[Music] [Music] at addiso we cherish the work we carry out with animals and thanks to us chickens and pigs grow with less feed and in a healthier way cows produce milk in greater quantity and quality for a longer period and fish and shrimp now have fewer diseases and grow faster these animals contribute to feed 8 billion people and have enabled us to become a global leader in the production of nutritional solutions for animal feed one of the leaders in methionine an essential amino acid and protected methionine for ruminants one of the key players in the vitamins and specialty products designed to foster animal health and welfare our story began in france with ali montessori in 1939 then in 1971 the company is acquired by hon pulak becoming its animal feed division in 2002 acquired by an investment fund and renamed adeceo the company regains its independence and in 2006 we joined the blue star group thus asserting opposition in asia and china especially adesayo is a unique company in many aspects our routes are in france but our products come from europe and china and 95 of them are exported to over 110 countries and have more than 3 customers worldwide we are a medium-sized company with around 1.5 billion euros turnover and almost 2 200 people working for adeseo our uniqueness also comes from our investments in both industry and research guaranteeing a competitive and innovative product and service offering 1.7 billion euros have been invested since 2007. our success is also due to our dedication to essential values team spirit creativity integrity commitment and results over the years our company has changed developed and grown putting safety and sustainability as top priorities safety of our employees safety of our plants and of our research centers safety of our products we never do any compromise with safety aware of present and future economic environmental and social issues adesayo has developed a growth strategy that is clearly in line with the un's sustainable development goals building on its strengths and success to be a partner of choice in animal nutrition with women and men who make the difference every day to feed the planet in a high quality affordable safe and sustainable way hello this is dr brian sloan today's moderator i am the global director of addy sales ruminant amino acid business on behalf of addiso i just want to extend my thanks to the dcrc for the invitation to participate participate in one of the key educational webinars today we will share with you information and the scientific background concerning the rules and importance of dietary methane and selena methane on health and reproduction we have two invited well-rounded well-renowned experts to talk on each of these topics without further ado i would like to introduce our first speaker dr phil cardozo is an associate professor at the university of illinois champaign urbana illinois he received his dvm and masters from the university federal rio grande in brazil and his phd from the university of illinois since 2012 phil has established a unique program that seamlessly blends his teaching extension and research efforts the impact of phil's dairy science program is in placing students in applied positions and academia phil and the students have published over 75 peer reviewed manuscripts and three invited book chapters to date the program builds from questions asked by dairy producers and focuses on having the dairy cow's diet as a medical prescription for performance health and reproduction this is achieved by understanding the impact of nutritional metabolism reproduction and health in the eddy cows as well as the mechanisms of metabolic adaption to stressors and forage quality without further ad i do feel the floor as huge thank you very much brian very good to see you hopefully soon we're going to be i'll be doing those in person and let's go ahead and share my presentation so i have this first 30 minutes with you guys so uh i'm not by the title please don't think that saliva and refining is less important than defining is just to highlight my session of this presentation the first 30 minutes right so i have this task of trying to connect uh nutrition in reproduction uh and we are trying to do that for a long time but i actually found a video that can exemplify and remind me back my years from practicing veterinarian back in brazil working with nutrition and reproduction so i want to play this video for you guys hopefully you can see it but i tried everything [Music] so that's pretty much what is saying there that warding was in portuguese actually there was a commercial for fiat back in brazil and guess what now i own a ram truck and fiat owns my uh reign at this moment so big company uh getting even bigger but that's some of the things that connects and that's your passion right so here we go passion nutrition and reproduction one thing that i want to call attention because we're going to be talking about amino acids and there's always this question about what are the amino acids they're limiting which ones are going to be the next one so what i'm pointing out here is that depending on the output right that we have for those species that limiting nutrient is going to be different and here i'm pointing out amino acids however we could think about trace minerals probably uh having wool as an output is going to be very different than having meat on those trace minerals and amino acid is the same thing right so if i think about for example papers talking about how important arginine is to reproduction into the offspring that is going to be very well related to arginine in for woven for lambs however cows since milk is the main output it's not saying that arginine doesn't have a hole or other amino acids it's just saying that hey the output at least since 2001 where the last nrc came out they already told us that methionine and lysine are the most limiting amino acids so that's why we need to kind of focus and direct our attention to those two amino acids and we are going to come back to those in a few another point is to consider is the timing so here you see an example of arginine uh of fall landing used right the that may mean that the spring the ones that fall in the spring that may be a little bit uh the lamb in the spring may be a little bit different and with cows also depending on which stage of lactation or that production cycle that may be a little bit different but we do know that the transition period three weeks before three weeks after is very challenging for cows but some cows are successful in that transition and they can achieve and we can can get more in in that thing to that but they can achieve a high meal killed and so saxophone reproduction as well however we through nutrition can alleviate that negative energy balance that we know very well and also that negative protein balance that is happening at the same time those first three four weeks after calving but we need to recognize that reproduction is not going to be at the top of the scale or the top of the pyramid in importance for that animal actually it's going to be one of the last so uh we do need to pay attention and configurate or strategize a diet that suffice all those nutrients right specific related to that output for that species in this case we're going to talk about uh dairy cattle now one thing that is very important i think you know this uh [Music] council has been very involved in a lot of the research out there especially on synchronization protocols and how to get the cow pregnant and i think uh with double of sync we really move to get cows pregnant and i think we still have that question okay the cow is pregnant how can we keep her pregnant now and you're seeing a lot of this embryonic loss and a very nice paper by santos while ago already he put together some of his trials and the good thing about this table is that he was checking or in his students uh pregnancy around 30 days after ai and also around 45 days so here is the difference between first check and second recheck and one thing i want to highlight is that anywhere from 10 to 20 percent was typical to see this loss of this embryonic loss so that's already one question that i throw out there is that you as a consultant veterinarian farmer nutritionist you need to know your number if you're doing 30 and 60 days 40 days and 60 i don't know but something in between there that you can have an assessment on that and of course there are multifactorial uh uh event that is embryonic loss but we can probably help that with nutrition and i'm to show you today one example of that of course there are several diseases that will require other fixes but nutrition can play a role so you make sure to cover your bases there and as you can imagine it has some costs associated with it that's why farmers are very keen into reproduction milk yield and also cows being healthy because they know profit comes out of performing well on those three areas right and the freeze out of albert out of florida determined that on that first month you can be losing around 150 to 360 dollar per pregnancy loss as times goes by that gets more