[Music] thank you for giving us the opportunity to present you those work that you are performing at the R&D chip so it will be a do presentation so it's the first time for us so it will be everything will be fine for that so go ahead so I will be the one starting and we will not talk this afternoon about how cell nutrition can influence peak that ability to face with winning stress that we just heard about and we'll be focusing on metering and selenium cases so we heard a lot about it this afternoon but going from lactation period to winning period we know that piglets are going to switch from the liquid maternal feeding to a solid feeding and they need to digest this field when into to digest the proteins in the in the feet and the digestive tract is not mature sufficiently mature yet in order to produce enough HCl to digest those protein and we know that protein when is not well digested can lead to pathogens proliferations and sometimes itself it can also result to this specific period of meaning can also with results in low feeding tech and consequently in low growth performance in addition to the performance part of that we just heard about we also know that the the gastrointestinal tract is going under significant development with the edge if the edge of the animals so at the birth of the piglets they the colostrum give them enough immune compounds of growth compounds growth factors in order to strengthen the ability and they also irritated from the passive immunity from the South but this immunity is decreasing with the age while the the gastrointestinal tract development is increasing with age and usually the winning alum winning usually happened between three weeks to four weeks and we can say that this period is a match with the decreasing maternal immunity and during the the lower development of the gastrointestinal tract and we know that gastrointestinal tract have many functions we have transport of nutrients we have the protecting function will have the immune system functions and having a well-developed gastrointestinal tract can help the animal fight with some inflammatory telling is that they can phase during the post winning periods so the question is since they only received the meek until winning how by acting on the sound nutrition how can we help to improve piglets ability to face stress during these post reading periods yes [Music] [Music] so we'll focus on those two elements sulfur so sulfur is very well known because it's connected to sulfur amino acids but arm attaining and sustain so maintaining is first of all an essential amino acid necessary for for protein synthesis and first connected to to growth of the animal to build both those muscles and protein and the other sulfur amino acid is cysteine this amino acid has more structural functions to connect it to protein folding for example and also signaling functions but thanks to some redox properties cysteine can transfer some electrons and it is also involved in some detox and repair functions we'll give some examples later on the other element is selenium so this one is a trace elements so this meaning requirements are much lower compared to the with sulfur element and but it's also known to be present in some amino acids that are the selenium amino acids and there are also two selenium amino acids but are certainly tiny and selenocysteine there are actually 25 genes coding for $0.25 proteins which makes selenium very important for many antioxidant enzymes I will give some examples so I'm coming back on some fundamentals of selenium because it's much less known in animal nutrition so selenium as I said is present in most two amino acids and there are specific properties connected to the chemistry of selenium which provide both amino acid enzymatic capabilities but are really unique because of their possibility to transfer specifically electron it to summarize it can be said that varies 25 genes connected to 25 proteins with Sarah no proteins so you see it's a very small group of proteins we are only talking about 25 genes and we know in the body we are about 25,000 genes so it's very very small maybe the most famous one is the glutathione peroxidase enzyme which is involved in glutathione system and this system is here to detoxify was reactive oxygen species that can be produced in case of oxidative stress so the glutathione system involve these enzymes and also other players connected to sulfur so if you have something to remember about selenium is that we often talk about selenium and but we forget that it's implicated into two amino acids for animal nutrition when as soon as we say sulfur amino acids we directly think about methylene and cysteine my dream in the future is that as soon as you talk about selenium you should directly think about two other amino acids that are surrender methylene and selenocysteine you should remember that first amino acid is a storage form it doesn't have any specific property but it is very useful to be stored in the body because it can be released in case of high oxidative stress and we were requirements in selenium may be higher so said in the methane in just a storage form however the other amino acid selenocysteine is a true amino acid which is genetically encoded there are both genes coding for Sereno protein or Sereno protein enzymes so it's not random incorporation is really on Falls because it's connected to protein synthesis so it's a specific incorporation of selenium into this amino acid an last slide maybe to detail a bit what's a Celina protein so here is a definition so first it is a protein I just said it's genetically and could eat like any other protein then this amino acid is contained in walls protein as at a single point so there is only one certain cysteine in each of a certain protein and it's not a mistake is really connected to the catalytic site of the enzyme connected to the properties of selenium the chemical properties of selenium that makes both enzymes very reactive and have a huge ability to