Iowa Swine Day 2017 Pablo Pineyro, Update on viruses: Seneca, PED, PRRS
Okay, so our next, speaker this, afternoon is. Dr. Pablo. Pineiro and he's, an assistant professor in the department of veterinary, diagnostic and. Production animal medicine at Iowa State University. He's. Also a diagnostic. Pathologist. At ISU, diagnostic. Lab and his research focuses on, infectious. Diseases, of swine and their associated, pathophysiology, and, also, Diagnostics, this, afternoon, he's going to talk about an update on a few, viruses and, swine production so. Join, me in welcoming dr., Panero. All. Right give. You few seconds of people coming vielen out. Okay. Thank you for staying, here this afternoon. I, work. In the the, junior technology lab here in Iowa, most, of you probably know we. Are one, of the busiest, diagnostic. Lab. Associated. With fun animals in the United States so, we handle. About seven. Five, cases. 25,000. Cases a year and about. 80%, of dinner pigs so, we'll get to see quite, a bit of diseases. Going through our, daily. Work so today I'm going to just, touch bases in, some of the things that we currently see in these three the system here PDB, first, and, cynical. Eggs. So. First we're, going. To kind of review, what happened with PDB during the last. Few. Years and show. You what, we are now, so. Just, a reminder. What. Is pretty. Pdb, the virus, that belonged to these families the corona virus coronary died and. I'm. Going to just show you three four things that we're going to use later on during, the talk. For. The diagnostic, side. So the structure of the virus is. This thing working. Okay, the, structure of the virus is the the corner wires mean that they have kind, of a crown, around, and that is the typical morphology, of the family so the virus itself has operating. Frames so this opera frames are going to qualify for a structure and knowledge tubular proteins and the one go we're, interested, are these for the, spy probe the membrane envelope a nuclear protein those are the structural, ones and I, won't probably, remember, those because we're going to use them. When. We talk about diagnostic. So. All this. Coronavirus. Family is. Dividing for different genus, Alpha Gamma virus, beta coronavirus. Gamma and Delta, and it. Seems like all of they have like a common ancestor, and sister that came from, a bad corona virus the Sham to the birth and they were spread to the different mammalian, species but what is important, for us which, ones are the one that affect pigs so, pigs, get affected with multiple, viruses within, this big, family.
So. If, the a. During. A, alpha. Corner by Reshiram we have three, viruses, that effectors, the pdb percent. Pyramid diarrhea virus the, percentage, material gastroenteritis CG, that we don't see that often anymore and percent respiratory, corner virus which is kind of the mutation, of the TG, so although threes effective. Then. The better coronavirus, percent, hemagglutinin. And settle on oil mal, passive we don't see that quite often but. Also. Another disease, that can be caused by coronavirus, intakes and finally. There's a corona virus so you can see we have three. Genes represented, affecting, the. Soil. Population. But. I, like. To kind of reorganize, this does this within. The genome. Species. But I like to see where, these, viruses, and what is the clinical presentation that we have I like to organize this in, this way so enteric viruses, within these in this family for signing the malaria T, G and Delta corona virus those are the ones that are gonna cause diarrhea. In neonatal. Team respiratory. For, some respiratory corner, virus, the. Clinical. Presentation is, not as severe, as any of the other respiratory pathogens that we know but, I still and cost see and this, one that I mentioned before the, P HEV basically. Is the CNN. Virus it cause necrosis of, the neurons and but, when we see it's a gutter enteric, clinical, presentation, basically all the my enteric places get paralyzed and those pigs vomit, a lot so I used to be bombing the, deceased, used to be called vomit and wasting disease so, but, it's, not because the virus subtract the GI tract is because it costs a neurological, problem. So. What is the situation of PDB that's, the. Topic. Here so as you can see is that kind. Of runs, on it's like here and this one tried to represent where we are now so