First of all, I'd like to thank the webinar that. 45 maybe. And to give the opportunity to show to you.

Yeah. Several things that we're trying to do in Brazil to try to, to, to control the possibility and rheumatoids in AS. As you know, that's a very important problem that we have in, in Airs.

And especially the special problem in Brazil because we are, we are doing a lot of transfers in here and the donors, they are more touchable for the prestain for building on the rights. I think that's because we when to use embryo transfer and we have to inseminate the mayor several times a year. And also we have a lot of kind of old mares, mayors with more than 18 years, I think around 20% of our donors, ember donors, they have more than 18 years.

That's a. That that's most of them they have this kind of. Persistent and the right plus AI.

And that's a huge ember that you can see that's not supposed to transfer this embryo. But, talk about the, the how they became a problem. Most of mayors, they became a problem or a bar mayor, because of the age, as I told you before, and also, because of the PKI.

And that's a, a, a bad combination for that Mary, she's old and she has probably the contrativity of the uterus to release, to clean the uterus after the insemination. When they are inseminating, most of them, they have an inflammation. They have to have inflammation that's a physiological inflammation to, to, to clean the uterus after insemination because after insemination, there is a huge inflammatory process that starts around 3 or 4 or 6 hours after AI, but, but this inflammatory process has to be released with 24 hours after AI.

The problem of this mares that normally they have an imbalance on the cytokine expression. They have the inflammation of the breathing, but because of the the regulation of some, some cytokine that's supposed to be expressed during the inflammatory process as the cytokine 61 and 10, and some of them, they are inflammatory cytokines, they, they, they ask the not to come to the uterus to clean the uterus and the other ones, they, they can, they, they, they are expressed. In the uterus to decrease the inflam, the inflammatory process.

But the first we have the inflammation, the, the inflammatory cytokines, and then in the, 12 hours, hours after the starting the, the, the inflammatory process. We have to have the, the expression of the, the anti-inflammatory cytokines. Based on the experiment that we did in Brazil and other experiment that people did around the world, they, most of the matter, they have a problem, the, the, the, on the expression of the anti-inflammatory, cytokine, the anti-inflam the cytokine and it allows in 10, and that's a, a, a problem that they're having.

Don't they? They, they don't, they don't, they don't, they don't express this anti-inflammatory cytokine, then they, they have a problem that the, the, the inflammatory process is more severe, precautious than supposed to be. Don't they have me that they, they start to inflammate with a few hours after 12 hours after inflamma after the AI or after breathing.

And we, and they have a, a later resolution of the process. Then when you have this problem, you have the, the, the accumulation of the fluid in the uterus and then we have some, some, cytokines that are released inside of the uterus, that, that, that can make it disturb on the ability of the to contract. And that's a problem.

The, the, the presence of the neutral fuel is a bigger problem also. That's why we need you most of the time to clean the uterus to have a better resolution of the process also doing lavas. I'm going to talk about this a little bit later.

And that's a, there is a very nice publication from Rigo Canizo from I United States. Yeah, we are working together, me and the team of the Uber and my team Brazil and, and his team in Illinois, and this a nice publication, nice review that I can send to you if you, if you want. The, the full paper that explain everything about what can happen in a mayor that's, has this kind of situation with delay on the the resolation, resolution, sorry, of the inflammatory process after, after AI or after breathing.

The standard protocol to treat me with the possibility the AI, is a combination of these three things. So the first thing that we used to do is to inject dexamethasone before AI. We used to inject dexamethasone, a large dose it's 0.1 milligrammes per kg one hour before AI and the some people, they use the, the moment of, of the I, I prefer to use 1 hour before AI dexamethasone.

With the combination of oxytocin after AII, I start to give oxytocin 123 hours after, you know, AI. That's very safe, not, not, not only one hour. I used to start at 2 hours after AI depend of the situation.

And also, we used to start making lavage of the ute 4 or 6 hours of the AI. It's very important to do the lavage also because when you're making the lavage, you are eliminating the neutral fus that produce that bad, bad, bad inioin. Now it's important to, to, to remove the, the, from the uterus, the, the, the neutrofus because they, they, they, they, they express the interloine that are in that are.

