Guests: Dr. Usman Arshad, ETH Zürich; Dr. Bill Weiss, The Ohio State University Professor Emeritus Dr. Arshad begins by reviewing the inclusion criteria in the meta-analysis he conducted. He wished only to look at studies where lysine was supplemented in a rumen-protected form. The meta-analysis did not include studies that infused lysine into the abomasum or intestine. In addition, only completely randomized design or randomized complete block design studies were included. Feed ingredients and chemical composition of diets for each experiment were run through NASEM to predict the metabolizable lysine content using the same model so all studies were on the same scale. (6:04)
Dr. Arshad begins by reviewing the inclusion criteria in the meta-analysis he conducted. He wished only to look at studies where lysine was supplemented in a rumen-protected form. The meta-analysis did not include studies that infused lysine into the abomasum or intestine. In addition, only completely randomized design or randomized complete block design studies were included. Feed ingredients and chemical composition of diets for each experiment were run through NASEM to predict the metabolizable lysine content using the same model so all studies were on the same scale. (6:04)
The bioavailability of the different rumen-protected lysine products used in the studies ranged from 22 to 90 percent. If the paper's authors reported bioavailability values, they were used in the meta-analysis. If the paper did not provide bioavailability values, Dr. Arshad contacted authors or lysine product manufacturers to offer them. (13:53)
Dr. Arshad discusses the percentage of lysine as a percent of metabolizable protein in the diets as well as differences among the prediction of the flow of amino acids to the small intestine from NASEM, NRC, and CNCPS models. (16:45)
Around 40% of the meta-analysis dataset was from early lactation cows where rumen-protected lysine was supplemented starting from 20 days in milk. The rest of the dataset came from mid-lactation cows. The duration of lysine supplementation also varied. Cows supplemented with rumen-protected lysine for more than 70 days In early lactation produced 1.5 kilograms more milk than control cows. Mid-lactation cows supplemented for less than 70 days produced 0.8 kilograms more milk than control cows. Increasing lysine as a percentage of metabolizable protein linearly increased milk and component yield. (20:11)
During the transition period, cows not only experience negative energy balance but also negative protein balance, estimated at one kilogram of protein loss from skeletal muscle during that time. Plasma lysine starts to decrease around 21 days before calving but bounces back after seven days in milk. This suggests that lysine utilization by the mammary gland and other tissues is high during the prepartum period. Supplementing lysine before calving and during early lactation should improve the efficiency of protein synthesis and may explain the difference between early and mid-lactation responses observed in the meta-analysis. (24:10)
Lysine supplementation improved feed efficiency. Dr. Arshad discusses potential reasons for this and also points out that somatic cell counts were lower for lysine-supplemented cows, suggesting that this amino acid may be important for mammary gland health. He also discusses some of the interactions with methionine found in the meta-analysis. Dr. Zimmerman and Dr. Arshad also hypothesize about the mechanism of action of supplemental lysine increasing milk fat percentage. (30:44)
Dr. Arshad describes how he would design the next rumen-protected lysine study given what he learned from the different studies in the meta-analysis. In particular, he would like to see more work with primiparous cows, and dose-titration studies to pinpoint the optimal amount of lysine to supplement and to further explore the impact of lysine on immune function. (42:42)
The base diet and bioavailability of the rumen-protected lysine product are critical components to determine the supplementation rate for a particular group of cows. Dr. Arshad details the differences between this meta-analysis and previous meta-analyses regarding lysine supplementation. (46:40)
In closing, Dr. Zimmerman was excited to see this meta-analysis and it validated observations from the field. Dr. Weiss underlines the strict inclusion criteria and regression analysis as particularly strong points in the paper. Finally, Dr. Arshad reminds the audience that creating a balanced ration should include essential and non-essential amino acids. He again emphasizes the importance of having an understanding of the bioavailability of rumen-protected products before conducting research projects with them. Lastly, he identifies a research gap regarding lysine supplementation of primiparous cows, which make up 30-35% of herd dynamics. (52:43)
You can find this episode’s journal club paper from the Journal of Dairy Science here: https://www.journalofdairyscience.org/article/S0022-0302(24)00499-5/fulltext
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Scott Sorrell (00:08):
Good evening everyone, and welcome to the Real Science Exchange, the pubcast where leading scientists and industry professionals meet over a few drinks to discuss the latest ideas and trends in animal nutrition. Hi, I'm Scott Sorrell. I'll be your host here tonight at Real Science Exchange. And we're back with my favorite podcast, which is our journal club with Dr. Bill Weiss, where we cuss and discuss the latest in dairy research. First, let's welcome back Dr. Bill Weiss to the table. Bill, welcome back and what's in your glass tonight?
Dr. Bill Weiss (00:39):
Welcome again, Scott. Tonight I've got a 50 West with the complicated name called Pilsner
Scott Sorrell (00:47):
Pilsner. Now what's a 50 West? Is that a local brew pub you've got there?
Dr. Bill Weiss (00:51):
It's local, it's a, yeah, it's a local brewery, 50 west of the major. It used to be a major highway, so they, they're on, on Highway 50 West.
Scott Sorrell (01:00):
Alright, good deal. Well I see that you've invited a guest researcher to the pub tonight. Would you mind introducing him for us?
Dr. Bill Weiss (01:10):
Okay. The guest today is Dr. Arshad. He's from University of Wisconsin. University of Wisconsin. But I hear he is now in Switzerland, so I don't know if he's updated or not. He'll have to tell us why he is in Switzerland. But this is a paper he did, I think, at Wisconsin. Are you a postdoc or on the faculty of Wisconsin?