with, the epidemiology. Diagnostic. Tools and the generic of the virus. So. Just brief a brief. History. This rest of you having in the, last few years so, back in 2013. Actually. I was working in Virginia. Way. Far from the Midwest and we got this thing actually that picture of still with my cell phone there and, we got this, the RIA with, this thin. Wall. Intestines, just loosen, or fill by gas, kind. Of atrophic, into riotous we ran the common pathogens that we knew Tajik or no viral, rotavirus. And all of them were negative so, we. Sell the sample for. Sequencing. And we found this. Virus. Close related, with the, PD week that had been reported in child at, the same time here in Iowa, people. In the diagnostic, lab they got about the same picture if you look at interesting. Kind of look the same this just loosen released in world in testing and the, collage occult picture is the same so, and they, came to the same conclusion, they did a sequence, and they, did, the first diagnosis, of PCB so, this is how everything came up but, here it's kind of a tie lab how, things. Progress so, the first cases were reported. Late. April early May in 2013. It's. Illogical II again, we see these. Atrophic. In terrace that, resemble. Like any other, enteric. Virus. Position, so, this year rotavirus could be but by, PCR they were both were made. Remember. That I took about this, coronavirus. Shelter so we did a little microscopy and, here we are we got the first evidence, that divider was affecting. Pigs. We, ran a pen coronavirus, PCR remember that I mentioned all these genomes, the Alpha Delta, so this tell us that is the corona virus doesn't tell us which one it is within, the genus but still telling that is the corona virus so we were in getting, close with, a full, sequence. In. Conjunction with NBS, cell and. We. Found that it was the closer mala G with a string, of PDB that was reported in China in 2012, so. By, mid-may. Not, even a month later use they came with the first official report of PDV in the United States and. About. A month later we, already have a PCR, in place to the ignores remember that I mentioned those genes at the beginning of the talk so our, PCR was biased, in the spike sheet that. Is what was important, the anatomy, of the of the buyers so.
But This as you can see it's just literally, more progression, up to this just. The, wheels came off and everything, was. A disaster, from there we, got this massive diarrhea, the pig died and every day Ritchie hundred percent mortality in, most of the farms. And. Our. Lab in the lab went really really, crazy but from. Both, first cases in, 2013. What, happened today what is the situation today, so. Use. The AML with a weekly update of PDB, cases and look at the. The. Pig. Produces this Iowa, Minnesota North. Caroline cancer. So in Iowa, we, have 1300. Premises. Reported. In the last three years if we think about, first. That can reach hundred, percent mortality think. About how many pigs we have lost in the last three years 13 premises, with hundred percent mortality that's. A big, number another, thing that we learn from the terminology, is just. Three years data, so, look, at this this is a weekly report we. Have a seasonal, pattern. So. This is is affecting. More. The winter, and give. Is giving us a break during the summertime, perhaps. The, virus can just received, in home where and that's where we get a break but it doesn't go away even when the scene that is clearing. Up during summer it doesn't go away cases. Came, back again and we have a river exit of pdb every year. So. That is kind of Epidemiology. And the situation, the current situation I want to show you probably. What it was the first experimental, a reproduction. Of the disease that was done she an Iowa State so. CDC. Peaks, sauce, salad, arrived colors to deprive they don't have any immunity at all they don't have any protection against, this virus the, word effected or. Graphically. With already low dose 10. To the 3 and. We. Took sample for a sickle, soft within, the crop the time course the cross he really. Sure because the duration. Of the clinical Sciences is really short no more than 3 4 days and, we need crop t and evaluate what region of the GI is most affected. So. To our force, infections 30 percent of the animals were already, showing clinical signs they already have diarrhea, dehydration.
