Cambodia: crocodiles, elephants and genetics

As part of the RSE investigates… conservation series, Dr Alex Ball will give a lecture on developing the first conservation genetics laboratory in Cambodia.

The Royal Zoological Society of Scotland WildGenes team have been developing the first conservation genetics laboratory in Cambodia since 2016.

Based at the Royal University of Phnom Penh, the team have been training staff in lab techniques to produce genetic data for conservation priorities in the region. They have developed a genetic test that can identify the critically endangered Siamese crocodile. This is important when selecting individuals for breeding and release programs as they are often purposely hybridised with other species within leather farms. They are also supporting the lab with genetic monitoring of the few remaining Asian elephant populations in the country.

This talk given by Dr Alex Ball will explore the most recent project updates and developments.

Please note transcripts are automatically generated, so may feature errors.

00:00

Hi there. Welcome everybody. My name is Helen Sen. I’m head of conservation and science at the Royal Zoological Society of Scotland. And I’m your chair for this Royal Society of Edinburgh investigates conservation series. This session is being run in collaboration with the Royal Zoological Society of Scotland and the RSE investigates conservation series is exploring the vast and varied work that’s being done in the area of conservation from the conservation of wildlife and the environment to the conservation of natural heritage. And throughout this series, we’ll be asking important and at times tricky questions about the ethics and conservation and why conservation matters, and how we engage the broader public in conservation efforts and the role of conservation in addressing the climate crisis and increasing biodiversity crisis in Scotland and globally. And the Royal Zoological Society of Scotland is a specialist in species conservation, working on the conservation of species as diverse as the dama, Gazelle and the small scabies mining bee. And we have a commitment to helping with the recovery of 50 species over the coming decade. Some of you may have already heard from our colleague, Dr. Helen Taylor, who talked a few weeks ago about the efforts to save invertebrate species that are on the brink of extinction through conservation, breeding and release here in Scotland. With this event, we’re going to be zooming over to Cambodia to discover how the rose illogical Society of Scotland has been helping with the recovery of the critically endangered Siamese crocodile and the monitoring of Asian elephants with the use of genetic technology, and the launch of the new Kunming, Montreal global biodiversity framework. Earlier in the year, the importance of conservation of genetic diversity has finally been recognised within global biodiversity targets. And this work that we’re going to talk about today illustrates why it’s so important to conserve not just species and their ecosystems, but the genetic variation which underpins them and allows them to continually adapt and evolve to pressures like climate change, and disease. So just to provide a bit of an overview about this event, this is an hour long event with a talk which will last for about 40 minutes, and then a question and answer session. Please submit your questions throughout the event, the q&a function, which you’ll be able to find at the bottom of your screen. This event is being recorded and will be available to view on the RSEs YouTube channel shortly. And if you require subtitles, these can be turned on using the custom live streaming service which is at the top left of the screen. And if you’ve got any problems with that, I’m sure you can pop a message in the chat and someone will be able to help you. Okay, so just leaves me now to introduce you to our speaker today. Dr. Alex Ball who is my programme here at the Royal Zoological Society of Scotland and who’s the programme manager for the RZSS whild genes conservation genetics lab. Alex’s job involves overseeing conservation work, conservation genetic work to support the recovery of 15 different species in Scotland and around the world. And the lab is currently working on cap Kaylee and scimitar horned Oryx find hoverfly and Himalayan wolves, and has a team of six staff working in the lab on analysis. And in the biobank. Using genetic analysis to help with the recovery of certain species, the fairly niche skill, and training is a big part of the work that we do. And to this end, we’ve been involved in a close partnership with the Royal University of Pnom Penh and fauna and flora International to establish the first conservation genetics lab in Cambodia. And this project is a project that Alex leads. And without further ado, I’m going to hand you over to him to talk about crocodiles, elephants and genetics in Cambodia.

03:44

Thank you very much, Helen. So I’m just going to share my slide.

03:52

Okay, so hopefully you can all see that.

03:57

And thank you to the RSE as well for inviting me to give this talk and for everyone attending. So as Helen outlined, I’m going to be focusing on our work in Cambodia, and I’m particularly going to focus on two projects, the Siamese crocodile and the Asian elephant. But to start, I’m just gonna give you a bit of an overview to the RZSS while jeans lab and the Royal Zoological Society of Scotland just for people who might not be fully familiar with us.

