Great, thank you, Anthony. They do call it the the graveyard shift, I think this one, don't they, straight after lunch, so hopefully I don't have to resurrect many of you during this talk, but we'll give you, there was a pun there somewhere, I'm sure. Thank you.

They only, they only get worse, I'm afraid the jokes are coming along, so, yeah. So yeah, I'm just gonna talk to you a bit about, my journey really and where we are, and I've got one incredible special announcement to make at the end of this. There's one or two of you in the room know about it.

So, that's really just to keep you awake really till the end. So, we, we will see, you'll see that towards the end, so hopefully. So really just an introduction, yeah, I'm going to talk about the work we're doing, a bit about Nature Safe and about what biobanking is and and so on, and about some of the new technologies that are out there that can be used in conservation.

. And really of the future, something that's a bit scary I'm afraid I'm gonna be talking about. They might be thinking, oh God, that's just maybe stepping over a line, but it's maybe make interesting discussions and sort of bio bank networking and the future goals and so on. So a little bit about my history, yeah, as Anthony said, I'm very much equine based, I've been, collecting and freezing equine semen for about 30 years.

I'm a cryobiologist, I've. I'm not a vet or anything, I've she sort of stumbled across it and we've, you know, we've collected from over sort of 1500 different stallions in the last sort of 30 years. And I've always liked my rare breeds, and I was a trustee of the Rare Breed Survival Trust, and I thought, you know, they came to me about something like the Suffolk Punch, you may, may have heard of them.

There's very few left and think how can we use new technologies. To save the stuff at punching these heavy horses through cryo technologies, and we developed some ways and then we thought, well, if we can save, you know, the heavy horse, why can't we save these amazing species as well. And to my amazement, There was very little out there of this happening.

The technology's been around for a long time, but actually the use of this technology for preservation is, is very, very small. So this is what I'm gonna basically talk about today, and that's came up with the idea about 4 or 5 years ago about setting up a living buy bank. I'm gonna talk about what the differences are, and it's called Nature's safe, the acronym for SAFE stands for saving animals from extinction, and it's a a tissue bank basically for wild animals working alongside sort of assisted reproductive technologies.

Our mission is to save animals from extinction by collecting and definitely storing reproductive cells and cell lines from this. So, you know, it's got all your DNA we always say in your skin cell, you know, when you cut yourself and it scabs over and heals over, that's why we always like the skin cell, but in that cell is all your DNA you're made up of about 3 billion DNA pairs, your eye colour, hair colour, and we can, we're preserving that down. Now there are lots of biobanks out there.

Other than that, but there's different types of biobanks. One of the first ones was at San Diego Zoo, and they're doing a lot of the sort of the zoo's animals out there, and it's a way obviously of, you know, bio banking these species so in the future we can in theory resurrect them or bring them back to life. There's different types of biobanks, you've got the human ones, you've got the plant ones like at Kew Gardens and the millennium Se bank and things like that, or we've got the animal ones.

There's always a bit of confusion between biobanks. What's the, there's lots of biobanks out there, but a lot of them are dead biobanks, what they're doing is they're looking at the DNA but they can't bring those DNA back to life. Whereas we're different, we can.

And these usually ones are -80 storage, and there's a lot around the around the country around the world, but we're freezing down at 196 liquid nitrogen. And the way it works is, just a bit of a biology is in a, in a cell. You know you've got your frozen pipes in the winter time, when you freeze, what happens when it gets really, really cold?

They swell, rupture and break, and the cell does exactly the same thing, and I've been freezing sperm cells for a long, long time. And we do exactly the same cell, and we thought well why can't we do that with a skin cell, and all we do is we have like a an antifreezeer or a cryoprotecting it, dehydrates that cell. So then when we wake it up, it just rehydrates and it stays alive for 1020, 30 or 1000 years' time.

So you can preserve the genetics for as long as you want. And we use different types of, we use something called DMF dimethyl for aldehydes, it's just a different type of cryoprotective, we use different cryoprotectants, and we just finished a PhD at Nottingham Trent trying to even further that, trying to use non-animal based products. We use a lot of egg and things like that, so there's further research to be done.

But as we know, you know, we've talked a lot about species going extinct, you know, there's something like 6 or 7 million species on the planet, there's 1 million at risk, there's 40,000 critically endangered. By the time I finished this talk, one more species would have gone extinct. So we can't wait.

We've got to do something now. And so the methods we use is we usually like the ear because the ear is one of the fastest regenerating cells. Obviously the animal has to pass away, although we're looking into other methods as well on that.

