[Music] welcome folks and thank you for asking me to be part of your program Brian so this evening we're going to talk about the importance and considerations of formulating diets for amino acids and so first of all a few comments about the importance of doing this number one amino acids are required nutrients some are typically more limiting than others and diet can have a rather significant effect on the sequence and extent of amino acid limitation so we're going to talk about all three of those points there first of all yes amino acids are required nutrients there's 20 of them and we typically would classify them according to their absorptive essentiality meaning they must be absorbed in the amounts as the animal requires them which we would call essential amino acids or nany said Jo all ten of those are still required for protein synthesis and other functions in the body however they do not have to be absorbed in the amounts as needed your reason for that is is that the body can synthesize one from another so there's a surplus of alanine for example it can be converted to do the mean or so on and also any of the essential amino acids that are supplied in excess of need can be used to synthesize non-essential amino acids so unless the animals are absorbing an exact optimum profile of the essential amino acids then there will be some in surplus and they can be used to synthesize non-essential amino acids now the primary function of these amino acids now is for protein synthesis however they have a number of other functions in the body as well so let's talk about protein synthesis first now as dairy folks we would think about protein synthesis as the amino acids here needed now to synthesize milk protein and that's obviously going to be a big requirement in terms of protein synthesis in the lactating dairy cow but every minute every hour of the day lots of other proteins are also being synthesized including all of the blood proteins and I've listed a few of them there all of the milk proteins ion channel proteins that be present in every cell within the body receptor proteins messenger proteins all the enzymes regardless of these enzymes have functions and protein or amino acid nutrition but fat metabolism carbohydrate nutrition whatever all enzymes are made up of proteins also tissue proteins okay now that represents protein synthesis represents a majority of the need for amino acids in the body and this is for the concept of ideal balance is really important because every time one of these thousands of different proteins in the body are synthesized it always has the same amino acid composition so a shortage of an amino acid in the diet or that is absorbed that is going to have a negative impact on the synthesis of protein so we never quite know where that's going to be is it just going to be on no proteins or is it going to be in terms of the synthesis of some other proteins amino acids also involved in metabolic regulation there's a lot of different functions of amino acids Theory will mention a little bit of that later and a number of the amino acids are also used to synthesize non-protein nitrogen compounds in the body of which there's about 60 of them and that doesn't represent a huge requirement for amino acids but it's still a requirement me no one else is then that are left over are going to be used for energy they'll be catabolized the ammonia or amino group will be released and the carbon chains will be used for synthesis of glucose immediate use for energy or the synthesis of fatty acids now over the years we've identified at least of the lactating dairy cow here and there are three amino acids that have surfaced as generally being both limiting for protein synthesis that would be lysine methionine and histidine now we really started to understand that about 50 years ago my PhD work was published in 1976 I ran my first amino acid infusion trial in 1971 and conducted five experiments at that time to sequence amino acid limitation in lactating dairy cows and our approach here was to infuse individual amino acid combinations of amino acids and into cows that were receiving just grain and forage diets no protein supplements of these diets were between ten and a half to twelve percent protein we wanted to know what the sequence of amino acid limitation was in terms of the impact on milk protein content milk protein synthesis without protein supplements and what we learned and was that lysine methionine were the first two limiting amino acids and in fact 42 percent oh the total response we got by infusing casein which would represent all 20 of the amino acids or the 10 essential amino acids 42 percent of the response we got either way there we got just with lysine signing so that gave us really clear evidence said hey yes lysine methionine are the first two limiting amino acids and then if you look at them you know as a composition of proteins you would know that once we start adding those to the diet we're going to extend the extent of the limitation of those because those protein supplements are typically alone lysine methionine or both and we'll take a little look at that in just a minute and then a more recent years histidine is surfaced as a potentially limiting amino acid after lysine methionine especially when we're feeding lower protein diets or lower RUP diets okay so let's see if we can make a little sense out of this this table shows the essential amino acid profiles in milk mixed room and microbes that would be a combination of particle and fluid associated bacteria and protozoa and some common feeds so the first line there we have the essential amino acid profile in milk now let's take a look at the essential amino acid profile in mixed room in microbes now if you add up those numbers across or within a line are gonna add up to 100% so we're looking at each essential as a percent of total no interesting they and as I guess we would expect right the amino acid composition of mixed room in microbes is pretty close to that of milk not exactly the same but it's probably pretty safe to say that at least on the lactating dairy cow the initial started trying to establish requirements would be to look at the amino acid composition of Melkor now I've highlighted in red those amino acids within mixed microbes or within feedstuffs here that are 20% or more lower than those in milk and that's indicated in a footnote there so I did that so it would be easy for you to identify those amino acids here in the table that are at least 20 percent lower than their concentrations in milk now if we look at mixed room in microbes there's two amino acids that come up here one is histidine and one strip to fame and as I said just a minute or two ago if we're feeding lower protein diets as they've done at Penn State in a few other places now where they've enriched the diet with room unprotected lacy a Mathai named fed lower bypass protein diets or a mixed microbial protein is providing a majority of the metabolizable protein to the animal histidine can come in as a third limiting amino acid so looking at that value there that's probably not surprising and a notice that that value for histidine in mixed room and microbes tends to be lower than in most feed stuffs one exception there with the feather meal at the bottom of your table so now let's just take a look at all the red values in the table and if you scan down each column you're gonna see more red values for lysine than any other amino acid meaning that the RUP fraction of all of these feeds it's gonna have a lower significant lower content of lysine than in mixed room and microbes today so it's pretty easy to see why lysine as a limiting amino acid because RUP from these fees will make up 30 40 45 hopefully not 50% of the total metabolizable protein that your college receive so there's a lysine issue if you