04:25

So the Royal Zoological Society of Scotland is a wildlife conservation charity that was established over 100 years ago, and so its main mission is to conserve species within the wild, but it also owns two zoos, Edinburgh Zoo, and the Highland wildlife park in the Cairngorms National Park in the north. So these are both based in Scotland and the conservation programmes are run out of both of these zoos, but they’re focused on projects within Scotland or native species, also on the conservation internationally have over 23 Different species across the globe. And one of the really unique things about the Rosie illogical Society of Scotland is the RZSS wild genes lab, and this is because we are one of only, or the only zoo based conservation club in UK. So I’ve just got a map here, just showing you the locations of the various conservation projects that we work on there to zoos. And these are a native species conservation project. And it’s just going to zoom out and show you the breadth of diversity of the projects. Whereabouts in the world. They’re focused. So we don’t use conservation genetics on all of these projects that we do on a large proportion of them. But other ones are focused on, on on using other tools to aid conservation priorities. And one of the really great things about the royal Zoological Society of Scotland is that we can draw on multiple different expertise, because we have those theories. And we’re also working on conservation projects in the world. So we have expert animal keepers and veterinary teams, that have a huge range of knowledge on a really diverse array of species, often lots of threatened species as well that we hold within captive collect, that can be really important for conserving the species in the wild. We also have the field project team that sits within the Conservation Department that’s focused on restoring species in the wild. And we have the conservation genetics team, or the RZSS wild genes team. And we produce genetic data that helps inform these conservation projects. And as Helen alluded to, in her introduction, biodiversity conservation is, is really focused on three main pillars. And these are basically enshrined now within the international community, by the Convention on Biological Diversity. And two of them are much more familiar concept people and to conservation practitioners. And that is ecosystems and species. So obviously, we want to preserve the diverse array of ecosystems that exist on that planet. And there can be large differences between them, between wetlands, deserts, Woodland habitats, and it’s important to preserve aspects of all of those. The most commonly thought of concept within biodiversity conservation is the species level protection. And that’s basically using species. The more species you have within an area, the higher by depth, biodiversity you have. And so it’s important to protect all of the species in those areas to maintain biological diversity. However, one of the most important aspects that’s often overlooked is genetic diversity. And this is diversity that can be with found within those species. And it’s really important to protect that if we’re going to preserve species in the long term. And it basically underpins both of these other pillars of biodiversity because if you don’t have high levels of genetic diversity, you can have, you can basically be left with species that are less able to adapt and survive into the future. And that reduces your species level, by biodiversity that fundamentally then leads to the degradation of ecosystems. So we’re going to focus on that third component, our genetic diversity. And we’ve been focusing on that for over 10 years within the Royal Zoological Society of Scotland. So we realised it was a really important aspect of biodiversity conservation, and had been working, working on it on site at Edinburgh Zoo since 2010. And it’s now being recognised globally and internationally as a key concept as Helen outlined under the treaty. And there’s now targets to restore and protect genic adversity. By 2030. All countries, or a lot of countries have signed up to do so in the lab at Edinburgh Zoo. On the right hand side of this slide, you can see a picture of what most visitors are busy with see, unfortunately, because of the sterile conditions that were required, and to reduce contamination, the public don’t have access to the lab. But if you were to go inside, you’d see a typical lab setup as it did on the bottom of the slide. Lots of white lab benches, white lab coats and machinery to extract and work with, with DNA. As I mentioned, we’re the only zoo based conservation lab in UK so we’re in a really unique position. And we’re one of only a handful in Europe. And we conduct internationally recognised scientific work. So I’m just putting up a few of our recent scientific projects and papers that we publish in peer reviewed journals. However, we don’t see that as The most important aspect of our work, we see ourselves as a bridge between the academic community and the research that’s going on in universities, and the applied conservation sector. So actually impacting conservation programmes and future management on the ground. And obviously, we were in that position because of the huge range of new technology, and new research that is, is being conducted on genetics within the university sector. And our role is basically to pick and choose and work out and develop new techniques from some of that, to work out the best way of tackling the questions that are relevant to conservation and using genetics to do that. So as well as producing our scientific papers, we also feed in our results to action plans, conservation action plans for a range of threatened species, and you can see a few examples of the ones we’ve been involved in on the right this slide. So I’m just going to very briefly introduce you to the team so we’re fairly small team within the Royal Zoological Society. We have two research two technicians in the lab that produce the genetic data on our various projects. We have two research scientists that then use that genetic data to analyse it produce results and feed into action plans and management reports. We also have Dr. Cecilia Langhorne, who is our biobank coordinator, and preserving genetic material from a huge range of species. So it can be used to answer conservation both now but also in the future. We have a new research assistant, who actually joins us this week, and various student interns and PhD students and you can see a couple of them.