And so we tend to get the ear sample of an animal passway and also the reproductive cells, the you know the testicles and the ovaries and freeze them down cos obviously you'll see in a minute the science behind that, that we can bring those cells back to life. This one picture, really, I could just put one picture up and take away all my other slides, cos it denotes everything we're doing. I feel like I've got to come up the front and explain a bit more about it, but it's from the the the revive and restore.

So you've got all your animals here going into the freezer, and at the moment we can do all this, we can do genomics, we can do gene editing and but a lot of the work we don't know. We don't know what is gonna be used. If I was gonna say, right, I'm gonna take a picture of you lot 20 years ago and send it to Australia, you said, oh, you're crazy, of course you can't do that.

But technology moves on at such a rate. But what we do know, if we don't bank now, we will have nothing for the future. And that's the important message, so a lot of it we just don't know what we need it for.

So this could be any type of animal, and. It's about inbreeding, a lot of inbreeding quite often is what makes species go extinct. So at the moment we've got a lot of our species we've got a lot of diversity, we just put the horse up here, that could be any species of.

And something like the Suffolk Punch, you know, they've got a 20 well the Clydesdale's actually got a a 27% inbreeding coefficiency. If you think 25% is like a, a brother sister mating. But this happens not just in applied sales, but what we can learn from them.

So we need to bank now, we don't, we can't, we don't want to bank down the line here because it's too late. We need 50 different genetic pairs really to make a species sustainable. So the task is is quite huge, but we can learn from our mistakes.

And it's called the extinction vortex. So when species get so inbred, they get into this vortex and they start going down and down and down. And this is where the Clydesdale horses, you most probably have heard of them.

They've got a 30% neonatal death. And what happens with that is when people try and breed these breeds, they die and they spend a lot of money and they said look we're not gonna do it anymore. And then it just implodes.

So with Nature Safe bringing these genetics in halfway up, we can hopefully climb back up and we're gonna show some examples of this at the end, how this can actually work. So some of you might say, well, how some of the, the methods can work. So epididymal sperm extraction, I'm glad you've had your lunch now because some of these pictures might be a little bit graphic, but it's where we can actually extract semen from from the testes.

I was working on a stallion the other, well, not the stallion science testicle that passed away at one o'clock in the morning a few weeks ago, extracting the seymour, and there's really valuable genetic lines from that, and we can freeze that down. And bring it out and we've been doing this for quite some time with horses. And whenever we do it with with semen, as it's from the testes, it's got none of these seal fluids, so we've actually had to develop something that wakes the sperm cells up so you can see here, they're totally immotile.

But we add something to it and it's exactly the same semen that we've added to it to wake it up. And we thought, and then a few years later, we got a phone call off a Scottish farmer and and he. And he said, oh, I've just bought this ball, and it's just for 10,000 guineas, it's just broken its leg.

Can you do bull's testicles? I said, well, I've only done horse's testicles. Went to Google, right, anatomy of the Bull's testicles.

I'll give it a go. So he sent the testicles down and they were, they were, 48 hours late. There is a bit of story to this, but it's probably a true one.

He, he was going down. I was at a show and I said, look, I can't get down there, you've gotta call me and let me know when you've dropped those testicles off. Anyway, heard nothing, heard nothing anyway.

About, you're all vets in the room, I think pretty much so you'll forgive me all this, but anyway, he, he drove, he drove down and I hadn't heard anything and I said, and I rang him up and I said, have you dropped those testicles off? And he said, yeah, I dropped them off sort of, you know, 4 or 5 hours ago. I've left you 4 messages on your phone.

I said, what's the message? He said, oh, I've dropped the testicles off in your kitchen, can you tell me how much semen you got out of them? Well, I said that's not been on my message, so whoever's picked up that message.

Oh they, they've got a rude awakening. So, but anyway, we did, we managed to get semen off it and believe it or not, we got the first calf, born from this from this method. And I was doing a presentation, it was about a few years ago and there's a vet sitting here and he said, I sent you those testicles.

And he said, you're not gonna believe it because I used to put up this calf picture. And he said that calf, now is this bull, cos I only knew of the one offspring, but from that one procedure we got 30 offspring from one that. And I don't know whether this is something we should brag about or not, but we actually made it on the front page of the summit as well.

So, so there was no Brexit or anything else going on there, so I think, but what I'm trying to say, there are other ways. So I thought if we can do this to a horse, we can do this to a a cow, why can't we do it on other species? And so now we we're linked up with lots of other zoos.