look at the values from athenian you'll notice a lot of red values there isoleucine also comes up in a few fees i barley like corn like canola meal like corn gluten meal like cottonseed meal and especially blood okay and then you're not gonna see very many other red values there in the table other than for tryptophane and tryptophane looks like it's it's lemon tea in there and mixed room and microbes corn silage is really low other corn products is really low as you'll see with DDGS and corn gluten meal okay now tryptophan is an amino acid that we really haven't taken a close look at yet in dairy cow nutrition but this data would suggest that it certainly could come in as being Cola meeting or limiting certainly right after lysine Matheny now other amino acids other than Laci Metheny and histidine can become limiting and lactating cows in ways other than his building blocks for protein and there has been a lot of work published in terms of amino acid metabolism of the mammary gland and other tissues over the last 15 to 20 years and dr. mark Hannigan at Virginia Tech he's been one of the leading researchers in this area really looking at what is the impact of some of these other amino acids on stimulating protein synthesis not only in the mammary gland but some other research groups looking at that in other body tissues as well so what these researchers have shown is that some amino acids like leucine isoleucine as well ESMA thiamine and histidine can have or do have potential roles in the body other than as building blocks for protein in that they are stimulating in various different ways protein synthesis within various cells or within various tissues or organs so many different types of in vitro experiments and using mämmi tissue have been conducted to try and better understand what's going on in the lactating dairy cow why do some of these amino acids tend to stimulator what other amino acids do stimulate protein synthesis to make protein synthesis more efficient so just in the last couple of months Peter Yoder just completed his PhD and down at Virginia Tech and conducted a really excellent study that really wants to take a look to see whether or not the cows would respond in a way to a few of these amino acids in ways other than as building blocks for protein so it was a really excellent study with eight high producing dairy cows had to buy at two four by four lamp squares and so for treatments for animals for experimental periods within a lab square and then that was replicated using a total of eight cows so the treatments were control where the animals didn't get any amino acids lysine and methionine and histidine then were infused as one of the treatments isoleucine and leucine were infused as a third treatment and then the five amino acids were combined and infused into the animals as a fourth treatment so amino acids were provided by jugular infusion and the daily infusion amounts are given here and they came up with these amounts of because they did want to feed a low protein diet as indicated in the last sentence there was 15.2% crude protein and according to the latest version of CM cps then with that diet the model was predicting certain degrees of deficiency of a number of amino acids including these five and so they basically back to fill those deficiencies by infusing these mounts of these five amino acids so note that the primary ingredients here at corn silage alfalfa hay corn grain wet Brewers grain soybean meal citrus pulp and soy health so a pretty simple diet but yet not an uncommon diet at least to us here in North America in terms of production results and this is all the only slide that I'm going to that I got here on the subject to show the results of but which is milk effects on milk protein content and milk protein yield responses so as we would expect these cows were beyond peak lactation and you Pro and you provide those animals with additional lysine methionine and histidine we would expect to see some sort of a bump in milk protein in fact there was a significant increase in milk protein as you'll see in the second bar graph there no effect on milky oh that's not surprising we often would not see an impact on on mochi oat but because of that significant increase in normal protein content there was a significant increase in milk protein yield now notice that with isoleucine and leucine had no effect on milk protein content so as we've been kind of looking at things over the years we would have said well isoleucine or leucine we're not limiting amino acids okay there was no impact on both protein content however there was a significant increase on milk yield so you multiply those two values together and you get a significant increase in about protein yield with the 5 amino acids now what's really exciting here is that when you combine those 5 amino acids together now you get the milk protein content response back you retain the increase in milk yield and now there's an additive effect in terms of the impact on milk protein yield so here we have two groups of amino acids that are independently and additively increasing milk protein you and as the authors have appropriately stated this then creates doubt about what we have been thinking as the single limiting amino acid theory in other words amino acids can be Co limiting but in different ways having different impacts on the mammary gland secondly the importance of formulating diets or amino acids is that achieving a more optimum balance of absorbed amino acids reduces the risk of a singular or a small group of amino acids being more limiting than other amino acids or other nutrients as well as increasing the efficiency of use of absorbed amino acids for protein synthesis and other metabolic functions so here we have we have essential nutrients we know it's important to formulate form to provide the animal of good balance and by providing a good balance we're doing two things okay we're minimizing the possibility that those most limiting amino acids are going to be limiting in the diet and we're allowing the animal an opportunity to increase efficiency of use of that of those absorbed amino acids so the result is going to be an improved animal performance and a reduced need for supplemental RUP now we understood the importance of amino acid balancing from the swine and poultry literature back in the 60s and early 70s so dr. rule ken in France way back in the later 70s 80s began to infuse lysine or methionine into the abomasum or into the jugular vein of lactating dairy cows and his goal was to establish what is the ideal profile or amount of lysine indigestible protein or ideal amount of digestible Metheny so then in later years as they designed a model that would predict amino acid flows to the small intestine and could predict the concentrations of lysine or Matheny or other amino acids in a digestible protein they took that information and run all of those diets through their model and so what we see here is a residual yield response plot and mathematically it was determined that the ideal amount here of lysine and digestible protein was 7.3 percent of that digestible protein and so they took their other set of studies and it wasn't necessarily that they conducted all the studies but they certainly did conduct a majority of those studies and and and here they looked at infusing incremental amounts of methionine across a number of different studies note in the footnote 265 cows were involved in this data set and the optimal amount of my finding then in in the digestible protein was 2.5% so just remember that 7.3 go back in 2.5 in some years later we at the University of New Hampshire ran a number of these studies ourselves some other people here in the United States did and so as a member of the NRC committee that was charged with introducing an amino acid model then you