expensive and of course the variation in depending on feed costs and replacement so there is some variation of it there is a cost associated with losing that so it's not a good deal right the other thing is if we are trying to suffice or we are trying to provide to the cow enough amino acid then we may say that's through protein feeding a lot of protein that goes back to high nuclear nitrogen it is shown to be negatively associated with that first breeding so if that first breeding is around 70 to 80 days that can be compromised in cows with a high mun or a high nitrogen circulating or feeding a high protein diet right they didn't find any effect on the super sequin ais so the second or third they didn't fight an association and they did in quite a bit of cows so they did over 10 000 cows on that data set and one good thing that they did here was to separate those cows in high producing cows moderate and low producing they also did separate them in in seasons spring or fall and just as an example here a high producing cow over 100 pounds of milk per day and if she is at 20 milligrams per deciliter on muns that means that she will have 35 conception rate that's on the average on this data set that talking about in the spring it's going to be a little bit lower in the fall but i don't get into the season effect but i want to get into the fact that if that cow was producing 56 pounds of milk per day now the same 20 milligrams per deciliter concentration of nitrogen in that milk that would lead the cow to have a 40 consumption rate so that brings the idea that we need to keep in mind uh that milligrams per deciliter is how much nitrogen you have in that milk as you have more milk you're gonna have more nitrogen circulating in the tissues of the cow including the liver including the uterus so that's why the effect is going to be much larger for cows that produce a lot of milk so farms perhaps that you are own pasture or milk production due to limited genetics are lower you may be a little bit more forgiving on that mum if you are tracking that uh but of course 20 is just an example here that's not you guys probably where is my little laser here yes you guys probably are going to be around 10 to 14 that's where you're going to be targeting that in your farm but the association is linear so if the cows are producing less you can be a little bit more forgiving with that mu but in a broader sense feeding a lot of protein is not a good idea right and why is that effect on reproduction there's this very nice paper here by rhodes where he got some heifers and one thing that he did was some of them he was injecting or infusing them with saline so pretty much a control right a placebo and he was following the p-u-n or plasmaria nitrogen that correlates really well with milk urea nitrogen circulating nitrogen in the cow and as you can see it just kept consistent as he did the the infusion and the ph in the uterus he was always measuring and that kept consistent now the other set of animals what they did was to infuse urea as a source of nitrogen to simulate cows eating a lot of nitrogen right consuming a lot of nitrogen and then you can see that as design plasmaria nitrogen goes up with time and the uterine ph goes down right so as you are changing this uh having that changing osmolarity in the tissues uh because of that nitrogen now needs to be um metabolized you're gonna have that exchange of hydrogens then you're gonna have the water pretty much being changed to make sure that nitrogen is diluted in the tissues and that's where you see this ph and then there's some other research on even in vitro data saying that that change in ph is not a good deal for consumption rates right so one of the reasons perhaps that high uh nuclear nitrogen high point affects pregnancies is because of this changing uterine ph and when we talk about pregnancies we identified wash identified at least seven factors that are important right to get that cow pregnant so we understand very well body condition score and how does that affect uh the cows health and reproduction and we understand the nutrition connection there but today what i would like to talk about is more about this uterine environment not necessarily prostaglandin but lutheran environment and this idea that a large embryo could have a better production of this interferon towel that's gonna signaling he's gonna sing on the cow about the pregnancy right so if you think about that cow is bred now that conceptus is gonna be traveling is gonna be reaching out and touching that uterine wall there needs to be a side signal around that 14 to 18 days telling the cow not to cycle again assuming she's doing that every 21 23 days there needs to be something saying her peg don't come in heat again and the signal is this interferon towel and there are associations between that and the size of the amber larger embryos more signaling to the cow so we are going to go back to this in the end of this presentation so this is one thing that we can impact with nutrition so what is the right diet to feed those cows especially those dry cows uh to impact positively the transition period right i go over in a uh [Music] symposium review with other co-authors so you guys if you want more details about it you can uh read there in general if there is signs but also and the dcrc is uh involved with that that book dairy cattle fertility from horse theremin i have a chapter there on nutrition and fertility and we talk a little bit about the in more depth what i'm presenting here today so feel free to go out there and reach to those publications but crude protein we talk about 12 to 14 of the dry matter and remember crude protein is nitrogen multiplied by 625 in your diet and here is a good point or a good data from dr clark and inacio uh in 2005 talking about why is not a good idea so two information that i get from this graph here so this is the dietary crude protein and how much milk those cows are producing right uh data from several trials so if we think about 18 crude protein i can have cows at 23 kilos per day so roughly 46 pounds of milk per day but i can find cows at roughly 80 pounds so i go from 46 to 80 pounds at the same crude protein and if i just invert this conception then i can have cows at 35 kilos or roughly 70 pounds that are eating 13 percent protein or they are eating pretty much 26 crude protein so the point is crude protein is not a good predictor of milk yield or performance right so that means that i'm not even talking about our what is the rup on those diets where is the rdp so crude protein is not good to prevent and if you wonder why we always talk about 18 20 crude protein for dairy cows is because that's where that curve starts flattening right so they said yep let's shoot there because that's a good point but look the variation in this data so we need to describe better our feeds our diets we cannot talk about crude protein anymore right so the suggestion is just what i talked to you metabolizable protein is going to give you a better idea on how much microbial protein plus everything bypassing the rumen and some amino acids getting to the duodenum of the cow so i want to know how much that pre-fresh cow is absorbing in grams and this is going to give me an idea of a good profile of that protein remember proteins are made of amino acids but i need to make sure that the two main limiting amino acids in that protein that is gonna depend on which sources are used in the diet are gonna be fulfilled right and the whole 1200 grams can give me at this point for us an idea that the other amino acids are taken care of if i don't feed the other ones and just feed methane lysine that usually is not going to go well as well we can talk more about it if we have time in the end now going to the health there is the group from italy uh trevisi and bertone in the past what they are developing was hey if i collect blood samples before calving and two times after calving and i have albumin cholesterol and bilirubin i can get to this liver functionality index or lfi and they identify that hey cows with low lfi they are the ones having trouble that means that the liver is not working properly and the cows with the high lfi they are having a smooth transition that means that they the liver is not overwhelmed and in this research by zizi what he found was that a tendency that cows that were receiving methionine room and protect and defining they were more in the high alifi group so let me share with you some data about it okay one thing they found so in white here you see the high alify those cows with a better liver functionality they eat more before calving they eat more after calving and they produce more milk after calving an indication now of metabolism like i mentioned before they have a higher concentration of methionine circulating the cows that are performing better in uh with the liver and they have lower trimethylhistidine that is an indicator of muscle depletion so that means that throughout that transition period from minus 10 to 28 they are using less body reserves from muscle compared to cows in the low lfi and some