And. By 24. Over. 72, hours hundred, percent of them were affected, so vomiting. Diarrhea, and we start seeing mortality, there so this is probably the first time someone. Really got the virus putting, the pig and reproduce, the disease. Here. Shut for compared, I know I already shoka this this picture we caught before, but this how a normal, gut will look like a really sick world we, can see the, base of the epidemic nestlé's compared, with this one is. Really thing just loosen, almost. Empty so, that is what. We normally see and a PD. Be affected I'll you how. This look is too logical is how we start, making the diagnosis, and this, is going to also help us to understand the Botticelli's, it if you think this is a picture of a normal, interesting. Think, about the normal entity like fingers. So, the, piglet, need all that of faith to, absorb, the nutrients if it, doesn't, forget affected, we start getting reduced on top of diarrhea the reduction, of the surface, that has gone observe observe there. So, keep. In mind this one, dividers. Can affect, the. Tip of the D light here and the, creek so these green cells, are affected. Cells so they'll integrate on top, of the B line. So. Here's where we start getting the closest of the intestinal. Mucosa again, the brown cells, affected. Cells all the way down to the crib and up, to the top of the data and if you compare this one we normally say that the cell T got to, have a ratio of, seven. To, one seven, long, the, be like comparable well to the crib here. They are getting really really short we are still losing that absorbed, the surface, of the guy. Finally. We have what we call atrophic so, if you compare the length of this one this is a very effective one with, the normal add so we don't have pretty much suffice, to get any absorb even if this pick survived then, I'm going to have a health gut to absorb any, nutrient, at all so we had the UH trophy, fusion, and blunting, so, the, detecting, tried to repair, the damage that the virus caused but. It's. Not going to be really effective and never going to get back to this probably the additional length that.
Is Kind of the progression of the disease and the pathogen see how things, happen, so. The clinical side that, we see in natural infected animal we mentioned that before diarrhea. And vomiting and. It's going to vary a little bit based on the age of the taste the younger the get infected up to a week is really severe if we go about two weeks we get the clinical science arrest less severe the same thing happen with mortality about. 100 percent in animals less than we, call more than ten days it, could be around, thirty forty percent the pain of the immunity of the farm. So. The, time of incubation can go from 24 hours up to four days and, the. Clinical science sudden, as we saw in the experimental, part is show really, really really fast and the other thing is we had in a far the virus, spread really, quick so that is why we had have, these high mortality affecting, pretty much every, single day this. Is just a picture of how infected. Cry look like so just one pic trying to eat, these are really lethargic, degraded. Or, covered by theses and this is what we normally going to see in, a visit to a farm affected, by PD be a really, high mortality with. Mostly, dehydrated. Pigs and covered, by pieces. What. They are knotted to would have if you remember, during, my time, curve presentation, but, back in the day we only have PCR that was and the morphology that was compatible with other diseases as well so now we have a more. Complete boot. Toolbox. For, the diagnosis, so we have PCR, as I mentioned already, we have a moose - Kimmy tree this is a bunch of immature picture so that is a direct method is going to show the virus in the gut so there's, no question about that so, sorry serological, tests, so indirect method to, detect the presence of antibody, if the the, pigs were in contact but doesn't tell us the presence of the dial so, IFA Elisha bias, neutralization. Those are probably the most common use. Technique. So, I put, here no virus isolation, it no because it no can be done actually it can be done it'd probably take time and when, we have an average like this we need an answer right away we can't wait for a week until the values grow and, we see the psychopathic, we can waive that so that is why we don't use that one as a common diagnostic tool and bio, say that.