conduct some detoxification or antioxidant properties the functions now the more precise functions are quite various here I remind the names of the 25 certain proteins I have been talking about the glutathione peroxidase there are actually five enzymes or five glutathione peroxidase Tara Dickson reductase they are very similar so those ones are very well known functions and we are all connected to redox or antioxidant functions so let's say implicated in the detoxification of the reactive oxygen species so about half of them are very well-known and very precise functions the others still do not have very well described functions we know genes are here we never was protein our synthesize some of them are known to be essential but the precise enzymatic reactions we are conducted conducting are not well described so remember most of them are still connected to redox regulations and reactive oxygen species detoxification so involved involves antioxidant systems there are nice comparison again that can be done between a selenium and sulfur so selenium is a biologically necessary for the amino acids and cysteine present involved Sarah no proteins sulfur is present in two of our mean acids but our metal in and sustains and there are actually interactions and connections between those two if I come back on the example of glutathione system we know this system is here to detoxify the reactive oxygen species that can be produced in case of oxidative stress so this is a reactive spaces you can inactivate or reduce it to water which is safe so for that we have the enzyme basura no protein enzyme glutathione peroxidase which is doing this reduction but the electrons are actually coming from glutathione which has sustained so of a sulfur present in the glutathione is in a sustained so it's the other amino acid that is here to make this system turning so where is relieved is connections between sulfur and selenium enzymes another example is a connected to methane in this amino acid can be subjected to oxidation in case of oxidative stress in that case if there is too much oxidative stress you can produce sulfur oxide or meta insult Fox I'd here is the the oxygen but as attacked the methane in and there is a system which is able to fix the oxidized metal in back to the original one it's it's a reductase and this enzymes is also a selenium dependent enzymes and his name is methane in self oxidoreductase so again there are two two connections between selenium enzymes and an amino acid in that case and selenium on James and a system that is able to detoxify reactive oxygen species so what are the other functions linked to sulfur amino acids we just heard some examples about detoxification but actually there is more than building blocks from metallian and sustain so today when we supply methane in the diet we use different sources gl methionine and methionine or interaction methionine and we have proven that those three methionine sauce is equivalent to sustain animal performance but we know that Demeter Union as well as relationship must be converted into an methionine before being used by the organism and meta learning as well as cysteine will be the one initiating protein synthesis by nutrients in learning functions and by doing so it will increase protein synthesis and reduce protein proteolysis resulting on into an increased protein equation in the muscle that's what we know and that what we see from the outside but apart from that we also know that methionine when it's being converted into sustained its give an intermediate which is s adenosylmethionine which will be involved in immune response immune system through metal donation it's going to activate or downregulated some genes related to immune immune function and on the other side just like Miguel taught about we have glutathione and toen which are synthesized from cysteine and we know the cysteine is supplementing the diet fro methionine so sustain being involved through those two components into the oxidative status of the animal all of that making different words for so family no acids today's more than building blocks that we are we are usually talking about we usually measure body weight and body weight can but there is more being that before that that that amino acids and now we would like to give you some examples of how adding more July methylamine in south dates can help to enhance to redock status at winning of piglets and the performance so like I said we perform trials about the equivalents of different sources of me turning so I'm not coming again to this subject now but I'm going to talk about what's happening when we add both suffer amino acid requirement when we had mama turning so we perform a trial in in the state in cells we started feeding them one month before following with three different diets the first one was a controlled ads which was adequate in all new trends and two other treatments were supplemented with 25% more amino acids through either deal methionine origentv so we did that during the legislation phase and also during the entire lactation period until winning at 21 days old and we measure different parameters body weight of South back-fat thickness brought something for different metabolites and amino acid and analyzes and Piglet performance at resin Mik Mik quality so what did we observed first of all on meat composition we observed for the second time this is the second time we are doing this kind of study we observed that fat was not significantly different even if it's numerical increased with the treatments to experimental treatments but most importantly in the colostrum and at day 14 of lactation we have higher protein contents in the milk and higher lactose I'm a given increase of methionine and what we can observe here is that violation CBA we got a bit more higher lactose in the colostrum and at day 14 this is significantly different from the Jianhua tyunin so like I said we have observed this kind of this kind of result in the past but the difference being