[Music] welcome it's a joy to be here this will be my first presentation here in Europe under the auspices of vadas ale was chemin before that so I got to see a lot of Europe starting back in 2009 actually 2007 I think a deseo hosted a amino-acid symposium in Belgium you know 7:00 in the spring of o7 so very quickly we're going to very briefly go through a number of topics here just to kind of set the stage for why ed Sao is doing what they're doing and why they have the products that they have and I might also just add that I've been involved with a deseo prior to that name back in the late 1980s and so I've enjoyed a long relationship with the company so today we're just gonna briefly touch on protein digestion in the rumen animal the sources of metabolizable amino acids the chemistry and classification of them which sometimes we forget that and I think it's just good to remind ourselves that they are unique nutrients we'll talk about the functions limiting amino acids and dairy cows unfortunately we don't have time to get into all of that research it's really exciting what is going on but I'll touch the high points the benefits of balancing for amino acids and again having been involved in this area for 47 years I've been able to witness the progress that nutritionists out in the field have made and it's pretty incredible what some of those folks are doing now out on commercial dairy farms and then just make a point about the predictability of nutritional models you're gonna have some other speakers that follow my heart goes out to them as they really struggle here to improve nutritional models to really clearly and make it easier to bring amino acid nutrition to the forefront and then just to comment about the future so I assume at least those of you that are nutritionists have seen this diagram or something similar to that for years and years and years we have worked hard to define requirements on the basis of crude protein unfortunately as I travel around the world it's still very important to people they spend a lot of time struggling and formulation to get their crude protein levels just right and I would just like to remind you here folks today that that is probably the most imprecise thing that you can measure in terms of trying to improve efficiency of nitrogen utilization and dairy cows so as you look at this diagram what happens to crude protein in the rumen some usually most of that protein is going to be degraded down to amino acids and peptides some of those amino acids and peptides are going to move to the small intestine intact but most of them include in the ammonia that is released as a result of the proteolytic activity of bacteria and protozoa and the rumen ends up being ammonia that ammonia needs to be captured fortunately most of that ammonia is captured in rumen microbes bacteria and protozoa primarily the rest of the protein and the rumen is going to resist or escape rule no degradation and pass through the small intestine in America we call that our U P rumen on degraded feed protein so we have two primary sources now of protein flowing to the small intestine both of which contain amino-acids the reason that I wanted to put this up again goes to the heart the issue of developing mathematical models to describe the availability of amino acids to the ruminant animal so just a review here there are three sources of the metabolizable amino acids one is microbial protein all of the models today assume that there is a fixed amino acid composition of room and microorganisms however everyone has understood that that is not true and that in fact there is a big disparity in the amino acid composition of various types the primary forms of bacteria in the rumen as well as protozoa and fungi a really good meta-analysis was conducted just a few years ago I'm going to show the results here and just another slider to to show that variability but the point is is that the amino acid composition of those microorganisms do differ and that's going to be impacted not only by cow there is within cow among cow variation but certainly will be impacted by diet and here are the results of that meta-analysis and what this slide shows is the average along with the coefficient of variation values but the average amino acid concentrations and fluid associated bacteria particle associated bacteria and protozoa and over on the right-hand side of that table are the contrast comparing bacteria versus protozoa and then comparing fluid associated bacteria versus particle associated bacteria not going to spend a lot of time on this because ultimately you're going to get a copy of this presentation but please focus on the lysine and methionine concentrations in room and microorganisms so if we look at lysine first notice that the mean valuing food associated bacteria 7.7 particle associated bacteria 7.4 sex protozoa 10.7 5 note the magnitude of those numbers methionine had two point four four two point two six and two point one three okay I want you to remember those values because when we look at their concentrations in feed which would represent what you would find in room and on degraded protein you're gonna see those values are significantly lower RUP on average that represents according to our models as well as experimental research 40 to 50 percent of the amino acid is flowing to the small intestine now wild amino as a composition a room and microorganisms are different the greatest impact and dr. honoree rookin here in france and in our own work when we developed the model for NRC 2001 we accounted for about 75 percent of the variation in the amino acid composition of intestinal protein coming from the relative contributions of RUP to total protein flow to the small intestine and the amino acid composition of RUP in other words the amino acid composition of feeds that was 75 percent of the variation so we could not account for 25% of that variation in the balance of amino acids in intestinal digesta and my guess is that a majority of that variation is a result of the changes in the relative contributions of microbes leaving the rumen across studies just a few things about amino acids amino acids are