Was Probably one, of the common, terms that we use at the beginning we're getting fixed material, we put the body back in Pig and if the pig reproducing, clinical sign that was our confirmation. Of the of the disease now, we have our tool to to, make, the diagnosis, so. Kind of the take-home. For this part of the talk so. It, causes severe malabsorption. See. Diarrhea remember, that I mentioned they don't have nor, be, like to do the absorption so that is animal absoultely diarrhea. So it's acute summation, in few hours will have the disease, so that is why halide, acute and high mortality, those are the main key feature, of the griga presentations a very atrophic entirely I show, you the slides how it's going to look like and I can go through. Affecting. Just one portion at the beginning of the disease duodenum, and did, you know the cranial Portia and can, get, a spread throughout the, whole small intestine, as the disease progress. Probably. The most important, thing about the disease these, are all this petition, and how I look like is economic, impact and as, what's mentioned in the previous talk all these diseases, they have have. A face and we have to remember that each one of these, farms that were affected. It's. Not only the loss of the pig there are other implications, and economic implication, is huge if we look at 300, millions a year shut by for PDB it's. An enormous loss but. Still. More compared with this so, as. You mentioned before this is still really really, important person till probably been, our most. Important. Economical. Disease but, is why the. Next part of the talk. Is going to be about purse what. We see in the diagnostic, lab and, what is required, from us of diagnosticians. This. Is just kind of the summary of, respiratory. Cases. As we know I'm going to show two pictures later, respiratory. Purse. Came at different, trails so respiratory, disease reproductive. Disease gonna, focus mainly respiratory. Problem so this is a percentage, of cases but different pathogens by, itself I don't even try.
To Touch bases in in co-infection, this is a purely infection, so per, still being probably the most important, 40 percent of cases, on a 40 percent of cases are associated with first followed, by SIV, mycoplasma, pneumoniae. Patella. Muto see another bacterias, and so on but. 40%. What. Does meaning numbers real number so this is just total. Case numbers, respiratory. Cases diagnosed it's only with birth nothing, else there's no infection, and, we're talking about an average of 2,000, cases a year, only. From Iowa. Industry. This. Is what I was mentioning before the, different clinical presentation, so abortions. Respiratory. Disease and, increased. Mortality in. Nursery, and grower. Pigs. What. Is the current, status, hopefully. We're, going to get the, new code pick zoom by with the stick up to their widdy's so this is how we borrow the receipt, we have vaccines. In. Our market, we deal, with this with Melissa, I live attenuated vaccine and as, was mentioned the previous talk this vaccine. Help, but, I think prevented. Totally the disease this, was in still getting animal sick so. Depend. Of of, the of the. Status. Of the herd could be just, a mild transitory, disease and we can see actually severe. Are works only just for, the present of the vaccine in Europe, they circuit, they, use the kill vaccine the project without is that the immunogenicity is. Not even comparable with with a modified life so the protection is, not. Enough to prevent an outbreak of the disease there, are other experimental, vaccines, are there by, the vector, DNA subunit. Vaccines all of them to be proved to. Be, thickest. II have, some efficacy somehow, but they, don't they don't. Work. As well as the most well live and unfortunately sport is the only tool beside, management, that we have to prevent the disease now. So. Diagnosis, this is what we do in the lab every day so, believe. Me we still rely on this we. Need someone to tell us that we have a respiratory problem if you send us just a piece of language tested. For. You. Know pathogen. We, can't help sometimes, we need a little description, persons, are animal, affected what. Is the clinical presentation respiratory. Means. A lot of thing means coughing, means distress, misses, me thing so when. We get our samples, we need a little more help to, give you a better answer so, in, this case we have the the. Typical blue here that, is. How that is used to be Kobe for whistling, Michelle cyanosis. They can breathe and they cannot, see sinead and start getting purple so this is probably the first clinical indication. Of respiratory problem, caused by my, purse then, when we open up the lungs, the, pig we see these these, larger, fail to collapse they are heavy in ways that, is kind of. Indication. Of a viral, infection it doesn't tell us that is purse still, a virus. Infection PCB can also cause kind of the same type of pattern so, still. Shell but is not that the, field. Which. Is what, I normally do is wear a doll, and pathology. So this is what we, look for in, a slide this is our tee shot pneumonia because it's thick ideally, this will be just one, single, rule a liar, yourself and here we have really sick alveoli this is will be the, air. Space space, so if, we look at the next picture there's no air space so it's, gone they don't, have space, able to breathe and the, confirmatory, probably for us is this one this is the molester chemistry, and the brown dot in there are.