11:47

And ultimately, as a team, together, we are using tiny tools to answer really big conservation relevant questions. And the reason we’re using tiny tools is because we we studied DNA. So the fundamental building block of, of biological organisms that is basically the genetic code or manual that creates the proteins that then build up an organism. And each genome or genetic material, and each individual biological organism is unique and is different. So your DNA will be completely different from from the person next to you. And ultimately, as geneticist, what we’re trying to do is convert those DNA double helix structure these molecules, really long molecules into four letter codes that we can read, and then look for differences in those codes between individuals. Just to give you an idea on how tiny DNA DNA is, if you were to take a single hair from your head, you’d be able to get 1000 strands of DNA across the width of that hair. So it’s really, really small. And basically, in the lab, we just work with colours and colourless liquids, we can’t see the DNA until we sequence. But although it’s it’s we can’t see it with the naked eye, if you were to take those strands of DNA from a single cell in your body, and align them end to end, stretch them out that stretch for about two metres length. And that’s just from one of your cells. And you have a copy of your DNA, or your genome in almost every single one of your cells in your body. So you’ve got a huge amount of DNA in your body. And that two metre long string of DNA and each one of your cells is about 3 billion letters long. There’s a huge amount of information in there that we can look at and look for variation and differences between different organisms and different things. So just a very quick schematic of that we obviously collect genetic material, and blood or tissue, or other forms of genetic material, we extract that DNA in the lab, we then sequence that DNA into the four letter codes that create that sequence. As our example, we’ve got a single individual that we’ve extracted in sequence here. And then we do that for another sample as well. And you can see that there’s a difference between these two different individuals, the G and the C there. And that’s what we’re looking for when we when we’re talking about genetic diversity because there’s a difference there. There’s a variation. That’s some diversity between these two individuals. And obviously, you can get different levels of diversity. So there’s only one variation between these two samples. However, if we sequenced another sample that is identical region, you can find more diversity and this is potentially maybe two different species in the sample rather than the same species. So that’s fundamentally what we’re doing Ashton. assist in finding that variation. And the reason we’re looking for that variation. And the reason it’s so important for conserving for conservation is that there’s a huge amount of variation within a single species. So on the slide here, we have one species, species of antelope. And the colours are basically depicting different genetic diversity that’s found within a population or within the species. And if we get a decline within the species, and we start losing individuals, you can see that we are also losing that diversity, we’re losing those colours. So although we haven’t lost the species at this point, we’ve still got, or in this case, three individuals left. We have under under the biodiversity, biodiversity categorization of species, so the species isn’t extinct, we haven’t lost this diversity. However, there’s been lots of mini extinctions of genetic diversity that have occurred, that we won’t be able to get that diversity back. So even if this population expands and grows, again, it’s just going to be made up of that blue and that white diversity and not the rest of the stuff, a lot of that’s already been lost. And this can be really important because genic diversity fundamentally can allow species to adapt to future changes, that happened in the environment, potentially due to new diseases, or climate change affecting the environment. So it’s really vital that we understand what we’ve lost, and, and that we can serve our genetic diversity. And another great thing about working with genetics from a conservation point of view, is that all biological organisms have DNA. And so we can use very similar tools to look at a huge diversity and range of species. And we work on mammals, birds, reptiles, and even invertebrates that you wouldn’t work. I heard about some of the investments in a previous court, and we’re doing genetic work, help me. But today, I’m going to focus on just two of our projects. These are an example of a range of different projects that we’re working on at the moment are recently finished working on that we’ve been using genetics in the conservation of all of them that if you do see anything of interest, feel free to answer ask a question about the end. But today, we’re going to be focusing on Cambodia, and I’m going to start by looking at the Siamese crocodile project. So this is a picture of the Cambodian cardamom mountain landscape. So the Siamese crocodile and the Asian elephant I’m going to be focusing on today, I found within the cardamom mountain, it’s an area where there where both species are found. And Cambodia is a real jewel of biodiversity within Southeast Asia. So it’s the biodiversity international recognised biodiversity hotspot. And it does have really large remaining tracts of forest and wetland ecosystem says the largest remaining tracts of forest within Southeast Asia. It has the largest freshwater lake in Southeast Asia. It has one of the biggest rivers in Asia, Asia run in fit Miko river. And so as a huge amount of landscape where elephants and crocodiles can still be, could still be found. However, what Cambodia has within its biodiversity and their species that is you’re hanging on to it lacks in terms of infrastructure, and tools to help conserve the species. And as before we started working with them, they didn’t have any conservation genetic facilities within the country. And what we have focused on since 2016, is trying to build that capacity. Over in the Royal University of Pnom Penh that’s in the capital city of Cambodia. We’ve been one of the universities and creating and developing the first conservation genetics laboratory in the region. And this means that not only can we help help inform conservation of these two flagship species at the moment, but we’re providing the tools and the training to staff in the university so they can then apply this to other species in the region as well as information from from a conservation perspective. And we’ve been closely working with fauna and flora, international oneness fauna and flora as of this year. And they have been part of a much longer term capacity building project with the Royal University of Pnom Penh, and they’re set up first conservation master’s programme within the country. And so we’re really linking up with that and I in in our training project as well. That’s been going since the early 2000. So this is just an example of some of the training that we’ve conducted. we’ve mostly been focused in on training the technicians and developing the laboratory, but we’ve also been involved in training master students on the master’s programme. And, and the first training exercise took place in 2016, you can see the team that was involved in that top left. We’ve been taking over new genetic techniques and tools that we develop in Edinburgh over to the lab so they can then use them for conservation. We’ve been doing that. And we’ve been using the Siamese crocodile and the Asian elephant in as training tools for that capacity building, but at the same time producing data that is relevant and being fed into conservation programmes on the ground in real time. And the reason we’ve been focusing on Siamese crocodile is because it’s a critically endangered species within the country. And it was thought to be extinct in the wild, it was a real a real concern in the 1990s. Until in the year 2000, fauna and flora international discovered a remnant world population in the cardamom mountains, there’s a map of Cambodia on the right hand side and the red star is the cardamom mountains.