small molecules there's 20 that makeup protein but they are all different they have a common characteristic of all containing nitrogen or an amino group but when you look at these structures and for those of you that have taken organic chemistry biochemistry and that sort of thing you know that differences in the chemistry of compounds affect how they are metabolized and how they impact overall metabolism in the body so Matheny is a nonpolar amino acid the structure is given there lysine and histidine are over on the right-hand side in green and they have a positive charge okay so those three amino acids most characteristically have been identified as most limiting and you can see that their structures are significantly different in terms of dietary classification we would and this goes back decades in the non-ruminant species and human nutrition but those 20 amino acids have been categorized as either being regarded as being essential that means dietary sensuality and the non-ruminant animal or non-essential all twenty are required for protein synthesis however there are really very few of any studies either in the non aruna species or in the ruinin species to say that we have to be concerned about the non-essential amino acids because they'll always be enough in the total protein flowing to the small intestine animals can make one non-essential amino acid from another or from surplus essential amino acids so this really simplifies things when we start thinking about how are we going to formulate diets for amino acids in roomis what it means is we can concentrate on the essentials so the ones on the left they must be absorbed in the amounts needed the ones on the right it's not important now let's talk about the functions of amino acids just very briefly we all recognize amino acids as being substrates or the building blocks for protein and we in the dairy area we tend to think about the importance of amino acids for milk protein synthesis which of course helps dictate no-kill well we really didn't think about a lot in our early studies going back to the 60s late 60s 70s when dr. Gookin got involved I got involved in 1971 as a PhD student we were focusing on milk protein now there have been books published many books published and I just show you two of them they're books published just on amino acids and the functions of the individual amino acids in overall health etc so we got blood proteins we got iron channel proteins we got receptor proteins enzymes tissue protease connective tissue the bottom line is nothing happens in the body nothing nothing happens until protein synthesis occurs first think about that so as we manipulate the mino acid balance in dairy cows and we see a spike in milk protein what does that mean what does that mean are we just affecting protein synthesis in the mammary gland or are we affecting protein synthesis throughout the body and the answer is the latter so amino acids are the building blocks for protein they're all joined together according to a genetic a predetermined genetic code what that means is that amino acid composition of a protein is the same every time that is synthesized and because that is a controlled event that means that the amount of protein that can be synthesized in the cow every day depends on the supply of the most limiting amino acid nothing more nothing less and the efficiency of use of those absorbed amino acids and now this is where hormones come into play this is where energy status comes into play etc etc okay so those are the two driving factors the supply of the most limiting amino acid and secondly the efficiency of use of those amino acids all the amino acids including the first limiting one that's absorbed here is a goat Mitchell and block a published a very often quoted paper that formulated a conceptual framework with this assumption that the most limiting amino acid regulates protein synthesis and that addition of other amino acids don't have an effect that has long been referred to as a single limiting amino acid theory usually depicted as a barrel stave theory in other words the efficiency of use of those barrels is going to be dictated by the length of the shortest stave right in other words it means protein nutrition that the efficiency of use of digestible protein or metabolizable protein will be determined by the content the relative content of that amino acid that is most limiting in metabolizable protein relative to need that's what's going to determine the efficiency of use primary factor determining efficiency of use of protein amino acids are also and this is another area that up until probably fifteen years ago we sorely neglected in dairy cattle nutrition and that is the impact that individual and combinations of amino acids have on metabolic regulation now we've known for a long time that the extracellular concentrations and profiles of amino acids impact concentrations and profiles of intracellular concentrations NS the differences in those concentrations in the intracellular concentration that regulates what we now would call post translational rates in other words the rate of synthesis of protein intracellular amino acid concentrations can change and the rate the speed at which that machinery operates and there's two areas that have received a lot of attention one is what we called mTOR and the other is the SI our pathways and the mTOR pathway is probably the overriding one in most cases but that's an important regulator of anabolic metabolism and there's a lot of factors that affect that and it has been demonstrated both in vitro and in vivo with lactating cows that if we basically jack up those pathways that is going to have a positive effect on protein synthesis as well as milk protein yield in different combinations of amino acids can do that now the amino acids that have been shown to have the strongest mTOR signaling properties or Metheny arginine isoleucine and leucine the latter two of which are two of the three branched chain amino acids and threonine I wanted to mention this because I think as we move forward in working with you folks and taking your input back to our research labs it's really important