Infected. Cells with parts so they're infected macrophages, with cars so that is kind of the easy, and, fast, way to approach. The, diagnosis. But. We have more. Tools so, we, have virus isolation. We. Can do sequencing, out of the virus isolation, that help us to confirm the diagnosis, of, course. PCR. And. We. Don't only, use a. Regular. Example, like tissues. And serum, we also use oral. Fluids we need to not only do. The final diagnosis, of a respiratory problem, but we need to do. Epidemiological, evaluation. We need to know if the virus is circulating so this, is probably one of the newest. Technologies. Use, oral. Fluids for PCR and get a sense of the dynamic. Of the virus in the population, indirect. Method I mentioned, before and I like to scratch. This one because indirect methods used to be sub, serological tests, I like. To say that it's an immunological tests, because, now we wouldn't, use serum, anymore we use more. Plataform. To evaluate the immunological response, ETA we use oral fluid so that is why shell is not a serological tests, anymore it's in the muna logical test applied. In the different platforms in Syrian and oral fluid we can use our for Eliza Munich. Florence embarrassment, relaxation, and we can still detect antibodies. Produced by. The virus, so this techniques, are going to tell us the presence of the wire here in this picture we shall see a pen with, a rope hanging there where we're going to collect the other fluid and we want to evaluate the. Dynamic. Of the virus in this population. Another. Question. That we we. Hear frequently in the lab what is the pattern, clinician. Did they. Need more information not just the presence of the virus they need more to borrow this disease so this is the tech needed have been, going. On for a while so this is a restriction fragment, length polymorphism. Pattern basically. We, put the, virus with different enzymes and then signs are going to cut the bottom recording, little pieces so when we sit here the shell of the pieces different size, and. Different, ways of the virus so it will happen the different virus that got, caught in different sections. So and which are named them by this. Defragment. So different. Restriction. Patterns for example the one that we used to differentiate, the body follows vaccine is the typical, palate - 5 - this, is the one that we see. With. Buccaneer a pig why, I'm a always that explanation because this is happening now we, are seeing shift, in this pattern, so, we typically see for quite. A long time the, 1 3 pattern, so this blue bars look at this case. Number here and now. We. Are seeing, more, than one seven four pattern and this is seem. To be more tough of sharing when, fans, get hit with these. Buyers. With, this pattern the. Clinical signs are more severe so, that is why, we, need, that, information. - we, need to provide that information to clinicians they need to know if they're they, have this severe disease because it's a new virus because it's a new, power so they need, that information, they. Also need this information when when, we get this piece, of land that will run a PCR and it's a is positive what, about my pigs were vaccinated. And as I mentioned before pigs, were vaccinated, with it with, the live virus so, we can't tell apart so. And that is an old piece. Of your admission that we need to provide so we have a statistic, PCR, that can detect in this, specific. Region in the old five. You. Can wait until your molar ship with sequence, and we do PCR with. The. Nonvaccine. Potential. Drug Puma, Park so in this case I demolish. It is about 80% we, can, certainly set that, is a wild, type strain but, sometimes homology. Is really really close and we cannot, tell we, can also so that is why maybe. This. Is going to be the future do the next generation sequence, so the full length of the wires just not that, little pissed off the or file we need to sequence the whole wires and see, if. The. Mutations, or the. Difference, are anywhere. Else more than your file so. Probably those are the answers. That, we need to oh, the question that we have every day in the diagnostic lab and we need to answer to the, diagnosticians. Finally. Cynical, virus I believe, the very words I heard about the disease in the last few, years, but.
