundle. You can see one tiny little egg there. And you'll see a pan out now. You can see many of the polyps that are producing eggs from bundles and they're being released and they're rising to the surface. The next thing you will see is a mushroom coral, which they're separate sexes. They're sequential method. They can turn from males to females because they have both organs inside. But this one is a male and you'll see the sperm being released from the mushroom coral. And the next one you'll see is a brain coral and it releases the egg sperm bundles all at once. So as I said, they go to the surface and they must fertilize a nearby neighbor. So here is my lab and they are many of the people who have produced a lot of the information you're going to hear today. So we'll thank them. So we can freeze, as I said, we can freeze coral sperm. We develop our own technology. We develop our own mechanisms to do this. And because what's available on the market for mostly human cryopreservation will not work for us. And so we really spent a lot of time developing the science and the tools and then going into the field to applying it and training people. And so the sperm cryopreservation is the most extensively done right now around the world. We have about four, here's some of our earlier sort of things where we actually try and make things very inexpensively, like our tools inexpensively because we want to democratize our science. We do not want people who do not have much money around the world to not be able to do it because you need a $25,000 piece of equipment. We make our own. And in this case on the left, it's flip-flop material with aluminum canes and cryoviles that float on the surface of liquid nitrogen. And we modernize that by doing a 3D printer, which is on the right, and we 3D print now our racks that go into the door, which is right here that is filled with liquid nitrogen. And then we monitor it with a probe. So this costs about $20. This is a little more expensive as the whole operation is about $80. So with our colleagues around the world, we have about 48 species now that have been, the sperm has been cryopreserved. And I don't want you to read the species. I just want you to see the oceans and the species that are in the, you know, that we've collected in those oceans. And the Great Berry Reef has been the easiest for us because we've been able to apply, they spawn all in one week. So it makes us very easy for us to go there and target that time when we're going to cryopreserve them. So since about 2012, we have been using the frozen anthod sperm to create new larvae, coral larvae. So we can create coral larvae from fresh eggs. We cannot cryopreserve the eggs just yet, but here's a coral, a baby coral that's, here's three polyps and you can see their tentacles and the symbionts here and it's glued itself down to the bottom. So the coral sperm works well, they develop normally, and they can go through the things that corals need to do to become competent and that is to settle and attach itself to the bottom and to take up its symbiont. We've also used coral sperm that has been frozen for up to 10 years. And here we, we froze sperm in Puerto Rico in 2012 or no, sorry, 2008. And we also had some sperm from Florida which we frozen 2016 and we took it in 2018 to Curaçao. So hundreds of miles across the arc of the Caribbean and we re-froze sperm in Curaçao and for the Elkhorn coral, which you see here, it's an endangered coral. It's one of the few that is on the endangered species list. And these were the major reef builders in the Caribbean. The Caribbean is really not in good shape at all, but these were so massive at one time that you had to use dynamite to clear a path to an island. They were so prolific throughout the Caribbean, but that is not the case anymore. There's only about 300 genotypes of these corals left in the Florida reef track, more in the Caribbean, but Florida is really not in good shape at all. So we froze here and here and here and we used fresh eggs from Curaçao and we asked the question, A, can we move sperm from these various areas as cryopreserved sperm? And can we cross-fertilize the same species? So we're not doing any genetic manipulations. We are just doing selective breeding as it were in that we are moving from place to place. And because ultimately what we like to do is say there was some amazing coral in Cuba or something like that, we'd want to cryopreserve there and help Florida perhaps, especially in the northern regions here, to have more resilient, you know, sort of heat-tolerant coral. And so the idea was to use this process to try and then do, you know, selective breeding. So there we go. And here's some of the coral, that baby coral that we produced. We produced thousands of coral settlers and this is coral settlers growing out of Acropora palmata and they're sitting in captivity in Florida right now and we'll get to the, we'll talk about this more towards the end of the talk, but we had hoped after about six to eight months that we could put these corals into nurseries or back out on the reef. And because there's only 300 genotypes and we have now in our, in our collection, we have hundreds of new genotypes that could really diversify the reefs in Florida and the managers are not willing to put them out on the reef or even let us put them in nurseries at this moment in time. And so we're still under discussion and we'll talk more about that. So the other thing we wanted, so the idea really is, you know, you go into a fertility clinic as a family or an individual, you want to possibly do sperm, you might, if you're a woman, you might want to freeze eggs or if you're a couple, you might want to freeze embryos, whatever. If you're undergoing cancer therapy, you might want to freeze ovarian slices. There's just a whole host of things that you might want to have done to make, make your family more stable in the future. And that's the kind of thing we wanted to have for corals as well. We wanted to have a menu of things that you could, you could use. And so the next thing we worked on were coral larvae, whether we could cry or preserve them. Now, this, this is a pic, a beautiful picture of coral developing. I think it's just magnificent and, but you can, they're filled with that. And just to give you some perspective, coral human embryos, we froze human sperm in like the 60s. I'm just using round numbers. We froze the first human embryos in the 80s and the first human eggs in around 2000. And yes, we could do it bet, you know, probably before that, but it wasn't used in, you know, clinics very well. But the thing about human embryos is that it became really well used and very good, like 90% you know, implantation rates and things like that for human embryos. After vitrification was devised by my colleague Bill Raw. And so the reason that it did better is because coral human embryos are half fat in them. And by vitrifying them, they, you form the glass and there's no ice crystals. And so we had to do the same thing for coral larvae. I started freezing coral embryos almost immediately when I started this work and I must have frozen millions of them and they all died. And so vitrification is important. But when you freeze, you have to warm more rapidly. And as tissue gets bigger and bigger, you have a more of a challenge to warm rapidly. So think of mercury versus Jupiter, the difference in the volumes of those planets is massive. And so it's kind of the difference between say a human embryo and a coral embryo. They're just, the coral embryos