And so we started doing this and on the go, obviously it's neck quid, so it's easy for us to do. But it worked really, really well. The problem is a horse's testicle weighs about 300 grammes and I had to do a mouse steer.

Did anybody know what a mouse deer is? It's a tiny little one very, and their, their testicle weighs 1.5 grammes.

It always astounds me. I, I've got the worst memory in the world, but I seem to know the weight of every animal's testicle out there. Weird sort of way.

So, but anyway, so you know, they've all got their different challenges. We did a cheetah not so long ago, and they've, they're very inbred and of course what happens with inbreeding fertility drops, but we did manage to get some sperm cells. I don't think it's working actually.

Oh yes it is, and so we pres preserved that down, so it's, it's working on other species and we've worked on lots of other species. I'm actually, going to, set a sail to do some more work actually in the next few weeks, well next week actually to collect off some different species down there. So other techniques that we can use is taking testicular tissue, because if you can get that.

You can actually harvest this work in the, in the lab and grow semen in the laboratory. Now I have to put it out there. This has not been done on rhinos yet, but it has been done on mice and other species.

So this work is ongoing work, all the time. same with the ovaries, in, in the ovaries you've got all the follicles in there, you can slice all that up, and, and get retrieved the eggs from that and then grow them in the, in the lab as well. And we've done this with some other species, but it is, every species is slightly different, has its challenges, but you know, every day is a school day at our place and must be.

You're always learning and that's what's so fascinating about this job. Cloning, side, so this is obviously done in the domestic side and we're just transferring that across. So it's where you've got a skin sample, it's got the DNA in it, you get an egg, you take the DNA out of that egg, and oh, it's most be better in in this picture, so you've got the DNA here, and you take it out of a host and you extract it and put the replace it with the DNA, the one you want clothing, and buy an electrical impulse.

That automatically grows and and divides and and carries on, you'll see a bit more of cloning animals in a minute. So the other work that we do is, this is like a cheetah a few years ago, just the proof of concept that they can work. So this died in 2019, I think, was it?

Yeah, 2019. We cryopreserved just the ear. We froze the ear down, we woke, if that's the right word, at 2 years later, and we got those cells to proliferate.

So after 3 days, as you can see a few cells here, and after a week there's millions of cells. So effectively, you can bring that species back. And this is what we're saying, by using this technologies, I find it quite amazing what we can do.

So, other, other ones, this is the Brazowski horse. Again, it's easier for the domestic side. I think he's a bit of a show jumper as well, this chap.

But effectively. This fold is 40 years old the day it was born. 40 years ago this so this just proves that this concept can work.

And you know, they've managed to produce this fault. Again, it was an easy win because horses are cloned and dogs are cloned, so it just shows what can be done. The Bruzowski horse, you know, it's a closed, closed herd, you know, there's 13 founders, but of course the inbreeding and inbreeding, and luckily they preserved this lost genetic 40 years ago and were able to bring it back and you know we're talking about extinction vortex, and that's one way we can get it climbing back up.

And they've just done another one, literally, er born in February. Just showing how well that, you know, works, they've done another one. But it's not just about the horses.

Or the dogs. I just love ferrets. There's something about ferrets, so the black footed ferrets, now, I don't really know much about black footed ferrets.

Back in the 1970s they virtually went extinct. There were so few of them, but when they were disappearing, they managed to freeze a few of them, and Elizabeth Ann was born, was cloned. It was on David Attenborough's programme, The Mating Game.

And I should have actually had his quote here, but I think his quote said something, now we have the ability to save any animal if needed by using it, which is an an amazing sort of statement to say. But this is what they did with the black-footed ferret and she was a, a genetic line was lost, and of course they could bring it back and do this. So this is the science that's really jumping even further, is now it's possible again, it hasn't been done in rhino, so I don't want to say it's been done, but.

If you can get a cell, You can turn it into what's called a pure cloak stem cell, in other words you neutralise that cell, it doesn't know what it is. And then by adding different growth factors, you can actually turn it into a sperm and egg crazily. It is expensive, but like anything it comes down.

So in theory, you can use this reproductive techniques to bring these species back. I know there's a lot more work, you've got to put it into a into a like-minded host and things like that. There's other work, but it, the possibilities are there, but I keep coming back if we don't bank, we cannot do it.

Some of the animals are quite difficult to do, amphibians. When we grow the cell lines from some of our, you know, the, the, like the cheetahs, they take about a week or two. These can take up to 6 months, but we're bringing in experts to help us do this, and there's a lot more technology and stuff that we need.

So it is quite fascinating. We're a million miles from from knowing it all, and that's what's so quite interesting about about this work, but the amphibians we learn an awful lot from. And so far we've collected from over 175 different species in the last two years.