What Is cynical eyes so, basically it's, a swine vesicular, disease. So. If. Someone. Tell us one particular disease, is, here in the farm what probably going to be the most important, thing this is that we're going to think about it. What. Is our, worst. Fear. Yeah. It's, indeed so. Sandy. Probably is going to be in our differential, for. Vesicular. Diseases, but it's not the only one so, we have some signal disease we have a cigarette stomatitis, I will have a cigarette sent Emma so, and so all of them some, of them within the same family of Seneca very similar, or not but. How they look they. Look all the same and. That's. Probably the most scary part of this thing when we got to the farm and we see these vesicles. So. Until. We don't get in any lab confirmation, we, can tell otherwise I mean there's epidemiology. Behind that, they behave differently but. The lesion itself it can be tell apart you can. So. I like, to give this picture because with this picture up here is, how everything start two, years ago I was. On call that week a producer, sent me this picture we have this animal, this is a show pig in. Wright. County could, eat I'm a right right honey. So it, came from a County. Fair our, County and we have these, vesicles. Up here, kind. Of blanching, in the coronary band hyperemia, here and that, was it so when, that being was ready to move to another County Fair so we kind. Of sushi eight why would you don't keep, for for a while and to see what going on so we got. Slop. From these vesicles of course we send it to MBS cell to Jaipur foot and mouth disease since, God will rule it out and. We knew that by the time there, was an outbreak of silica, virus in Brazil the. Seal like a month before these have reported another, hotel of the secret of this is a neonatal, mortality that is why I won't have that picture here to remind us that they have two presentations, the basic representation and the innate immortality. So. Knowing. That that was going on in Brazil we request, to test for Seneca virus until now that this one was possible so these Hanneman here were the first animal a word agnostic here, in Iowa State. So. We. Start getting, more and more cases we were. With. Her black and both of the producers help us keeping animals and allow us to do some tests to learn more about this it's because we didn't know much so. First, thing is in a finisher farm in North, Dakota they. Send a Fivefinger pig, with vesicles, so we open those pigs and we, check. Head. To tail anything and we couldn't find any other lesion more, than the vesicles, so we learn something that nothing. Else more than a basic, euler disease was going on. So. If you see the lesions here basically in the austrial alteration, of the plant Alicia, this is an acute vesicle. Again, ulceration. Of the plant operation, and we leave me if you walk into the farm, one week after this, fabric you won't find anything, the. Delicious. Had resolved, and the, animals are just fine, so this has to be detected. In the really, really acute stage. This. Is the older presentation, that I've mentioned the neonatal mortality so, you, go to a farm and you see these healthy. Leaders. Here and the next one is affected. So for. Awhile people be described in associated, with this presentation, diarrhea, so. Far we couldn't make any cost. Association, with the diarrhea we detect the virus until the hell diarrhea but, when you try to identify the virus in. The intestinal. Aisle they they don't show up so. Yeah, its association, with its associated with diarrhea most. Of the time but, Seneca virus is the cause I don't, believe so so far. This. Is the element that I mentioned, that we got from South Dakota we have five, finishers here in the diagnostic, lab we were able to check every, single tissue so, if you look at the variety. Of lesions, just kind, of hyperemia, and the coronary band most, eration branching, here, is kind of ulcerated. In. Love the hood completely, a - deucer here acute. Ulcer in the planter region so for. Example if you get this animal no one goes through the farm shot you, know picking up every leg so you will miss that one and so's, are all day leading, down so that you know they don't show lame is that easy so that, is why so. Sometimes. Is the clinical diagnosis, is so hard to make this is. A the. Nostril. Remember, that the picture that I show you with the different bacillus, is this eye this one doesn't look any different with, the other ones this is what happened this one was like ten, days pause break, so, it was already repaired, this is car tissue so you will barely, gonna see this this lesion, so the, timing, of detection, is really important for the clinical diagnosis, here so now we're a scar tissue a little bit of hyperemia, but, nothing, else in.