that we started feeding the South from day zero of of lactation and we got the same increase of protein and lactose which are connected to the increase of some amino acids in the milk some of them being isoleucine leucine which we know that can influence protein synthesis and can explain the exert on protein and we also know that there is a relationship between Mik healed and lactose content and we've the increase of those amino acids in the milk and the plasma of cells we come to the hypothesis that probably the me killed even if we didn't measure it was increasing with this house resulting in a higher lactose contact with Asian trivia so as as a result of this better meat quality when we look at the piglet performance on day 1 and day 7 there is no significant difference so one information that I should give you is that I add a zero of lactation we we homogenized all of the cells to have 10 piglets pass out so there is no effect of of the number of piglets and we have from the 14 and 21 of lactation we have higher body weights for piglets which were fed with the nick of the South we're saving more methionine on the two two different forms so look at here there is no significant difference but I'm sure it can make the difference for farmers still an increase of body weight so this can be explained by the better milk quality observed with the increase of methionine and in addition to that when we measure in that last much different amino acid and metabolite and the one connected readers control which so metallian was not was not different between treatments but we can see that cysteine is a much higher increased evasion TB so this is the result we have observed in the past we know that the trans configuration from methionine to cysteine is enhanced in the presence of smtp a comparison to Almaty onin so we got more sustained from a shim to be a more sustained meaning more glutathione and more towing and that what you can observe here motto in it has been higher than the GL methionine and in addition to that we we have higher calcium in the in the piglet plasma and calcium is also connected to a scavenging and he does control so all of that contributes to give you some approach about how we can influence the redox statues and repeated performance at weaning and had them go through the the winning period so if you want to sum up on this part of the presentation just to remind you some of the results so we increased the metal in consumption by cells during the legislation phase and the lactation period and we got those interesting results in terms of quality but also sour conditions I did not show the results but for sales we have a lower body weight losses and back fat fitness losses we wish them to be in comparison to the two other treatments and when we analyzed the plasma of those house we observed that some biomarkers for protein degradation are higher in the control group valuation tibia treatments which means that the SAU in the control group are mobilizing more protein more amino acid from the body reserves which not the case for a team TNG helmet ionian when they I increase for example so this can make us think about maybe the revision of the amino acid recommendations for South specifically during the legislation phase there is a lot of publications saying that we should increase the nutrient supply during these periods to support maternal health and and piglets growth and in addition we can we also observed that this increase of metal in his house resulted in a Bella piglet growth performance and enhance redox statues that can happen during the post weaning period so what about yes so we have seen Matheny is very efficient is it the same for selenium so in my introduction I've been coming back on the function the biological function of selenium which is involved in both redox system and cellular protein enzymes now from a nutritional point of view we are some requirements in selenium historically selenium is supplemented to the animals with those mineral forms but are mainly a silly night since something like twenty years now there are organic forms the first ones were was silly new yeast and it is well very well established but we the reabsorption and the transfer of selenium to the animal is much better with was organic forms like say no yeast so this was very big revolution let's say when was organic forms appeared more recently now other organic forms available on the market that are certainly methane in or derivatives like hydroxyl and tiny so adhesive developed one of those product the hydroxyl in the mutiny known as Silesia and it has again an addition efficacy mainly based on the fact thanks to is higher purity because because was yeast only contained sixty percent said in the Mitanni were a bit less efficient than a pure form at a 100 percent settlement on e so this has been evaluated now for many years and so here is one example in in the pigs so when you feed the animals different selenium forms either a negative control not supplemented or selenite some yeast and and Cilicia and when you look in the tissues the amount of selenium that is transfer you see that selenite as a as a limited efficacy it's not nothing but it's very limited when you take wall Sereno yeast comparing the point 3 milligram per kilo of selenium you see that there is a huge improvement in the transfer of selenium in the tissues and finally with this over organic forms hydroxyl and methylene Perez again an additional transfer of selenium to the animal so this is very well established it has been demonstrated in in pulled free in pigs and in cows so in most of a farm animals and it's very very strong now what was known also is the transferred to piglets it's quite well known that organic forms can be fed to to the South during the gestation and lactation and there is a transfer to the piglet that is will see mainly through during the validations so this was known for organic forms with selenium and we wanted to confirm it with the hydroxyl metal in form