We've got a lot of zoos on board. Chester Zoo is our foundation zoo. They've been quite brilliant to us.

You know, we've got ZSL and we've got so many other zoos, but now we're wanting to set up hubs all over the world cos with the CITES things we can't move tissue around, so we're setting up subs, all over. But we all think of the lions and the cheetahs, I was trying to do an epididymal sperm extraction off a mountain chicken frog about a month ago at about 11 o'clock at night and boy that was difficult. I mean, it was about this small, but what stimulated the sperm to swim was actually water crazily.

Every other species say don't put water near it, but it was amazing and we actually photographed that semen under an electron microscope and we could really look in detail about that as well, so it's yeah, we'll present it and quite often we don't know what animals we're gonna get until the last minute, so. So future technologies, there's there's lots of different ones out there that we can use and we're learning a bit about what's nature's safe and I'm a great believer we've got to use these technologies, you know, years ago it might be saying we're stepping over a line. But I think the line has been crossed with us.

We're at a cliff edge. You know, and we've got to do something about it. So one of the things we're doing at the moment is sexing technologies or using sex semen.

There's about 4% more DAA in the X-bearing sperm, the female sperm, so we can home in on that by putting a fluorescent stain in there and of course it shines 4% brighter. So then we can zap that sperm cells and it can be extracted. And it's amazing they do about 20,000 sperm a second.

And there's about 7000 go in the pot that you want, but you need millions, so it's a, but this has all come from the bovine industry. And I thought, well we can do this, why can't we do this with a Suffolk Punch? So the Suffolk Punch, they have about 30 folds a year, and for some reason it's always male gender heavy, about I think last year they're 20 males and 10 females or something like that.

So we thought let's see if we can use this technology and this was a quite a, a groundbreaking moment. It it's gonna work Ruby was produced at a 304. And it's not just any fault.

It's, it's a foul worn by a sex semen. This bleeds, the first ever in the world for a rare breath and also do sex semen. And using these technologies can really help us breathe.

It's only about 70 or 80 minutes left in the country. It's only 300 left in the world. They need all the tools of the industry, which is quite incredible now what we can do with science and technology, so using this technology, maybe, just maybe we can help stop these res from burning things.

This is hold the future of the idea. And I tell you what, with a gestation period of 11 months, I've never looked between a horse's back legs when it cos there was a 5% chance it was going to be a cult. So, as soon as you look, yeah, oh, it's a fully, which was just, it was just brilliant, so, and we're still that work is carrying on going, but we have had literally just more success the other day.

The problem we we did that, that was with fresh semen, so what we do, cuts off the stallion, sets the semen, seminate the mare in the evening, but, There's so many things that can go wrong with that, er, in other words, that the sanding didn't be good enough quality semen or the mare didn't ovulate early enough, so we've just had our 1st, 1st successful fo born by using frozen ISI AI and this bo was a a product of that. And so that really opens the door up now using ICSI, so you'd only need one sperm cell. So this has made a huge difference in, in taking this forward.

And now the same work, because we have to use it on the domestics, we can't just throw it across to rhinos cos where's the research? So now it's actually been used on rhinos and and and and work like that, which is quite fascinating. So our next slide is, yes, we are looking to see if we can actually, on the, on the frozen, starting to to freeze down these embryos as well.

Other work that's been doing is the oocy recovery, which is for ICSI, so we take the take the eggs out and then using ICSI one sperm cell, and again this is starting to be used on some species, and this is a time lapse camera taken every 2 seconds and you can see the actual embryo developing. This is actually the first one for a horse actually showing how an embryo does so. So we're saying it's from the ear, it's a fairly simple process that we're doing when we are bio banking, so we tend to use the ear, most of the time, and, we tend to get, take about 10 vials from the ear, they don't need tiny little snippets.

Put in 10 vials and we put 6 in one tank, that we can do, try and grow those cell lines up, and then the idea is to put them in 3 other tanks and put them in 3 different locations. You know, in case anything happens, so, so, but it's a fairly, I say it's fairly inexpensive to do. There's a lot of, we use a a company that does all the IVF stuff for the for the matching systems because we want to track all this as well cos track tracking these samples is really important so they're all barcoded and so we know exactly where these, these samples are.

So some of the other genetic side that we need to be looking at is sort of identifying these, the architecture, the genetic traits of of maybe these genetics as well. And and when we're freezing these animals, we want to be making sure that we're freezing the right ones. We don't want to be freezing closely related ones.