21:05

For that region, there was one remaining population down that was thought to be less than 100 Siamese crocodiles remaining in the wild in the world. So since then, a few other remnant wild populations have been found. It’s not very many, and it’s very few numbers. And this is a species that used to be found across the whole of Southeast Asia within within the freshwater landscape. So huge, massive drastic declines. However, there are 10s of 1000s of crocodiles in Leather farms throughout Southeast Asia and not just in Cambodia. And this is one of the reasons for the declines. They have been poached from the wild and taken into these farms for the leather industry. And there’s over 500 Crocodile farms there’s about 500 legally registered crocodile farms in Cambodia at the moment, most of these around the areas. So this is the largest lake in Southeast Asia you can see on the map here, and most of the farms are cultivated around them. And so what fauna and flora international wanted to do was use these crocodiles from the farms in a breeding and release programme, so set the breeding release programme using these crocodiles and then reintroduce them into the wetland landscapes that have gone extinct from across the country. But one of the problems within the leather funds is that crocodiles issues are purposely high by a hybridised together. So there’s two native species to Cambodia, the Siamese crocodile and also the saltwater crocodile. They occur in very different habitats. So as as the name suggests, the saltwater crocodile is found in the coastal regions and estuarine habitat and the Siamese crocodiles found in inland in freshwater. Now, both found in the farms and they both hybridise purposefully together. And they’re actually really difficult to tell apart visually, especially when they’re young. So the saltwater is one of the largest and most aggressive crocodile species in the world, the Siamese is one of the smallest and most docile, but when they’re young and juveniles, they look very, very similar. And the hybrids are even more difficult to tell apart from from each other from the parental species. So that’s where the the genetic lab came in, and where we can use genetic data to try and help identify the Siamese crocodiles within these farms so they can create their their breeding and release programme. And we developed two tests that the lab over in Cambodia could run. So this involves the teams collecting blood samples from crocodiles in the lab, extracting the DNA, and then running a mitochondrial sequence and test and a snip. Snip stands for single nucleotide polymorphism. It basically just means there’s a single change within the genetic code between the saltwater and the Siamese crocodile, that unknown region. So we work out these snips by looking at known Siamese crocodiles, and known saltwater crocodiles, and looking for the variation in the differences. So we’ve got 25 snips we know for example, in this one are a C in Siamese crocodiles, and our T in saltwater crocodiles. So then we can just look at these 25 locations within the genome at the farm crocodiles to work out how much Siamese and salt water genetic ancestry the individuals have. So just the important point with when running a test like this, is we’re not going to identify every single crocodile that has hybrid ancestry within it. So just to provide you an example. I’ve got basically a crocodile family tree on the slide here. You can see our hybridization event has occurred up here on the top, on the top right. So Siamese crocodiles are in green and yellow. There’s been a breeding The hybridization event between them here producing an f1 hybrid. This is basically a crocodile it’s 50% salt water genetics and Siamese 50% Siamese crocodile genetics. However, if this crocodile them, and our test would would identify it as a hybrid crocodile, and we’d exclude it from the breeding programme. However, if this crocodile then mated with a Siamese crocodile, which we termed as the backcross, because it’s been a hybrid meets with one of the parental species, its offspring is only going to be 25% salt water genetic ancestry and 75% Siamese ancestry. So in this first generation backcross, our test would identify as having hybrid ancestry and the same is also the case for a second generation backcross. So again, if this first generation, individual mates have a Siamese crocodile that will produce an offspring that is only an eight saltwater crocodile ancestry test will still pick that up, and it’d be