that we identify what is the ideal profile of absorbed amino acids for lactating dairy cows and in order to do that we're going to have to think outside of the box a little bit and not just think about which amino acids are most limiting for protein synthesis I had the pleasure of reviewing a really excellent manuscript out of Virginia Tech here a couple three months ago it was just accepted for publication probably will appear in the Journal of Dairy Science in the next month or two this was an excellent study large group of cows high producing cows cows were assigned to this study post peak you can see their dry matter intake sup they're for dietary treatments control the second one Metheny lysine and histidine infused into the jugular vein in concentrations to make up the shortfall or the deficiencies relative to the predictions of both NRC and CN cps and in this case a reusing amts the third treatment was leucine and isoleucine and then they added those five together now they picked leucine and isoleucine because they have been shown to have the greatest impact on mTOR now let's move to milky methionine and lysine and histidine together had no impact on Mo Kio however isoleucine and leucine did and you add the two together and milk yield at least numerically went up a little bit more not surprisingly to either the researchers or certainly not to me after being involved in this area for 47 years the impact of Matheny and Laci and histidine was on milk protein milk protein concentrations whereas isoleucine and leucine had no effect whatsoever you put the two together you get their response again interestingly and dr. rookin had dated to support this as well is that the branch chain amino acids influence mammary blood flow notice the significant increase in mammary blood flow with leucine and I salute okay Metheny and lysine histidine did not have that effect even though dr. Gookin did have a study that he reported in journal dear ESI as many years ago that histidine had a positive effect a mammary blood flow so I think this is a really neat study I think this is a going to be a study that's going to be quoted many many times and is going to serve as a foundation for future research to marry these two primary functions of amino acids together in defining ideal balance of absorbed amino acids amino acids are used for the synthesis of NPN compounds now there's about sixty of them that are synthesized in the dairy cow they're all going to be synthesized from amino acids does not represent a big demand by the way for amino acids but nevertheless it is a demand now if we go back and especially the swine literature we have no reason to believe anything other than this that an ideal balance of amino acids exists to meet these three sets of requirements amino acids not used for these functions that are going to be used for energy just a few comments and on limiting amino acids lysine methionine we've known this now my PhD research certainly supported that made it very very convincing like seeing a methionine are going to be the most limiting and lactating dairy cows fed conventional feeds we observe 43 percent of the total response we got with all 20 amino acids with just lysine and methionine in five different studies where cows were fed conventional feed cells just like you feed over here without any protein supplements in them you start adding protein supplements on top they're going to be deficient in either lazy or methionine or both so that's a no brainer that's an absolute no-brainer now we've got studies indicating that histidine getting follow especially as we start lowering protein in the diet now I wanted you to remember what those lysine methionine concentrations were and rumen bacteria and protozoa remember on average for bacteria 7 sex protozoa 10.7 if we look at those concentrations in milk at 7.7 so based on this knowledge if we could have the best of all worlds we would like to see lysine concentrations and RUP not protein 7 percent on average however if you look at the rest of that table you'll be hard-pressed to find any protein supplements with a lysine concentration of 7 percent ok it's not there one exception is blood meal second exception is fish meal and as I understand it here in Europe you don't feed either okay but you do feed soy products you do feed canola meal and so on look at the values for Metheny sort of a similar situation if we select feeds that are high in lysine such as our soy products note very low concentration of methionine so after working in this area for 47 years I have yet to see a situation where we feed a conventional diet to dairy cow supplemented with soy products that was not Metheny and efficient we can always get a response with Metheny histidine look at the histidine values in bacteria and protozoa relative to melon now we've known this for a long time - hasn't really been a big deal when we fed higher protein diets with a lot of supplemental protein but as you folks other folks around the world are trying to feed less protein less RUP that means microbial tene now makes up a greater proportion of the total protein flowing to the small intestine that means now that the histidine the average histidine concentrations in intestinal protein tend to go down as microbial protein makes up a greater share of that total protein thus it's not surprising to me at all that once we formulate diets for higher levels of methionine and lysine that histidine now come in comes in as third lemming team so these are the amino acids that have surfaced as being the most limiting probably and not too distant future there will be another company we'll probably be coming out with a source of protected histidine I show these structures to let you know remind you that we're dealing with three uniquely different amino acids and another thing that I want you to remember here is when you think about functions look at the large number of functions in the body for Metheny as compared to lysine or history they're all three involved as building blocks for protein synthesis but look at that long list of functions for Metheny and a lot of the things that we see with respect to Matheny come