So looking, doing the profile of this species and we're looking with Nottingham Trent University to be doing this as well. And the price is dropping and dropping on this, so the idea is to identify the right ones to buy a bank. We don't want a freezer full of related ones, we want a freezer full of different traits.

In this next slide I might sort of think, God, this really is out there, but the next thing we're looking at is digital DNA producing through digital 3D printing. They can already do yeast, and they're looking at this at Nottingham. So I buy banks in the future, you know how you have the profile, you can get the profile basically on a computer.

So our pro, so in the future, our buy banking might not be in a 196 tank, it might be crazily on a computer. I'm not saying whether this is good or bad, I'm just saying what is it's round the corner. I think it's about 5 to 10 years away.

You could say, God, are we creating life with this, but this is the, but we've got to use it for the right reasons, obviously, but this is what they're looking at seeing maybe in the future of 3D printing and possibly DNA. But it's all about knowledge transfer for us, we've got to get other parties in, and we do. We try and get as many people in to help us with this.

We can't do this alone, so we have a huge bank of scientists, zoo people coming in. I was speaking to somebody just the other day about snakes. I don't know this video's going to play or not.

Maybe not. Anyway, it was collecting semen off a snake, maybe he didn't want to see that, but they knew about how to extract semen from a snake, but, oh yes it is, yeah, but they didn't know anything about cryopreserving it. So we said, look, we'll have a, have a go and they and they literally just by massaging it, they managed to get the semen to the semen snake to give semen, so which was quite interesting, so there's lots of new.

I wouldn't say it's my favourite part, I'm sure there's certain snakes that you might want to not do this way, so this is snake semen that you you can see here, so. And also coral, corals that we're looking at. I didn't, I must admit, I'm quite naive in this, so we went down to the Natural History Museum in November.

There was a coral meeting on. Coral is one of the fastest clining species on the planet. We've lost 50% of our coral since 1950.

They're the lifeblood of our seas, and they produce sperm and egg. Once a year on a full moon, so you've got a bit of a short window, but down in London they've managed to artificially make coral spawn, it's quite a, a crude way of doing it, you know, you get your your dam lighters and they cut a cut a tennis ball in half, put it over that, measure the light, the lunar light from the from the moon. To get the right light that comes off it and through that sort of crude but very good way of doing, they've managed to get the coral to to spawn, but they're not too sure about cryopreservation, so we're we're developing a, we actually did a 3D thing that we've made in our lab that now we can hopefully freeze coral, sperm and eggs.

The problem with coral. And again I learned this the other day, is that coral be feed off the bacteria that they're around, so you have to feed the freeze the bacteria as well as the sperm and an egg, but it's yeah it's really, really fascinating. So, you know, we need all your help and I, I some great people.

I spoke to somebody before about, you know, stuff we're doing stuff with livestock now cos we need to buy a bank all our livestock, but that's another story. But if anybody in this room wants to reach out, we'd love to hear from you. And one of the things that we really are trying to champion now is are you.

Native wildlife and this is where a bit of a loss with this, so I'd love somebody to come and help us with this, where it's easy for us to get samples from zoos, but this is an area we've just been given some money from the postcode lottery to focus on this. This has always been our remit to do this. So, you know, we would love to anyone to come forward and help us, we're champion that to get these samples in because we don't get the samples.

You know, us as a charity we can't move but we also want funding, so, but that's a very quick shout out, but so that's a really big important part and Anthony was saying there's a great platform here really to to champion that. So, just this is my other work, I do. I'm going out to South Africa fairly soon, hanging on to an elephant's penis for about an hour a day.

Seems a bit weird, but I have to say it's the most amazing experience I've ever done. and, we did about 8 bull elephants. We're about to go out in 2 weeks' time to do the same with rhinos in the 1st 2 weeks in rhinos, and it's just incredible.

I feel like I'm the luckiest person alive. And we've just recently been given the Queen's Award for enterprise for this, now it's the King's Award. So I was very fortunate.

I'm very lucky because I stand up and do this, but I've got an amazing team behind me of scientists and researchers to do this. But the big news today, we've already mentioned this one, that was a press release went out yesterday, but we've just been shortlisted for the Earthshot prize. Which is incredible.

Which, again, I'm very lucky to be here, but it is, we have an amazing team and that is super exciting. We won't know until the 31st of August whether we're through to the final, but I think there's 2 or 3 1000 do get the first bit and then we're through to the next, but we we think we're down to the last 100, we think, but we will see, but we keep our fingers crossed because that will be absolutely amazing. But anyway, but thank you for listening.