Dr. Usman Arshad (01:32):
No, I was a postdoc at Wisconsin.
Dr. Bill Weiss (01:34):
Okay, okay. And it's a meta-analysis, it's called “Effects of Feeding Rumen Protected Lysine during the Postpartum Period on Performance and Amino Acid Profiles and Dairy Cows”. And I, I'll let you discuss some of the co-authors before we get into the, or name the, the co-authors before we get into the paper, but welcome to the, to the podcast.
Scott Sorrell (01:56):
Yeah, welcome Dr. Arshad. And this is not your first time to the pub. I think you've been here once, maybe twice before as a student spotlight at the ADSA, is that right?
Dr. Usman Arshad (02:06):
Yes, it is correct. I guess when we met at ADSA, in Missouri we recorded two podcasts there on calling, so it's not my first time. You're right.
Scott Sorrell (02:19):
Yeah. Good. So welcome back. Thank you. Now in, in, in the theme.
Clay Zimmerman (02:23):
Hey, I wanna add also, it's not your first time publishing a meta-analysis, correct?
Dr. Usman Arshad (02:28):
Yeah, it's, it's, I think it's now fourth in this field, so, is that right?
Scott Sorrell (02:33):
Yeah. Good deal. So kind of in the theme of, of the, the real science exchange. I understand that you're originally from Pakistan, so if you had something in your glass from Pakistan in the pub, what, what kind of a drink would that be?
Dr. Usman Arshad (02:51):
It'll be milk and you put soda in the milk. So this is a very common drink we, we used to drink in, in summer.
Scott Sorrell (03:02):
Oh, well that's interesting. Yeah. Well, I always, I like the milk part, that's for sure. Yeah. Alright. And I don't wanna forget my, my trusty rusty co-host Dr. Clay Zimmerman. Clay, welcome back. Good to see you again, sir.
Clay Zimmerman (03:16):
Yep. Thanks, Scott.
Scott Sorrell (03:20):
So what's in your glass tonight, Clay? I, you know, and I've had this before, but I'm having a rock valley spirits single malt whiskey. It's a barley scotch, you know, I didn't know I was a scotch drinker, but I'm really enjoying this one. It's, it's quite tasty. So it's, it's made very close to here. It's made by the milk family, which I, I found kind of interesting. Maybe that's one reason I kinda like it. So that's what I'm having tonight. And before we get started, gentlemen, here's to a great podcast. Cheers.
Dr. Usman Arshad (03:51):
Cheers.
Scott Sorrell (04:00):
The economics of feeding ReaShure Precision Release Choline. ReaShure is fed during the transition period. And because it's fed for such a short period of time, it costs just $15 per cow. And yet the benefits will continue to generate income throughout the year. Cows fed ReaShure produce five pounds more colostrum, which pays for your ReaShure investment on the very first day of lactation the cows are fed. ReaShure also produces five pounds more milk per day every day. That means after the first day, everyday is payday, invest in ReaShure during the transition period and recoup your investment on the very first day of lactation after that. You got it? Payday. Alright, Bill, to begin, can you just kind of give us an overview of why you selected this paper?
Dr. Bill Weiss (04:55):
Well you know, li amino acids are, are big now, and the Naum really emphasized amino acids. So I've, I've learned a lot on amino acid in the last couple years. This was just out in 2024. So it's new, a meta-analysis, and that's kind of good if you don't know all the, if you haven't been reading the last, the research for the last 20 years or so, this kind of summarizes it, but then it also quantifies the relationships between it. So it's a good way to get caught up on old literature plus new, new results from that old literature.
Scott Sorrell (05:30):
And has there been a lot of research bill on, on lysine and dairy cattle?
Dr. Bill Weiss (05:36):
Well, he cited, or the, the authors listed 43 publications. They actually have a question for USAN Usman. You know, you have 43 pubs, but you ended up only using 10. And I think w would, which, you know, the inclusion criteria is important in meta-analysis. So can you tell us why you ended up with 10 publications out of that pretty big list of pups?
Dr. Usman Arshad (06:04):
Yeah, no, that's a good point. So the way I learned to do meta-analysis from Dr. Santos is that the meta-analysis should be hypothesis driven. And since there is already huge variability, you are collecting data from different worlds from different regions where people do these studies, there is already quite a variability in studies, you know, every study with a different, with a different setup of diet, with a different setup of environment. So you try to reduce the minimum. You try to reduce the variance whatsoever you can during designing the inclusion criteria. So the reason why we have only 12 studies and 30 experiments, because the inclusion criteria quite a rigid one, because we wanted to enroll only those experiments where people supplemented lysine in a room protected form. Many of those studies infused lysine either in the ABO or in the intestine.
Dr. Usman Arshad (07:08):
But that is a whole different scenario, right? Because the idea was to see what happens when you supplement a product which is encapsulated in the diet, on the performance of these cows. So that was one of the major criteria. The second criteria, which I choose, and this is my personal preference, that we only included completely randomized design studies or randomized complete block design studies, because the, the rationale to not include Latin square design studies was that Latin square design studies are relatively short period of time. And given the fact that amino acids they need to be supplemented for a given period of time, which is not like two or three weeks, it should be a little bit longer than that, which you can have in CRD or RCBD based design studies. So that's why we only chose those studies which are CRD or RCPD based designs and have a control group where no supplementation of lysine was was a treatment group, and one group was had tru protected lysine group. So at the end of the day, we, we, we came down to 30 experiments with the data set over 600 dairy cows and 40 treatment means, and out of those 40 treatment means 17 treatment means were without supplemental human protected lysine. And 23 treatment means were with women protected lysine.
