Joining together for the second episode of the New Revelations in Transition Cow Nutrition from the 2022 Cornell Nutrition Conference four-part mini-series to discuss animal nutrition requirements are remarks from Dr. Jose Santos, University of Florida and topical insights from Dr. Clay Zimmerman from Balchem.
Guests: Dr. Jose Santos, University of Florida
Joining together for the second episode of the New Revelations in Transition Cow Nutrition from the 2022 Cornell Nutrition Conference four-part mini-series to discuss animal nutrition requirements are remarks from Dr. Jose Santos, University of Florida and topical insights from Dr. Clay Zimmerman from Balchem.
Bringing forward the most recent research on choline supplementation, Dr. Santos began the second in the series focusing on nutritional mechanisms and their essential benefits in animal growth and performance. 3:45
Choline was first introduced in the 1700s by chemists and pharmacists. Still, it wasn’t until about 40 years ago that Derek Lindsay from England discovered that most phospholipids in ruminants are synthesized de novo.
Dr. Santos mentioned that studies show more than 90% of choline in feeds doesn’t show up past the rumen, adding to the lag in understanding when the essential nutrient shows up in the small intestine. 5:32
As a required nutrient, Dr. Santos shared that choline is required for the structural integrity of cell membranes, neural tissue and the components of phospholipids and sphingolipids. 7:01
As cows approach calving and during the first two weeks of lactation, Dr. Santos suggests that it is the optimum time to provide choline as a building block for phosphatidylcholine. 17:35
Sharing a heat map study on the effects of choline in hepatic tissue, Dr. Santos said it ultimately shows that supplementing choline reduces glycerol and increases the synthesis of phosphatidylcholine. He added that as studies focus on hepatic triglyceride, acid basis or dry matter basis, the benefits of choline at different dosage levels benefit the same. 22:30
But do low-body condition cows react to choline supplementation the same as high-body condition cows during the transition period?
Dr. Santos shifted directions, adding that he’s found low body conditions cows responded to choline with more milk and energy. He added that studies show supplying choline to nutrient-deficient animals enhances their ability to transport and absorb nutrients from the gastrointestinal tract. 41:55
Dr. Santos mentioned not only does choline facilitate phospholipid synthesis and plays a large role in the transportation of fatty acids, he believes it’s an unquestionable supplementation that fits the requirements as a required nutrient. 46:35
Wrapping up the conversation, Dr. Zimmerman highlighted key points from Dr. Santos and summarized consistent responses the industry continues to see in the meta-analysis of added choline in a range of production levels. 49:15
If you would like to review Dr. Santo’s webinar from the 2022 Cornell Mini Symposium, you can view all four webinar series at balchem/com/realscience.
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Scott Sorrell (00:08):
Good evening everyone, and welcome to the Real Science Exchange, the podcast we're leading scientists and industry professionals meet over a few drinks to discuss the latest ideas and trends in animal nutrition. Hi, I'm Scott Sorell, one of your hosts here tonight at The Real Science Exchange. Tonight we're continuing the new miniseries of podcasts recorded at the 2022 Cornell Nutrition Conference called New Revelations in transition, Cal Nutrition. I'd first like to welcome though my co-host, Dr. Clay Zerman. Clay, thank you for joining me here once again and as always watching your glass tonight.
Dr. Clay Zimmerman (00:44):
Tonight. I, you know, being the world class athlete that I am,
Scott Sorrell (00:53):
Oh, very nice. Well, good. Well, tonight I'm having something called so I wanted to have something that I thought Josiah would drink, but I don't know what he drinks. So he's from Brazil, so I'm having a, a
Dr. Clay Zimmerman (01:22):
Forward to the cheers.
Scott Sorrell (01:23):
Yeah. Conversation tonight.
Speaker 3 (01:25):
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Scott Sorrell (01:47):
So, as we begin tonight's discussion, we'd like to note that this is the second in the series of four podcasts taken from the mini symposia Alchem sponsored at the 2022 Cornell Nutrition Conference. If you'd like to see the full presentations and slide decks, go to alchem.com/real science and search for the 2022 Cornell Mini Symposia. This podcast features Dr. Jose Santos from the University of Florida. Clay, would you like to give us a quick intro to what the our audience is gonna see when a look at the presentation?