are massive. And so we had to use really, we had to develop new techniques to be able to do this. And we did this with laser warming. We use standard vitrification protocols, but then we had to use lasers to warm them at millions of degrees. Because as you freeze during vitrification, you form ice crystals or ice nuclei form in the solution. You cannot see them, but they're there. And if you don't warm fast enough, you go through this process called de vitrification. And those nuclei bloom into crystals, ice crystals, and then destroy your tissue. But if you can go fast enough, then you won't, those crystals won't form that, those crystals will not form. And so that's what we did with lasers. We warmed up millions of degrees per minute. And here's what happens when you, you just, you know, just when I said, I froze millions of coral larvae. This is what it looked like. Oh, there's a coral larvae on the left. And if you just bring them down to zero degrees, their membranes are so chill sensitive. They just want the power. So you have to vitrify them. You cannot slow cool. You cannot use slow freezing. And then the vitrification process, you know, if you're just using bench top warming was not fast enough. So John Daly, my postdoc John Daly, who's now in Australia, helped us develop laser warming for coral, along with our colleagues at the University of Minnesota that ensue is working with in the ATP bio program, John Bischoff and kind of Koshla. So here's a little video of the process. So you take your pipette and you put a drop on, onto a cryotop and it's filled with coral larvae and gold particles. Now the laser cannot warm the embryos directly. It has to warm a radiator and that will radiate heat to the larvae. They go into liquid nitrogen. They go into liquid nitrogen that freezes them and vitrifies the droplet. Then we bring the cryotop up and a laser hits it and warms it at millions of degrees per minute. Again, with the laser warming the gold and the gold warming the larvae. And this is what it looks like about 50% of our coral larvae about two hours later start swimming around. So we know we have we've had success. And the nice thing is the we get so I'm left to see the cryotop with your coral larvae in it. And you get about 40 to 50% of the coral swimming. And we also know that they settle and they will take up their symbionts. And so their symbionts are here in the coral larvae. But the great thing about this as I said with sperm, if you have frozen sperm, you can diversify shrinking populations with sperm. So any kind of sperm frozen for a population is really important. Even if you can't freeze the eggs of a species, you will be able to diversify the population, which is the critical thing for managing populations. And here's the same thing, but it also prevents species extinction. If you have frozen larvae, you can actually just regrow them, you know, back out on the reef and re and and reseed the reef in the future. But we want to be able to do this faster. So this is this is our restoration process where we are actually printing droplets and then putting them into liquid nitrogen. And what this allows us to do, if you look back here, let me see, yeah, we only did on in a day, probably 300 larvae, because we were doing everything by hand. But once we we started this mechanized process, it's called the cryo wheel, we were able to freeze tens of thousands of larvae at a sitting and now remember reproduction only happens a couple days a year. So you have this very tiny window of time to be able to do this. And you want to take your cryopreservation process and make it a restoration process so that people can use it in not every person might be able to do it, but you might be able to do it at a resource center like Australian Institute of Marine Science or something like that. But the cryo wheel that we devised was all made with 3D printed processes and balsalwood and liquid nitrogen. So if you wanted to, you could make this. It's not again printing droplets of this application media with Barbie. Okay, additionally, we can freeze the other complete half of this story are the symbionts and here they are here, they're about 10 microns in their brown and photosynthetic. And they are critical for this whole process because some species only accept one species of symbiont. So it's not like, you know, the symbionts are out there and it's very egalitarian. No, they're very specific about the type of symbiont that they want. And most coral must take up their symbionts after they either before they settle or after they settle, they don't get them from their parents like we get our mitochondria from our parents. Some do, but not all. And so in order to help larvae, if you've got cryopreserve larvae or whatever you're doing, you must have the symbionts. So my postdoc, Jess Bowmeister, is in the process. She's using lasers as well for the cryopreserve symbionts. And recently, although our numbers weren't massive, we, if you look down here at the bottom here is a coral larvae that's smashed. You can see it on the, and the only, we do this so you can see the brown symbionts, the coral larvae are brown, these white cells. And they take up their symbionts at the species at about day four. And, and then it, we look at them about two days later. And here's a cryopreserve one. So they're taking up quite as many, and it's, we're still in the beginning process to make this better. But we're very pleased that the coral larvae will take up the coral, the cryopreserved coral symbionts. So this was a huge proof of concept for Jess and in the whole process. And we'll, we'll nudge this forward and make it better and better. Okay, we also cryopreserve coral reef fish. Now, my kind of, my colleague, colleague, kind of Coachella and John Bishop, and I cryopreserve the embryos of fish. They're even bigger than corals. And that's where we first used laser warming. I worked on it for over 20 years. And then when the lasers became available, that's when we had the success because we could not do it with just bench top processes. And so kind of got that to work. And, but there's another process where you actually cryopreserve the testes of fish. And the good thing about fish is that their stem cells are pluripotent. And that means that they can turn into, they still have the capacity to turn into ovarian cells or testicular cells if you have the stem cells. And so if you cryopreserve the testicular, like you just cryopreserve the testes of a male fish, you, the stem cells can then be re implanted into, so here you're cryopreserving the cells. And it's very simple. It's like, I can teach anyone to do this in a half hour. It's so simple. And you can do it in the field. You then can thought out your cryopreserve cells and inject them to the embryos and are a new or sterile host or embryonic larvae that have had their reproductive cells destroyed. They will then grow up the host will grow up the cells from the donor. And then when they, when you spawn them, you get sperm and eggs and you can produce thousands of new fish from the donors. Now this is good for almost every species we've tried it for. And this is the work of Yoshizaki et al. in Japan. He worked out all of this and we've now started applying it to coral reef fish. And we're hoping that NOAA and other groups will start to include this into their workflow so that when they collect fish, they cryopreserve them and store them in places like the Smithsonian because you do not need the original species. This, this species can go extinct. As long