Dr. Bill Weiss (08:39):
Yeah, I think, you know, a lot of people on med analysis, they kind of skip over the materials and methods, but reading that inclusion criteria, I think is critical. It finds out what your inference space is in your, in your experiments. I think that's a really critical point, even if you're not a statistician to read that, that
Dr. Usman Arshad (08:57):
Section. Exactly. Because if you see now that every paper has two groups, one control, one room protected lysine, the experimental design is same. All papers supposed to present the feed ingredients because some papers will be conducted using CNCPS models, some NASEM, some NRC. There is a huge variability in, in, in formulating diets for that experiment. So one of the idea was to collect the data of feed ingredients of each individual experiment and put those diets back into NASEM so that we can predict the metabolizing content using the same model so that all the studies are on the same scale. So that was also one of the reason that we only chose those studies, which provided the chemical composition and the feed ingredient composition of the diets.
Scott Sorrell (09:52):
Mm-Hmm.
Dr. Usman Arshad (10:14):
It's a great point Scott, because when it comes to the lysine, the encapsulation is not an easy job because there, there are a couple of things which are specifically associated with lysine. One of that is lysine is highly water soluble. So it, it, it, it takes to make an effort to make encapsulation of such products, which are highly soluble in water. Second point is the lysine, the, the encapsulation of lysine. It varies when you mix the diets. So, for example, there was a study conducted a couple of years ago where they had six different products of lysine, and they they performed the mixing of those diets. So by the time those diets were mixed, two out of six products, they were already releasing lysine. And the third point is, it, it, it matters a lot, the stability of the product within the TMR.
Dr. Usman Arshad (11:13):
So out of those six products, three products are, are already releasing lysine even before cow started consuming that product. So I guess the encapsulation technique like clay always says, all ru ru protected products are not same. The very famous sentence is, is correct. It's not an easy job. And the second point, as a scientist, I, I believe that we should test a rumen protected product well ahead of time before designing an experiment. We spent so much money, so much resources and so much labor on conducting an experiment without knowing that if it, if it, if it would really show up in the blood or not. So I guess when we are designing the experiment, a pilot study should be designed on the cow level to see if the room, if this product is truly bioavailable or not. So another thing is, you talked about bioavailability.
Dr. Usman Arshad (12:10):
So let me tell you this. If you, if you, if you go to the literature, the bioavailability of lysine products, which are commercially available is from five to 80 percentage. So it's a huge variability. So, and it matters a lot to account the bioavailability factor while formulating the diets, because you might be formulating it to match to the NASEM standards, but if the bioavailability is less, you are feeding less amount of lysine. So I think it's a, it's a major factor in the, in all, not just lysine in all those encapsulated products, that we should have a clear idea of bioavailability. I, I, I would, I, I would like to add one more comment here. There was a study which was conducted by Dr. Alex Histrov from Penn State University Group. So what they did there they established a criteria, a protocol, I believe, to measure the bioavailability. And then they compared three different lysine products, which were available in the, in the, in, in, in market. So what they found is whatsoever lysine bioavailability was claimed by the manufacturer, they observed half of that. So, which tells you that yes, it is, it is true that many of those bioavailability studies are conducted using in vitro studies, but I guess it is time now to, to test the bioavailability of those products in a pilot study using a cow model. So that's my take on this, this one.
Clay Zimmerman (13:46):
So, so Usman, what, what values did you end up using in the meta-analysis and for bioavailability?
Dr. Usman Arshad (13:53):
So I guess the range of bioavailability of the seven products, which were, which, which, which were based in this meta-analysis was somewhere from 22 to somewhere close to 90 percentage. So the values, which I used if the, if the, if the authors reported those values in the paper. So I was, I was bound to use those values because the diet was formulated according to that value in case if the bioavailability was not provided. So then I contacted authors not every time you get a response back. And then eventually I contacted the manufacturing company to get to know what was the bioavailability and concentration of licensing in their product. So those were the values I chose to, to, to formulate the diets.
Clay Zimmerman (14:44):
Okay. Thank you. Hey, Usman, I want to take a couple steps back here to earlier in the conversation. I think I'm aware of this, but maybe the others on here aren't. So congratulations on your new position. Oh,
Dr. Usman Arshad (14:57):
Thank you.
Clay Zimmerman (14:59):
So tell us why you're in Switzerland.
Dr. Usman Arshad (15:01):
So I finished my PhD from University of Florida, working with Dr. Joseph Santos. Then I moved to University of Wisconsin Medicine, and I did little over 13 months of postdoc in Dr. Heather White's lab. I was also working closely with the Francisco Pan Nano there. Francisco was also my committee member in my PhD program. But then I I got to know this position in Switzerland. This is a position of research scientist and study director. So I applied for this position, and here I'm working under the supervision of Dr. Muhi n so the role is to, it's, it's basically the role is like a faculty. So you get students I already have four PhD students to to supervise them. So the role is mentoring, teaching, and conducting research. So they, they, they named this portfolio as a study director. So that's where I landed up in zero. Yeah.
Scott Sorrell (16:04):
Congratulations. Yeah, thank you. Yeah. Cheers.
Dr. Usman Arshad (16:06):
Thank you.