about because methionine is the molecule that is used to synthesize things such as homocysteine such as Torrey such as glutathione which are really important in transition cows so we have this information we know amino acid balancing is important we know amino acids are important to the dairy cow and what are we going to do about it well we can do one of two things we can feed a lot of our upa just use a shotgun approach and the more RUP we feed the more lysine Metheny that's going to get to the lower gut but the amino acid balance just gets progressively worse and worse the more you feed so that does nothing for efficiency in fact efficiency goes the other way a literature summary was done nearly 20 years ago which indicators showed very clearly that feeding more RUP to cows does not increase the predictability of animal response and there was two reasons for it as I said you feed more RUP to cows via feeding higher protein diets those animals become less and less nitrogen efficient also when dairy farmers do that or you as company people push more and more RUP as you might have years ago you increased your risk of shortchanging the requirements of room and microorganisms for room and degradable protein and so that works against you as well so it just makes sense that we take a smarter approach to this and we start balancing for amino acids and when we balance for amino acids if you remember what we're doing is we're balancing the amino acids in RUP you started feeding smarter meat some are to mean and medicine mark the protected lysine supplement what are you changing you're not changing them you know as a composition of microbial protein you're changing the amino acid composition of RUP and you're increasing its efficiency of utilization so when we think about amino acid balancing we are talking about mixing and matching protein supplements and amino acid supplements in a way to achieve and improved amino acid balance and we want to get a balance that comes as close as we understand it to meeting that optimum balance for the dairy cow here's one aruch in studies and he and i started communicating back in the 70s and he was way ahead of us in the US he had a lot more data quicker than we had and here's what he showed in terms of the relationship between milk protein yield and increasing concentrations of lysine indigestible protein notice how tight those those red dots appear to that quadratic curve what does that tell us that toes that a lysine is important in that the primary function of lysine has to be as a building block for protein remember lysine did not have a long list of functions here's a relationship in milk protein yield response to increasing amounts of methionine and digestible protein notice there's more scatter there's more scatter there I wonder why might it be that methionine and some of these trials was using for was being used for things that the cow need is for methionine remember Matheny has a lot of functions and it's not all just going to be related here to changes in milk protein yield so when I had to develop the protein and amino acid some bottle for NRC I wanted to develop a similar curves that dr. rule Ken had developed I didn't have as much data to draw on but here's a relationship between changes in milk protein and model predicted increases of lysine and metabolizable protein notice we basically found the same thing here a plateaued off at 7.2 percent lysine and MPs or we got the maximum milk protein yield response and here's the plot for Matheny so after the publication of NRC 2001 there were a few changes to the NRC that involved just some small differences here in the predictability so you see the optimum levels for lysine methionine and MP and the optimum lysine and methionine race that really didn't change from the 3.0 to 1 ratio in what was in the publication we took that exact same dataset then and we developed those curves for CPM theory at the time and then various versions of CNC PS and you can see over the years as as a biology if you will of the CM CPS model was improved that those values have changed the point being here is we got some models here that worked very well for us to help us zero in on diet variation formulation to get our levels of lysine methionine as close as possible to the optimum so what are the benefits of amino acid balancing for cows oftentimes we don't think about this but I would suggest to you that you do think about it and that is this to avoid the risk of the most limiting amino acids being the most limiting nutrients avoid that risk you don't there's no need to have any of them you know asses being more limiting than other nutrients and here is a list of what we have learned probably in the last 12 to 15 years now when supplemental Matheny is provided to transition cows that are fed totally normal diets with protein supplements like soy soy products in some cases some blood meal to bring the lysine up to more optimum levels I'm not saying up to optimal levels but up closer to optimum levels and then filled the shortfall that is created for methionine with supplemental Metheny everything that you see on this slide was created by a deseo supported research initially look at that long list of functions should that be a surprise no it's not a surprise we just hadn't looked before and I think what is really exciting is the impact am athenian has on birth weights of offspring growth rates of offspring now data accumulating on the production of those offspring sir evidence that fetal programming is occurring her so my point that I said before we see an impact of amino acid balancing on changes in milk protein content I cannot tell you how many situations over the last 20 to 30 years where we've come in with a protective Metheny product in lactating dairy cow rations and within a couple days we start seeing a rise in milk protein now all of this good research that has been done what does that tell us that tells us that those makalah's were indeed Metheny deficient and methionine was the most limiting nutrient in those diets because Matheny was having an impact on things below the surface of the water in this slide we've known for