know within NRC I needed to do something similar to this so we took the data that we had and we went through the same set of we used the same procedure to establish these residual plots but being able to combine the data and we came up with an optimum value of lysine of 7.2 very close to what dr. rule cannot come up with in terms of the European or the French model using a different data set and using a different model for predicting content of lysine in mp and at this point we did this with a beta version of the NRC model and we took our methionine studies and we came up with this plot so 7.2 and 2.4 value is very similar what to what dr. rule Ken had come up with then after the NRC came out and there was no more changes being made to the model we redid all of our analysis here and notice our values for lysine of athenian dropped a little bit from seven to down six point eight two point four down to two point two nine but still very close to a three-to-one ratio and then there were some revisions made to the model a few years later that really didn't change things a whole lot and then several years later and we shared that at a DSA meetings and 2009 we took the very same studies the very same studies and evaluated the diets and everything was CPM dairy so we could arrive at these optimum values for CPM dairy notice that the values for both lysine and methionine were higher than NRC so that represents model differences folks that does not represent any differences in the data set that was used and then as the more recent versions of CM CPS came available we did the same thing with those models and you'll see the values here so anyways those are the target values that we've come up with using the same data set now what's the benefits and of using that information what's the benefit if we try and optimize lysine Metheny nutrition using those values that I just gave you well what we've learned is that we definitely can reduce the risk of a deficiency of the most limiting amino acids which would be lazy animal finding here piggy every shorter responses and early lactation cows in terms of milk eel would be about two to four kilograms more milk some studies have shown more than that number of studies less than that but for early lactation cows where we start supplementing these amino acids at the time of calving or before calving that's what we can pretty much expect to see obviously that's going to depend on what the diets were are diets that were fed prior to the initiation of amino acid balancing increase milk components especially milk protein point 1 to point 2 percentage unit increase sometimes we see point 3 we see in point 3 increases both in research and in several dairy farms across the US and 60 70 percent of the time an increase in butter fat and lastly healthier and more productive transition cows and if you watch the presentation by dr. Osorio he was sharing data or they fed diets that tended to have higher concentrations of lysine in metabolizable protein and then supplemented those transition cows and with a supplemental Matheny and if we were to summarize not only all the data that dr. asorio presented but other data as well this is what happens when supplemental Athenian is provided to these transition cows so these cows would have been Metheny deficient they were receiving diets that had higher levels of lysine and metabolizable protein not as high as most of us would like to have seen so we really feel that some of these benefits of Matheny supplementation could have been greater than what they were but nevertheless these are most of the benefits all the way from increased feed intake to having an impact on health immune function inflammation reduced oxidative stress large him a larger Imperial to decrease pregnancy loss in a few studies there is more studies going on in this area and what's really exciting I think is the impact here on the offspring offspring born of mothers in which supplemental defining was provided to the dams over to the mothers okay so that gives us some evidence here that improve Matheny nutrition and probably as we look deeper in the future on improved lysine nutrition history and nutrition we might see some of the same things there the tactic can have then on fetal programming there was some older data that existed most of these studies would have been conducted in the earlier 1990s as a Genna modal was evaluating one of their early versions of a protected lysine supplements so they were using that in line or along with smart M&M just to see if they added some Lacy and Matheny two very fresh diets what would be the impact on lactation Oh performance and across those early studies the average yield and milk protein was 3.8 kilograms now lacing has I think really become increasingly more important in dairy cow nutrition we're feeding more corn silage unless a human grass forages certainly here in the US over the last 10-15 years corn silage is low and lysine often replaced soybean meal products with canola meal because of price well canola meal tends to have a little lower concentration of lysine and what soybean meal does blood and we always fell out of favor for some producers which has a really high content of lysine often we like to feed distillers grains and other high-protein corn products in place of other protein supplements because they're less expensive okay they're low in lysine and we're probably getting a little more aggressive in recent years with feeding room and protective Athenian supplements so we may very well be hitting the ceiling quite quickly in terms of not having an adequate supply of lysine to really make use of the RP methane supplements that we have available to us we've also got a greater the desire to feed less RUP than before we're also learning that lacey may not be as available in our feed stuffs as we thought in other words maybe some of our feed stuffs has a lower content of lysine than we really thought they had and that could very well be true for something like our corn our corn that we feed our animals and soybean products I was just talking with dr. Tom Stein University of Illinois a couple days ago and he said a trend that they've seen is reduced lysine an amino acid concentrations in both corn and soybeans and he thinks it could very well be a result of the increased yields that farmers are getting in terms of bushels of soybeans per acre bushels of corn per acre lowering the amino acid content of those feeds and we know that milk and milk protein yield is very closely tied to absorb lysine supplies lysine basically only has one function in the body and that's protein synthesis it does have some other functions but they represent a smaller part of the total need for lysine so just remember lysine as primary function is for protein synthesis and just to kind of drive home this point back years ago when I was at UNH way before I ever left you an H we had a few students that wanted to do senior project or they started a master's program and wanted to do an initial study and so they so they took a look at supplementing diets was smart I mean ml that was the original smart immune ml that had to say I had put some smarter mean m in the diet as well to make sure that smarter mean M was that supplemental math I mean what's where we thought it needed to be and so that really created a somewhat greater deficiency of lysine but these were well these cows went on trial at of a HANA average 106 days in milk and notice the impact of that supplemental lysine had on Mielke a really push milk eel as well as increasing milk protein content the next experiment was a smaller experiment only involving five cows but nevertheless we saw the same effect we set the study up the same way adequate Metheny in at least what we thought was adequate I mean at the time and then brought in incremental levels