classed as a as a hybrid that we would exclude from the programme. However, once this second generation backcross makes it a Siamese crocodile, the resolution of our test is no longer able to identify its offspring as a hybrid or a Siamese crocodile. And we wouldn’t be able to screen those individuals out. So what we’re doing with this test is getting rid of the most problematic individuals from the breeding programme, and then using the other individuals to set up and using the best doctors. So just a really important point to be aware of when we’re conducting hybridising. And I just want you to show some of the results on the 354 cocktails that we have screened. So this will have actually screened over 400 Crocodiles now. And we’ve just had another batch of 80 Crocodiles arrive in the Cambodian lab last week, the team are busy extracting DNA from now. But these, this shows the first 354 cocktails with tests, and it’s a structure plot. So this is a really common way of visualising genetic data within a genetic community. If you’re not familiar with one of these, I’ll just try and talk you through it. So each of so in this plot here, each of these columns is a single individual that’s been tested. And the amount of green in that column is the amount of Siamese crocodile ancestry within that individual. And the amount of yellow is the amount of salt in that individual. So that’s how you can you can think about it visualising a big takeaway from this is that there’s a lot of green. So that’s really good news. Now, the majority of the samples that we’ve tested have Siamese crocodile ancestry. So that’s been really great for the breeding and release programme. However, you will notice some blocks of yellow here. So this one here, don’t worry, these well known saltwater crocodiles that we use as positive controls, there’s about 20 individuals here that all show 100% saltwater crocodile ancestry, and then known wild saltwater crocodiles. So that’s, that’s great. We’ve also got known wild Siamese crocodiles of in these tests as well that come out as as green, these columns. But what you’ll notice is that there’s quite a few crocodiles that have both green and yellow in their columns. And these are the ones that have mixed ancestry. And so these are the hybrids that we identify, and then we can update for them for international, this shouldn’t be used for the conservation breeding programme. And so far, we’ve found about 10% of the crocodiles that we’ve tested, have come back with hybrid ancestry. So we’re screening those ones, those ones out. So that is the first test that we conduct in the lab, you’ll I did mention that we do two different genetic tests. The second one is the mitochondrial sequencing as so it’s, it’s slightly different in that we’re sequencing one region within within each crocodile. And we have a database of saltwater and Siamese sequences. And so we can compare and check whether we’ve got sort of two assignments, a sequence of mitochondrial until we get a readout like this from this test, and what we found is that there are some individuals that didn’t match the salt water or sinus crocodiles within within our database. And in fact, they actually matched this species here. This is a completely different species called the Cuban crocodile, and as its name suggests, it’s not found or it’s not from Cambodia. It’s from one island in the Caribbean, Cuba. And it also happens to be a critically endangered species. And somehow, it’s made its way to the leather palms in Cambodia. And we’re actually we’ve actually picked up a few samples with this mitochondrial marker. That’s That suggests that they have Cuban crocodile ancestry. And so this is a bit of an unexpected result. And we’re now focusing on trying to develop markers and include this within our screening. And it’s, it looks like the hybrid tests that we’ve been running, a lot of that hybrid signature is coming from the Cuban crocodile hybridization that’s occurring within within the pharmacy as well. So that’s continuing work and something that we are now at work with the lab on as well, going forward, as well as continuing new, the new fund blood samples. So that is a very brief overview of the crocodile project that we’re involved in in Cambodia, I’m going to focus now on the Asian elephant work. This was actually one of the main projects that we first started working with the lab on and we’ve had various iterations, various different focuses for this project, I’m going to focus just on one of the most recent