as you have the testes, you can put them in genuses that are related and you can get that species back up. So it's, it's this incredible sort of resurrection science that, that Goro Yoshizaki developed. Okay. I think I told you that coral, coral reefs are having problems in terms of reproduction with climate change. It's, it's having an impact on them. When coral bleeds, now think about this, just let's go back to humans again. You know, if humans go through any kind of stress, it could be cancer, it could be any kind of disease. Generally, they're not going to have, you know, enough reserves to have children in the near future. You have to build up your reserves again. And then, you know, then you're ready to go. It's the same with coral. When they go through a bleaching period and they are stressed, they use up a lot of their fat reserves and they don't either don't reproduce or they reproduce poorly. And we can see that we use a process called CASA, which is used in human fertility clinics as well, where we actually look at sperm motility using computer assisted sperm analysis. And you can see that after a bleaching event, the sperm is about 50% less modal. And you can't cry preserve sperm if it's if it's below 50%. Generally, we try not to. It's true for humans as well. That's the cutoff. And because you won't get much fertilization, cryopreservation process is damaging, you know, nothing's for free in life. And so, you know, you have to you have to have a really healthy coral to cryopreserve. And so, we're not giving up on reproduction, we just want to have an additional tool. And so, one of the things, and this is coral reproducing from the Big Berry Reef. And we want to do, we want to preserve the asexual fragments of corals. Remember, I told you, they can reproduce sexually, but you can take their small fragments and they can grow into a new coral. It's a clone, just like when you take a cutting of a plant, it's a clone of the parent. So here's what they look like. They're tiny little fragments about thumbnail size. And on this coral fragment, there's about 30 or so polyps. And this ring, they started gluing themselves down and growing. And this ring of tissue is stem cells. And when they're this tiny, they grow like gangbusters. And so, when we cut them into these small pieces, they'll grow, they'll grow into a new coral in just a few years. Whereas, you know, if you just take a larvae and want to grow into a mature coral, it can take seven years or five to seven years. So this micro fragmentation process stimulates stem cell growth and stimulates the overall coral to grow and actually can get reproductive in say two to three years. So it's a process of, you know, sort of turning on growth processes by this stimulation of stem cells. And so we're using a very sophisticated process. And this process is probably the future of organ preservation for medicine. And so I'll show it to you today. It's one of the first processes. We're using it for vitrification. And it's my colleagues in the engineering school again at ETP, but bio partner, Boris Ravinsky and Matt Palm Powell, who are at the engineering school in Berkeley, and they use this thing called isochoric freezing. Now, here's the isochoric chambers. It's just an aluminum chamber. And in this case, it's about, you know, this big and a little bigger than a cryo vial. And it's made of rigid aluminum. Now, I want you to think about when you were a kid, maybe you put a soda can in the freezer to get it cold really quickly. And then you forgot about it. And then it you came back and it was the soda can was, you know, enlarged or it burst in the freezer. And your parents weren't very happy with you, probably. But um, but what happens this the the liquid state of water is takes up less volume than the the frozen state. And so ice crystals form, they take up more volume. And so if it's in a vessel, it will expand like the coke can or the soda can. And the this process is has rigid sides. And so it it stops the the development ice crystals just by the rigidity of the container, at least ideally, that's the case. And so you put your your vitrification solution in here. And because we are, we can freeze at at much faster rates and warm at much faster rates, we can actually use slightly lower concentration. So it's really, really good for human organ preservation. And I know the the the number of people in the ATP bio group or network are trying it on a variety of human tissues and organelles. But this is a tiny little fragment of coral again thumb side thumbnail size that is sitting down in here, we add the vitrification media, close it down so there's no air bubbles, so it's not compressible at all. And then there's a strain gauge out here on the side or this little and it goes to computer that tells us if the volume expands, you'll see an increase in pressure. And so it's really just telling us if ice crystals are forming and we're we're knowing that by looking at the pressure. But if the pressure stays static, we know that it's gone from a liquid into a glass without that ice crystal formation. Again, a great way to preserve tissue. And so here's our first trial at this, which is for me very, very exciting. And here's a coral that has not been cry preserved. And you can see the polyps are really beautiful, the morphology is intact. And this is 124 hours later after freezing and fine. And you can see it looks, I'll tell you it looks fabulous, you may not think it looks fabulous, but it does. What we tried freezing fragments for so long, and it looks like someone put it through greater after, you know, that you thought them out, they just look terrible, and they don't stay intact. And so we're using a wide variety of tools, such as particle imaging and looking at green fluorescent protein, to try and understand stress for these individuals, as we take them through this process, and, and to get them from this very stress stage to a healthier stage. And so for us, it's very much about sort of, you know, this is the, this is the hospital phase when they're like 24 hours to 72 hours. And we're trying a number of things we don't want to use antibiotics on them, because we were concerned that the the corals have on their surface bacteria, it is, it is their microbiome, just like our microbiome is in our gut, it is here on the surface. And if we put antibiotics in to stop exogenous bacteria from attacking them, then we could kill their microbiome. So we're trying to use probiotics and also using antioxidants, as well as just really enhance care for these to get them through this very sort of stress period to the point where two weeks later, we know that they're alive. And again, corals produce green fluorescent protein in their tissue, they can auto fluoresce. And we can look at the organization of that green fluorescent protein, you see these beautiful rosettes of their polyps right here, forms a beautiful pattern of green fluorescent protein on the confocal. And as they're stressed, where if they're dying, that green pattern fades over time. And so we have like a two week period where we're monitoring the growth and health, the health of these individuals using that, but we also have other tools to look at their symbionts, the health of their symbionts. And we will probably look at the health of their microbiome going into the future. This is cutting, this is just like hot off the press, we've only done this for the last couple of months. Okay, so the, you know, science has said we must intervene and help to help restore corals. And