of supplemental icing and in this trial here we did not use smart Ameen ml but here we did use infuse Metheny to create those concentrations those increased concentrations or supplies of lysine to the animal but the diets did a smarter mean m in them so just making a point here that one of the reasons here for amino acid balancing Naza to improve lysine and methionine status in particular we'll be focusing on other amino acids in the future I'm sure but another reason for doing it is to increase efficiency of use of metabolizable protein and here I've gone through an exercise to show you that there's different ways of achieving a hundred and eighty grams of metabolizable lysine and 60 grams of metabolizable Matheny so if you happen to be using the NRC model and your cows are given roughly 90 pounds of milk with three one three two percent milk true protein test that's about how many grams of lysine and methionine those animals are going to need now you can achieve those hundred that 180 grams of lysine of 60 grams of methionine several different ways you can feed a high corn DDGS diet which would be the first diet in your table low content of predicted lysine of metabolizable protein fairly high content of lysine of metabolizable protein at 1.9 no me know ask the supplementation there that is a high high corn diet now we know that a diet like that works out in the field however we also know that you have to feed more RUP the dietary crude protein content of that diet is going to have to be higher and if you go through the math as I've indicated here in the footnote notice that that diet according to these calculations require about 8.1% of dry matter as RUP now you add that to an RDP requirement of about 10% of diet dry matter and you're up to an 18 percent protein diet or you can go the other way and say you know I really want to push lazy and athenian i want to optimize my diets i want to improve efficiency of use of metabolizable protein so I'm gonna shoot 46.9% lysine an MP and 2.3 percent meth I mean an MP notice that the MP requirement now fell from 30 157 grams in the second column all the way down to twenty-six hundred and nine grams that is a huge saving on metabolizable protein and that's all that saving is going to be on RUP you don't want to do anything to negatively impact the animal's ability to provide metabolizable protein to herself from mixed microbial protein so now if you go all the way over to the right hand column you go through the calculations there that allows lowering RUP in the die 25.4% so what do we have we have a diet here of 5.4% RUP our DP is gonna be around 10 maybe a little bit higher than 10 depending on what works best for you and you're in a 15 50 and a half percent protein diet so a tremendous opportunity here to save feed costs on the purchase of supplemental RUP and we've done it through improved efficiency of use of MP now we we knew this to be true years ago after the NRC came out we put a number of diets through the model and we just looked at the difference between actual milk in these cows and MP allowable milk and interestingly as the model predicted increasing amounts of lysine and metabolizable proteins over time and so we pick studies here that had different types and amounts of protein supplementation notice that if lysine an MP was really low on average cows gave a little less milk then what the model says was MP allowable but as we increase lysine in metabolizable protein now the cows began to give more milk than what the model said via MP allowable milk okay and the same thing with Matheny so when Matheny was low cows tended to give less milk and what the model said the cows oughta give via MP lol no but as the amount of predictive Athenian MP increased the cows gave more milk than MP allow and I'm just gonna show you one study out of many examples from the literature of reducing dietary RUP by amino acid balancing and this work was out of the University of Wisconsin they fed their control diet what they call the traditional protein diet corn silage alfalfa silage hi boys your corn and soybean meal distillers grains some expeller soybean meal okay so some complementary proteins and their diet was sixteen point nine percent protein NRC model predicted that diet would provide a hundred and sixty grams of MP lysine then they formulated in a lower protein diet they took the distillers out expeller soybean out increase almond extract of soybean meal put in a little high moisture corn Lord protein down to 15.7% notice that the model Oro was calculating the same number of grams of MP lysine 161 grams but the predicted flows of MP methionine fell a little bit and the lysine of athenian ratio increased so the whole goal was here now was to backfill that deficiency of Matheny with some supplemental finding sources in this case he's met a smarter smarter to me now well they use both as two separate treatments and increased the methionine supply from the original 48 or what was in the low protein diet 45 up to 54 Sol from an Amazon from an amino acid standpoint we have an improved diet don't we we did not lower we did not lower model predictive flows of MP lysine but the cow is getting more meth I mean via an increased lysine of a thiamine ratio and if we go right to the results here and I'm just gonna show you a few of the results but just go to the bottom of the slide and notice MP balance so with the traditional protein diet that was designed to be basically have an MP balance of zero it's very close to that but then when they pulled out some of the protein those animals then became negative in terms of NP so the mom was saying these cows are gonna give less milk because they had a negative MP balance well the model doesn't correct for NP requirements based on amino acid balancing okay so now if you look at milk protein yields we or if you look at mo kio there was no differences amongst those treatments milk protein content was actually higher with the last two experimental treatments there we've got a smart smart amino turns a yield of protein and America Lee those values were higher for those two treatments as compared to either the traditional protein diet or a low protein diet nope nitrogen to feed nitrogen risos improved so the bottom line here is the animals on the lower protein amino acid balance diet here did just as well if not better than that with the traditional protein diet even though the model says would say that those cows would not do as well so there's some take-home messages here reduced RUP feeding saves cost for purchase protein supplements reduced RUP feeding provides an opportunity to feed more the rest of the diet which would be expected to increase microbial protein synthesis and really if you think about it amino acid balancing is the only option we have for selectively increasing efficiency of use of RUP so you have to decide as a dairy producer as a nutritionist do you want to try and improve efficiency of use of RUP or not is it to your benefit and the last point here just to remember is most recent formulation models do not adjust model predictor requirements for empty as a result of amino acid balancing which means you cannot pray you cannot trust your model alone on predicting milk yield based on supply of metabolizable protein and if you're using a good model you should be able to consistently run negative MP balances as a result of amino acid balancing so how can we be successful here and this is really where considerations come into play number one you've got to let go of balancing diets recruit protein and instead balancing for rumen degradable feed protein rdp and the most limiting amino acid as a dairy producer as a nutritionist you should know in every herd what is the rdp content of your diets is it ten percent nine and a half percent of diet dry matter with the model that