Dr. Bill Weiss (16:07):
What, what university I didn't hear. I might have missed it. Which university are you with?
Dr. Usman Arshad (16:12):
It's, it's DH Zurich.
Dr. Bill Weiss (16:13):
Okay. Okay. I'll eat some chocolate while you're there. Yeah.
Dr. Bill Weiss (16:21):
On this paper we're gonna talk a means a lot, but the, the difference in between control, and I know there's variation, every control and, and treatment was seven, 7% of the MP is metabolism, lycine and control, and 7.6 for the treatments. Is that a, a good reasonable range? Is that excessive, or
Dr. Usman Arshad (16:45):
So if you, if you see the variability between control and treatment, yes, you are correct. So the, the lysine supplemented diets, they had 7.6 percentage of metabolizable lysine as percentage of mb. But if you, if you, if you see the if you see the the, the diets with ruin protected lysine group, the variability was from 5.5 to 4.7 to, so 5.7 to all the way to nine percentage 0.9 percentage of metabolizable lysine as a percentage of mp. So I would say the majority of the data, it existed somewhere between 6.5 and 8.5. And that's why those were the two values, which I chose to predict response variables, because we wanted to predict values where most of the data relies and not predicting outside the range of the data limits. So what I could tell you would, I did not find many studies with the very high concentrations of lysine. So only one of the, one of 13 studies, they supplemented close to nine percentage of metabolizable lysine, another percent of ami, which tells me that not many studies were performed using high dose of lysine, which, which is a ga which is a research gap to me to identify if we should supplement what more than whatsoever is recommended by NASEM.
Dr. Bill Weiss (18:16):
You use NASEM and you gotta use something, but do you think you'd have gotten similar overall results, say if you use Cornell or some other nutritional model? I don't know how the two two MO models compare in predicting amino acid or mp, but you think similar results that
Dr. Usman Arshad (18:34):
There is a, there is a recent paper which came out from Dr. Mike Vandenberg lab. They conducted a meta-analysis and I guess they have a very large dataset where they compare three different models, NRC, NAS and C-N-C-P-S. It came I think couple months ago in 2024 in general of data science. According to that paper the, the, the, the model predictions for the outflow of essential amino acids, they, the NRC and naso, they are, they are very close enough for some amino acids. CNCPS did not predict as NASEM or NRC were predicting. So it was, it was below then their prediction. And for some amino acids, naso or NRC, they were over predicting some value. So they're, I would say they are not equal because if you go to C-N-C-P-S, they recommend that 7% of metabolizable lysine as a percentage of MP is sufficient. But when it gets to NRC, the value gets to 7.2%. And when you get to the NASEM, the value gets close to 7.5 to 7.7 percentage of mp. So there is a variability, depending upon what model you select to predict those values, I would say.
Dr. Bill Weiss (19:56):
Okay. I guess we would go ahead and get into the results. What, we'll start with production, what kind of response do on that, we'll start with the average, and then we'll get into the regression type stuff. But on average, what was the difference in, in milk production?
Dr. Usman Arshad (20:11):
So we conducted analysis in two different ways. So first of all, the analysis was done very straightforward. What happens when you supplement or you don't supplement women protected lysine? So we only had the, the both treatments or the categorical variable. So in that scenario, the response is dependent upon the stage of lactation. Interestingly in our dataset, almost 40% of the dataset was generated using early lactation cows where rumen-protected lysine was supplemented 20, starting from 20 days in milk. And the half of the dataset where rumen protected lysine was supplemented during the mid lactation. So, interestingly, when you see the data, the cows, which were supplemented in early lactation, they produced 1.5 kilograms more milk as compared to controlled cows. But when you supplement unprotected lysine in mid lactation, the response was still there, but the magnitude of response was less.
Dr. Usman Arshad (21:23):
So only Roman protected ly supplemented cows, they produced 0.8 kilograms more milk than controlled cows. So I would say the milk shield in response to supplementation of lysine was increased, but it is dependent upon the stage of lactation. That was one of the factor. Another factor which we, we explored was the duration of women protected INE supplementation is also important. So the studies where people supplemented women protected lysine 70 or less than 70 days, the response was less as compared to those studies where women protected lysine was supplemented more than 70 days, and the difference was almost 50% less. So if you, so the whole, if you ask me in one sentence studies where people supplemented women protected lysine starting early lactation, and for longer duration, they produced 1.5 kilograms more milk as compared to studies where people started supplementing during mid lactation and for less duration, they produced close around 0.8 kilograms of more milk.
Dr. Usman Arshad (22:35):
So that was one of the analysis that we did. What happened when you feed or you don't feed room productive lysine? Another analysis, which we did, was we wanted to draw the relationship to see what happens to milk yield or other productive performance variables when you have lysine as a percentage of ambi. So in that case, we found a linear relationship. So increasing metabolizable lysine as a percentage of MP increased linearly yields of milk, fat corrected milk protein yield fat shield, or energy corrected milk yield. So increasing metabolizable lysine from 6.5 to 8.5% of MP resulted in increase of roughly close about two kilograms more milk. But you never know what happens beyond 8.5 because there are no data points or there are no studies exist, which, which can tell us that if there will be an additional benefit of supplementing beyond 8.5 or more than 9% of metabolizable li percentage of amp. So that was, I think, the, the major results that we observed in this meta-analysis.
Dr. Bill Weiss (23:49):
Do, do you think the, the bigger response in early lactation, this is, you can't really, you don't know the answer to this, but is it because they don't eat that much or because they should be producing more milk than the later lactation? Probably on average eating less. So which do you think it is? Or is it just early lactation cows respond more?