there's a great paper in AAAS or in PNAS that a group of scientists wrote saying, you know, we have to try a wide variety of things. And many of things that they talk about are in situ or in place, in the place. But they're still impacted by global by warming. And these are marine protected areas, selective breeding, ocean based nurseries, outplanting of coral fragments to the reefs, larval seeding of reefs and genetic manipulations, all very important, all necessary. But the caveat is, they will be impacted by warming. Then there's exciter out of the place that are not impacted by warming. And these are land based nurseries in Aquaria and cryoproservation and biorepositories. Now, one of the things that is, is that we are doing is we, a consortium of institutions around the world have decided that we must add a layer of security to what people are thinking of already. And that is we want, we form this thing called the Coral Biobank Alliance. And we are bringing all 1000 plus species of coral into captivity. And the reason we're doing this is because if you look at distributed agriculture for corals, people have been able to maintain corals and tanks since the 1980s. And then if we pair this with cryopreservation and reproduction, we get not only sort of, you know, sort of clones in tanks, but then we can take them and we can cryopros, put them into reproductive systems and we can cryopreserve their sperm or their larvae or their fragments. And this is just kind of a timeline. I mean, we didn't even know coral were produced until the mid 80s. So that's, it's very, very recent in terms of, you know, them as animals and knowing about the reproduction. And, you know, the cryopreservation wasn't, we didn't develop until, you know, our first paper was in 2012. So as I said earlier, we are building the plane in flight and, you know, in terms of developing the technology, applying the technology and getting it so that people use it for restoration. So here's the network. We're a group of biobanks that want to preserve species for ecosystems restoration research. And if I didn't mention it earlier, the reason we want biodiversity is so critical is that it's obviously essential solution to climate change because ecosystems are far more robust when they have many species in them. And they recover better from natural disasters. And, you know, a biodiversity is just essential for our ecosystem services. So here we are, we have, our group is going across the oceans in terms of the areas that we're interested in. And to date, we have about 150 or so species that are in captivity. And you can take these fragments when you collect them, you can cryopreserve them, you can bank them, corals can live for tens of years in culture. So maybe 50 or to 100 years, they will grow, they can be sent to other institutions, we can genetically identify them. And then we can put them into reproductive systems, we've got corals that can reproduce in captivity. And then those offspring can go to managers and tradition, you know, for restoration. So I think, and as I said, here's an example of one of our coral nurseries that we have in Maria. And this is what it looked like when we first started it. And a year later, a year and a half later, these corals had grown out and started reproducing. So there are ways to really help corals to get them to grow faster. And we just have a lot of tools now to really help them move forward. And so I'm going to come back to these questions. And just, you know, part of the part of the problem we're having right now is people really just don't understand this process, the cryopreservation, the power of the cryopreservation process, and their risk averse. I mean, when you think of the assist, you know, the assisted gene flow corals that we created across the Caribbean for the Florida reef track that have, you know, Puerto Rico and Curacao, you know, genes crossed with Florida genes, it could do wonders. But the managers are just really not allowing it. And we're afraid we're going to lose them and lose this amazing resource. So I'm going to stop here and say, thank you for your time and open the floor up to questions. Well, terrific. That was so interesting. I realized at the very beginning I was distracted by a technical issue. So I didn't even introduce myself. I'm in Suhia and I'm on the faculty in the Center for Bioethics and Director of Research Ethics. I'm also the director of the new Center for Life Sciences and Public Learning at the Boston Museum of Science. So I'm going to pose a initial few questions while I curate these questions and get to the audience questions. Could you speak a little bit more to that issue? I mean, I heard you say once in the past that what you're facing, what you're finding is that the science is the easy part and really the problem is more of a social or decision making problem. Could you speak more to that? Yes. Thank you for bringing that up. I mean, conservation is about people, right? It is about animals, but it's about people and how people interact with animals and their ethics. They're moral code as it were. And if you do not find it important to have a diverse ecosystem, then conservation is not going to be important to you. And so I remember watching this NOAA program on climate change and they were interviewing people at a gas pump and they said if you had to pay one cent more per gallon of gas to apply to climate change around the world, would you do it? Everyone said no. Every single person said no. And so you can kind of understand that it's kind of the maslow triangle. People are really, if they don't understand it and they're really trying hard to keep their families going and money is tight, you're not going to pay one cent more for gas so that you can have this ethereal thing, even if it's something that they really love like birds in your backyard. And so most people don't see coral reefs. They don't think it's important. They don't understand the importance. And so it is, and in the case of the managers, to sort of weigh in for them, when we started the assisted gene flow project, it was just proof of concept. We had no idea it would work. And that it would work as well as it did. And we really, in an ideal world, would have gone to our stakeholders and our managers and said, we're going to plan this. This is what we'd like to do. Will you be our partners in this? And what are some of the challenges and things we have to think about for us to move forward to try and see if this will work, even in a nursery in open ocean? And then we could have kept that in mind. In Hawaii, unless you talk story about a process, you will be blocked. And it is that human process of bringing people together and having them understand the process, the challenges, and some of the goals. And because it was, we didn't think it would work really. We didn't do that process. It is the most important part of the science I can tell you. That's fascinating. So questions come in that I want to actually preface and frame a little bit with some further content. So the gene flow, assisted gene flow project is where you take prime preserved sperm from corals from one part of the ocean to another locale in, say, Florida. And you're trying to introduce, in that same species, more genetic diversity. And is it true then that you're also trying to select for this breeding corals that seem to be a little bit more resistant to heat? Is that part of the plan? In that proof of concept, we didn't go to populations that were really heat resilient. We wanted to prove