you're using is it 11 eleven and a half but that's the value that you should always be watching okay because that's a requirement for room and microorganisms and the cow novel requires amino acids our DP is rumen microbes amino-acids n in terms of metabolizable protein and amino acids contained within that that's what the counties now we've known for a long time that the relationship between the crude protein in milk or milk food protein yield and crude protein intake that relationship is very very poor now we've known that ever since I went started graduate school in 1969 I was a very poor relationship and then over the years as we introduced the metabolizable protein concept you can see here not that the relationship between milk protein yield and model predicted MP supply is somewhat better than with crude protein so some improvement there now if you go to the bottom of the slide you can see the relationship between actual protein yield and model predicted flows of methionine per model predicted flows of lysine notice how much tighter excuse me those relationships are then with metabolizable protein also notice that the relationship here between MOU protein yield and model predictive flows of lysine is even stronger better than it is with Metheny that goes back to the point that I made lysine has one primary function in the body and that's as a building block for protein the animal doesn't have enough she cannot make that no protein you supply her more and that's a limiting nutrient she will produce more and so we knew that and because the NRC model did not predict requirements in terms of grams per day we after the NRC was released we ran a lot of studies a lot of diets through the NRC model initially we ran three four hundred diets through with different production levels different amounts of protein in the diet because we wanted to look at the relationship siri between actual milk and milk protein yields relative to model predicted flows of MP and pipa thining MP lysine and then some years later we added another four five six hundred diets and we improved our relationship but I don't have time to get into that other than to say just like you saw in the previous graph protein yields were more accurately related or predicted from predictive supplies of MP Matheny and MP lysine then from metabolizable protein and again this all makes sense because what the cow requires are the individual amino acids the cow does not have like an absolute requirement for metabolizable protein okay it's the individual amino acids that she needs achieving success also requires us to understand that the digestibility of RUP is important or that the digestibility of amino acids in the RUP fraction of feedstuffs is important and that is not a hundred percent dr. Mike Vandenberg talked about that he made a big deal in his presentation and he actually showed you a study where they depressed our euphy digestibility of blood meal and showed us the impact that doing that had on animal performance so RUP digestibility here is important and we need to model our protein supplements and our feed stuffs make sure that we get our RUP digest abilities that's close to what they actually are as possible okay but we also need to understand that there's differences in the mean acid digestibility there is differences in the digestibility of the individual amino acids that are in feedstuffs and just very quickly I want to show you some data that indicates that and I got this data and I'm giving you this information this is dr. Tom Stein's website at the University of Illinois and if you go to his website you'll see that he's got an in feed in a feed ingredient database so if you click on feed ingredient database you're going to find lots lots of spreadsheets there lots of spreadsheet join the nutrient composition of different feed stuffs as determined in swine but the reason that I'd like to have you go to that website and just take a look at what they have reported there on ileal digestibility values of selective feed stuffs as determined in pigs and they have the same set of data for poultry now why am i showing you this well if we look at dry Brewers grains here notice the digest abilities of the individual amino acids are all pretty close together I think I see a high there of 92% or 95% for arginine and a low of 82% for Laci now notice that lysine tends to be a little bit lower on average the average here for dried Brewers brains is 87% no what does lysine is 82 go to color or corn yellow dent corn now on every digestibility as determined in pigs is 82% across all 10 of those amino acids but notice lysine as the lowest the values that I've highlighted in red here are at least 10 percentage units or 10 percent lower than the average so that 74% is 10% or more lower than the 82 okay so I did that just so you can see very quickly here just perusing the table which amino acids tend to have the lower digestibility values corn distillers grains lysine is also low again seventy-six instead of 81 but the value the amino acid having the lowest value is tryptophane okay so you can just look at that table the point is here is that there are differences in amino acid digestibility as across feeds and there's differences within feeds for the ten essential amino acids this table shows a comparison of digestibility values for canola meal rapeseed and some soybean meal soybean meal we've recognized over the years tend to have very high digestibility as you'll see the average digestibility values therefore those two soybean meal samples it's 87 and 86 percent and pretty uniform in terms of digestibility across those ten amino acids however if you look at canola meal you will see the average digestibility values a little bit lower there then for the soybean products and that would include rapeseed meal as well okay threonine tends to be pretty low in that one canola meal sample the one on the left but that's really the only amino acid that stands out here in this table is being but I would say significantly lower than average and here's a comparison of some digestibility values of animal protein supplements so so blood cell spray dried blood cells blood meal feather meal hydrolyzed feather meal with blood or without blood and then a really high-quality fish meal high-quality fish meal as we would expect would have very high digestibility as we wouldn't expect to see much of a difference in digest abilities amongst amino acids excuse me the blood cells are sprayed right okay heat was applied it was a very low digestibility value therefore isoleucine the next blood meal sample also lower price of leucine than the other amino acids a feather meal tended to be a digestibility value for histidine and lacy on average we're lower so if you go across all these lysine kind of comes out as that one amino acid that tends to have a lower digestibility value as compared to the average of the other amino acids I'm emphasizing this in part because we know that heat now I can damage have a negative effect on protein digestibility and amino acids availability in a lot of the feeds that we feed in the room and Industry including dairy cow who have gone through some sort of a drying process here in the Upper Midwest last year almost all of the corn that was harvested for dry corn would have gone through a dryer what effect does that have possibly on lysine digestibility we really don't know we do know that there are three different types of reactions at a current feed as a result of heat processing the protein cross-linking reactions would be the type of reaction that we would see in something like a blood meal product for example that is primarily just protein doesn't have a lot of carbohydrate or any carbohydrate in it so any of the depressed digestibility there would be as a result of protein cross-linking reactions and