Dr. Usman Arshad (24:10):
I guess there, there can be several factors which can be discussed and taken into consideration. So for example during transition period, cows are not only experienced net energy balance, they also experience net protein balance. If I recall Dr. Data from Dr. Bell's paper, so he, he, he mentioned that transition cows experience almost one kilogram of protein loss during, during that period from skeletal muscle. So that's a huge amount of protein, which early lactation cows, they, they, they do that. Another factor is what, what Dr Lou’s group from Illinois, they pointed out. So what they did is, so they made concentrations of different amino acids during that transition period. So what they found is so let me talk about just two amino acid lysine and methionine. So, so they, they found that concentration of lysine in plasma, it, it starts going down, starting 21 days before calving.
Dr. Usman Arshad (25:16):
And at day one, or one day after cavi it, it reached at another point where the smallest concentration of lysine was observed in plasma. But that concentration came bounced back at seven days in milk. But in case of methionine. So methionine also started to go down and as coming approaches. And thus, the, the smallest point was observed at 10 days after coming, which they came back on 28 days in 28 days in milk. So suggesting that, so the lysine, the utilization of lysine by the membrane, gland or tissues is expansive during the prepartum period, let's say two to three weeks before calving. And later on it's less. On the other hand, methionine, the use of methionine by the body tissues is probably more in, in, in right after right after calving. So like you said besides these factors, it, yes, I agree with you that trimester intake is less and cows are mobilizing lots of protein.
Dr. Usman Arshad (26:26):
So if you supply amino acids or those nutrients which are going to be utilized by the membrane gland the response will be way better. Another reason is the lysine, it is not extracted by the hepatic tissue. Majority of the lysine is, is extracted by the mammary gland and mammary gland. The uptake of lysine is way greater than the output of lysine in milk protein. So I believe the lysine is not only utilized for the synthesis of milk protein, but it also utilized for the synthesis of branch chain amino acids or other non-essential amino acids. So if you supplement lysine before calving and during the early lactation period, so I would say the response would be, and the utilization to improve the efficiency for the synthesis of protein will be, will be, will be way greater as compared to the mid lactation cals.
Dr. Bill Weiss (27:28):
I think one, you know, we can learn a lot by, by outliers. And I looked at one of your graphs, and if I did my counting correctly, just about a fourth of the studies and milk production, didn't, they didn't respond or they were even a little bit negative. Did you look at why those four? I had seven studies, but why, why those studies? What, why the cows in those studies didn't respond? Did you try and find out maybe to identify when you're more likely to get responses to these to, to RP lysine?
Dr. Usman Arshad (27:59):
I, I'm not sure Dr. Weiss, if I get your answer question. Okay.
Dr. Bill Weiss (28:03):
Well, I said you the, if you look at some of your graphs, seven studies or seven comparisons did not respond positively. And did you look at to why those mo, you know, most of three fourths of them were positive of this, of the comparisons, but about one fourth was not. Did you look at why, what what might have been different between those 25% of studies where the cows didn't respond?
Dr. Usman Arshad (28:29):
I guess one of the one of the possibility could be the experimental unit used in those, in those studies. So out of 13 experiments only four experiments, they had treatment groups where the experimental unit in their, in their individual studies, but more than 15, so 70% of the data, which is generated from the individual study, does not have 15 or more than 15 experimental units. So I guess one thing, the way, I think one of the reason we do meta-analysis is to integrate findings across all the data points to, to see the overall response. So one factor could be that majority of the experiments were not conducted by having lots of cows. So probably those studies were underpowered. That could be one of the thing. The second thing, which Scott pointed out earlier, the bioavailability. So I guess the, it, it is possible that the bioavailability of Roman, of, of the Roman protected products was not enough to elicit the response as it, it is expected. So this could be the second factor, I would say. That's why you see the the 25% of the studies, the comparisons, you don't see the response there.
Dr. Bill Weiss (29:59):
If you didn't look at maybe some unique diet aspects. And some of these are, are even to the, to the left of the line, which wouldn't be power related. They're just, they've responded less. But you didn't look at, is there something unique on those diets or things like that?
Dr. Usman Arshad (30:15):
No no. Okay. I didn't look into that, but it's a good point.
Dr. Bill Weiss (30:21):
Yes. The next thing is I'm, I'm enter, always interested in energy, and you've got increased feed efficiency with RP lysine.
Dr. Usman Arshad (30:29):
Yes.
Dr. Bill Weiss (30:31):
You know, more milk and about the same intake intake. So what do you think is going on here with suggests they're getting more energy per unit of of feed? What do you think is going on there with that?
Dr. Usman Arshad (30:44):
So we found an interaction between metabolizable lysine and metabolizable methionine for fat deficiency. So if you see that, if you see the plots in those f in, in the paper where we have in where we plotted metabolizable methionine on ax axis, and we have those five lines with different levels of metabolizable lysine, those lines will tell you that whenever the diets were fed with eight, eight or 8.5 percentage of metabolizable lysine and 2.5 to 2.6 percentage of metabolizable methionine, the efficiency was greater. In other words, those diets, when the methionine level was less, the, the, the, the increased supplement, the increase the, the level of metabolizable, lysine was the magnitude in the response to feed deficiency was greater when there was any increased when methionine in the diet as well as lysine in the diet.