idea on the inflammation is that those cows in the high lfi they have a lower acute phase protein like hypoglobin in this example and they have higher pro-inflammatory molecules like peroxidase is just pretty much hey if something happens here we are ready to attack but that doesn't mean that we are out of sense right so that's kind of idea of the good inflammation bad inflammation uh that's what we are uh playing here with peroxinase so very interesting i wish we would have like a calcite test that could tell you if a cow just capped what is the chances of she being in one way or another but i think we're gonna see much more coming from this to talk about the health of the cow this can connect to well-being so welfare so i think we're gonna be seeing more of this type of research here very interesting one thing that we've done with cows that were uh fed roman protected methionine for is this discovery of this uh neutrophil extracellular traps or nat by short so here we did some histology with those cows right so it seems like this control cows they are having a little bit more activity on the 30 right so you can see the number of cells per gland in the uterus the perimeter the epithelial heights are a little bit harder to understand here since we didn't measure all the hormones but it seems like for some reason that tissue was more metabolically active but one thing to be concerned is that since that control cows at the 73 days in new york so when you're getting them ready to be uh bred they had more of those pmns or those neutrophils supporting more for nuclear or a tendency to have higher and that's probably not something we would like to do to interference with that conception right and this is something just to be aware of that i think we're going to be seeing more in the future is that we were able to identify here this next right so that can be activated for us several factors that was not associated with methionine or not so we just saw it happening in the uterus then you can see these little green things here called that's the elastase that enzyme that realizes this trap so it's may imagine by spider web trying to trap all these pathogens like a bacteria for example and we don't know if that's good or bad but we know that that's happening in the uterus now we knew before that that would happen in the circulation of the account but so if this is happening here and if i get another information and give you from atlus's lab where they identify that cows that were repeat breeders or cows that were having problems uh uh breeding they had a higher number of this lymphocytic foci that they call so all this mass that you're seeing here uh does accumulation so they saw that hey uh a small or intermediate full sign of histologic had something related to embryonic loss so that could be something that it starts as a net so those neutrophils are extrapolating on their defenses and now you have all this foci and now you have parts of the uterine tissue that is not secret secreting anything and then you may have this problems with placenta development and embryonic death so something to watch in the future but at this point we couldn't find any association with methionine even though we didn't have a big number of cow a large number of cows right the other thing then we did in a larger pool is to again use that papasmir that brush collect some cells from the body of the uterus and measure those pmns and count them as a gold standard for endometritis and we saw that at 15 days in milk cows that were receiving methane they had higher pmns but that changed throughout the lactation so another concept that is important to know when in the life cycle of the cow which daisy milk she is because at 15 days in milk higher pmn with of course no fever no loss of intake of milky that means that the cow is perhaps the mountain uh process of inflammatory process to do the uterine regeneration for example at 73 days in milk that's not not something you want to have happening because of all this uh issue with conception 15 days of milk you you don't gonna have any worries about trying to breed that cow right just to make sure that cow gets recovered as soon as possible and in 2014 of conference in ireland actually dr leblanc from guelph he suggested that that the healthy cows when i'm talking about the uterine inflammation are the ones that can mount a response but then they quit that response because they clear the inflammation now the problem cows are the ones that keep responding and the cows that don't even respond at all because they perhaps have not those uh enough of those nutrients to build a response right and more recently we just had this paper accepted that where we collected we did some gene expression of the psychology of those cells that we collect in the brush but also we did a uterine biopsy and we want to see how are those related so i'm not going to get too much in details here but you can see that yes you have higher expression of that methionine receptor in that cell there is some proliferation going on higher in rpm less inflammation happening in those cells in um in the uh rpm or methionine group compared to control and in the endometrium you have some similarities or yes you still have more proliferation on that uterine tissue you have more metabolism going on on this uh homocysteine you have perhaps better capacity for nutrient like glucose conversion of cholesterol and also a polyp of protein related to fat transport and less inflammation so very interesting that that amino acid that we know is required can have such an effect on the uterine tissue as well compared to not being fed room protective refining those cows at 73 days in milk we actually harvested we super ovulated them collected the embryos from them and summarizing here we found that the embryos from cows on methionine they had more lipid than control it's not saying that the embryo is fat it's just saying it had more lipids right then trying to understand why more lipids is something that we are doing it at this moment one thing we did was to use this moldy technique in partnership with purdue university where you can put each embryo shoot a laser on it and see how it reflects and then you have this detector that it can map what is the fatty acid profile of that embryo so what we get back is just this heat map that is going to identify what type of fatty acids we are having on those individual embryos so i don't have to pull the embryos i can do one by one and this is what we have we have so not sure totally sure yet what does this mean but definitely i think i'm going more on the sign of that embryo have a type of fatty acid that can communicate or be involved more in signaling rather than accumulating energy uh for uh for the future right so there is this shift in having higher poly saturated fatty acids inside that embryo and we also had the biopsies from the uterine tissue now with more tissue we can go more in depth a little bit so you can see that at 15 days in milk there is no difference if the cows get meth got methionine or not on the fatty acid profile but as we go on the lactation overall we are having more saturated or unsaturated uh fatty acids in that tissue so refining being able to perhaps be part of that particle protein remember that fat to be transparent in the body it's extremely insoluble so we need to be attached to that protein so perhaps that methionine is needed it's the required for that protein a protein to form and now be able to transport that fat around and be able to get to the uterine tissue perhaps now to nourish and to play a role in the nutrition of that embryo and later on feed us very interesting connecting different worlds right so we go north of here illinois where i am we go to wisconsin uh milo wilt banks group in collaboration with dr shaver they harvested embryos from cows fat methionine or not after calving and what they found they did this transcriptomics and say hey tell me which genes are up which genes are down tell me everything so they found 30 the 3-0 that were different and guess what with methane two were altered one we don't know what it is yet you know you just go fishing what's going on there but this the second one that was up was a polypropylene tree like so they also identified some association with lipids and in this case the embryos from those cows receiving the finding had higher expression of protein and tree like and just reminding you this is the whole complex that we need to be transporting fat so imagine a cow during a transition period how much fat needs to be transported not having methionine can be a big deal on the liver and also other tissues like the uterus right [Music] and coming more here to the end of my talk you can see that uh we also saw some of the effects of methionine on the placental tissue so just remember that in nutrient nutrition of the fetus is happening through the placenta so there are receptors not only for amino acids but for glucose fatty acids vitamins and we then harvest the placenta of those cows as soon as they can we collect the placenta did some work on protein abundance and