the Maillard reaction as being the most important reaction that would occur in other feed stuffs so lysine is the most vulnerable here because it has that reactive epsilon amino group and that's the group then that's going to get tired up either with sugars reducing sugars in the case of the Maillard reaction or in the protein cross-links in higher protein containing feeds and just very quickly sir Boucher when she was at the University of New Hampshire for PhD work she used the rooster si out of the University of Illinois and actually did that work down at the University of Illinois with dr. Hans Stein down there and she wanted to take a look at the impact of heat treatment of two different sources a solvent extracted soybean meal expeller soybean meal and then four sources of distillers grains those are the feeds that she collected and then she took one sample of each of those and exposed them to some additional heat the conditions of which are described here in the slide okay and the reason for doing that was not only to look at the impact of heat treatment on depressing digestibility in the RUP fraction of feedstuffs here but when RUP digestibility is depressed does that have a greater effect on lysine than on the rest of the amino acids and that's exactly what this slide shows ok the if you go to the right look at total amino acid is the first 4 bar graphs or that would be the protein supplements without any additional heat applied then you apply some heat you get depressed digestibility in that one soybean meal sample but notice if you go to the left placing digestibility was depressed more than RUP digestibility okay and then she took the other sample there applied he got a depressed amino acid digestibility value but lysine digestibility was depressed more and she also looked at four different samples of DDGS and she took one of those samples and applied additional heat and noticed an really pretty serious negative impact on lysine digestibility one of those DDGS samples already had a depressed lysine digestibility as you can see it was not necessarily picked up if you just looked at total amino acid digestibility but if you looked at lysine in particular yes it wasn't eat rest I'm making this point because we tend to feed protein supplements that have been heat processed and there's always that possibility here that lysine digestibility is less than the other amino acids or for the other amino acids in that RUP so just be mindful of that and and this is a figure that I'm sure that most of you have seen some work by dr. Norman st. Pierre years ago where they looked at the influence of depressed RUP digestibility in blood meal and the impact it had on our pea lysine digestibility so far yupi digestibility of the blood meal is really really high than the digestibility of lysine is high but as RUP digestibility goes down in blood meal samples notice that lysine digestibility continues to be depressed even more so if you focus at an RUP digestibility of 40% there the lysine digestibility on average was 20 okay so these are just things that you want to keep in mind if you want to have success in terms of the amino acid formulation always be thinking about what is the lysine availability are the lysine values for my feeds in the feed library is that correct does that represent the lysine levels that I have in my feed stuffs number three in terms of amino acid balancing really important here to use reliable estimates for the bioavailability if you will of amino acids in our our you in our room and protected amino acid supplements very important because if we're going to be successful here with amino acid balancing we're obviously going to be employing the use a room and protected lysine and methionine supplements and so do you know do you really know what the bioavailability of lysine or methionine is in the products that you're feeding is it what the supplier tells you that it is or is it something less than that and we became we started getting concerned about this and back over 20 years ago at the University of New Hampshire we really felt that this whole concept of amino acid balancing and mic dating dairy cows was gonna progress and we'd probably always be behind that a swine or poultry but nevertheless there would come a time when people would be interested in feeding less bypass protein lower protein diets and so amino acid supplementation is gonna be getting more and more important so we kind of figured that more and more products would be coming out on the market what methods are available to evaluate these products well I've listed a number of the methods here and I've listed the shortcomings and we considered these different methods but nevertheless they're in vitro or there and said two methods with the exception of the last three the plasma free amino acid area under the curve well this is where you put into the room and a pulse dose may be 50 grams of signing or a hundred or 120 grams of lysine and then you take a lot of blood samples and you look through the spike in lysine or the spike in Matheny and the blood plasma and then you calculate the area under the curve so you're doing that with either and fuse lysine or Matheny or by providing a product and so the area under the curve then that gives you an idea of then of how much of the lasing of methane was being absorbed okay but you're relying on the blood so it is an animal bio acid however that's not the way we feed these products and so we really didn't feel that this was a good method going forward for the future we could use something like The Milk Protein dose response method but the problem there is we have to make sure that we maintain amino acid deficiencies and all cows over the entire range of the dosage is used and that's always impossible to confirm so what biomarker can we use in which increased absorption of an amino acid will reflect a change increased amino acid absorption may not reflect a change in milk protein of the cows are not deficient in that particular amino acid however we learned early on that those animals will respond with an increase in either lasing or methionine or whatever we're you know providing those animals so what's the shortcoming of that approach well really not a whole lot other than the fact it takes room and cannulated cows and as long as we can demonstrate beforehand that the responses between increasing amounts of the absorbed lycée of Athenian histidine or whatever results in a linear increase in blood plasma concentrations then we've got ourselves a biomarker so we really felt that a robust animal based method was needed we wanted a system where we can put the products right into the diet we can mix it in with the diet whatever not because that's an important part of the whole evaluation process and we know it's just difficult and we realize way back then it's very difficult to protect these and these amino acids from ruminal degradation and yet have them be available for intestinal absorption not just avail but be digested and be absorbed so the decision which product that you folks buy should be based on knowing not only the cost of the product and not really what the supplier may or may not be saying as a vine availability but what is the actual bioavailability value as determined under experimental conditions using a standardized protocol and the method needs to be one that you can use for liquid products like element for example or liquid notice more now I'm sort of a surprise I think to the industry because nobody had really done this kind of a study before and this was work out of minor Institute where they looked at the effect of on-farm feeding practices on the bioavailability of lysine and room and protected lysine products I'm just gonna show you the results here of the first study and what they did in this experiment and they had