Producing the inflammatory process, in this man and all the things that's gonna happen also because of the presence of neutrophin. That's very important to remove mechanically the, the neutrofus from the, from the uterus and to have also a better, a better action of oxytocin that they use and to, to have also a faster resolution of the problem. But, when you use, dexamethasone, we have some side effects that are not desirable, like, mess that they can develop, lainides.

All the mess that they have, a metabolic problem, they have, they, they can have a, the situation can, can, can be worse when they have metabolic problems that we're looking for a new therapies to control the, the inflammation, the post breathing. Persistent inflammation, using, think more safer than dexamethasone. There are new papers also using now, .

Pyrocoxide, that's OK. I, I, I don't have time to talk about yooxy now with you, but, it's interesting also. It's more safer than dexamethasone.

But this paper is published by Stephanie Booker from Italy. Doctor Estefan, she showed that, on this paper that, I think this paper was published in 2080, OK, 2018 in ology. And that, most of men, they have increase on fertility, when they are treated with, dexamethasone.

That's OK. People are using, routinely, but we have the side effects that are not desirable, especially for, for old mares. And then that's why we decide to look for a new strategies to treat this condition, the, the pre-binding endometritis, and we, we look for different treatments, possibilities, and we decide to play twisted the PRP that's a concentrate of of platelets.

And also we we did, we did some research related to using the stem cells and also now more recently, we are using also the condition in medium from the, the, the stem cells that the media that when the cells are in culture, that theme that your normal trash that we are trying to see what's possible to do with this medium because this media has a lot of factors that are good factors also. And then we start to, to just trash this media, we trying to use them as anti-inflammatory or the rational to treatment. And then PIP is the one that we are more focused because PIP is a concentration platforms as I told you before, that's the best they can prepare at field situation.

And the other thing, the stem cells and the, and the, and the And the, the other, they, they, we, we, we, you don't, you, you don't, you don't, oh sorry, and the other, like the stem cells and like the, the, the medium condition from the stem cells, we cannot, we cannot we cannot . Use it routinely. It's not possible for a veterinarian to prepare our field situation.

That's why we decide to use, the CIP, as the, our first, option to try to control the, the situation. And talk about this, PIP, it's important to see that PIP is cheaper. The PIP is, has a, a gras sight.

And these black things that that you can see on this a later microscopy, picture. And these grammes, they have a lot of factors that are factors that, release. That are facts that can work as antiinflammatory and they have a, a, a, a lot of, that it is very interesting for the way energy process after inflammation, the damage of the tissue, and people are using a lot, this PIP for sport medicine.

And there are several publications right the PIP is a good thing to use in athletic horses, horse riding competition, and they have lesions in the tendons, on the joints. It's, it's very common to use that to use, on sport medicine. And now, we are trying to, to see if it, OK, PRP is good also for to treat end rights.

Then that, it's not not so the idea was not my, the first idea that the first idea was from Elizabeth Metcalf. Elizametreau she's from USCA. Thank you, Elizabeth for your idea to use by PIPAs.

And the first was an abstract from, a meeting that I was there in the United States. And she showed that the, when she did the infusion of PIP in MERS, she showed that there was a double regulation of most of inflammatory interlozin, but it was just, the first paper, she just evaluate the expression of interosse, but it was OK, you know, it was showing that, OK, we can use PIP and PIP decrease the inflammatory, the inflammatory expression of this inosins. Then in the 2nd publication.

That she did. She did the infusion, it was in 2014. She did the infusion of the mass 24 hours before AI.

Of PIP And she observed really good results, using this images that had a persisting on their rights. She was a very select group of mayors and she had a shortage in Portuguese, but you can understand the treated group of PRP. She got 67% of pregnancy rate I guess with the control group.

19% of pregnancy rate. And then she also observed, decrease on, on the fluid accumulation of the uterus. That's a good thing, that means that if she had less neutrofus inside of the uterus also in the same year in 2014, we decide to do, an experiment and that was published in 2016, on tele, in 2016, yes, 2016.