Dr. Usman Arshad (31:51):
So I guess it is a cumulative effect, which is increasing the efficiency overall efficiency. So it means that not only lysine but methionine, which is also an essential amino acid, they both are involved in, in improving the efficiency. And those two amino acids, as we know that the, the diets of us in US dairy cows, they're mostly on corn and soybean diets. So it met, it, it, it matters a lot when the, when you balance the, those amino acids, which are critical and essential. So that is, that is why you can improve the efficiency. Another aspect is the, the immune function, which is linked to those amino acids. So, like I said, methionine or lysine, they are the precursors for the synthesis of different different antioxidants, let's say like glutathione or, or, or some other, some other ones which are necessarily needed to reduce the reactive oxygen species.
Dr. Usman Arshad (32:58):
So in order to understand the link between immune function and amino acid in this meta-analysis, I guess half of the papers, they also presented data on somatic cell score. So what I observed is the cows, which had increased supplementation of lysine, they had less somatic cell count in, in the milk suggesting that those amino acids, they also improve the health of memory gland. And there is a, there is a, there is some evidence in, in human literature and in mono gastrics that those amino acids are also involved in the, in the proliferation OO of cells, which tells me that probably the supplementation of amino acid, especially well ahead of calving, they, they can mobilize they can not only synthesize the milk components, but also they might proliferate some epithelial cells, which are, which are necessarily required to increase the synthesis of milk. So I guess those points might be involved in improving the overall efficiency of those cows.
Clay Zimmerman (34:04):
So, so Dr. Usman on, you know, on the, from a feed efficiency perspective you know, depending on how much metabolized methionine is in the diet, so is as feed deficiency improves with, with lysine supplementation on the higher methionine diet, does that, does that mean that we're seeing a, a higher milk response to the lysine? If we have adequate methionine in the diet, then?
Dr. Usman Arshad (34:37):
So I guess if you have adequate methionine then the response to supplemental lysine will be, will be there, but the magnitude of data response will be smaller. I would say that's my take on that.
Clay Zimmerman (34:56):
Okay.
Dr. Bill Weiss (34:59):
Because the overall intake, durometer intake wasn't related to, in your regression to MP lysine content, was there an interaction with, with methionine, with intake? No. And lysine. Okay. So it had to be just more milk then? Not now. The efficiency had to be driven by milk, not intake. So
Dr. Usman Arshad (35:19):
Yeah.
Clay Zimmerman (35:25):
So, I'm curious sort of a little bit on, on that topic still. So I don't know if you, if you, if you looked at this value in Nasim or not, I'm curious, what's the lysine to me ratio on these diets? Do you know what the range was for that metric?
Dr. Usman Arshad (35:44):
No, it's no, I, I don't remember now. What was the range. I
Clay Zimmerman (35:51):
I'd be curious to know that at some point. Yeah,
Dr. Usman Arshad (35:53):
I'll send you an email on that because I have to go back to those NASEM sheets and look into that, but I, it's totally top of my head.
Clay Zimmerman (36:02):
Yeah, no, thanks. I noticed it wasn't reported. I was curious though, 'cause that is a metric we look at a lot in these diets. Yeah, commercially, yeah.
Dr. Bill Weiss (36:14):
Yes. What surprised me a little bit, and I'm not an amino acid guy, you know, milk protein was related at a lysine as you'd expect, but so was milk fat percent not, not yield, yield was two, but milk fat percent increased with increasing mp lysine. What, what, do you have an idea on the mode of action or they said, I don't know what lysine does on fat synthesis, but it increased it in this, this meta-analysis.
Dr. Usman Arshad (36:43):
Yeah, I, I really don't know what could have been there because I, I try to look for what for some potential pathways or mechanisms which can be part of that. I found some information that lysine is a ketogenic amino acid, and it can be associated with the synthesis of some ketones. But I, I really don't know what's going on there with reference to the, with the milk fat synthesis.
Dr. Bill Weiss (37:13):
I don't know either. I don't know if these guys know, but I don't.
Clay Zimmerman (37:16):
So I, I, I would say commercially we do observe that a number of times, you know, that increase in milk fat percentage when we're supplementing lysine. There may be, there may be a mechanism there, I think, related to carnasine perhaps.
Dr. Usman Arshad (37:34):
So, yeah, well related to carnitine, I know that lysine, if, if, if it gets a methyl donor if it gets a methyl group from acid cell methionine, it converts into trans methylene, and which eventually is used for the synthesis of carnitine. And now carnitine can be used for the beta oxidation in the, in the hepatocytes. But I really don't know how, what would be the role of carnitine at the level of memory gland which can be used as a pathway for the synthesis of milk fat. But yeah, it's a good point to explore
Dr. Bill Weiss (38:17):
It. The response on, on milk fat was actually greater as a percent of the mean milk fat than the protein. I mean, it, it was a huge response on milk fat percent. I think the slope was 0.5, so really big.
Dr. Usman Arshad (38:32):
Yeah.
Dr. Bill Weiss (38:35):
I guess, but one question, you know, and on NASEM, we, we kind of got away from ratios you know, the, not in NRC, there was the, the lysine methionine ratio. But in Nas o you know, it pretty much was not discussed at all. Do you think there is an optimal ratio between lysine and methionine?
Dr. Usman Arshad (38:58):
Well, I would say if you have a ratio between lysine three to one, it should be, it should be, it should be enough. But like you said, I, I couldn't find also the ratio in NASEM, they only focus on now balancing the amino acids and not on the ratio.
