 presented an abstract for a contributed talk and please go on, he's speaking about Plasmids in Qatar, so very fascinating, please go on, you have 15 minutes. Yes. Okay. Good afternoon so thank you I would like to thank the organizer for giving me the opportunity to, to give a short presentation of my work. So the title is about Capsula genomic epidemiology. So about some of the data set we generated in Qatar. So recently I have relocated from Qatar to Singapore so now is the midnight here in the South East Asia. So I'm going to talk about some of the results study generated by some of the junior physician when I was working in Qatar in the city in Qatar. At the moment I joined Tantosing hospital and I am the principal scientific officer in the National Center for Investors disease, but the data presented is related to my work previous work in Qatar. So, before presenting the story, I, all this data. I would want to add more natural the scientists and physician and so the, or the sum of the collaborators involved in the data discussions from Hamad Medical Corporation, University of Pittsburgh and also University of Melbourne. Thank you for cats and also Kelly's discussion and suggestion. The story began with a small molecular epidemiology study to look at the general the molecular epidemiology of Kappa panamase producing in the 1980s. Some of the study, there's a, there was study published last year on the epidemiology of the CRE circulating in Qatar so based on the statistics we learned that Capsula pneumonia was the most prevalent or most common material and carrying the Kappa panamase in Qatar. And so you can see from this table indicating based on like 150 isolate recollected almost 80 of the eyes that belong to Capsula. And then also for the age and also the MDMs are the most common Kappa panamase circulating in the country. So, and KPC is very aware in that part of the world in the Middle East region. So we decided to look into the genetic context of all these Kappa panamase producing Capsula. So the idea just trying to look at the to do a whole genome sequencing of the Kappa panamase producing Capsula. We cover from all these clinical samples basically from blood from urine and different content from the patients, and then we're trying to understand the mechanisms and how. Yeah, just how these particular become resistant to Kappa panamase. So, yeah, we did whole genome sequencing so and this is a very general protocol so I think it's pretty similar I think pretty standardized among different parts of the world. So we use in the minimum next seed and then we use a standard types pipeline just to look at the to assemble the genome and then to call the variants and then to, to search database for all the ML genes and also we look for the fibrillance genes as well using a software developed by cat host group in Melbourne. Now he's in UK. She's in UK. So this is one of the summary diagram just to, to illustrate the epidemiology of all genomic epidemiology of all these capsules isolate mostly capsula pneumoniae and also capsula quasi pneumoniae. So this tree just shows you the composition the presence or absence of AMR genes in blue and also the presence of absence of fibrillance genes. So you can see from this tree. The most prevalent ST types straight sequence type in capsula ST 147 and also ST 231 and capsula quasi pneumonia is very homogeneous so there's two dominant sequence type ST 196 and 1416. I'm really interested in the ST 383 because all these isolate we have only four eyes that belongs to ST 383, but they both, they carry both and DM and also for the eight, and at the same time in terms of the presence of absence of fibrillance genes. Isolate belong to ST 383 they also carry the RMP a and also the aerobatic which is very important we will inspect us in terms of forming mucous and causing infections in capsula. So, we decided to go deeper to look at the the genomic composition of all these ST 383. So then we pick up one of the isolate belonging to ST 383. And then we did a sequence again using short we and also long read by me and we did hybrid assembly. So we find the sample contain one chromosome DNA and plus five plus mid including an INC H I one B and also an I am a con L. So what's interesting is, we find that the end DM five gene plus eight other major CT exam 15 and also nine day and extracts are they are located on the first past media, the H I one B, and then the also for the eight plus the same CDM CD exam 14 they're located on the INC L plasma. And then we did a blast search of all these past these two major past me. And then we find it is highly similar to another ST 383 reported in UK just like last year. So, one of the, the first, the largest plus me, it is like a hybrid plus me it contain both AMR gene and also the wavelength genes. So this is just a diagram to show the genomic comparison between the, the plus mid of fq 61 and also the plasma belong to isolate isolated in UK. So you can see there's lots of concert region. So basically, we can do that I said the, the, the, the plus mid into three major region so I call it the AMR one region which basically contain both of the AMR genes, such as MDM five and also, and see the exam 15 and for in this AMR one region is basically over 99.9% identical between our plasmid with to the UK plasmid in that particular region. And also in terms of the difference region also is almost over 90% identical in terms of sequence identity as you can see, even the arrangement of the difference genes in purple there. Also in Cincinnati with each other. So the major differences between our plasmid with the UK, it's only in the what we call the AMR two region which contain around six, seven AMR genes that there's some sort of recombination and rearrangement happening here, just that is fair or is there some sort of rearrangement between when we do the comparison is very office. So, then, we kind of know that, like, our, our isolate, our kapsella STV 83 is the first plasmid the biggest plasmid is very similar to the one actually reported from UK. Well, and also we did the comparison to of the second largest plasmid of our eyes of the two to the UK to the one from UK is almost 90 over 99.9% 6% identical between the two so I would say also was the same that's only like a couple of mutations between between among the two plasmid. And then on this plasmid they carry the Oxford 48 and also the CD exam 14 gene plus like two additional AMR genes. So, and this plasmid is like carrying the Oxford 48. It was first reported in Australia around 2013 but it's like if you look at when we did last search like the multiple hits, and then this plasmid actually was carried by different ST types of kapsella pneumonia and also E coli and also Proteus. So it's been widely circulated in different countries and since I think since 2013. What's interesting is what we're trying to look at