And it was a student, a master student of mine, and Maria Fernandez is showed that the fluid accumulation was decreased after PIP infusion, but in this paper, we did PIP 4 hours after AI and Elizabeth, she did, 24, 24 hours before AI. It was a little bit different. And then when the men were treated for hours AI on this, they observed also that a severe decrease on the percentage of neutral fuels.

I'm showing again that, OK, PIP is, has anti-inflammatory effect and a good anti-inflammatory effect when you make the infusion, on this mesh which persistent possibility the endometris. Then, on the, the last publication that we did about PIP was a PhDT is from Lorenzoquesegainnazio. Lorenzo is a, a professor now at the Ro school and, and.

In, in USA, United States, and we compare his PhD thesis and, the, the protocol of police against our protocol, OK, like Lisa Metcals used before AI and they use after AI. The PIP then you compare if you the, the CO2 expression and the inflammation and also the pregnancy rate of MS3 to these two protocols. But it was the same.

The results were the same that we could, we could not observe any difference between the treatment of PRP before and treat it with PRP after the AI. One difference in, in our protocol that the it is a about medicals used to, to prepare the, the concentrates of plates in Villo with the zero. Our PAP was prepared with a more simple technique.

I'm going to show you later. We just centrifuge and you concentrate the plates. And we, you didn't feel, but you can see the, the, the, the, there was a, a, a huge decrease on the inflammatory cells I can see the left the table and, and the, the, the, the figure on the right, as you can see that the pregnancy rate was 30% when on the control cycle.

We use the same males as a crossover experiment. We use the same mass in all groups and As you can see, the consumption rate or the pregnancy rate of these mares after treated males were double than the with the control group, the cycle that they were treating around 65% against 30%. It was a really good thing.

It's important to say that this mare, they, they, they, they didn't have bacterial contamination. They are mares that are classified was classified as having a, a precious thing for endometric, but with no bacterial contamination. They just inflammate very, very much after AI that's important to say.

And that's another paper that we, we did in combination with the group of Ivo Canizo in Illinois and, and me, our group in Brazil and where we compare. OK, how important is to have a lot of plates, you can have just a little bit of late it's gonna work or not and just to see the, the, how important is the, the, to have more lates. On the system.

Then we, we compare the, the, the, the, the, the poop, the, the fraction of the sum, the plasma, sorry, we feel more latter. Plates in the, the part of, less plateau. We used to have more late because the, the weight of the, the, the plateau, because of the, the, the, the precipitation of the plateau, sorry, in the, in the part of the, the plasma that is more close with the red cells.

It's important to say that to do the preparation of the PIP. It's important to use surgency. We cannot use another kind of anticoagulant.

And we observed in this paper that both PRP and the, the plasmari plateau in the pan with poor plateau number, a decrease on, on, on fluid, a huge decrease on fluid accumulation. As you can see on this graph, which is 20 or 4 48 hours after, AI, there, there is no more fluid in there which of most of the mens treated which should be poor plasma. Platters number of plain or reach number of platters.

But that's OK. But what's interesting that, on this same paper, so, the English is not really good, but you can see that one thing that we observed was that on me with, with PIP, doesn't matter with poor or rich plasma plateist. There was an increase on the protestant levels.

Talk about plasmatic level of 8 days after ovulation. We don't know really why, but probably what's happening is that these mares, they, they, they, they, they have a better SCLs because when the formation of when the, the, the, when the still start to be formed in the beginning of the dioces, there is no inflammation. But when you have inflammation, you have More prostagnant being released in there and this release of prostagnant can interfere on the formation of the CL but it's something that you have to look for.

OK, the level was good, 9 nanograms is OK, but the level with the control group is much the treaty group was much higher than the level of person with the the control mass. I was interested that the in recovery rate was higher. The me treat with PRP when you compare with the, the mas with a small number of plates, means that something else has happened.