Dr. Bill Weiss (39:20):
I guess on the, one more question on NASEM, did you, you know, your response to, you'd have to convert this to grams of, of metabolizable lysine, not proportion of mp, but did you look at the, the relationship between change in grams of, of our, of metabolizable lysine to milk protein and how that slope compared to to NASEM slope?
Dr. Usman Arshad (39:43):
No, Yeah, no, I didn't do it, but it's a good point. I should do that analysis.
Dr. Bill Weiss (39:49):
I'd just be curious to see how it fits. Hopefully it does, but yeah,
Dr. Usman Arshad (39:53):
No, I didn't do it.
Dr. Usman Arshad (39:58):
I guess I, so I guess, sorry. When I was, when I was planning to do this analysis, there were a couple options. One was to, to, to, to do the relationship between metabolizable lysine as grams per day or metabolizable lysine as percentage of mp. I guess the reason we chose metabolizable per protein as a percentage of FMP, because it's a function of trimester intake, right? So in order to avoid collinearity, we chose to use that factor as opposed to use the grams per day. But it's, it's a, but I would like to see that relationship as well.
Dr. Bill Weiss (40:42):
Now it makes sense what you did because of, you know, intake drives everything, so you never know, is it an intake, a nutrient response, or a, a lysine response? I don't disagree with the way you did it.
Dr. Bill Weiss (40:56):
I guess one, one last question for me. On the duration greater duration, do you think this is just, again, because on protein studies, the response just takes a long time to get maximum response, or do you think it's something else? And I, you started this by saying you don't like Latin squares for protein experiments. I agree 100% with your statement, but do you think it's just a, a time effect or, you know, just it takes that long for cows to fully respond to any, any protein treatment, basically, whether it's slicing or methionine or just crude
Dr. Usman Arshad (41:34):
Protein? Yeah, I, I, I guess, no, I think I agree with you that duration matters a lot because proteins or amino acids, they are not something which, which a body can store a lot for, for the needs of the, let's say for the milk synthesis. I get it, it takes a while when you supplement those amino acid and they get part of the body and get into tissues, and then they start mobilizing and get the count, get into the play a role in the synthesis of milk or milk protein. So I, I don't have any other reason other than that, that it takes a while to, to see the effect of amino acids.
Scott Sorrell (42:19):
Yeah. Kind of one last question for me Usman is that you've had an opportunity to look at multiple different studies and kind of dissect them and look at 'em. If you were design the next lysine research what would that look like? And this is a two part question, then the second part would be, do you, do you anticipate that you'll be doing any kind of amino acid research?
Dr. Usman Arshad (42:42):
So yeah, back to your first question. So I would let, let, let me give you an example from one of the meta-analysis that Jose did I think three years ago. So in that meta-analysis, what they did, they wanted to see what is the effect of metabolizable protein prepartum on the postpartum performance in primiparous and multiparous cows. So in that meta-analysis, what they found is between the range of the dataset that cows, when they were fed 800 to 1100 grams of metabolizable protein in the prepartum period, the milk yield was response was linear. And if you just get the percentage of lysine based upon that meta-analysis, so that percentage was 8.2 basically. So the 8.2 percentage of metabolizable lysine as a percent of MP is an optimum one during the prepartum period to, to, to improve the performance of those cows.
Dr. Usman Arshad (43:50):
And based upon the results of this meta-analysis where you have a linearity until 8.5 or close to nine, I would like to design a study with increasing levels of metabolizable lysine as a percentage of ampi, starting from the, the, the one which are recommended nowadays from with nasm. So let's say one group is at 7.7, another one is at eight, and the third one is let's say 8.5, or probably fourth one at nine. So I guess the, the, the, the finding from this meta-analysis was that only four studies out of 13, they presented in what happens in the primiparous cow. So we really don't know if those responses, which are observed here, but would be also valid for the primiparous cows, especially when those cows are creating more tissues towards the memory gland or towards the, towards their growth.
Dr. Usman Arshad (44:47):
So I guess there is, there is gap to do studies with primiparous skulls, and also there is gap to do studies with more dose titration experiments. The second thing I did my post-doctoral work with Dr. Heather White majorly focusing on rumen protected lysine. And we not only focus with different levels of lysine, but also we evaluated the immune function of lysine. So basically related to the immune function we should be submitting that data soon. But we wanted to see what happens to the bioenergetic measures specifically of tail lymphocytes when you supplement lysine or you don't supplement lysine. So yes, I conducted already two experiments with ru protected lysine. And in future I hope so, I will be working with rumen protected histidine, rumen protected arginine or some other amino acid work
Scott Sorrell (45:50):
Very well.
Clay Zimmerman (45:52):
So, so Usman bank just talking about supplementation levels that were in these experiments. So and maybe, maybe hitting on a few points that Bill had brought up earlier was, was blood meal used in any of the diets that were used in the meta-analysis? Either, either in the controls or in the supple, the lysine supplemented diets?
Dr. Usman Arshad (46:21):
I have to look carefully again, but as if I remember correctly, I don't see that if I added a paper in this meta-analysis, which used blood mail specifically. But I, I will, I will need to look again, double check this.
Clay Zimmerman (46:40):
So the, the mean, the mean lysine is supplemental lysine level was, was 19.3 grams a metabolizable lysine, correct?
Dr. Usman Arshad (46:53):
Yes. Yes. I think the range was somewhere from five to 40, but the mean was close to 20. Yes.