but various visa with transfer of selenium to the progeny so for that we use different groups of solves that were supplemented either point 15 0.3 ppm of different source of selenium and was selenium additives were fed during the last months last months before the stations and during the all lactation so we have been assessing verso and piglets nem status in the blood or in some tissues and when we have also be conducted a complementary tests to assess different biomarkers and if some immune parameters passive immunity could be transferred and modify thanks to this selenium status and source so here are the results of the transfer of selenium from the so to the piglets so some piglets were blood collected directly the day of of birth so postpartum day 0 or the day of winning so postpartum day 21 looking it was blood samples what is the selenium level we see that at vnf of a lactation period you had a significant improvement of selenium in the blood in the on the day of birth there is no difference it's not surprised because it is known that it's mainly through a transfer by the milk but the the selenium is given to the piglet so you have to wait both VNV lactation 'he's looking in the tissues so in the muscles of those piglets it's even more obvious there is no difference on the day of birth and looking at 21 days so at the end of lactation period we see that there is a huge increase of selenium inverse muscles and the dose is also influencing with selenium status so selenite is indeed as some efficacy it's increasing a bit but organic forms are much more efficient for to improve centium status of voice piglets then we have been measuring different redox biomarkers here we have been measuring the glutathione peroxidase activity so remember this enzymes is a sureno protein so it needs the selenium to have this selenocysteine amino acid to be to have a good activity quite surprisingly and I've shown in the previous slide but there was no salem status differences but the activity is slightly increased so maybe the total selenium in the blood is not explaining everything and that some and somatic activity may be already activated more than others remember 25 enzymes so it's not because the total selenium amount is the same but some activity some enzyme activity are not more activity than others and it seems to be the case for GPX at the end of lactation we see this difference but it's less obvious it was only in the miracle in that case we have been measuring over Sarah no proteins this one is named Sarah no protein P it's actually used to assess the selenium status in the blood it's a human standard to know if people have an optimal CM status so we have been measuring it by elisa kit in the piglets we see improvements but there are quite high variability so what difference your not significant but only in a miracle but we see again that there is an influence of the organic form of selenium 2 in to increase this biomarker so it can hypothesize that the antioxidant status of was piglet has been improved with the hydroxyl in the meta inform and finally with selenium and vas certain proteins are highly connected to redox and antioxidant functions but there are also a lot of results showing that it's also necessary for a good activation of the immune system here we have been assessing the passive immunity so the cells were challenged with a specific antigens but induce the production of antibody and we have been measuring it in the piglets to see if it in if the selenium source influenced this transfer of immunoglobulins so on the day of birth there was no consequence the human or globulins was almost not detected then two days after so after a time to have access to some colostrums containing those immunoglobulins we observe an effect in the piglets of most transfer what we do not know today is a due to a higher amount in the colostrum or a better absorptions and unfortunately there was no collections of the milk so we cannot determine it we just know that in the in the piglet there is more of was antibody but we do not know if it is through a better absorptions or earlier predictions so this to say that it could influence the innate immunity of passive immunity disarray of other piglets so other conclusions we have been confirming that organic forms of selenium with hydroxyl maintaining can be efficiently transferred to the piglets and so we can influence the milk quality let's say and it's a good way to have an optimal selenium status for follow piglets and thanks to some biomarkers like glutathione peroxidase activity we could determine that it can improve the antioxidant status of was piglets then some complementary tests have been demonstrating that we can influence the transfer of some immunoglobulins via mechanisms which is still quite elusive what would have been the term mind but sell in Yemen and some cellular proteins well known to have some influence on immunity activation so it still me now will we did complementary tests more connected to two inflammatory challenge so and more on after winning phases to see if if Matheny and selenium could also influence the rest of the growth of those piglets here in this trial yeah the protocol was a bit different because we started with wind piglets so they were they are the same selenium status from the very model because it was not a control it was only supplied by selenite so we could could consider that we have a quite low selenium status and we started to fed different level different forms of selenium on that day so either organic or mineral forms then we did some do some challenge using some inflammatory challenge so we use both lipopolysaccharides which is a can mimic a bacterial infection and induce inflammation to the piglets and we follow growth performance and collected some blood to see what where the risk response talking about oxidative stress biomarkers but also inflammatory biomarkers first we also collected some tissues just to confirm that the product as an efficacious on the