This Farm so, the, the. Owner helped us and he keep temping. For three, weeks, when those guys were ready to go to market so, we keep those animals and we, didn't know much so we try to learn as much as we could and fast, as we could so, we took theorem three, for. Three weeks here, should set day. Post object because we don't really know when they were infected so there's no date post infection they should break, 36. Day later we start sampling, then every. Every. Week so, what we learned here first, the serum there, was no detection to me this is the total your life and even let me mention did can you see there - lies, in the city. Escort because we didn't even know which was the cutoff so. We, these are the further assault I mean we run everything up to 40 cycles. But. We didn't know which one was it kind of so. Serum. Was not a texture watch what. You means for us that the bimah didn't last that long so we were, 35. Days in those. Clinical. Average and there was not virus detected, make some swabs rectal, swabs and total swab I cannot. Touch on the tenders after shedding so, by, oral, road or fickle Road so we should see animals, positive. At least. 36. 43, days post. Break so means for us that the virus is still being shed. For. Several weeks after the clinical presentation. So. For. The diagnosis, of the disease one we should start with this we have only the PCR, with, the full length sequence, and the worst evaluation, in this virus from the previous, reported, I mean this value had been around. For a while it's not a new virus, the, difference. That was having see critical presentation, but, the original buyer, of the Seneca Valley virus, the SBB so, we. We compared, our signals, with the the, original as reveal and see here is and, I. Was about 10, percent Amala see if we look at the BP one which is the the main structural, protein, and three percent we would look at the whole genome so there was a little bit of difference but we didn't know that the clinic representation, was. Immunization. Of these new strains or just. To happen really another. Diagnostic tools that we develop really quick to make the diagnosis, was in, situ realization, and music, images this is an immunity chemistry, so, basically, we are labeling the virus in this vesicle. Section so we cut through the skin put. Them to boy there and we detect the virus. Another. Thing that we learn from that clinical. Experience, so, we were testing basically. Basically. But, it's. A neural tissue useful, for diagnosis we didn't know that so those peak that we necropsy, and check every single organ of course, we PCR every single organ see if we, can. Come. Up with the consensus, with the parent which one would be the first the best handle. And we found by us all over the place by consistently, the, bicycles, were of course the best ample, inguinal. Inferno, was useful. And tonsils. Basically give us the option to use other fluid if it wasn't a total to, have as many Indian fluids. So. Again. The the techniques that we've been using so. We develop administer geometry as I mentioned before in situ realization, here, instead of use an antibody we use a nucleotide, probe, that bind to the virus is a little more specific, virus, isolation, here. We got a nice mono layer of particular. Percent, a tubular cells and here is an infected, one so we have a really. Strong. Psychopathic. Effect in, 24-48. Hours this. Is the electron microscopy to confirm the diagnosis, so those were the first techniques basically. Techniques. That have been used for, a, while we didn't develop anything, original. We, need those two to make the diagnosis, now all we have indirect methods to make the Dinos we have Liza we have developed an Eliza base or targeting. The BP one this protein this structural protein that I mentioned before and we. Also used, in IFA, here is a picture of the IFA. And. We also have virus neutralization. So those are the indirect, method that we use for, antibody. Diagnosis, a. Little. Bit of neonatal. Mortality associated. With an equalized we. As, I mention before people, have described, this. Diarrhea, and chest, mortality, but.