When you have more lateters, it's better than to have a, a, a small amount of platters, not just the control of the, the fluid or the inflammation, but something else is happening also. One, another that was interesting for us on this paper, is that, there is no bacteria where on the mas treated with the really PRP, the, the, the, the, the, the plasma which are more number with the plateis showing that somehow. The PIP is eliminate the bacteria from there.

As you can see, we're working with 1212 mas. These 12 mares on day 8, they have most of them, 6 of them, not most, most of them, but 6 of them, they have, some bacterial contamination in the uterus on day 8. In the treaty of PRP they were, they have no contamination.

I don't know if it's a, an effect on on the bacteria bacter see the effect of the PRP is killing the bacteria could be or it only because the PIP is helping the, the, the, the. The, to eliminate the bacteria from the uterus, not because the straight direct action in the bacteria, but helping to, to eliminate the bacteria to clean the uterus from the bacteria. It's something that you have to look for.

We are looking for this now, doing several experiments to see how PLP is controlling, controlling the bacterial contamination. Something to see in the pictures. Interested that there, there are people from other groups also working with PIP.

That's an interesting paper that was published last year, from a group from Egypt. They, in combination with a group from Spain, I think, and they put from terror and also it's a, it's a, a very nice, publication, that the, the inclusion of PIP, also decrease the contamination of the, the uterus of the mares. As you can see here on the, on the, this table, there was no bacteria on, on Mayer Street with PIP when they compare with with the first day of the, the, the, the, the site, the, the first RAI of this man.

Then they were contaminate before they, no day before the insemination, day before the treatment, sorry. And 70% of them are, they have septococcus betamolitico, and on day 14, after treatment, there was no bacteria anymore on day 21. Then showing that the PIP, it was the same for the, for the, for the streptococcus epidemic was PIP is helping them to eliminate the bacteria.

How is happening, we still don't know. And also, in another paper publish from this group, the same group in 2022 also, showing that, showing that 2022 last year that the, that the, the liophyly PIP was good to control the inflammation and to increase the pregnancy rate. That can seem stable.

They, they, they, they, they liophylly the PIP and then compare the liophyla PIP with the, the, the PIP, the fresh PIP. That's important to see, to say, sorry. We use the fresh PIP.

We have to collect the blood from the main prepare and straight use the PIP. We, we don't use to froze or to cool because we don't know if you, if you. If you freeze or if you pull the PRP, it, it can make and distribute the pledge for why we are using just a fresh PIP, but this, this group, they compare the liophylis.

That's a good idea because you can store longer, you can prepare and they, they don't use from, from the same as the, the. The autologous PIP blood. They use PIP from several mares.

They're not from the same man, but another interesting thing, because we used to use also PIP from the same man has to be autonomous. If you're going to treat a man, it has to be from the dead man for a while, that's we are doing only autonomous. They use all of this from other males, but the results are really good.

As you can see, the pregnancy rate with the liophylysis was 66% against 6% from the contra group. And the PIP also there was an increase in pregnancy rate, there was no difference between. PIP lilies or fresh PIP on the increase on, on the pregnant pregnancy rate.

It's something to to look for that and because it's gonna be easy to, to make something that's commercial talking about a product. And also there are papers with PIP in cows. This is a paper made in Italy, and it was published in 2016, 6 years ago, and showing that PIP in cows, that when they induce the inflammation, it was really good to control the inflammatory process also.

They, they observed a decrease on, a huge decrease on depression of some cytokines, inflammatory cytokines, and they, and, but they, they didn't, I don't know why, but they didn't publish a further experiments to compare pregnancy rates. They just observed that it was good to, to, to decrease the expression of the inflammatory cells, but they did not compare the The pregnancy rates. But one, another interesting observation of this paper is that they observe an increase on the receptor, the, the number of progesterone receptors or the expression of the progesterone receptor on the uterus of the this cows treated with PRP.

It's interesting, interesting, because if you have improvement on the, the receptor of before, it means that you are going to have a better action of the before, because the circulating before is going to bite on this receptor and probably a better uterine environment for the embryo to survive and to develop. After, when you have more receptors. It's something that we have to, to look for.