selenium transfer so here is the total selenium content of the muscles in the six groups so there was a non supplemented selenium group a group supplemented with selenite at point 3 milligram per kilo and the last group with the hydroxy share in the meta 9 also at point 3 milligram per kilo and half of those piglets were not challenged and we all have were subjected to the inflammatory challenge with the lipopolysaccharide those results are a confirmation of the good transfer of selenium in the tissues of the animal by measurements of the total selenium in the in the long muscle now we have also been measuring different biomarkers and the best response were on some cytokines particularly the il-10 cytokines which is known to have anti-inflammatory properties so here are the results of the different measures that were conducted after blood sampling on day 7 and day 21 either one hour and a half after the challenge or three hour after the challenge comparing the possible and the challenge group we see that there is a huge increase of these cytokines showing that there is indeed a response of the model and now looking about the treatment effect we see that at some point particularly on day 21 there was a numerical increase of these cytokines it was not statistically different at this point again three hours post challenge we also observed this numerical increase and finally on the last day and the second timing we observe that this numerically increase was significant so it's those biomarkers are always very variable so it's always quite complex to interpret but having several times numerical increase and sometimes also tested statistical increase would indeed indicate that you can influence the production of those cytokines thanks to different selenium Zeus and potentially make animals more resistance to different information status yes so what about what about sulfur amino acids so we know that when there is an inflammatory stress these are with always on innate response which is cost a lot for the animal and under that an adaptive response which came later on so there was some trouble from by closing and on brothers and wifey deities was the control group and they submitted some a group of of chickens to to equal I and very observe that the body weight gain decreased this is the same same thing here we've with the polysaccharide and the perfed another group with the same amount of heat from the eco light group and they observe that there is a little bit increase of body weight so is it's not all all related to the decrease of intake but with the same amount of feed intake there's also an increase of body weight gain in the in this group non challenge group of animals so they they concluded that there might be some other elements involved in the response to on inflammations apart from the feed intake so what they did is to measure in the in the liver of the challenge animals in comparison to the non challenge animals very measure the amino acid levels and this is this is the difference the ratio between the challenge and the non challenge animals and this is the so the the gap between between those animal in terms of amino acids and you can see that cysteine is the most important amino acids in the the biggest difference is on insistence in the liver so what they demonstrated here is that actually when animal has submitted to a challenge protein degradation increase and those amino acids coming from the protein degradation and directed to the liver to produce cement inflammatory proteins some cytokines and what we analyzed is the acute phase response protein and they observed that those acute phase response protein willy-willy higher in sulfur amino acid content which may explain why do in the liver does challenge animal there is 40 percent most sustained in this liver so they concluded that probably cysteine is related to the response that we observe when we perfect animal in the control group and based on this hypothesis we know that like I said if you slide before SMTP a is better true self you eighty in to sustain from Jian methionine so here is the metabolism a simple metabolism of methionine you give me turning you got sustained at the N and taurine and Rita town and there is a truck that was studying to invest your bachelor now where they measured in different part of chicken instead intestine to sustain level and taurine level coming from machine tol me turning and they observe much more higher system for this metal in source so using those facts we decided to run a trial to compare these two metal in sources when animal are challenged by leepu polysaccharides in the post winning period and to see how they will respond to this destroyed pants and and challenge so it is in the same trial when when i just show you to dispatch we just discussed about it just for information we kept the animal win at 21 days for 14 days after winning we fed them with the same kind of treatment as the south and we confirmed here that the advantage we had in body weight is still maintained at 25 days old so i won't show this result but just studying the results and what we did here is that we chose animal having iran approximately the same body weights in each treatment group and we submitted them to the LPS challenge and kept them until 63 days of age so as as a just equipment so we have the GL med treatment which was increasing some amino acid and the egypt smtp a treatment which also was increased in some family no acids in comparison to the control group so when you observe the performance we can see that daily feed intake decrease in the challenge group in comparison to the challenge grouped and tell you it can also decrease which confirm that actually i was our challenge did work and we can pursued with the result looking only what it's happening in this group okay so this is the some measurements of some pro-inflammatory cytokines and you can see that a few hours after the challenge the increase in the in the challenge group and comparison to the in