what we were able to see is that the cows that received methionine they had a higher expression or realization of that phosphorylated mtor over mtor so that means that protein synthesis was more activated in the placenta of the cows receiving methionine during the dry period and not a coincidence but also reflected on those calves when born from the methionine cows a little difference around two to three kilos there but a difference right playing a role in this nutrition of the calf within the uterus is still no differences in this dystocia anything like that but a difference that we were able to capture statistically versus a tendency here in the protein synthesis and my last one here coming back to the state north of us from the dr wild banks and shaver work is that they fed cows from 30 days after calving to 128 days feeding the finding or not and i'm bringing this paper here because it's very important they were able to do that in a commercial farm right so a lot of work you have to make sure the cows are eating the top dress with the with the amino acid so a lot of attention needs to go here and what they found of course a lot of cows 300 cows they were able to recruit primpers and multiparous cows but the main thing that we're going to bring to you is that when they palpate those cows or when they check for pregnancy at 28 days and later on at 61 preemie paris cows no difference but look the difference here in multiparous so went from 19 percent embryonic loss to 6 percent if cows were fed roman protected methionine and here mainly remember that i told you in the beginning this is typical it's not off the charts 10 to 20 percent we said that's pretty typical so we need to know and seems like me finding can impact that but the good thing here is that they went a little bit further and actually when they were checking those pregnancies right they were able to measure the volume of that embryo and guess what preemie paris no difference in the size and the volume of that vascular in that embryo however multiparous cows the ones that were receiving rumen protect and defining the embryo was larger than the control so probably meaning that signaling of pregnancy was more present in here and there is research that i didn't show today that shows how important the finding is for the embryonic growth so all the media that you guys use for android transfer ivf i will bet you you're gonna you're gonna have to fight not to find methionine in those media right but there's another area of opportunity to manipulate that media to impact that embryo survival when you are transferring an embryo or doing that fertilization but different story there so take home message here uh i think it's very important for us to focus on giving the farmers what they need as a veterinarian or nutrition they're really gonna focus on those three things i think making sure that the cow is eating the overall diet is important to prevent any of those problems and here is a little bit on how we see that so we think that that 1200 grams at least of metabolizable protein you're gonna need a softer for that a model right to tell you how much is that uh we here are using cncps associating that grams of amino acid with energy where is that energy coming from we're going to be telling more in the future if it is from carbohydrate sugar fats for now it's energy right and here a little bit of the reasoning of the best where these numbers are coming from and it's pretty much is going to be your best efficiency of utilization how much you feed and how much the cow can use even though sometimes you don't want to be efficient right you want to focus on performance so you may play with that number there is some variation there that you may want to play but you have to start somewhere and here's my suggestion for you and with time you're going to be learning and you're going to be getting your own perspectives on some farms may be a better number a little bit higher some farms a little bit lower right but we need to start somewhere and we are plugging this from lactating house into dry cows and that's how we are doing right now so a lot of the papers you saw here that's the concept we are using so with that i would like to thank the dcrc thank addiso and i'd be more than happy to to chat about it now or in the end whenever you guys want to pull me in thanks phil for a great presentation i'm sure this has provoked some very good questions from the audience if you have questions for phil please feel free to enter them in the q a we have a few came in but we're going to set the bar at 10 so that nobody gets any credits unless we get 10 questions at the end maybe that will motivate some of you to uh ask some more questions but uh without further ado i'd like to introduce the second speaker today dr darren juniper worked within the university of redding's animal science research division he gained his phd from reading in 2003 entitled diet and endocrine responses in beef cattle he's been involved in animal production research for over 30 years and in this time has worked with most classes of domestic plant livestock both ruminant and monogastric covering a diverse range of research topics but more recently over the past 15 years he has worked on the area of trace element nutrition primarily selenium and iodine in animal diets his selenium-based research has focused on the effects of selenium source on aspects of animal and human health as well as a deposition of selenium in the products and post and postmortem tissues of both ruminant and monogastric livestock darren the floor is yours thank you very much brian and thank you to phil for uh an interesting introduction i'm not sure how well i'll actually live up to his introductory uh video i don't have anything nearly as flash as that so what i thought i would start with is a brief introduction as to what selenium is and then lead on into maybe the benefits of selena methionine and certainly in terms of the effects that she has on health and fertility of dairy cattle so um very simply um first question sometimes i ask students when asking about nutrition is what is selenium many of them have never heard of it none of them except if they watch the film evolution uh if you go to wikipedia you'll get this very simple explanation a non-metal with atomic number 34 discovered by john jacob basilius in 1817 from an animal nutrition point of view it's an essential trace element which deficiency has been associated with things such as impaired growth impaired fertility and possibly poor health within farm animal species and this health and the physiological effects of selenium are mediated by these things called selenoproteins so a cilanoprotein shouldn't really be confused with the selenium-containing protein so we have selenium methionine so selenium refining can be incorporated non-specifically into body tissues into body fluids and that is a selenium-containing protein a cilanoprotein is a protein that is functional it contains a selenocystine functional core so it's the catalytic site of the enzyme it's not free roaming it's not something that's actually found um floating around in the body it actually requires a genetic sequence for incorporation into a selenoprotein so a cylino cysteine insertion sequence so the cello proteins 25 have been identified to date most well-known i've put up here on the screen so we have the glutathione peroxidases so these are some of the most important cellular antioxidants thyroid oxygen reductases so they're involved in the reduction of thyroidoxine which is a cell redox signaling mechanism the deodorants is they serve a role in thyroid function the conversion of t4 to t3 so they can affect metabolism and then there is solano protein p um cilantro protein p is involved in selenium transport approximately just over 50 percent of endogenous selenium is found linked to cylinder protein p it's not only involved in selenium transport but it has actually shown that it um also serves a role in phospholipid or reducing phospholipid hydroperoxides um so what is the purpose of selenium supplementation well primarily it's actually to support animal health so many people when they actually look at their animals i'm not necessarily aware that they are selenium deplete or they are actually requiring some form of supplementation and that goes probably most trace elements so this is a slide taken from larson or an image taken from larson and very typically when you have an adequate supply of a trace element there are no problems and then you pass through a phase which is subclinical subclinical because there are no clinical manifestations of it if you were to take a blood sample or you look more intensively at the animal you might notice that there is a deficiency so this is where you might expect to see in the terms of selenium changes in saline protein expression as that selenium status or trace element status continues to decline then you see the visual manifestations of deficiency and with selenium when you see those deficiency symptoms you are probably quite chronic and we'll talk a little bit about what types of symptoms