I'm trying to see if it's some the same thing happen in Mars after PIP treatment. And if you can, if you, the, the improvement of fertility that we observe with PIP mares could be relay with a better expression also, also of receptors before in the uterus, but we still don't know the answer for that. How to prepare PIP?

I already told you that it's important to concentrate the, the, the plates, and it's, it's easy to concentrate because they stay in suspension when you collect a blood from air in the, in the, in the. Plasma, they stay in suspension and when you make a a centrifugation, they are going to down, they are going down, then you can just aspirate the, the plates that are in the, the. The, the, the, the part, that's closer of the serum that's closer to the, the red cells.

It's important to as you, as you told you before, so citrate as an anticoagulant normally use this kind of vacuta tube we use plvacuta for HMA. You just centrifuge. There are also available in the commercial commercial things that are to use to concentrate PIP, but they are expensive.

And these are this commission methods to, to prepare PIP. They are important to use when you need to eliminate completedleocytes or to avoid some contamination. When you are using for uterus, there is no problem if you have a little bit of.

Termination of leukocytes. What is important is to concentrate the plates and to not damage the plateau during the centrifugation and to observe the, the, the, the, the right, the, the right to speed to centrifuge and the time to not disturb the, the viability of the, the plates. And then you see trade tubes around 20 tubes, each tube has around 40 mLs.

That's gonna be. ATMLs, around 80 mLs of of of blood, then you are going to recover around 30 mLs of PIP. We just sent you a huge 200g for 10 minutes and And you, you, what you used to do is just to aspirate the fraction that's more closer with the blood to To, to, to do the infusion.

Then you pirate the, the part that's more closer with the blood and, and normally we try to, to now we are using, when we start, we use that distraction. We took this, the, we, we, we just stretched the eliminate the, the part that was in the, the higher part of the, of the serum, and it just spread the, the fraction that was closed of the blood, trying to avoid to the lekocyte that is close to the house. But now we are everything.

We just all the plasma, try to respirate more close with the . The, the red cells trying to avoid respirate them, no. And because in there, we have more, more, more plates.

And then, and that's the, the, the. The another system that people are using just to make it easy, the process to prepare the PIP is a system where, where we take the, the, the bags that we used to collect blood that has it's important to have sodium citrate. Normally they have CPAG.

There's a combination of salt citate and glucose, and we collect the the the dent from inside of these bags. 10 mLs for the CPAD, just the anticoagulant to just aspirate with a 7 inch 10 mLs. Then we put in a in a in a.

50 mL centrifuge tubes. Then we're going to use just one tube or two tubes for HML in to to have 1220 or 20 tubes of vacutenis. On this situation we're going to use just one.

Some vests they, they like to use just one because which one they have already had to not PIP and for me that's OK. We use it for each Falcon 150L 5050 tube of Falcon tube. We just put 10 mLs of CPA day and just collect the blood of the mares with a syringe, then you put in the.

And the centrifu didn't even make the centrifugation. Make it easier to prepare. I'm going to show you a video that's doing that, preparing, preparing the, the PRP at field situation.

That's a bit, . That's a vet he's preparing the PIP that's 1st, 1st he's putting the in the tube, so citrate. And then he's going to the sodium citrate that he's going to prepare 2 tubes, for corn tubes.

And then he collect from the, the jugular, the blood. In a syringe without anticoagulant has to be fast to avoid the coagulation. And after the, the, the collection, they just put in the token tube that has, has shot and rate.

In this process they use 300g for 20 minutes. You can use 400g. It's very safe for 20 minutes also and then make the infusion, this map, the samangalaga man, a very typical Brazilian bread.

And, and when to treat the me that's an important thing to say. No, we, the, the, the, the standard protocol is to, to make the, the infusion 24 hours before AI. That's been the moment that you make the ovulation inlusion because a good moment because the vet is in there.

In the farm, checking the mare, and when she, she, the mare is gonna be treated with ovulation induction. Agent, the, the, the, the vets prepare the the PRP and make the first treatment of PRP. That's gonna be 24 hours before AI.