that we could use the coral, the cryopreserve sperm that we could so the sperm had been living in Colorado for 10 years, that we could ship it all over the world safely, which we know. I mean, we do this for cows, right? That most cattle is produced this way. But getting it to, it got lost in the Caribbean actually, it was flying around in the Caribbean and a dry shipper. So there were some challenges. But to just work out all the processes and see if we could actually get it to work. When you have a population that's so distant, and there actually was a genetic barrier in Puerto Rico. And so we weren't sure that the even reproduction would work over such vast distances. So we wanted to try all these things out because some places in the world there are more heat tolerant corals. But in this experiment, it was to try everything around it. And then the goal would be to go someplace where there are more heat tolerant corals and combine them with say Florida eggs or whatever the case may be, and then try and put them out on the Florida reef track. But even so, I mean, even though they're not, we don't know if they're heat tolerant or not, but they're more diverse for sure. And with only 300 individuals left in the population, we have thousands of individuals, genetic individuals that are different. Just from a genetic diversity standpoint, it would be fabulous. So this gets me to the question that somebody posed is related. Are all coral reefs disappearing or will some species be resilient to ocean acidification? If so, can we just facilitate the proliferation of the resilient species? So I asked about heat tolerance. This is having to do with ocean acidification. Are there some that are more resilient to that? Because that's that's going across all of the ships, right? Right. So, you know, I can't, I can't produce the future. I'm just going to give you my take on this, right? And I don't want to say never, say never, and strip hope away from people. But for now, I think we should be, we should be doing everything we can. Every conservation aspect should be thrown at coral reefs right now, everything from cryopreservation to restoration, however you want to do it. But as we become intransigent about climate change, our window for getting, for getting coral reefs and other ecosystems back to health is closing, right? And so our ability to do this, because as corals warm and warming becomes more and more constant, like if, if we have warming every year, which is predicted by the end of this century, if we don't do anything, corals will stress every single year, and they will not reproduce. And without reproduction, we cannot get adaptations. And but, but you forget, acidification is going along as well. So this erosion of the skeleton is happening. And once we get over 554, something like parts per trillion or parts per billion, I forget what it is, you know, of, of CO2 in the water, that in the atmosphere, this will, corals will start to dissolve. So it's a, it's a chemical process that we're up against, as well as sort of a biological reproduction and adaptation process. And they are hard facts, you know, they're, they're not open to debate in any way, shape or form. And so, you know, unless we can really move towards a more proactive lifestyle around the globe, then no, corals will not exist anywhere. I mean, it's just a fact of the ocean. There may be, there may be some deep ocean refugia. I mean, we do have deep water corals, and it's possible there could be some deep water. That's why it's so important that we have these cryopreserved things, because we can keep them for hundreds of years. And so they can stay in captivity, or we can keep them frozen for hundreds of years. And, you know, our goal is to think about the oceans of the future and the people of the future, if their ethics are that they want coral reefs back, and they have returned the oceans, I mean, think of some of the rivers that were on fire in the 70s and 80s in the US, now have trout running up them, you know, in Cleveland, where you are right now, I think it was Cleveland, I don't know, it might have been Pittsburgh. I mean, we can help ecosystems back to health. And but our timeline for the ocean is probably much longer. And the cryopreserved samples can sit there for hundreds of years and be fine. So that's the important thing about them is they have a huge timeline that allows us to talk, allows us to change our policies, and our, you know, our structures and our ethics, really in some respect, and really work to making a better ocean for us in the future. Great. Okay, so some sort of questions have come in that I'm trying to put into category. So here's one. There's a follow up question related to this and that's how is the pharma industry involved in such research initiatives, particularly those who may use medicinal product properties of corals for chemo, etc. Is there interest in pharma? There is some, I think, you know, I know NIH has really banked a lot of corals. They were working in Guam for a very long time bringing corals and, you know, and just, and, you know, obviously you can just put them in, you know, a minus 80 freezer because it, you know, the chemicals are going to stay intact. The problem is it's a really long time to go from, you know, you know, a marine product to something that you could use, but there are, there are a few things. There is a chemo drug and that is, is from coral, but I think the one that's going to be most amazing is corals fight with each other, you know, like because they're, because they're, they're trees in a sense. So they're fighting for light, which gives them their nutrients and growth. And the way they fight is through, through a type of antibiotic. It isn't exactly an antibiotic, but it's this chemical that dissolves the tissue of the other coral. And so, and the way they do it is they kill the microbiome on the surface of the, their competitor. And so there are going to be some amazing chemicals, you know, there are amazing chemicals and corals that I think will help us with, you know, you know, antibiotic resistant bacteria and, and maybe the new antibiotics of the future. Interesting. So a related question is, how do you see the role of the private sector or broadly to engage in conservation and restoration efforts to protect coral reefs? It is critical right now, you know, because when I started this work, like NSF and NIH was not at all interested because it fell in a crack between technology and conservation, right? And NIH said, oh, no, that's conservation. Go to NSF. NSF said, oh, no, that's, that's technology. Go to NIH. You know, they're like, what do I do? And it was the private sector that stepped in and funded this work. So I think, like, you know, the Gates Foundation and other big foundations are going to be, are critical in helping us with this. And right now, we're approaching audacious, which is a TED-like thing. And we're hoping that that will help fund this global network of bringing fragments into captivity and allowing us to really secure the biodiversity and genetic diversity of, you know, all coral species on the planet. So you said in your talk, and I really find this so admirable, that you're trying to make the tools you use very affordable and sort of, you know, democratic in their accessibility. But with laser rewarming, does that sort of make it difficult for others to fall? I mean, are these lasers expensive or are they hard to manage? Yes and no. We are in the process now of even trying to make that easier. I mean, so each stage