We Couldn't find a causal association, and actually, this is the case where. We've, been called because of this there was an increment, of mortality. In that week if you look at the numbers I mean there's. No really spectacular so, nothing, like PDB. We, haven't slime diarrhea. An increase in mortality but. We. Don't have to really worry about it like PvE, in in. The enteric side of the disease mostly. We see associating with e.coli rotavirus. So who, knows really, sanika, viruses, is the Costner, we. Got a little we did a little teeny investigation, because, what. We saw is that this, neonatal mortality was. Associated, with, Britain but there was not a real association, with clinical, affected, cells or no clinical affected cells so what we did we walk into the farm we look for animal to have vesicles. And we are ready to follow an animal that didn't have vesicles, so we have eleven, and eleven clinical, and non-clinical effect. And we follow the, road trip for a couple of weeks so, percentage. Of positive souls in. Blue here the clinically affected, and in. Red the non-clinical, affected, so at least we, have have half, of the percentage. Affected, animals. That, were positive to silica wire that didn't show any, clinical sign and when we look at their, piglet, the, same thing so, half of the piglets coming, from self that didn't have clinical, side they were positive, for the buyers so. We took this experiment, a little further and we follow. Those animals for a period of six weeks is as, long as we could go so, salt, again. We. Did three, platinum. Thesis tungsten swallow and theorem if, we look at the serums in South. We. Detective IRA for probably, two weeks after, the break so again, we support the original idea, that by lineae dozen last. Really, long and now experimental, inoculation, have frozen the, viremia doesn't last more than ten days if. We. Will look, at this group here pieces, and total swap maybe, we. Can get the same idea that we have before just, shedding of the virus the ship dividers get shared on a little bit longer up to six weeks we still detecting the virus now, we're going to move to the piglet look, at the pigeons so there, are buyers. In serum that decay four. Weeks, after. Birth and they. Also also. Virus, elimination, that also they gain about four five six weeks, so basically, up to the time of our wing, so. These, guys didn't. Get affected, but, they were shedding Meyers well we look at the antibody, response. The. South where. Sample, while we post break and there is no different antibody responding, clinically, I'm not cleaning that affect the soul so. Something. Is going on the. Also getting infected, but they are not showing clinical, disease or perhaps the. Caretakers, are missing, that clinical. Disease that we mentioned before because it's so brief and so acute that we don't if you don't check you know the the soft really closely means the clinical presentation and. The. Piglet they, have a decane antibodies, they go four or five weeks, after. The birth so this is really consistent with maternal, antibodies, so perhaps the gun antibody to the south and they cleared up the divorce in five weeks, possibly possible. A. Little. Bit of the PME ology in a seguir with PDB look, at this one this is our case. And a weekly case we, keep, track of cases, and it. Has again a pattern, and this is really interesting I was looking a, report. From the producer, of association. Back in the sixty and they. Took about this, linear. Non, seasonal. Vesicular. Disease that, go through the summer of. Course they didn't talk about period, they should talk about seasonal, signal disease which. Filled. With this with, a seasonal, pattern that, we've seen out during the summer in, so in summary what we have seen for the last two.
Years Is an increment in the secure disease and neonatal mortality, both. Associated with the presence of technical errors with. We, identified, this new or contemporary. Stray we like to call it that is a little. Bit different from the historical. String of technical. Errors because. These. Diseases new, for us we still learning a lot and there's a lot of questions that need to be answered, so. What, is the origin, of this viral mutation we, don't know if the decision Eric pressure or. Obviously. We don't know exactly the host-pathogen interaction. And what. Is, the. Molecular. Part. Of the interaction that we need to understand probably, to prevent the disease but. It's posing factors, we know that virus it, kind of tagged animals as you see and they don't get clinically, sick so probably the Halle predisposing, factor than magnetic. Clinical. Serological and viral prevalence, there are ongoing, studies, so we going, to learn so how, widespread, is. The virus to, the United States. And by. Our persistence, in environment, and by real activation should probably close this out in really nice paper come out from the. University, of Minnesota where, they evaluate, the. Environment. Persistent, and dividing, activation. So. With that I'm, glad, to take any of your question, thank. Questions. For dr. Pinner. For. The sequencing, of the, first. Buyers what, percentage, of the, what. Percentage you look at now and how much more processes. Into the entire sequence so. Now. With a or five so, we look. At the whole over five which. Probably, that, where we see the more variation. That is why we use the duration. So, now with this technology the NGS, the next generation sequence we can get the full length sequence, in, three. Four days that's. The timeline, so. But. Let, me let me take a back three, four days to get the sequence then we need few, more days to get the analysis which, is a different, part so chef the sequence itself within, guarantees up to three four days yep. Honestly. I know that our experimentally. Viable. But I don't, know if anyone, have even tried to apply it in the market and if they don't go to the market we don't see it's really going to in that soil I don't. Have much experience with, that sorry. Any. Other questions. Well. There's no questions, and let's thank dr. Fierro one more time thank you.