31:05

iterations of the project work, which is focused on population monitoring of some of the agent within the country. And the reason that we’re focusing on Asian elephants is that it’s classified as endangered, and it’s undergone huge declines in the last few years to be classified as endangered, it’s gone, it’s undergone a 50% decline in the last 60 years, the massive decline and as you can see from the map on the right hand side, in the historic rain, light red, and the current rain. So you can see, you can see there’s been a huge contraction in the range of Asian elephants. And there’s actually sort of three subspecies of have gone extinct. So this decline has been happening for a really long period of time, perhaps over 1000 years, leading to, to the extinction on the range edges of this species. And as you can see in Cambodia, so and it’s led to habitat fragmentation for this species that I’ve seen, see, the dark red is, is separated into lots of separate fragments now, and that, again, from a genetic diversity point of view, that’s, that’s not great, because you can’t have the exchange of genetics between those populations very easy anymore, and it can lead to inbreeding and other genetic diversity decline. And as you can see, in Cambodia, there’s been huge declines as well. So there’s population just really remaining at the edge of the country. But I’ll just zoom in on Cambodia and and show you some of the work that we’ve been working on. This is a map of Cambodia, we have is thought to be between 400 – 600 Elephants remaining in the country. But this is a really rough estimate. It’s a lot of it’s based on very anecdotal evidence. The Asian elephants, although they’re the biggest land mammal in Asia, they’re really, really elusive, they’re really hard to find and to count on to estimate population sizes. From this, they live in really dense jungle. And so you can be really near near them, but you can’t see them or count them, or, or survey them in using traditional techniques. And the two main areas that they’re thought to remain within IS THE EASTERN PLAINS landscape in the east of the country and the cardamom mountains. So the picture that I showed you where the Siamese crocodiles are found earlier as well. These are basically two Highland regions within the country, where deforestation has not been as extensive as in much of the rest of the country. And more recently, we’ve also been focusing on this northern landscape. So it’s called the prey long, extended landscape. And in all three of these locations, there is elephant signs, and there’s anecdotal evidence of elephants. But there’s no real, nationwide robust estimate of population sizes. So that’s what we’ve been working with the lab in Cambodia to do is to basically help inform the government and the NGOs that are working in the country, how many elephants we need to detect areas. And we can use, we can do that using genetic because instead of counting and finding the elephants, we can get genetic signatures from Donald that the elephants leave behind. And then those unique genetic fingerprints can be used to count up the number of individuals that are found within protected areas. This is the prey long, extended landscape and satellite image that I’ve overlaid on here just to give you an indication of where the forest is still found within this region. And again, this hasn’t been studied from from an elephant perspective in the same way that Eastern Plains in the cardroom mountains have been previously. There’s unknown number of wild elephants remaining within that region. We have been involved in Project previously in the other two regions to help estimate population size. So this is zooming of that satellite image. And the study I’m going to talk about, again, is really close collaboration with forum for international focus, focusing conservation efforts can be pretty long area. And in 2021, they collected elephant dung samples from three different protected areas within the region after doing surveys to, to basically survey for any kind of indirect elephants signs within the region, so they also surveys. So it’s another area, as you can see on the left of this map that they didn’t find any signs in. And so the collection was only focused on these projects. Yeah, and so the field teams of FFI were out in the field collecting swabs, or genetic swabs from these drum samples that were then sent back to the lab. They did six different survey expeditions over about a six month period to collect hundreds of elephant dung samples in the region. Yeah, and these were transported to the lab, where again, we transferred techniques to study and have been transferring techniques for various projects. Since 2016, this is just the most recent training that we’ve done. And again, we have used two different techniques in this project. So one was similar to the crocodiles, and that we were using snips. That’s to get an individual ID, or what you can think of as a genetic fingerprint of each single elephant that has produced the dung sample. And obviously, you can collect on samples that have been produced by the same elephant. But if we can individually ID each of those, we can tell which ones have been produced by by each elephant and count up how many elephants we have within the environment, we also ran a genetic sex test. Again, this is really important for the conservation of all conservation planning for elephants, in particular, because the sex ratio can have a really big impact on breeding productivity and the long term prospects of a population. So we ran a jack sexing tests, we could identify the sex ratios within each of these areas. And the teams in the field also look at dung size and bolster confidence to get an idea of the ages of the elephant as well, or whether there’s any young juvenile elephants in the population is, is breeding successfully. And so it’s multi faceted approach that we’re using to buy and build up, get information to build a national Elephant Conservation plan. So I’m just going to show you there’s obviously a lot of genetic work involved in getting those genetic fingerprints and running those tests. Second Test, I’m just going to show you the results of how many genetic fingerprints we we found and the estimates are the population size areas. We found 20 elephants in the northern wildlife sanctuaries and 31 elephants in the prey long Wildlife Sanctuary. That is a really, really low numbers of, of elephants compared to the other two regions in Cambodia that we’ve been looking at helping with previously. The reason that I’m giving you both of the figures separately is because we didn’t find any evidence of these elephants moving between the prey long wildlife sanctuary and the prey rocket and Chip wildlife in the north, over there was evidence of moving between those two northern sanctuaries, though it’s likely that there is some fragmentation occurring here. And there’s limited movement of elephants, potentially, between these two areas. So we analyse those, both separately. And we’re also we also produced diversity data as well for both of these elephant groups within this environment. And that’s now being stared into a bigger picture of elephants nationwide across Cambodia, looking at gender diversity between these three different areas where elephants still remain. So as well as that we’ve also been focusing on getting the database together, compare ivory samples to as well. But this is the main monitoring work that we’ve been doing recently. And we’re just about to start a project in the southern part of the eastern plains as well to fill in a gap in some of the population work. So that’s a brief introduction to the Cambodia projects. Obviously, we have much wider impact at RZSS wild genes. Since 2010. We’ve been working on over 70 different species. And in our biobank, we have data from over 300 species that’s being fed into current programmes, but also, it’s there for researchers around the world to use for conservation questions in the future. And if you do want to find out any more information about our projects, you can obviously go online to our website, we have lots of updates about our various projects. We also have some YouTube videos as well that delve into a few of our projects in a bit more detail. So the links are on the slide here, if you want to go and have a bit more of a look at those. And that just leaves me to thank everyone for listening today. And to thank all of our partners on conservation charity projects. So conservation is a huge collaborative project and vision and you need input from so many different players and stakeholders. And obviously, in the projects that I’ve mentioned today, the key ones were fauna, and flora International, the Ministry of Environment in Cambodia and the whole university of content. So I want to thank all of the staff and partners involved in those projects. Yeah, and I think we’re going we’ve got time for questions.