Dr. Clay Zimmerman (02:22):
Yeah, I mean, ba you know, basically Jose was, he was making a case for for coal being a required nutrient in and for all dairy cows. So he he spent a little bit of time, you know, just giving a little history of coal in, in diets and so forth, and, you know, did, did a nice job walking through the, the history in of coal and dairy cows, really, which really goes back to the 1980s and, and, and ended with some very recent research that has been done in his lab to really make a, really make a great case for colon being a required nutrient for all dairy cows.
Scott Sorrell (03:11):
Excellent. I look forward to the presentation, and I think you'll enjoy it as well.
Jose Santos (03:15):
I hope that I will present to you and make the case that co colon fits those criteria and it is a required nutrient for dairy cows. So during this new revelations on transition Cow Nutrition Symposium, I'm gonna spend the next 40 minutes discussing with you some aspects of CHO nutrition for dairy cows. So in my presentation, I'll give you a very brief history on cho, the role of calling on intermediary metabolism, then some new data on the effects of CHO in reducing fatty liver in dairy cows. I'll spend a little bit of time talking about how CHO and what are the impacts of CHO supplementation during transition and subsequent performance productive performance and health. And then I hope to make the case that every cow really, or most cows respond to cho. And the example I'm gonna use is body condition score in response to cho, and then I'll finish up with some potential mechanisms.
Jose Santos (04:20):
And I hope by presenting this data, I'll make a strong case that Colleen is a required nutrient for ruminants as it is for other species. So Colleen was discovered the first studies in the 17 hundreds by chemists and pharmacists, but it wasn't until about 40 years ago that work in England by Derek Lindsay here who showed that most phospholipids in remnants are synthesized Denovo. And the reason for that are synthesized Denovo from calling that's being synthesized primarily through transmit methylation reactions. Okay? And the reason for that is because most dietary colon that's present, either in feeds or if we were to supplement the cells are degraded by the gastrointestinal microbes. And this was then clearly demonstrate by Dr. Richard Hardman at the University of Maryland who confirmed those findings and showed that more than 90% of the colon in feeds do not show up past the rumen.
Jose Santos (05:37):
So they don't, it doesn't, they don't show up in the small intestine cho or those ingredients containing coal don't deliver coal for absorption in the small intestine. So this required a development of room protected products. And upon the development of those products, then numerous universities in the US and Europe, in Europe conducted experiments primarily with transition cows to evaluate the impacts of supplemental coal as room protected coal on performance in health of dairy cows. Now I think it is a required nutrient because it's critical for, since phosphite choline as well as shingle lipids, which are needed for integrity of cells, either cell membranes in every cell, or an integrity of neuro tissue. The colon also plays a role in this one carbon metabolism because CHO can be oxidized to bein and bein can donate met groups in trans methylation reactions that affect gene transcription.
Jose Santos (06:46):
Choline is also a substrate to be acetate to acetylcholine that is part of neurotransmission it's important for lipid transport and absorption. So in multi, multiple species, the recommended the daily allowances have been established such that for humans such as Dr. Zimmerman here the amounts of coal that are suggest to be consumed daily vary according to physiological state and sex, but anywhere between 400 and 550 milligrams per day. In the case of lane hands depend upon the intake the amount can be approximately 0.6 grams per day. Young checks it's less a hundred micrograms per day, 500 milligrams per day, I'm sorry lactate in cells depend upon the intake somewhere between four and six grams per day. But then when we get to the adult dairy cow there is this question mark. We just have not established the recommended daily amounts to be fed.
Jose Santos (07:52):
And I think part of the reason is because the microbes destroy choline. So we have to feed as a room and protected product, which creates some uncertainty on the exact amount that's delivered for absorption. So that's one aspect. And the other one is that we probably don't have a lot of those response experiments demonstrating the optimal amount of calling to be fed to there it cows, but because we don't know the exact amount, it doesn't mean that it's not a required nutrient for cows. Ok. So obviously colon has major roles, but I'll show you some of the biochemistry of the major metabolic pathways involving colon metabolites. So, colon, when absorbed from the gastrointestinal tract, can be a satellite in nerve cells into acetylcholine. That's important for nerve transmission between neurons or in the neuromuscular junction. Colon can be oxidized to this bean, which is a trimethylglycine product, which can donate methyl groups to homocystine, to regenerate methionine.