goes through a process of being hard and technically hard to do. And then we make them easier and cheaper and etc. And so the folks at the University of Minnesota now are in the process of making it much easier. But on top of it, we're using it like you probably are thinking of lasers that cost hundreds of thousands of dollars. The laser we use are found in malls in jewelry stores. They are welding lasers and they're expensive. They're about $18,000. So you could imagine that a country could have one or two, a poorer country, and could have a couple that are not so beyond their means. And it could be at a resource center. But we are trying to make it even easier. And there are other ways we might be able to do this. There's another kind of warming called radio frequency warming that uses just a coil with radio frequencies. And that would be another way we're looking at it. So we're very cognizant of the fact that we're using very sophisticated tools. But we will make them easier and we'll make them so that every country on earth that wants to will be able to afford them. Great. Now I want to start moving into some more general ethical questions that have come in. So one of them is very interesting. So our local communities, for example, Hawaiians, Tonganese, are they involved in these preservation initiatives? If so, which elements of local culture's knowledges have been utilized? So yes and no. I would say no in that there's a lot that local communities can do to help their coral reefs that they're not doing. And since I live right here in a coral reef, I would say sediment is the biggest issue. I mean, local issue. And it's so easy to figure out ways to block sediment from getting into coral reefs. And we are here in Hawaii are doing a very poor job of it. And I would say yes. In that recently, we had a biodiversity assessment here in Hawaii. And we spent a year talking with the community, talking story, having dinners, talking about how they, I mean, some of the folks and the families that have lived here in the Bay have been here for hundreds of years and have extensive knowledge about the algae, which they call limo, the fish, amazing fishermen, and the coral reefs. And so we really took advantage of that. And we had a sort of our cultural ambassadors who were our Kapuna. And even though we talked to them for a year prior to it during the biodiversity assessment, we had them with us. We did it on Coconut Island. And they were there to act as a resource for us while we were collecting. And it really was a wonderful collaboration. And it still is going on. We did the DNA barcoding of a lot of the animals in the Bay. And they're helping us with some of the naming of the new species, like sponges. When we went into this biodiversity assessment, we only had 10 species that were known. We now have 200. And so a lot of, a lot of our aunties are very excited about being able to name, you know, name some of these species. So it's a great collaboration. And they're not involved, particularly in the cryopreservation, but in the spirit of preservation of the reefs, they're very much involved. Yeah. So let me pose one of my questions at this point then. So in your attempts to make the tools more democratically accessible and sort of a low tech approach, maybe, you know, the need to kind of involve some local people, do you think there are opportunities for citizen science in this area? Oh yeah. No, absolutely. And especially, you know, we were just great question. Thank you. And the next two weeks, we're launching a training, because it's been so difficult to train people during the pandemic, we had to go virtual. And we engaged a group called Adventure Scientist to help us come up with just a series of videos and very detailed of how to make things. We even have, you know, the computer source sourcing for the printers, lists of where you can buy things and just very, very detailed tools of how you do this. And there are places in the world, like Mexico and things like that, where like dive shops and stuff like that want to be part of this. And, you know, the freezing of the sperm will be so detailed. And so, and I'll just put this in quotes simple. It's not exactly simple, but they will be able to do it. In fact, we've had groups that have done it on their own just by reading the papers, which was good. But this will be much, much better. And I think it, what we want to do is standardize things. We want people using the same tools, the same operations so that if we go into a bank, we know how it was done. And so that that standardization is really critical, I think, as we move into the future. And these kind of virtual trainings and things like that will help with that. That's great. You know, I've had the opportunity to visit the Great Barrier Reef and Reefs in Florida and Mexico. And I know there are many other people who just feel like they want to do something, they want to participate in some way. So that could be currently empowering if you get help and they can help together. Absolutely. This common goal, that's terrific. Okay, so several other questions have been coming in. Dr. Hagedorn, your work is incredible. I'm interested in what you can share about diversity, perhaps extrapolate a little on different areas with different requirements. Are there impacts to consider positive or negative on the surrounding ecosystems regarding our choices in the areas we may choose to expand the reefs? Yeah, very good. Okay, so think about, so the area of emerging diversity in the world for coral reefs is Indonesia. So Indonesia, when you look at it, it's considered a hot spot and it has, like you go the reefs there and you just swim around, like in 50 meters, you'll find 100 species. There are 800 species of corals more or less in Indonesia. And when you go to the Great Very Reef, there's 400, still a very lot of, you know, when we say 1000 species, we're not really sure because they hybridize and we don't have really good genetic tools for corals right now because they are such an amalgam of algae and bacteria and coral tissue. So it's been very challenging, I think, to get, you know, distinct tools, but we're getting there. So there are many, like Hawaii, we only have here in Kaniyia Bay, probably only 10 species, you know, so it's not a very diverse area. And the problem with the Caribbean is that it only has 60 species. And so I'll use the Caribbean as an example to answer that question. Because there's only 60 species going on, there is a disease now that's going through the Caribbean called stony coral tissue loss disease. Most coral diseases affect one, in the past have affected one or two species. This is affecting 22 species in the Caribbean, they only have 60. So one third of the species are, and they're being obliterated in weeks, you know, and it's traveling relatively fast. And so my, if we can get this fragment cryopreservation to work, we cannot cryopreserve in communities that have the disease, because we will cryopreserve the disease. Right. And so if we can get this fragment cryopreservation to work, the very first thing we will do is we'll go to probably back to Curacao or someplace like that where there isn't a lot of, there is the disease front has not gotten there and try and do this and get that population, you know, banked down, because to see if it will work. And then the other thing that we will hopefully do in the future, I'm sure many people saw the CNN video of the new coral reef that was discovered off of Tahiti. I mean, just beautiful. And my colleague helped