actually two experiments in that particular study but in this experiment their goal was to look at well how much lysine is released from six different room and protected lysine products when incorporated into a total mixed rations that differed slightly in dry matter content and those dry matter contents are indicated on the right hand side of your slide ok so what we're looking at here is the loss of lysine in the total mix reaction okay this is not lost in the rumen this is lost in the totem extraction how much of the lysine and those products were they able to recover in the TMR after those products were exposed to the TMR for 618 over 24 hours and these results were pretty shocking to some people but the point is it really demonstrates here is something and that is we have to consider in evaluation of these products the exposure of these products to the diets that we're feeding our animals if you feed one group tmr's and you're putting your amino acids in those one group tmr's then you need to have some understanding of what is the stability of those amino acids in that TMR all the way out until the cows are fed again which would be 24 hours later so this is very important data so with all of that behind us then I'm just going to explain very quickly the dose response method that we use at the University of New Hampshire and so we use high producing dairy cows and Nancy Whitehouse at the University of New Hampshire she's been involved in all of these trials and over the last 20 plus years so we've adopted a very standardized protocol what's important here is that these supplements are put into the TMR before feeding so they're exposed to the TMR eight hours before feeding cows are milked at eight-hour intervals their milk at 12 hour intervals I mean they're fed at eight-hour intervals and then blood samples are collected last three days of each period so very standardized protocol so the experiments are set up in such a way that we've reached steady-state the animals are are consuming these products three different times over the course of a day the products are being exposed to the TMR here's the results of one study where we evaluated the first generation project product AG Pro L so in this particular study we had six treatments animals and this would have been six by six land square okay and so one treatment the animals didn't get any lysine one treatment animals receive 30 grams per day of infused lysine third treatment 60 grams per day infuse lycée and that's continuous infusion over the course of a 24-hour period for seven consecutive days and the fourth treatment animals receive 30 grams of lysine from a G Pro mellow 45 grams and 60 grams so three different levels of a G pro were fed their notice linear increases in plasma lysine for both feeding the product as well as for our infusion treatments so if you take the slope of the dose response line there for the AG Pro treatments that slope is point zero zero six eight divide that by the slope for infused lysine and you get a bioavailability of 37 percent so this is the approach okay use the same approach for every amino acid that Nancy has evaluated so this is an evaluation of the 2g and the third generation product and infusions were also part of the treatments here we evaluated smart Ameen ml this was the old smart amine ml that at assay or head so we wanted to evaluate the lysine and that up against infusion and another supplement and you see the bioavailability is calculated at the bottom of your slide this shows the bioavailability of lysine and the new smart amino ml product and got a bio availability of 80.8% so Nancy summarized for me all of the studies and the average by availabilities of all of the lycian studies so what you see here the number of products that were evaluated or the number of tests made in some cases one product might have been evaluated more than once and on the x-axis we got the calculator or the predicted bioavailability values using this technique so notice that the bioavailability values for most of the products is pretty low not surprising the two values there of 80 and 85 percent is the bioavailability of lysine and the new smart amine ml product same approach used to evaluate room and protected Metheny products and this shows an evaluation of smart amine m and bioavailability here eighty two point eight percent notice the very low standard error bars so as nancy has gotten better and better in terms of refining everything that she does more and more precise estimates of bioavailability here nancy evaluated smarty mean m coming out of their older plant their new plant no differences up against me prawn and amino sure room and a more recent trial here doing a comparison of bioavailability z' between smart amine am amino sure m XM i should say betty pearl and Timnath and a comparison of bioavailability zone Matheny and smart Amina and ml again getting bioavailability values of eighty or slightly higher in those two at a sale products the technique can also be used to evaluate the analog products first thing we had to do was demonstrate that we got cleaner increases in total sulfur amino acids plus the analogue itself as measured in the plasma as we infused increasing amounts of the analogue okay and here's a first tab at the evaluation of meta smart bioavailability here in this trial of 50.3% and here is a summary of the studies the Matheny studies that Nancy has done so at the end of the day why are we doing this well let's use methylene products as an example so currently in the US we have a number of different Matheny products okay we know what their methionine content is we know that the question is what's the bioavailability values do you want to see independent determinations of those bioavailability values well if you go back some slides you'll find values for most of these products then you can put them into this table then you can take column 1 and column 2 and multiply them together and that will give you the metabolizable refining content within that product okay and then from that you can determine what is the relative feeding rate relative to smarter me down I don't think anybody would disagree smarter mean them as the highest bioavailability value of any of the protective Athenian supplements so we'll give that a 1x so do you have to feed two times more three times more let's say of me prawn to deliver the same amount of methionine well the data is here so you can go through those calculations and see what the relative feeding rates are so if you know the relative feeding rates and you know the cost of the product you can make some informed decisions here about which product you wish to feed so take-home messages here don't feed any of these products we would suggest you don't feed any of these products unless you seem confirmed estimates of bioavailability using the amino acid dose response method as described make sure that when you see those results that there's confirmation that those amino as the supplements was mixed and consumed what the rest of the diet during the study do not accept comparative milk production data as proof of claims of amino acid bioavailability and suppliers really like to show you that data show that both products their product and a superior product resulted in the same level of milk protein and so therefore the product is just as good however such data always favor is inferior products why is it well because cows respond to increased absorption of an amino acid only of that amino acid is the most limiting factor for production otherwise the cows won't respond okay therefore if you're going to use the production data and a comparison of different sources of that amino acid all cows fed the best product must remain deficient in that amino acid after supplementation and no other factor be it heat stress some other dietary