You see, if it's frozen semen, you have to, to make the infusion, to, to make the, sorry, the, yeah, the treatment 12 hours after ovulation induction. Now it's gonna be, it's just gonna be . Cooled semen, and you can do the infusion at the moment of the ovulation induction, but it's gonna be, if it's gonna be frozen semen, we have to be, to make a little bit later because the semen is gonna be a little bit.

Don't we make the ovulation induction in the morning or the night, depends on the situation and you make the infusion 12 hours after the ovulation induction, if it, if you are going to use frozen semen. And we, it depends on the situation. If the vet is available, we used to do another infusion for hours after AI.

That's an easy protocol because normally the vet is in there in the farm. In the moment of the evolation induction and the vet is in there and the, the moment of insemination. That's a protocol that I like.

Same time people ask Michael, can I use the, make the infusion for 3 hours after, yeah, OK, you can do that. But in my experiment, we decide to do 4 hours after. I can combine the oxytocin in lavash.

Yes, you can do that. No problem. You can make the lavage and then you can make the, the injection of oxytocin.

And after this, you can make the PIP also. Never before oxytocin or lavage. And in my experience.

Experience, sorry. Other things that are stood in here trying to eliminate this inflammation nerves, as I told you before, is the instead to use the platelis, and to use the congestional media from the, the stem cells that are in Q and normal when you, you make the culture of stem cells, they stay in culture for 1014 days, normal 10 days. Then when they are in control, they are releasing several factors and some factors are growth factors, but some factors are anti-inflammatory factors also.

And then we, there are paper also using this, this, this conditional media for other things as anti-inflammatory or tissue regeneration repair, and we decide to, to use to see, and it's OK. One thing that's interesting with the, the conditional media that you can also freeze and you can in the future probably lilies when I think about this. And yeah, you, you can store and you can probably in the future make maybe a product can be developed using the conditional medium for stem cells.

The stem cells are is more expensive. Then you can have the, the conditional meat from stem cells that are in culture for other things. When you prepare the stem cells for treatment, they are more especially this is difficult, it's gonna be difficult to use routinely in emails.

We are used for a specific case. I'm going to talk to you about this, for what we are using the, the, the stem cells. And the, the, as I told you, the, the the the, the conditional meal can be likely, easy to store, and it's kind of expensive if you prepare the, if you make the culture just prepare the, the, the conditional meal, but you are doing the, if you are doing the, the culture of the cells, it's just something that you can have that you're supposed to, to, to, to discharge and you are just using for another thing.

We did the first publication in 2021. We have more, more, experiments in progress now about this, but in the first, experiment that we did, which was in 2020, we observed a decrease on, on the special, of inflammatory cytokines, especially the cytokines that, a decrease on the . In interleuin 6, that's inflammatory cytokine and, and what's interesting we observe an increase on the expression of the interleuin 10, that's anti-inflammatory cytokine.

That was really interesting to, to, to have this result because we are, the, when you use the, the, the condition immediately from the stem cells, we had two good things, a decrease of inflammatory cytokines and an increase of anti-inflammatory cytokines. Then we have the experiments in progress now to see which amount of, of, of, of conditional conditionally need you need. To treat the mass, which is the the frequent to use, if you can, if you have to wait, how long you have to wait in the cells in culture to have more of these factors, but, all of these things are, things are, experiments that are in progress and I'm expecting to have the the results to be published in in 2 or 3 years.

One thing that you did, talk about the stem cells. That's a paper that we published, in 2010 in, in June of the quantitative and science, was the, we tried on the, the idea of this experiment was to use stem cells, not talk about the mess with inflammation, post-breeding inflammation, with prete positive inflammation, with severe inflammation post-breeding. We use these, the stem cells trying to, to decrease the, the fibros of the mare.

Then we use 12 males that have like this mare, a sever fibres in the uterus. And we try to see, OK, let's see if you, there are papers showing a horse, treating the tendons that, there, there is a, a change on the collision of this, of the, the tendon. It's, it's very common to use stem cells for tendons to, to replace the bad collision for a good collageion.