you might see with fairly chronic deficiency so in terms of the symptoms so in very severe deficiencies the clinical symptoms that might manifest themselves are things such as muscular dystrophy cardiac myopathies liver necrosis and at very extreme levels you might expect to see impairment the thyroid function at this moderate mild deficiency so we're talking about the clinical symptoms there are no as i say no symptoms that have manifested themselves but you find that if you actually look at some biochemistry in hematology is that there is impaired antioxidant status as you find there is down regulation of the expression of glutathione peroxidases are the fire reducting reductases you also might find that there is some reduction or some [ __ ] growth some impaired fertility and that's could be attributable to down regulation of cylinder proteins p n w and glutathione peroxidase iv so cilantro protein p i've already mentioned is involved in saline transport solar protein n is involved in protein folding we're leaving endoplasmic reticulum stress selenoprotein w deficiency has been linked to muscle degeneration and glutathione proxidase 4 is quite important as it's very important in maintaining cell membrane integrity it protects the phospholipids um carrying on with that reduced or impaired immune or inflammatory responses and they've been attributed to down regulation and selenium proteins k and s salino protein k is involved in calcium flux in immune cells it's also been shown to have a role in t cell proliferation and so on protein s again like saline protein n has been seen to have a role in protecting the endoplasmic reticulum from stress so again affecting protein folding so although those things are things that may not necessarily manifest itself it might be something that's affecting an entire herd and because there are no clinical symptoms associated with it it might be accepted or might be accepted as being the norm so the thing about selenium deficiency is the symptoms that you see are somewhat reflective of selenium status and selenoprotein hierarchy so a little bit more um information about selenium hierarchy so three important points firstly not all cell and sorry not all selenoproteins are expressed to the same degree so the first thing you have is what's called tissue specific saline hierarchy so there are some tissues in the body that hold on to selenium for dear life and those tissues are the brain and the testes so the example i've given here the figure at the bottom of the slide shows the effect of actually selenium depletion admittedly it was poultry but poultry brains are easier to get hold of than cow brains but what you find is that in animals where you deplete selenium yes there's an impact in terms of what you see in muscle tissue but it's conserved in the brain and similarly you find the same in testicular tissue but the same does not apply to ovarian tissue there is no reserve selenium is not conserved in ovarian tissue but a reduction in selenium within ovarian tissue does have an impact on steroidogenesis so secondly there is the hierarchy solenoprotein expression so not all cylinder proteins that i said are expressed in the same degree they're all very much dependent on the selenium status of the animal so as previously mentioned the deodorants they're fairly bulletproof you need to have really fairly low circulating concentrations of those to have any really serious impact on deodorants activity and then as selenium status improves then what you find are the glutathione peroxidases and there is a hierarchy within the cell within the glutathione peroxidases so the one that is more resistant or more tolerant to low selenium would be glutathione proxies iv and at the other end of the scale would be glutathione proxies one so as selenium status of the animal improves you find there is better expression of those solo proteins as status increases fire reducting reductases they sit within a niche or within an area similar to expression of the glutathione peroxidases when you're talking about selenium status then we come to solenoid protein p and cellular protein p actually requires a much higher level of a better selenium status to have maximum expression of that particular celano protein and certainly in humans um the recommendation i think in humans is an intake of 65 milligrams per day um or micrograms a day sorry that fairly fatal but micrograms today where it's been shown that to actually get a maximum expression of cylinder protein p you're looking at an intake of about 100 micrograms per day so there's quite a marked difference in how that is expressed and finally where selenoprotein expression was often thought to be just the function of selenium status so if you had adequate selenium you had adequate expression well actually the oxidative stress or stress on the animal impacts how cena proteins are expressed um so a couple of studies listed here so polyunsaturated fatty acids and inflammatory cytokines added to cell culture were shown to upregulate the mrna for gsh px4 and the addition of peroxides to again tissue culture have been shown to increase mrna for gpx1 and gsh ex4 so an indicator of oxidative stress up regulates mrna expression so this up regulation or expression of gshpx mrna is affected by oxidative stress and that is independent of selenium status whereas the actual expression of the solenoid protein itself is dependent upon saline state so the example in the graph shown below soon to be published this is looking at peroxidizability index so we looked at the fatty acid profiles of muscle tissue and from that we calculated the peroxidizability index of the tissue and we looked at glutathione proxies activity and what we found is there's a positive correlation between glutathione proxies activity and increasing peroxidizability and that happens in animals that are selenium deplete but in selenium deplete animals is that you have a similar peroxidizability index but you have no response so a very good analogy is that the mrna is if you like an effect of turning on a beer tap and wanting beer to come out but unless there's beer in the barrel then nothing will come out so mrna is the tap selenium is the beer so if we come back and we look at um reasons why we might cull dairy cows so phil has already touched on a lot of this so i may be repeating things already said or touching back on them but primary reasons for culling dairy cows is a failure to conceive mastitis and poor production there was a study conducted by hanks and kusabayati they were working within the university of reading and they did a survey of 500 uk herds and within 75 of those surveyed herds the culling rate was 24 and exit was 3.7 lactations which is you know it's a fairly short productive lifespan so when you actually look at this failure to conceive mastitis and for production is that they are symptomatic of this thing called the dysfunction inflammatory response typically during the transition period and this has been attributed to oxidative stress um there's also been reported that it's an inability of the animal to actually adapt during the transition period results in systemic inflammation immunosuppression so oxidative stress as i already mentioned up for regulate selenoprotein mrna that's selenium independent but selenoprotein expression the proteins required to deal with oxidative stress is selenium-dependent and what maybe what you are seeing is that in this clinical phase is where these proteins are not necessarily being expressed to deal with the problems or the physiology that the animal is encountering so i apologize for how this slide is laid out it did have animation but i took it away because it became too confusing but very simply if we take oxidative stress it's been linked to things such as retained fetal membranes and that is a typical indicator of selenium deficiency it's also been linked to elevated somatic cell counts mastitis and metritis and certainly elevated somatic cell counts mastitis and botritis they are leaked in turn to reduce immune function and reduce immune function has been shown to be dependent to some extent on the selenoprotein expression or the expression of certain cyanoproteins particularly those that are involved protein folding cell signaling and calcium regulation so one question we might want to ask is the selenium supply that we give our animals is that actually adequate to meet the selenoprotein expression requirements of modern productive genotypes and the reason why i say this is that at the bottom of the screen there are um two graphs these are admittedly based on poultry but it tells a very interesting story is that according to the nrc these two species of poultry have identical selenium requirements but when you actually look at how they deal with selenium they actually deal with in markedly different ways in that one actually manages to accumulate it one doesn't necessarily accumulate it doesn't deal with it particularly well which suggests that as we have moved on with our dairy calves most recommendations are quite dated so are they fit for modern genotypes so that the