discover that. It's just beautiful. And so we're already making plans to, you know, I can't go down. I don't dive it, you know, 150 feet, but we will use rebreathers and the dive teams and go down to some of those reefs and, you know, potentially take them, you know, and cryopreserve their fragments or whatever. So, you know, I think, you know, in areas where they're, where they're, you know, genetic diversity is low and we're starting to get disease or there are other problems, the cryopreservation really is going to be a godsend. And, you know, I think it will be very important in helping us maintain some of the diversity in some of these ecosystems like the Caribbean. Yeah. And several questions have come under this similar themes. I'll try to kind of put it together. So as background, just so the audience knows, you're not proposing to do any kind of genetic editing or manipulation, you're just moving populations in one spot to another, maybe or introducing more genetic diversity. But the question is this. So of course, people might think that genetic editing is unnatural and kind of ethically problematic, but is there a sense of kind of worry that what you're doing is also somewhat unnatural moving, you know, genetic populations around that there could be unknown risks to that. And, you know, kind of, it is kind of a natural activity, right? Like cryopreserving and moving things. And, you know, so what's your response to the kind of like, well, is it really not natural? How do we know that might upset? How do we know that won't upset the ecosystems of these areas? Yeah, very good question. And so it's, you know, when you think about it, all of our beef cattle around the world, they are produced through cryopreservation. Turkeys are produced through cryopreservation. Many, many things that we eat are through cryopreserve sperm being moved around. Now that's very different, like putting it on a farm, as opposed to saying, okay, I'm going to put it in an ecosystem. And yes, there is a risk, but there has been a genetic analysis, especially in, well, again, stick with Florida. There has been a genetic analysis of the patterns of flow in sort of the Florida reef track. And the flow of genetics is actually into Florida, and there's very little that flows out genetically. And so the risk that a NOAA genetics CRC group did that has a lot of coral geneticists on it was that the risk was less than two or 3% that there would be a problem with genetics. And one of the NOAA managers was like, Oh, one of the risks is we'd have coral, what a horrible risk. And, you know, and I think that you have to get the risk of doing nothing in my mind is far worse than the risk of it's, it's the same thing with the pandemic, right? The risk of not getting a vaccine is far worse than, you know, the dying, right? You could get sick. But the risk of dying is far worse. And so, you know, I think it's really how we understand and manage risk that really is at the heart of all of this. And I'm not trying to say that people are bad because they're risk averse. I'm just saying it's, it is something that blocks us and it's something we need to talk about. And, you know, when you think about, you know, driving a car even, you know, the thousands of people die each year, young people die each year. And we, we accept that as a risk when we get in a car. And so, and the other issue is that, you know, when you think about ecosystems, the ones that have gotten the worst rats are the, you know, you've introduced an animal and I'll use Hawaii again. We have mongooses that were introduced to try and kill the rats here. And rats are nocturnal and mongoose are are, are, are diurnal, right? So they never meet each other, but no one figured that out before they let the mongoose go here. And so I don't know, we have mongoose and we still have rats and it's just like, you know, do a little bit of research. So, you know, clearly there needs to be, you know, and that's why using, you know, off-shore off-shore nurseries are really powerful because you can put them on, you know, onto coral trees or whatever, and they can be monitored. And if, you know, a storm is coming or whatever, you can take those in, you can remove them from the water pretty easily. So I, that's what I would love to see as a intermediate step is to say, can we put these out on coral trees and can we look at them and would they flourish, would they be good? So. Yeah, thank you. So a few more technical questions came in. So let me address those and then I want to finish by kind of zooming out and looking at the bigger picture. So one question is, has it been shown that the corals formed from prior preserve sperm live as long as regular corals and their progeny are as healthy? Good question. And we're working on that now. So, and I hope the coral that we produced for the Cystegene flow are now three years old, or four, it's going to be four years old soon. And hopefully they will start to reproduce. You know, since we only started doing cryo preservation in 2012, and curl don't reproduce for eight or 10 years, we don't really have the answer to that. But they seem normal in most ways. You know, the only beginning part is that fertilization is not the fertilization success with prior preserve sperm isn't quite the same as it is with fresh, but we've actually, we've overcome that hurdle as well by using CASA and understanding what we had to do to make the, the modal concentration in our in vitro fertilization of fresh and prior preserve sperm to be the same. And so we think now we've even overcome that. So, you know, it's difficult to know we've not, there's over 10 million humans on earth that have been produced with prior preservation. And we've not even asked that question about them either. You know, I think people honestly don't want to know the answer to that question. Interesting. So there are two questions that are related. So let me start with the first one. Thanks so much for the fascinating, fascinating presentation. I wonder if you might speak a bit to the question of ownership of and access to bank specimens. Really good question. I, I try and make sure that no private organization owns any, you know, I don't give my, my cryo preserved samples to any corporation or any private organization, even like a private zoo or something like that. You can go out and get your own, right? You could, you can, you can cry preserve your own, but I don't do it. And, and I, is I'm very sensitive to this idea that it should be a public good. It should be owned by the people. And, and like for instance, we went to Maria, and I tried desperately to find a bank in Maria or in Tahiti, French Polynesia, they weren't any that would take wildlife. Even in France, you know, which is French Polynesia, there was no, there were no wildlife banks that would take the frozen material. And so we had to come up with a letter of agreement for the material that we took out of Maria, that this is their material, and we're just holding it for them. And until they're ready to move it to their own buyer repository, we will hold it for them and not charge them anything. And so USDA, which is my partner in this has, has been doing that. And it's, it's really been very terrific, but I'm very cognizant that this is, it's, it's like when people tried to patent the human genome, you know, it was such a, such a huge viewer. And rightfully so, this, this should be something that is in the public good. Right. Another question. Where, where in Florida are your corals being kept? And what Florida