41:10

Great.

41:11

Thank you very much, Alex, for that really informative overview of the work that the lab is conducting in Cambodia. I was wondering, do you want to just stop sharing your screen so that we can go to full screen mode? And I can see that there’s questions coming in in the chat already, which is great. And encourage you to post any questions that you have in the in the chat. And we can we can answer them as we go. But we already got a question here from Roger. Which is really good question. I suppose that crocodile hybridization in the wild is just part of natural selection and evolution. So why avoid it in captivity?

41:56

Yeah, no, that’s a really good question. So there is there is some evidence of some that introgression so basically, that’s another term for hybridization between saltwater and Siamese crocodiles in the past, so there is that remnant of, of introgression of migration that has occurred in that species in the wild, hundreds of 1000s of years ago. And so it is something that happens, and it is a big component within that it does lead to potentially new species occurring as well. It is a natural process that occurs in the wild. However, these who the Siamese and salt water have been separated, and don’t commonly hybridise in, in, in the wild, especially now, when they they don’t overlap in their ranges at all. So the only found in the kind of mountains of saltwater crocodile would would not be found in the cardamom mountains. And also about the different behaviours between these these two species. And the different environments that they’re found in it can potentially lead to us release in very aggressive large crocodiles into very small freshwater streams, if we didn’t screen out the saltwater hybrids from using these this genetic process. And I did I use the word purposefully hybridised in captivity as well, because there’s there’ll be very different levels of hybridization both in the wild and in captivity. And in in the leather farms, they purposely hybridise them to try and improve the leather quality basically use the different aspects that they want from each of the crocodiles to produce different kinds of leather. And so there’s much higher amounts of hybridization within these farms than there would be found in wild. So hopefully that answers your your question, but it’s a very good one. Great, thanks,

43:56

Alex. I’m just wondering if you could tell us a bit more about the challenges of doing this sort of work? And why why do it in Cambodia and not not in the lab here in Scotland?

44:14

Yeah, no. So there’s a yeah, there are a huge number of challenges involved in international projects of this, but one of the reasons we are we do it in Cambodia is because we really the only time the only way you’re going to get long term conservation, by an in progress is if you provide the tools to the people that are living alongside these species. And so providing them with the tools they can then apply to the species within their within their country, and also to other species that may become threatened or need, need conservation don’t work in the future. So it’s a really big part of our approach. And we don’t just do it in Cambodia. We’ve done it in other countries as well. And one of the reasons that we focus on Cambodia is because it is a really under low economic country. So it has very few resources of its own to put into into projects like this. So we can help develop techniques. But then we take them over there because also there’s lots of difficulties with move in genetic samples around the world as well. So having them not having to travel halfway across the world for this work is a great bonus. Great, thanks,

45:29

Alex. Just wondering is well, if there’s any parallels to the work that you’re doing in Cambodia to other projects here in Scotland?