years that if we add protective Metheny and lysine supplements to lactating dairy cow diets at the beginning of lactation we see pretty dramatic effects on milk yield well it makes sense the science can explain that back over 20 years ago we ran two low studies at UNH one was an undergraduate project one was a master student project and we made sure that those dyes were Metheny inadequate but we wanted to see what is the impact if we add some athenian on top of those otherwise Matheny and adequate diets got an increase in milk yield got an increase in milk protein content increase in protein yield Wow remember lysine has one primary function as a building block for protein and if it's limiting it's gonna affect milk yield and it's going to affect milk protein synthesis in the mammary gland here's another study same thing same thing the last thing that I'm going to mention here today is the impact of amino acid balancing on sparing RUP in the diet again you're gonna get a copy of this presentation here I've just gone through an exercise saying how does changes in the lysine content in mp and changes in the methionine content in mp affect how much MP the cows need to deliver 180 grams of lysine and 60 grams of Metheny and if you go way over to the right-hand side of this table note that the requirement for RUP as a percent of diet dry matter drop from eight point one two five point four percent that's calculations but the cows tell us the same thing I want to just mention two studies involving smart Amin M and meta smart this was a study that was published in 2003 I mean look at those diets you see there those are pretty normal diets high RUP diet low RUP diet low RUP diet plus up on mel Matheny Matheny came from both smarty me and having a medicine Mart go to the right hand side of that slide notice the RUP was lowered such a crude protein went down from eighteen point three to sixteen point nine percent and MP lysine supply dropped from a hundred and eighty three 274 lysine was six four of MP methane years one point seven the Lexington Messiah narration was three point eight to one I'm both positive control and the low RUP diet came in with a combination of meta smart smart to mean em to get a ratio three point three two one not as aggressive as we would do today not as aggressive as fuel nutritionists would do today but if you look at the results of this slide compare the production data animal data from the left-hand column to the right-hand column notice that those animals performed as well with the low RUP diet that had supplemental Metheny in it as compared to the high RUP diet they did just as well or if not better but look at the last two lions milk nitrogen as a percent of nitrogen intake went from 31.1 up to 35 and if you calculate excreted nitrogen divided up by nitrogen intake which is really what we would be concerned about in terms of the environment those those values dropped from 2.25 down to one point eight nine study out of the University of wisconsin-madison very similar a traditional protein diet in this case it was sixteen point nine percent protein diet they lowered the protein they they took out some of those protein supplements like distillers grains expeller soybean meal just left soybean meal and it increased soybean meal a little bit notice on the next slide here that the animals performed just as well on the lower protein diet with either meta smart or smarter mean M both perform equally well and that was assuming a bioavailability value of 80 percent of Metheny in smart amine ham and 50% for medicine art it was those calculations are those values used for determining inclusion rate just as a preface that I think what you're going to hear throughout the rest of the conference or by a few of the speakers and that has to do with models improving the predictability of models has been challenging frustrating and slow for those of us that have worked in this area a long time and work with dairy producers and working with dairy nutritionists they want better models and change has come slow and overcoming these changes is really important as a first step to accurately predicting amino acid requirements and this really relates back now to the complexity of protein digestion in the rumen the complexity of amino acid metabolism and not understanding that has also gotten in our way and fortunately in the last 10 to 15 years there's been a lot of good data and thirdly as nutritionists out here we cannot forget that the complexity of protein digestion has created challenges for predicting our DP requirements boomin microbes require our DP we cannot forget that and as I travel around the world I see so many examples of commercial dairies they want to feed less protein but they're starving the rumen for room and degradable protein and simply add in your ear to some of these diets improves feed intake improves milk yield so fortunately a lot of progress has been made as I've already said the do dairy NRC we have been promised is coming out in 2020 which is next year I've been privy to a lot of what's happening there I'm not on that committee and I think it is truly exciting so to wrap things up amino acid balancing is recently being accepted there's several reasons for this but most important for people out in the field achieving success requires letting go a balancing diet for crude protein and instead focusing on our DP for bugs and amino acid requirements for the cow and if you look at some of the published data the relationship between milk protein yield and dietary crude protein is horrendous it is bad look at it in the upper left the relationship is better trying to relate milk protein yield to metabolizable protein but look how things are improved when we begin looking at the relationship between protein yield and the first limiting amino acids this is revolutionary for the dairy industry we also have to use reliable estimates of amino acid bioavailability I'll talk about that tomorrow enhancement of region formulation programs with optimization capability that's going to be great and wherever possible use Kyle feedback so the future is bright and thank you for your time [Music]