Jose Santos (09:02):
And in this process, there is an interplay with vitamins of the B complex, such as folic acid and methionine can be a precursor for sense of acid Enzo methionine, which is the universal methyl donor. So during this pathway of Rey methionine and trans methylation reaction, we can have acid methionine that can donate met groups, and this can be sy the synthesis can be enhanced by providing colon to oxidized to the cho can also be phosphorylated into phosphocoline. And phosphocoline can now combine with C T p to form c d p colon. And c d p cho is substrate to be phosphorylated to phosphite choline, which is the predominant phospholipid in cell membranes. This pathway of de novo sense of phospholipid takes place in almost every cell in the body. But there is a second de novo, a second pathway for de novo sense of phosphite choline through transmit methylation of phosphate ethanolamine, or the P m T pathway in which met groups are donated by acide to c d p ethanolamine to synthesize phosphite colon.
Jose Santos (10:30):
So we can synthesize phosphate colon through donation of methyl groups from acide, which can be the abundance can be enhanced by oxidation of colon into bat, or we can synthesize PST through this Kennedy pathway. This the novo synthesis of phosphocoline through phosphorylation of absorbed cho in gastrointestinal tract. So there's two major pathways for Synthes of phospholipids in cells. Now, when cows undergo transition, we know very well that as cows approach Calvin there is an increase in nutrient needs because of the growth of the fetus, concurrent with synthesis of colossal and optic of nutrients by the memory line. At the same time, unfortunately, particularly when cows are in the last week of gestation, dry matter intake decrease in most cows. And combined with the hormonal change that take place in that period, there is a up upregulation in lipolytic signals to accommodate the needs of the memory gland as cow's beginning lactation.
Jose Santos (11:43):
And this re results in extensive lipo mobilization. Okay ad PostIt tissue subcutaneous or abdominal aose tissue is mobilized to supply nutrients to the memory gland, particularly long chain fat acids for CINs of milk fat in colos and CINs of milk fat in milk. But those fatty acid can be taken up by other tissues, including the liver. But if in excess, the liver will find other pathways to dispose of the excess of fatty acids, primarily through ketogenesis, which can export keto bodies now as an energy peripheral tissues, including the memory gland for the novo synthesis of short of median chain fatty acids. But when excess of fat acids are taken up by liver, they can accumulate as fat droplets as droplets of triglycerol, which are quite dynamic quote unquote organelles within the liver, not just a storage site.
Jose Santos (12:46):
One of the impacts of accumulation of fat in liver is that it can induce inflammatory response in the hepatocytes. Okay? So eventually this count might develop some, some degree of hepatic lipidosis, and unfortunately, it takes time for cows to dispose of those fatty acids, okay? Because ruminants evolve without the same ability to export triglycerol as very low density lipoproteins as other species. But this hepa triglycerol can lift liver as very low density lipoproteins, which will deliver fatty acids to peripheral cells, including the mam gland for milk fat symptoms. But when in excess that can damage, the hepatocytes can induce a proinflammatory response. And in vitro studies have shown very clearly that access triglycerol in the hepatocyte can suppress urea jenesis and gluconeogeneis important functions for the remnant liver. So this can affect potentially the health of the cow and influence animal performance.
Jose Santos (14:00):
So remember that one of the pathways for synthesis psid is through trans methylation reactions, okay? Through that PMM T pathway in a potential donor of meteo groups is metin through the action of acid enzo methionine. Unfortunately, when cows undergo transition, period, methionine is limited because now there is increased uptake by the memory blood for synthesis of milk proteins, including synthesis of immunoglobulins that are transferred into colos. So this probably limits the supply of methyl donors for synthesis of phospho tido choline. So I believe that there is probably an increased need of colon in this period, not only to provide more methyl donors by being oxidized to bein, but also to provide choline to be phosphorylate for since the phosphocoline and eventually since the phosphoryl choline that would be critical at this point. So when animals have deficiency of choline, or when they have extensive lipo mobilization, they can develop fatty liver, as in this example here shown in this picture.