challenge group so what is the result in term of performance only for the challenge group i don't i am not showing the wizard for the own challenge but only for the challenge group we can that the increase of sulfur amino acids at the 35 we got basically the same body weight and at 14 days after the challenge we have a tendency a huge tendency with 0.06 of p-value between the SMT be a treatment and the control treatments and at 63 days take the significant difference of of body weight between duration T on Duke and the control group so the increase of of methionine in those those animals fruition TPA allows us to supplement probably enough cysteine to compensate the production of the acute phase protein and the decrease little bit the protein degradation that that cleansing it is talking about and when we look at some of the two biomarkers of the inflammation so we know that albumin must decrease in response to inflammatory stress and this is what happened here compared to the challenge group and another another biomarker aspartate transaminase increases with inflammation and you can see here that in this basically it's increasing the control group in comparison to the others with the delamination sepia treatment it's at the same time similar to the control and the unchanged group and this is a 12-hour post challenge and 24 hours post challenge we can see that with the essentially a group we kind of had the animal cope with this challenge by giving them some building blocks to form the inflammatory proteins that we talked about and somehow it's helped to decrease a little bit those transaminase produced and in addition to that these are some some metabolites of metal onin so sitting here is a bit increased relation to being comparison to to the control and not significantly different with deal medallion but when we observed Turin sostiene is going to give Turin also we can see that it's much more increased visitation TPA treatments and if you remember Michael told about greeted I on measurements as as a measure antioxidant compounds and you can see here that the ratio between the oxidized glutathione and reduced with that ion is decreasing with the with the treatments it is not significantly different but still we can see that if the same GPA we got lower oxidized greet at home over the reduced one and comparison to the control so all of that allow us to conclude that we might with the increase of methylamine in this post weaning period and under challenge we made given some some blocks in order to produce inflammatory proteins and to hurt the animal cope with this oxidative and inflammatory stress yeah so the take-home message so but we have seen that first part was more on the focus on the soul and how we can influence the piglets and so we have seen that indeed with a different meta in source can influence the performance of those piglets through by a milk quality and milk composition certainly and that @vn it's also connected to the growth performance and also that the supplementation of VR drugs see MIT ihnen above requirements can influence his ability to fight an inflammatory challenge and maintain a better growth performance even under was was inflammatory challenge with selenium we we have confirmed that indeed we can influence the milk quality the transfer of selenium to the piglets and using different biomarkers we know that is connected to the antioxidant status and also to some immune power measures so by those two nutritional solutions we know that we can nicely influence the composition of the milk which are the consequence on the ability of the piglets to fight the winning phase which is really a challenge and it can add vient maintain or promote better growth performance of those piglets and thank you for your attentions and I will try to answer very different questions [Applause] thank you very much for your nice presentation we got some questions here the first one is why and how HM GBA is producing a more sustained comparing to deal and ill methionine so that's a good question and that's in 1 million dollar question but we got some some hypotheses in the conversion pathway of SMTP into el methionine we have hydrogen peroxide which is a co-product and thinking is that in my act act as a signaling molecule to initiate the Cascade of trans configuration in order to give most sustained to have the animal having a better oxidant statues but this needs to be to be verified in an experimental condition that our hypothesis based on the theory of the conversion and that's all I can say about it thank you and the next question is what was the protein level of the treatments I think it was the first in the trial you show maybe I can recall exactly the value but the one that asked the question I can give you the information later on but I don't recall exactly what was this potential okay and the next one is what is the interest of saline methane and versus other chelated forms yeah there are different selenium forms and actually for selenium very no real chelate on the market this you have it for corporal sank manganese but not for selenium you you cannot from a chemical point of view you cannot chelate it there with one converted or product which is a with sank but it's actually a certain limit Aenon which is chelating zinc so it's a clay which is set in the methane source actually so it's a chelate of a Celina meet I mean chelating Zhuang but you really have own evil was mineral forms and was organic forms and the only one truly efficient is very serene retaining on the mineral forms you also can found some protein aid or that are a mix of silly night some neon foam with some amino acids or PCC Nate extract but at Viennese it's mineral forms and did not show it was results but the efficacy is similar as silly night thank you and the last question is did you see any difference in picker diarrhea with the high size and in level no we did measure it but there was no no difference and there are we are we've met down in levels [Music]