Clay Zimmerman (46:59):
Yeah. So, so ni you know, bill had asked the question earlier, so you know, that mean level, that's a pretty good difference from control, you know, that that level of, of additional MP lysine, and of course the, that, that five gram dose, that's a really minimal dose, I would say commercially, that that 40 gram dose is a huge dose. Yeah. Commercially,
Dr. Usman Arshad (47:26):
Yeah, you are right. But I guess it, it, it matters a lot because so for example, in in Switzerland they have mainly forage based diet in, if you talk about us, they have mainly soy and corn-based diet. So I think that the, that the ingredients of the diet matters a lot if you use, if you have to use 10 grams versus 20 versus 30 grams of metabolizable lysine. So I guess at the end of the date really matters. What are the food ingredients and what is the bioavailability of the source of lysine, which has to be used for the proper formulation of those amino acids.
Clay Zimmerman (48:05):
Yeah. So there, you know, there have been other lysine, meta-analyses, you know, conducted before. What, what were the basic differences between this meta-analysis and previous ones that were done?
Dr. Usman Arshad (48:21):
So let me tell you one thing specifically. So the one meta-analysis, which was done in, back in 2010 by Dr. Robson at that time we only had seven studies. So very few number of studies with RU protected lysine. But the, the, the issue with that meta-analysis was on the record, I'm not discrediting their work, I'm just pointing out what were the limitations. So basically in their meta-analysis, they included all the studies which had rumen protected lysine or where the lysin was infused in the albinism, or where the lysine was directly infused in the anine. So basically, they did not segregate the studies with only women protected lysine or with the lysine, which was supplemented at the level of intestine or albinism. So that was, I think, one of the major difference between that meta-analysis and this meta-analysis is that we only included those studies with RU protected lysine product and not included those studies where you infused lysine at the ASO or intestine level.
Dr. Usman Arshad (49:35):
I think that was one of the major difference. Another meta-analysis, which was done by Dr. Mann from Maryland. I guess in that meta-analysis, they also did not differentiate between hormone protected or if the lysine was infused at the amass or intestine level but also the only reported data on the milk protein yield. They did not report data on the intake or milk yield or milk components or amino acids or anything. The only response variable was milk protein yield, which was evaluated in that meta analysis. So I guess if you talk about the differences, this meta-analysis included all the studies whatsoever available in the literature with tru protected lysine, and also covered majority of the response variables as compared to those ones.
Dr. Bill Weiss (50:36):
I got one, one last question. I'm looking at my notes and I missed this. In, in your regression analysis where you looked at the effect of milk or protein or whatever on change in, in MP lysine, did, did those data points include the controls? In other words, you know, there was a lot of variation in the mp, the lysine as a percent of MP and control, or did it just include the, the treatment ones in that regression?
Dr. Usman Arshad (51:05):
No, those regression points included all the 48 data points. Okay. All 40 treatments, yes.
Dr. Bill Weiss (51:11):
Okay.
Scott Sorrell (51:15):
Alright. Sounds like a good time to call, last call then. If you guys will just you know, as we close this out, kind of give us a, you know, 1, 2, 3 kind of key takeaways that you guys take away from this research. And, and Clay, why don't you start us off?
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Clay Zimmerman (52:43):
Yeah. Yeah. So I, I was very excited to, to see this meta-analysis. Obviously we, we have a lot of interest in in amino acids lysine and methionine. So I think this really, you know, this validated a lot of the things that we, we've talked about for a number of years that, you know, ear, the earlier in lactation you start supplementation and the, the more likely you are to see a response and you're not, not so likely to see a response to the lysine if, if you're methionine deficient as well, right? So, you know, it's important to, to really cos supplement these, these amino acids. So I, you know, I think there's a lot of, lot of good practical takeaways here that that we can utilize that we can utilize in the field. And I, I would be curious
Scott Sorrell (53:54):
Yeah, great paper. Bill, any final comments from you?
Dr. Bill Weiss (53:58):
Well, I like you know, the, you excluded a lot of data, but for good reason. And I think that makes the actual conclusions more applicable. Like you got rid of infusion data and all this, and it makes it much more applicable. And I, again, I urge people who read meta-analysis to read exclusion data. And the other thing I like about this is the, the, the regression part, the quantifying the response, it again helps people determine is this gonna be economical? What's their expected response? So those two things, I think make this a a, a really good paper.
Scott Sorrell (54:33):
Yeah. And Dr. Arshad, well, any final comments from yourself?
Dr. Usman Arshad (54:38):
Thank you for the appreciation. I guess there is no question about that cow needs a balanced Russian, which also includes the balancing not only essential amino acids, but overall amino acids. And also it is very important from research perspective that we should take account the bioavailability component strictly and we should do some pilot studies to confirm the bioavailability of the product before we start utilizing the resources. And the third thing is, I would say there is still need to identify the optimum dose of lysine for the performance in cows. And we, we, we still have a research gap to understand the role of supplemental lysine in primiparouscows. Because majority of the herds, they have 30 to 35% of the cows, which are primiparous, so it'll be, it'll be very important to understand the role of supplemental amino acids in the primiparous cows, which contribute 30% of the herd dynamics.
Scott Sorrell (55:50):
Hmm. Thank you. Well, I gotta say this has been very enjoyable. It's been a great way to spend an evening. So I want to thank you guys for your, your time and your knowledge and spending some time with us here this evening to our loyal listeners. As always, we thank you for coming along with us as well. We hope you learned something. We hope you had some fun. We hope to see you next time here at the Real Science Exchange, where it's always happy hour and you're always among friends.
Speaker 7 (56:17):
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