all what we're trying to compare the, the available genomes of all the ST 383 in the gene bank and trying to look at the evolution of this. ST type of capsula pneumonia over time and then we discover something very interesting so most of the caps, most of the apps ST 383 they were first reported from Greece and mostly they carry beam, also CM or KPC. And, or CMY. This kind of beta letter maze, it's only until recently that's this ST type pick up different plasmid so if you look at the tree based on similarity I mean this is tree generated just by mesh is based on genome percentage similarities not based on evolution just the distance matrix. So, but just look at how similar they are. I mean, we have four isolate from Qatar belonging to ST 383 and then there was one isolate from England. And also, in terms of the riverlands genes and also the AMR genes, all these sides of the reason I saw it in addition to two Lebanon isolates, they both carry the OXA and also the OXA 48. And also they, because of the presence of the hybrid plasmid, which in addition to MDM and OXA they also carry some of the real and genes like IMP which is make it could be potentially considered to be a hyper villain capsula. So it's in addition to multi drug resistant also is hyper villain so let's make this ST 383 kind of interesting based on the comparison of all this data. So after all the comparison, and we are going to compare more because I think there are lots of reason in the gene bank that we can create and then trying to see maybe there's a bigger connection of ST 383 in the gene bank and then trying to look for the evolution of all these sequence type. So this is a small scenario so I think based on the literature we know that the ST 383 was reported in Greece the first time in Greece in 2009 and then it have a VM and KPC and CMY. But then there was in, I think since 2015. This is a report from Germany. There's, there's the ST 383 pick up OXA 48 plasmid and it's been circling around, and there are lots of eyes that have actually reported in China, also containing OXA 48 belonging to ST 383. So basically in the Middle East region in Lebanon in UK and also I think there's an informal reports from UAE in Dubai also having ST 383 bearing to the OXA 48 plasmid. It also bears that hybrid plasmid contain those AMR genes and also virulent genes. So I think it's worrying the fact that like there's a spread like there's an acquisition of type of panemase producing bearing plasmid and also the plasmid with hybrid containing multiple hyper virulent genes. So this is one of the interesting findings from our comparison. Another just trying to an ongoing studies because we're trying to what we are looking at is the desolate the the Capsula quasi-pneumony isolate in Qatar is kind of very conserved and then we look at them and in particular the ST 196 they are actually connected from different hospitals and but they are genetically almost identical in terms of SNPs and also in terms of the composition of the AMR genes. So we kind of believe that there's like a non-socomial transmission among the ST 196 Capsula quasi-pneumony within Qatar within this country. So in literature and also from the Jinban database we knew that like ST 196 was has been reported in UK and Denmark only there's only a couple of sequences or assembled genomes in the Jinban and they mostly contain carry KPC so somehow some of these isolate when they came to the middle is they somehow pick up the NDM bearing plasmid and then being circulating around within the country. Yeah, and this is one because we were trying to do the long the long re sequence but based on the sort with assembly we know that one of the content containing all these four AMR genes. They are, we can find context of highly similar context also in other Capsula pneumonia as well so there's might be some sort of fragmented evidence suggesting that these the NDM bearing plasmid in Capsula quasi-pneumony and Capsula quasi-pneumony also being circulated to or acquired by other Capsula pneumonia species. So I think this is the summary of my talk so we based on some epidemiology studies we know that Oxford 48 and NDM enzymes are prevalent in Qatar. Based on when we try to dig in and look at concentrate on one ST383 we find that we carry a hybrid plasmid with NDM and multiple surveillance genes and also it has a second plasmid bearing also for the AIDS and then I think just based on the evolution of ST383 there's some sort of evolution from long species to become hyper-vivalent on the pathway to become hyper-vivalent just by picking up acquisition of this kind of plasmid. So, and yeah, the final note is the ST196 is circulating, it seems to be like an endemic cone in the Middle East region. And thank you, that's the end of my talk. So, thank you Clement. It was a really interesting data set. Unfortunately, we have no time for questions. If you have any questions for Clement, please write them in the chat and discuss them. I mean, the connection will be left open so you can discuss while the people can take a five minute break please because then there is Yana and then there is Olivia and we are running terribly late. So, it's a break. Thanks Clement, it was really interesting. Thanks a lot. How long the break? It's probably five minutes. Okay, okay. Well, let's say 25, let's say we restart. Okay, seven minutes. Seven minutes. Okay, thank you Alice. You're very kind. Sorry. I show you the CISO. It's very nice. Similar to yesterday, sunny and everything. One thing Alice, for your this talk, this introduction, it was very nice. I liked very much this example at the beginning and everything. But next time, for next time, you should think about how to measure. The importance of point mutations versus recombination versus horizontal gene transfer. You have many, you have, you have many modellers around you that can. But this is one of the reasons why I organize this because you need also data sets because to measure the importance you cannot, I mean, probably because I come from physics now, but you cannot measure the importance if you don't have data. So you need to have data in which there is both point mutations and structural variation, let's call them, and then trying to match them. I mean, one thing that I'm missing and that I wanted to ask was, if somebody, probably you or Cornelia, who are wisest around, can suggest me on how these things happen on the mechanism presiding. Because like point mutation is something intrinsically random and intrinsically Poissonian because there is the, the polymerase and the polymerase makes a mistake because it gets the wrong one. And this is intrinsically Poissonian, while I have no understanding of how Plasmid, I mean, the talk, Adam's talk was really interesting on this because he probably can suggest me something because he too, I should stop the recording, we don't need to record this.