nurseries are potential hosts? So, thank you. Florida cram has some and moat marine lab, which is down in the Keys has so for a cram is in, in Tampa, and moat marine lab is, is down in the Keys and the coral restoration foundation, which stretches all along the coral reef tracks has volunteered to take our, our assisted gene flow coral and to test them out on their coral nursery trees. So we're hoping, I mean, so we've had the willingness to do this for, since we, we got them. It's really just at the state level that's blocking it. So we're hoping that you know, and it's been very difficult, the pandemic. I mean, add, take one more risk to the pandemic, right? This is not something people really want to even talk about. So hopefully, after the pandemic, I'm hoping I'm going to be able to give some in person training in Florida this summer and maybe, you know, bring up this issue one more time and nudge it forward, if possible. Right. One more technical question. What do you think about efforts to treat disease corals by application of microbiota, biotica targeted methods? My colleagues at, in, in North Carolina and in Florida at the Smithsonian Marine lab there are using probiotics. They, they've like gone to corals that survived and they've harvested the microbiome. But then you have to, not all bacteria is good bacteria, right? So you have to go through all the multitudes of bacteria that they played out and see which ones are going to be good. So it's, there's a time lag there. And they do have some probiotics that are helping the corals, but there's a question of whether it sticks, you know, whether they help them for several months and then they get reinfected again. So I think it's still very much in process. But I think there's, there's definitely, you know, a future for, for having it's really, you have to bring them into captivity to do that. And, you know, how do we, how do we do it? My colleagues are actually putting bags around corals, you know, on the reef and injecting the probiotics and things like that. But it's still a very, how do you make it, how do you make it again, reef wide? And, you know, it might be something that could be pelletized, you know, put in pellets that could dissolve and you spray them with, you know, a, you know, a crop dust or something like that. I don't know that I don't know the answer there, but I think we will have some very, or even robots who get underwater robots that deliver it. So we are going to, we're going to move forward on it, but, you know, right now it's, it's still in its infancy and unfortunately the, you know, the corals don't have a ton of time. Yeah. So I'm going to finish with a couple of questions as kind of zoom out and get some bigger, bigger, broader issues on the table. So one person asked specifically what changes and ethics would resolve the problem? So let me try to address this first. I don't know if changes to ethics would resolve the problems, but definitely I think changes to the way we think about ethics would be necessary. So ethics from the time of the Enlightenment, secular ethics has been very individual responsibility focused. What makes one person responsible for some bad outcome? Who bears responsibility to correct the problem? Where now I think we need to talk much more about collective responsibilities, collective duties to, to act collective duties and responsibilities for bad outcomes. And so it's kind of what I think some philosophers have called the many hands problem. How do we kind of address these many hands problems where many people have some bit to do with, with both the solution and the, and the problem. So I would maybe encourage a reorientation of the way we think about ethics and also include under ethical responsibilities, responsibilities for toward ecological systems, not just other people societies. But I don't know if anything, if you would want to add anything to that, kind of like rethinking the way we do ethics here. I, you know, I agree with you entirely. I think that's, you know, that's sort of, sort of more socialistic approach to ethics in a way, you know, in terms of responsibility, but, but also, you know, you know, we have seen an evolution of how we think about animals in particular, you know, where we never thought of they're good before we do, you know, PETA, we treat farm animals hopefully better than they used to certainly zoo animals and research animals definitely differently. Many people have pets that, you know, are part of their families now, which 100 years ago in the US, you wouldn't see that. And certainly, you know, it's an evolving ethos in the rest of the, in the rest of the world. But I think the question of, do we, can we, can we really, in good conscience, wipe out species with no, with no, you know, sort of consequences to us? The answer really is no. And, and, you know, it shouldn't be just because they do think good things for us, right? They give us accident, we like to breathe. They protect our homes. I mean, there's some really big ecosystem services that corals do, but it's really a moral thing, you know, we should have consideration of other species. We are not the only species on this planet. Right. Yeah, I wanted to introduce your topic to my speaker series because I wanted us to rethink what we mean by doing bioethics, what the scope of bioethics should actually include. Let me conclude with just the following final question to you for me. What do you think is the biggest challenge facing your work going forward? It may be technical, it may be social, and what gives you the most hope? It's a combination of time, running out of time, money to sort of get solutions into people's hands and train them. And, and yeah, so money and time are the, are I think the biggest issues in, you know, my work. And what gives me the most hope are is the, is the, the students that are coming through the, you know, the younger generation because they are changing. You know, they, they, they culturally they're advancing in ways that, that, you know, people who are still stuck in saying, I only care about profit and, and, and I don't, I'm not as part of this larger good of society, you know, having some financial stability is a great one, but there's quality of life and there's quality of, of, of, you know, people love gardens, they love, they love to go to the ocean, they love, they love seeing the great breweries, they love wild places. This is of enormous value to us, and as, as a species, as individuals and, and, and getting that so that's more in the public realm, you know, just as we're trying to change how we do business, so that one, a group of small group of people are not making billions of dollars while the workers are making next, next to nothing. All of that I think needs to be, you know, addressed and I think in terms of hope, I think that, that people are seeing that, you know, these very narrow, sort of more capitalistic perhaps, attitudes are changing and they're changing not only towards our finances, but also towards, you know, animals and ecosystems as well. Hoping, hoping for a different type of climate change. Well, with that, I would like to thank our guests, Mary Hagridorn, for sharing with us your fantastic work and your insights. I also wanted to thank Ashley Trotman and Helen Stephanidis for their help in organizing the session. And I want to thank you, our audience members, for joining us yet again for the series. I will see you again next month. This concludes today's session. Have a great weekend. Thank you, everyone. Pleasure.