nrc recommendation says it's somewhere approximately 0.3 milligrams per kilogram the legal u.s limit is 0.3 milligrams per kilogram so you can't actually feed any more than that anyway but glutathione proxy's responses have actually been reported up to 0.4 milligrams per kilogram now that tends to suggest that the recommendations the limitations do not actually reflect the needs or the physiology of modern high-yielding dairy genotypes so what about how do animals respond when you give them selenium supplements so the response that you see will be dependent upon the selenium status of your animal prior to supplementing it is actually showing clinical symptoms therefore you might see a marked improvement are the symptoms subclinical whereby actually what you need to look at is some marker of selenium status and the marker that you choose is going to be affected by the selenium status of the animal and what remedial action you may have taken are you measuring in blood are you measuring milk are you measuring plasma are you measuring total selenium or are you measuring a cylinder protein or another biochemical indicator so they will have an impact on the decision that you make the status of the animal itself is going to be largely influenced by your geographical location or the selenium soil content so in the us i understand there are areas where there are fairly high selenium contents equally there are areas where um saline content is quite low here in europe um which we were once part of i believe um selenium contents in the soil are fairly poor we're known as old soil types so saline says in the uk is generally poor anyway so the management system that you have is going to affect the selenium states of your animals particularly the geographical location so how much access do your animals have to graze past you so if your animals are quite extensive and they're grazing quite large amounts of pasture is that if your soil selenium is fairly low then their selenium intake is also going to be fairly poor in terms of the diet types you're feeding are you feeding a forage based diet or are you feeding a concentrate based diet if you're feeding a forage-based diet that's going to be locally conserved forages most likely and that will reflect soil selenium conditions if you're feeding more concentrates are those concentrates commercially sourced they may already contain a selenium supplement or are they made on farm and are you adding a silly supplement are you feeding your diets as a total mixed ration or are you feeding them as a forage on the floor and then top dressing your concentrates because the difference between the two is that a tmr for a comparable dose of an identical source there can be 30 difference between the responses that you see between feeding it top dressed and feeding it as a tmr so how you feed it have quite a profound effect then we come to this question about supplement use of supplement source so principally selenium supplements come in two primary forms shall we say so there are the ones that are termed inorganic and those are typically selenium salts sodium cylinate sodium cellulite and then we have the organic and these are typically the ones i'm talking about sileno-methionine-based now um in terms of um how effective those sources are it's not really the topic here but generally the responses to organic forms cylinder methane-based forms give a two-fold difference in terms of responses and they actually occur in a much shorter period of time and the organic sources tend to be considerably less toxic than the selenium salts so in terms of the fate of ingested selenium so this is fairly simplistic i borrowed this from margaret raymond but very simply it doesn't matter what the source of selenium is whether it's organic or whether it's inorganic is that regardless is that all selenium to be used as a functional selenoprotein needs to pass through the um selenide pathway um for inorganic sources when passing through the selenide pathway they either use for incorporation into selenoproteins if they're not used for the incorporation into selenoproteins then they are methylated and excreted from the body so they're either used or they are lost uh they're not retained in terms of the organic forms so these are those that are predominantly selenium pathology is that yes selena methionine still has to pass through that selenide pathway in order to be used but one benefit of selenium methionine is that it is um non-specifically incorporated into body proteins body tissues and it acts as an endogenous selenium source during times of of deficit so as the animal undergoes natural protein turnover that releases selenium back into the systemic circulation and provides a buffer if you like for animals that might be extensively grazing so this is a rough and ready graph this isn't published data but this is um a collection of data which i think worked could be about just over 300 dairy cows and this is the post supplementation period so the dotted red line at the bottom represents animals that were unsupplemented throughout the period of the study so generally poor and this hatched red line that sits just above it is the level of selenium that is required to actually um have reasonable gshpx1 expression so when we look at those animals that received um inorganic salts or selenium salts yes it's elevated over control animals there is a decline um i know i said you use it or you lose it but what you have in there is that within the diet there's background selenium and that's organic in nature so there's a little bit of inorganic and organic tied up in this particular line in this green line this is organic saline saline fine and so if we actually look at glutathione proxies and admittedly this is a an extrapolation is that if we look at how long post supplementation that um glutathione proxies or whole glutathione prostate levels are maintained to a point where they might become a concern is a hundred days whereas when you feed solar methionine-based supplements what you find is that's extended by a further 75 days so it gives you a total of approximately 175 days coverage when you use it or when comparing it to to no supplement at all so those are probably the benefits for the animal on the ground certainly protecting her but then there is this question of well what about um the implications that the damn selenium status has on the calf so it's not unknown it's been fairly well published that there is a significant correlation between the solenoid status the dam and the cylinder status of calf and there are a number of studies that have shown that with improved dam selenium status you have improved calf selenium status certainly in terms of total sodium in whole blood and plasma in a number of studies that's translated into better or higher glutathione processes activity so that shows better cylinder protein expression and in some studies they said that calves that have received or calves born to dams that have received selenium during the the latter part of pregnancy have increased bigger scores um but more recently uh so this study conducted by myself and others that looked at hydroxyl cylinder methionine um which is a selenium methionine analog and what we found was yes it increased um the selenium selenium status of the dam and of the calf glutathione proxy's um levels were enhanced um but we stopped we actually looked at the selenium content of colostrum and what we actually found was that with animals that were fed selenium methionine from dams that have been fed hydroxy selenium methionine is that we had elevated colostrum selenium well again that's not anything new there are a number of studies that have said that this selenium content is higher in first milk than it is in following um and when you might ask the question well what is the benefit of that again there are a number of studies that tend to show that actually intestinal transfer of iggs or of albumin tend to be improved when the selenium content of the milk is actually elevated or higher and this is well confusingly whether the selenium is enhanced naturally either via the dam or whether you actually add selenium to the colostrum itself and then feed it to carbs what was of particular interest is that generally when you feed selenium methionine to dairy cows is the predominant salinized amino acid that you find in the milk is selenomethine but when we did speciation on the first what we actually found was that the predominant selenized amino acid was selenocystine not selenomethane and if you recall from my very first opening statement is that selenocysteine is indicative of a functional selenoprotein um so digging around trying to understand a bit more about this admittedly this was not in cattle but this was found a study done in mice is it was reported that celano protein p so that is a selenoprotein is the predominant selenium transport protein in urine milk and it was postulated that it was actually an important mechanism for the maternal transfer of selenium to the neonate in that early postpartum period and if you think about it is that the consumption of