45:45

Yeah, so there’s a project that people might be familiar with. Within within here is the Wildcat breeding and release project that’s currently undergoing in the Cairngorms National Park. So that just occurred on the festival. Yeah. So the first releases are going to be occurring this year, for that project. And there’s been a hybridization screen that has happened in Wildcats as well, because of hybridization that has occurred between feral and domestic cats and European wild cat. So feral domestic cats are originated from the Near East and not African Wildcat. And there’s been a lot of hybridization between those and the European Wildcat in Scotland. And so that is really a parallel between the Siamese crocodile work we’re doing. And we’re using very similar tools to look at the hybridization question both of those projects as well.

46:45

Great, thanks, Alex. And another question from Mary, do you feel that in Cambodia, people, local people are supportive of your projects? And maybe whether you’ve got projects, whether you’re doing this sort of work in other places as well. So

47:09

yeah, one of the really great things about especially about the Siamese crocodile project in Cambodia, is that one of the reasons that that species is still in the wild with us is because of the local community there. So the local community have safeguarded the crocodile species in the Kado mountain. So the indigenous people of that region basically, since saw them as sacred, and did their own patrols to try and protect this crocodile even before we, the Western world, and scientists knew that it was still remaining there. And that’s. So I think there’s a lot of buy in from the local communities from this project, which has been really great. And I think FFI and  in Cambodia will probably be able to answer this question a lot better, because he works on the ground with those with those communities all the time as part of the project. I work with the University, which does it caters to students from all over the country. And there’s just there’s a huge thirst for knowledge within the university, which is been really great to see. And so yeah, I think we get in the excitement about this novel technology can also get people on board with conservation and feeds into those conservation messages as well. Because obviously, it’s just it’s really nice to see how focused and dedicated people are to improve in the national situation in Cambodia. And they want to just they want a load of tools and different ways of trying to improve that improve that national situation. So that’s been a really great video. Great to be involved with local people.

48:52

Great, thanks, Alex.

48:55

Just checked it ticking another question. Many critically endangered species exist in tiny, often fragmented populations. So loss of genetic diversity must be a very wide problem, sort of more generally, how can it be mitigated?

49:14

Yeah, is is? Yeah, it’s a huge problem. And it’s it’s a common theme of, of the species that we work with, because they are, they are small, just by definition, they’re threatened, they have declined, and they’re often in small fragmented populations completely right. And we don’t know, all of the long term impacts that this loss of genetic diversity is going to cause in each of these separate species. But we know that inbreeding is a massive problem. And that lack of genetic diversity could mean that we don’t have all that that species doesn’t have the ability to adapt as well as if it had more diversity. So one of the ways that we can make To get that, or the most obvious one is not to let the declines happen in the first place. And that will retain the diversity is also really important to move or translate K individuals between populations, if it’s if they can no longer move naturally. And that helps to sustain like, you can think of it as like a metapopulation. that’s larger than the sum of each of the parts, and how you can transfer genetic diversity that may only be found in one of those populations, to those other populations and help safeguard it into the future as like a very general general, general way of trying to conserve. Great,

50:44

thank you, Alex, I think that might be all of the Oh, there’s another question that’s come up in the chat. From Ian Lewis, with respect to translocation of individuals, are you also creating a stomach?

51:03

Yeah, that’s a really good question. So we currently don’t have a studbook of the wild of the wild crocodiles or the releases, but we could create one using using genetics. And that’s actually a focus of attention. The next phase of our Siamese crocodile work is to try and work out which individuals are surviving in the wild post reintroduction, and basically by creating individual genetic fingerprints of all of the individuals that like we have for the elephants do it for the crocodiles, and that can build up not a studbook but like a relatedness network that we can then use in this scenario. It is something that we are looking into for other species as well that we work on that have been warped potentially been brought into captivity from wild populations, and to keep really accurate scrapbooks of them, which is really important from a conservation perspective. And this is again, one of the great things about being part of the rosewood, logical scientists, Scotland is that that’s routinely done for all of the European captive populations that are kept. And that can allow us to mitigate loss of genetic diversity by making sure we have found a representation being maintained through generations within the captive populations.

52:28

Great.

52:29

Well, thank you very much, Alex, on behalf of the Royal Society of Edinburgh investigates, conservation series very pleased to have you speaking about conservation genetic issues in Cambodia, and really an example of how genetic technology can be used to help with protecting the genetic diversity for threatened species.