factor can be limiting production well that's absolutely impossible to achieve so that isn't the case and you can't do that with any degree of certainty then the superiority of the best product won't be seen an inferior product relative to the superior product will look better they are and I'm just gonna show you one example here are four products okay so let's look at Milk Protein we got our negative control and then we're evaluating four different supplements looking at effects on milk protein without product a has a value of three point four four percent no to protein product C three point four six you would say those two products are not different right no protein concentrations or the same 3.44 3.46 however if you go to the bottom of the table take a look at plasma Medina concentrations and plasma total sulfur amino acid concentrations product a the value is 102 point 5 yes higher than the 95 point 4 but that's an increase of only about 7 units of total sulfur amino acids in the blood now go to product C notice the value there is one hundred twenty two point six that is significantly higher than the 102.5 value and if you just go through the math here you would say that the bioavailability of product a is probably somewhere around 25 to 30% that of product C okay so if you look at the milk performance data in this case milk protein content you say those products are the same when in fact they're not the same achieving success also means using correct model inputs and dr. Mike Van Amburg he explained that it's really important to make sure that all your production and animal inputs are correct also make sure that all of your feet' inputs are correct if you want your model to be as close as possible to being right you got to make sure that all of your inputs are correct and lastly it's really important as we get more and more into them you know asset balancing to use raishin formulation programs with optimization capability especially those that will give you model compliant solutions so this is kind of a big deal you still want to make use of your guidelines or a targeted profile of absord amino acids in mp as well as for targeted Graham supplies relative to some model predictive supply of those amino acids up against available energy and dr. Mike Van Amburg talked about that as well and you see that in the right-hand part so you want to keep these values in mind so just a few quick comments and then we will wrap up your comments regarding andrey formulation of diets through optimization too many people think of me no asset balancing is too costly and therefore that's why they don't want to do well the reality is it doesn't have to be that way we now know that implementing amino acid balancing by using ruin'd protective Medina supplements in combination with higher lysine containing protein supplements or are using a combination of room and protective lacing and methania supplements together does not always have to significantly increase diet cause it really depends on what you're already feeding in fact you may be paying for the cost of immuno acid balancing but you're not amino acid balancing and you're not going to know unless you look at an optimized reformulated diet so the approach here would be know exactly what your current diet costs are know that then reformulate that diet to optimization to get a least cost or optimize reformulate a diet that supplies the same amount of the most limiting amino acid which is usually going to be Matheny now when you go through that process make sure that you consider all the available feeds and supplements maybe you'll find that as a result of optimization that hey the model is pulling in some feeds and little higher amounts or maybe a little less of this or that then what you thought was really optimal and you're able to save some money maybe you're feeding a surplus of something and now you realize oh maybe I really didn't need that much rdp in the diet I can take a little bit of that out save a little bit of money okay now we would typically call call that the cost window diet now that diet now should be less money than the diet that you originally started with and it should be just as good if not better now the difference in cost between those two diets as the money you have to spend for amino acid supplementation and amino acid balancing so that's the approach that we've used here in recent years it really all depends on the diet that you're already feeding if you're not already amino acid balancing and some cases that cost window' diet has saved producers or that cost window diet is actually less costly by 4050 we've seen as much as 75 cents a cow a day that gives you a tremendous opportunity than to go forward with amino acid balancing but typically that cost window diet is going to be closer to 2530 sounds less but sometimes higher than that and lastly use Kyle feedback to fine-tune your diets in other words correct for model prediction errors no models that you folks are using or that we use are perfect so regardless of how good we think the model is we still need to rely on collars to give us some feedback and there's a number of things of course we can watch for we look at the influences on dry matter intake milk you whatever we're looking at in terms of room and function and you see the list here but the questions you need to be asking yourself after your feeding your improved eyes are you really feed enough RDP are you sure are you sure your mom was right you definitely want to make sure that you are feeding it enough and you won't know unless you challenge the cause with a little more RDP that's why it's so important in terms of tracking the amount of RDP in your diet are you feeding enough supplemental Metheny you're achieving the right lasing of Athenian ratio but are you sure the cows won't respond to a little bit more you won't know unless you challenge the cows okay and you got no protein concentrations to look at well your cows respond to additional lysine well I think as the years have gone on I think there's a lot of cows out there in the world today that would respond to additional supplemental lysine but we haven't challenged those cows we haven't challenged them we're just looking at the lysine and methionine ratios and maybe we're not even at the optimum model predicted levels of lysine and metabolizable protein so if you're not already there challenge those cows what are you going to be looking for they're an increase in milk yield increase in milk protein yield possibly an increase in milk protein concentrations are you feeding more RUP than you really need because you're looking at your model and the model says well I need this amount of RUP well if you really amino acid balancing you don't need as much probably as what your model is saying so take some RUP out of the diet see what happens if there's no negative response okay the cows are right the model is wrong RUP is more costly obviously than our DP so to summarize accept the basic aspects of protein amino acid Nutrition let go of balancing for crude protein instead focus on balancing for our DP and the most limiting amino acid make good use of your cows use cow feedback to correct remodel prediction errors in so far as you can but all means use reliable estimates of my amino acid bioavailability for ruined protected amino acid supplements you're going to be really disappointed if you use some of these products and you're not using the right estimates and use raishin formulation programs with optimization capability hopefully your program gives you that opportunity if not encourage your software provider to include a more robust optimizer in your model that will get model compliant solutions that's fast and easy to use you and your cows deserve that with that that's all I have to say thank you for your time and attention [Music]