immunoglobulins in that very narrow window postpartum that's through pioneer's pinocytosis across the gut wall well the cylinder protein p is a fairly large molecule and it might be that also passes the gut wall through pinocytosis and that must confer some benefit onto the calf in turn of selenium status i suppose sort of moving on and this heralds again back to a little bit of what phil had to say is that there have been a number of studies conducted in a number of farm animal species and the ones that i'm most familiar with are those that were conducted in sheep or in pigs that have shown that any nutritional insult in that first trimester of pregnancy um so an initial nutritional insult of the dam has an impact on the offspring so one it has an impact on placental development so poor nutrition can actually enhance placental development ironically but that is driven by the fetal side of the center and it's also shown that when you actually look at the offspring postpartum is that in that first generation there is an alteration to energy nutrient partitioning in fact what you find is that the neonate actually finds it very difficult to deal with things such as glucose so that was admittedly in sheep and pigs but more recently alzheimer i listened to a talk by him a year or so ago and they're now looking at this in dairy cattle and they've shown actually that these pre and post-natal insults can actually have long-term consequences for the growth fertility milk kiln and longevity of dairy cows so the dairy cows are following on as the next generation and these are reflecting very much the observations that were made in sheep and pigs 10 or 20 years ago so what i offer up now is maybe another which is intergenerational effects so where we mentioned that we have this impact fetal programming so that you apply a nutritional deprivation um to a pregnant dam the f1 generation shows altered energy metabolism but if you take the f1 generation and you breed from them you don't need to induce any form of intervention but the f2 generation has the same altered energy metabolism so what you have here is where you have a macronutrient that has affected not just the first generation but it affects the subsequent generation and if we think about how we breed replacements for dairy hurts you do have grandparents daughters and the population moves on so if that's the impact of a macronutrient we're only just discovering impacts that micronutrient deficiencies are actually having on animal um performance animal physiology so is it likely that sort of these small deficiencies are having an impact on animals in the longer term in multi-generation steps so um i'm going to draw close to it a time is moving on so i um thank you for your attention and i move back to brian thank you so much darren for that excellent presentation um so and now we will open the floor everybody will be glad to know i have at least 11 questions so i know these credits are at risk here so um but keep the questions coming in particularly for darren because um i see that um we have quite a few fulfill but keep um i'm coming in for both speakers but but in particular if if possible so let me go to the q a so um a first question for phil is why muns only affect first ai but not subsequent ai in the study that you show sure um i think that's uh as we move cows move through lactation the new components they change and i think as well the concentration of uh that nitrogen circulating or pretty much cows are going to get into peak so around that first breeding that's where we expect that cows are going to be in their high highest milk or or just after that and as times goes by that milk yield probably goes down right as lactation curve goes down and then maybe that's not as representative to change that uterine environment for example so i think one of the things is associated with that and perhaps the liver status starts getting better so you have more apologize free to deal with the nitrogen clearance and uterine tissue as well so i think it's more related to that i think but they don't have any conclusion any hard conclusion because it was just a epidemiological type of data not so much doing the controlled research brian you're muted yeah big mistake i always do it once at least so and bear with me and one for darren what would be a good or normal level amount of selenium and what would be considered too low or too little so a very good question so what would you consider to be too little or too low so we've mentioned that for productive dairy cattle is that selenium response has been shown to be 0.4 milligrams per kilogram of dry matter which is above the the us recommendation i would suggest that maybe what's too low again what is the purpose of the animal how metabolically active are they so if you're talking in terms of high-yielding animals i would probably say that you need to feed whatever the maximum recommendations that you can feed to meet those requirements of those animals thanks darren um phil another one for you um you talked about profiles um according to your observations or at least your um reading of the literature um do you think poopers are essential for early embryonic development yeah that's a very good question and i think we uh definitely have a lot of information out there i think there is an overall recommendation to include some pufas to diets of cows we're not sure one pound there's a lot of anecdotal information about it but point being is that some of those party insider fairy asses especially omega trees right so we can design products that have this specific types of fatty acids and we can or we have to protect those because if not remember in the rumen everything is going to get saturated so you're not going to end up with anything uh being absorbed by the cow uh but there are some research saying that you know if you're feeding more of those pufas you can have some of that prostate gland in production being prevented and that would lead the tissue to convert more into cholesterol and that could lead to more progesterone so just by itself could lead to a better embryo survival so there are classic researchers in florida but i didn't see that being that that have been repeated uh again but i think that's kind of where it is if there is that chance of that lipid profile to be more unsaturated in the uterus that could lead to a better survival based on this theory of cholesterol and progesterone if you remember right i showed you data that the cows uh that consumed methionine they had a higher expression of that l-cat enzyme that is the one converting cholesterol so that would kind of go along with this theory that i'm just talking to you but as far as so far is hypothesis but there are a lot of products out there playing with that uh amino acid fatty acid profile and prevention of prostate landing production so yeah i think you've already really answered the second question is here but i'll just ask it anyway you can add to it if you want so is there any interest to um do you think to feed methane or more methane with specific fatty acids supplement supplements to get um let's say uh the best effect that's a good point um i'm not sure i i want to assume that if we are feeding amino acids that profile is changing it's because it's it's needed i don't think it would change just by chance right or just to store that's not the type of fatty acid would be stored so i think some of the signaling that's needed but we are always comparing supplementation with no supplementation so hmm yeah i don't know believe we'll leave you to think about that okay so okay okay so and another one for baron um do you know if selena metanin and methane would affect other animals health and reproduction similarly for example horses instead of dairy cattle [Music] i was hoping you could lip read but there we go um so um going back to your question is that um horses is a good example to choose and the problem with horses they're actually quite sensitive um to selenium um in fact i think anecdotal evidence says that custer's last stand was because of celenosis in pack horses um there is evidence to show that actually selena refining in horses can have a beneficial effect i'm going to use horses as the example is that the study which i mentioned where we looked at 300 dairy cattle and removed the supplement and looked at post supplementation the effects on glutathione peroxide's activity is you get the same effect in horses so you do have that protective effect so i would say yes you would see it in other animals okay thank you darren um phil do you have any evidence whether methane supplementation could be beneficial to rule out the negative impact of heat stress on fertility very good point and um i'm trying to think about it here and uh i don't have that uh answer straight i have some things that may connect to that and help but i think we we do need that data a little bit more specific so we do know we know that if we have heat stress and that was done in canada with metabolic chambers they induce heat stress and they saw that plasmaria nitrogen goes up and that the profile of amino