Then we what we decide to do is, OK, let's see if the scar tissue is gonna be replaced for a good tissue and me treated with severe severe chronic deris which a lot of, of fibros. If you could have a, a, a decrease on the. Or replacement, a bad collageion for a good collageion.

That was good for farmers, that's 30%. The, the, the problem of the stem cells that are expensive. They are not easy to use that few situation because we decide to do something different than other groups, we inject the stem cells.

Because we worry about the, the absorption of the stem cells, we should just make the, the infusion. Then we decide to do the injection, we use a long needle that was a needle. That human people will use for a colonoscopy, it's called scleros needle.

It's a long needle that's connected with a syringe. Then we connect this needle, you just put the needle inside of an endoscope, a flexible endoscope, and it just connected for or with a finish as you can see here. And you'll make the endoscopy to see the utin as you can see on the picture now, that, the, the, the moment of the, the sequence of the injection.

It's a needle that you can just control when she will show up and when she disappeared. Then there you just make the endoscopy, the hysteroscopy, we inflate the uterus with air, obviously, and then we make the injection. I have a, a small movie here that I, I can show you.

OK, you got some. Then you can see here, . You can see here the injection, the moment inject we played the the a lot of here to see the looming.

Then you can see that the, the, the, the stem cell. The injected? And, and now you can see the, the bifurcation.

They used to inject in, in 5 different places in each horn. And it's important to say that this man had no, they had no bacterial contamination also just fibrosis. It's an easy process to do.

The only problem is to have an endoscope. It's not easy for a vet to have also, and most of them, they don't have an endoscope, and I used to to. To clean very well the endoscope before use because the endoscope can be contaminated with bacteria from because people use for other things.

But I can see that easy to see that the, the stem cells going after injection, no. We inject around 1 mL in each point, we feed 20 million cells in each one. And they just go, just go inside of the uterus, injecting at least in, in 5 different places until the, they close the tip of the horn, OK?

And it's a procedure that, that takes around no more than, than. 50 minutes to do everything just the mayor is sedate, you know, with silenzine or the Tomiine doesn't matter and they stay quiet. That it's an easy process to do.

And it was for the 3, former from 12 and they we are also the me that not we that it was not observed. We not, did not observe a huge, a huge decrease on firos but. They start to relax by the cervix.

Then, after this observation, why this matter that they are relaxing better the the the cervix, no. It was observation from the vets because they are, they were client mares and the from Marco this me after treatment with stem cells, they, they, they, they were able to clean better the uterus. Then we decide to use the same treatment on the saps of the, some of these matters.

It was interesting because he really, most of these matters, they start to relax better the, the cervix. when they were in heat, because that's a big problem. We have mares that they did not relax the cells and when they don't relax the cells, they cannot clean the, well, the uterus after AI or after bleeding.

And, and the one, another thing that interest for this protocol that the best they can do by themselves, the infusion, the, the, the injection. Use the same needle, they don't need to use the endoscope. They just go with the needle with their hands inside of the vagina and they inject in the, the several parts on the, on the er.

That's 11 thing that's interesting for the method to try, using the stem cells, inject without endoscope, but they have to use this special needle that's . this, the need that you use for endoscopynes, I can send you the, the, the protocol, the, the, the, the, the number of this needle where to buy if you want. I'm going to give you my, my direction soon.

. OK, I think that's all. I'd like to thank you again, the webinar seminar for inviting me to give the opportunity to show you how around the world what we are doing here in Brazil, and I'd like also to thank the Ivo Canizo and Lorenzo Segainnai. They are good friends and we are doing a good job working in the combination with you.

I, the Sao Paulo State University NSP in Brazil. Any large State University in United States and now Doctor Lorenzaga, that's a professor in Ro school, is also working together with us. I also like to thank the, the Brazilian Foundation of .

They give us, give me the money. To try to make my dreams happen, that's the speed. And also the, the, the Brazilian cons of Council of Research for the scholarship also and for to support our experiment.

And and here, as you can see, it's my, my, my Instagram it's perilliad Equina we have to, to receive messages and if you wanna follow me at Instagram. I, we have several information in that. Thank you very much.

See you.