acids including methionine goes down so cows in heat stress they seem to have a lower concentration of some amino acids including methane and the circulation we have done uh one trial here with the heat blankets where it can induce heat stress and we saw some positive effects when cows are under heat stress they are par fed so that means we take out all the intake uh confounding and it seems like when they have the room and protect methane in their diets when we did uh mammary gland biopsies we see that the alveoli when you see which ones are proliferating which ones are dying that number and the association if they get methionine they have more um more proportion of those of all eyes that are functioning so i think there is some effect on for sure on the mammary gland but i don't have any idea specific to the uterus or to the follicular dynamics but i would guess that yes there is something there thanks phil perhaps just another one on uh for you and on the embryonic losses and the embryonic size data you shared what is the mode of action on methane and why is there no effect in premium virus okay and uh i think one um logical one is for us to think just like hypocalcemia why hypocalcemia happens more in multiparous cows than creamy paris so i think it's just a intake and output issue so multiple cows are going to be producing more milk so you have more methionine being excreted so i think that's where you're going to see more of that methionine being needed for the milk protein and not so much getting into the uterus and if you really look to that number uh when you compare multiparous with premie paris the number of the volume is pretty similar it seems like the control multi pairs that was lower that means that they were suffering and not getting enough of that amino acid so i think it's just milky old that is justifying how much is being partitioning i think that's it thanks another one for darren do you know what the relative safety slash toxicity is for the different selenium sources inorganic versus you know yeast versus saline with ironing versus uh the hydroxy analogue of cinnamon that's a real difficult question i imagine it is a difficult question brian um so very simply is that um for inorganic sources there's quite a lot of information about the ld50 and typically ld50s are measured in lab animals such as rats mice rabbits in terms of the ld50 for um inorganic sources ranges between five and eight milligrams per kilogram of body weight um for solenoid finding data is a little harder to come by but generally speaking is that the you know the organic sources sorry are actually less toxic um so i know something about the solenoids is that in terms of i think european is that i think the uh inorganic forms are category two tops toxins whereas the selenium yeast were classified as i think category five so considerably safer so in fact it is this ability of the body to actually store selenium methionine actually makes it less toxic because the body doesn't need to methylate it and get rid of it thanks darren um i think we'll take her and do another couple if that's okay with the audience and with um with christy um how much methane should be given in feed to protect from bad inflammation so i think that what we've seen now is and it's kind of interesting to see the papers and everything that was published and how we design and formulate diets maybe 10 years ago and how we do it today it's a little bit different right but i think we keep to the recommendations that i've mentioned about that association with methionine to energy that we are feeding i think the models they predict energy pretty well uh i'm not sure about protein requirements and amino acids yet but i'm pretty sure that if we tie that grams of methionine to the energy that the model is calculating then we're going to be doing a better job than we did before on percentage of mp or before on just the ratio of lysine to methane so i think we've been growing on how to do that but i think this bad inflammation i think the cow can take care of it if she has the pieces like amino acids to handle and produce the antibodies and all the cytokines and self-proliferation that she needs to do uh if we fit at this point but who knows 10 years from now i may be saying something different but for now that's how we're doing that's how the results you're seeing how we're getting phil i'm sure you'll be saying something different so uh maybe a last one for darren um can inorganic selenium satisfy selenium status and permit sufficient de novo synthesis of selena protein like some data suggests oh no sorry some data suggests that this may be up to 40 percent of selenium proteins end up in milk so the short answer is yeah yeah it's fine so the short answer is that yes inorganic saline can satisfy selenium status the point that was trying to put over is that inorganic selenium sources are actually quite short-lived they're either used or they're lost the benefits that you have with selenium methionine-based ones is one you have this endogenous supply which you do not have with inorganic forms secondly there is this issue of toxicity is that selena methionine is less toxic than selenium salts so although you can say that um yes you get a similar response in terms of de novo synthesis of cylinder proteins is everything goes down that selenite pathway is that in terms of the longer term protection and probably response efficacy uh that's where you see differences between the resources so responses that you see in um organic forms is much more marked much i would say much more rapid than you see from inorganic sources okay and one last one to finish the webinar and phil the levels of methane and lysine you presented relative to me or as a ratio have they been developed solely to optimize milk response and if they have will the recommendations are the recommendation recommendations different to optimize reproduction i think that's uh i think that's a very very good point and i'm not sure if that was or but uh that's grams to me not so much the ratio we still have the ratio but uh i would focus on the grams to the me right and then you adjust your lysine based on on that ratio and make sure you are within that grams to me uh and that's a very good point because uh as i show that little graph there that talks about the efficiency how much i feed how much the cow is going to be using and that's the most efficient that translates to how much i should feed however if you guys follow formula one or any race uh car race we're not talking about efficiency we don't wanna be efficient those guys are going to the limit of performance right so sometimes you may only not want to be on that most efficient you may want to go to the better performance and i think that's where any model fails a little bit to tell us how does that happen we are requiring the minimum what the count needs to perform or to survive right so to be honest i don't have your question right now we are using data from lactating cows to feed dry cows that's what we have now on that ratio perhaps it should be more perhaps it should be a little bit less what you're seeing is this huge difference between control and what we feed and it's seems to be doing very well right but to optimize performance maybe we should be feeding a little bit more or less but we i don't have that answer yet but i think for sure we need to get everybody to the same level where we are feeding that needed methionine that's not a full reality yet in my perspective so we need to get comfortable there and then we can start talking about high performance low performance reproduction and try to see what is that difference there with everybody being fed me finding see who are the highest performers lower performance and we have some more epidemiological data to get started maybe that's a good suggestion good point well thank you so much uh phil and aaron on behalf of the audience i'd like to thank you both for the excellent um webinars you gave and the excellent answers you gave to the the questions um just to point out that if the question was not answered um they will be answered subsequently and so i'll just explain what the next step is in terms of providing additional information there has been a recording of this webinar which will be made available before or on october 24th on the dc rc website in addition it will also be made available on odysseo's feed channel online where it will be posted with a document that contains of today's questions and answers so um without unless egg christie has um anything to add but i just want to remind you all that of the upcoming annual general meeting of agm uh sorry dcrc on the 9th to the 11th of november it's a must i must say you must do so um for all of you that are members please um please if you have not signed up please uh sign up in the very near future so again thank you all for participating and if there's any questions specific that you want to pose to add a sale please feel free to contact christy and she will give you my contact information so thank you again and good morning good night and good afternoon bye [Music]