Jose Santos (15:18):
And obviously, fatty liver is a common phenomenon in dairy cows, approximately 40% to 50% of the cows in literature have shown at least a 5% concentration of triglycerol in hepatic tissue in the first two weeks of lactation. So fatty liver is very prevalent as far as we can tell, and we believe that this has implication to subsequent production and health. For the sake of time, I'm gonna show you data only on incidence of disease. But recently we addressed, we asked the question, what is the association between the degree of triglycerol infiltration in the tissue in the first week postpartum with production health and survival? And here is the health data of those cows. And what we found was a linear association between hepatic trig, gly and incidence of disease, either clinical or subclinical disease. So we had about 330 cows in which we collect liberation on the eight postpartum, and we evaluate incidence of disease for the first three months of lactation.
Jose Santos (16:30):
And what we found was that was that incidence of metritis increased as the triglycerol increased from two and 5% to seven and a half percent. So if we look at morbidity, we saw a tendency for increase in morbidity in those cows. We saw a tendency for increase in the risk of those cows leaving the herd by 300 days in milk, so increased from 8.9 to 13.7% in cows that had increased. Hepatic triglycerol concentration in the liver had were associated with an increased risk of hyperemia and hypocalcemia. And this is subclinical hypocalcemia. And this is hyperemia in the first week, first two weeks of lactation, I'm sorry. So cows have increased. Hepatic leaks are at greater risk of developing disease which compromise survival, and that has economic implications to the producer. Now, why do we focus on this transition period? Because this is the time when the concentrations of phosphite choline in plasma are the smallest in the lactation.
Jose Santos (17:41):
So we can see here that when cows approach Calvin, and during the first two weeks of lactation, the concentration of phosphate cholines in plasma are reduced in lactating dairy cows. This is why we think this is the optimal time to provide colon to cows to improve synthes of phosphate this critical time in lactation. Now we think that choline, by being a building block for phosphorylcholine and phosphorylcholine, by being a building block for synthesis of cell membranes, this becomes critical to the ability of tissues to export triglycerol. So choline has this lipotropic effects. So here are two experiments, the first one from University of Wisconsin, in which cows were subject to a fatty liver induction protocol. So they were limited fed to consume only 30% of their net energy for lactation needs to develop fatty liver.
Jose Santos (18:48):
And half of those cows were assigned to a treatment that received room and protected colon. Half of the cows remains untreated controls, and what the altar showed was that in every cow, there was an increase in he pad triglycerol content. But this increase was reduced in cows that received room protect colon. So we can see this marked reduction in lyce when cows on pre pregnant pre partum cows receive colon and were subject to field restriction. Marco Zabi here at the University of Florida addressed the same question, but then he asks, what is the response in terms of Hepa triglycerol when we increase the dose of choline ion fed as reprotect colon? And we can clearly see that when cows were induced to develop fatty liver, the triglycerol increase, but there was a linear decrease in the concentration triglycerol as the intake of choline ion increased up to 25.8 grams per day in a room protected form.
Jose Santos (19:58):
So again, two experiments showing that choline has lipotropic effects in dairy cows. So in Marco Nobis work, we collect liver tissue, and then we decide to explore our genes involved in the effects of colon and the par tissue out there by supplying colon to those cals. So let's just look at this heat map here. So if you look at the bottom of the heat map, we see the treatments, which are the dose of doses of ion from zero up to 25.8. Each row here represents a gene that was differential regulated by the level of colon fed. And as we go from the red to the green, we see an increase in expression of that gene. In other words, an increase in the message to synthesize a particular protein. As we go from green to red, we see a decrease in the expression of that gene, and we can clearly see that genes involve in antioxidant defense of the cell gene involves genes involved in sentences of phospholipids such as polyprotein, B 100, CHO kinase, microsomal transfer triglyceride protein.
Jose Santos (21:19):
They were all upregulated. They go from red to green as we move from left to right, in other words, as we increase colon ion intake. Whereas genes involved in synthesis of triglycerol or reification of triglycerol genes involved in acute phase protein synthesis, or in proinflammatory response, they were down regulated ascal consumed more CHO ion. So this tells me that by supplementing choline ion, we reduce he al CG glycerol, and either the increased intake of CHO or the increased synthesis of phosphatidylcholine or the combined effect with the reduced triglycerol outs gene regulations such that ites genes important for export of triglycerol in hepatocyte by the hepatocyte. And it reduces the expression of genes involved in reification of trio of fat acid into triglycerol and genes involved in the proinflammatory response. Now, we decide to readdress this question with an experiment that we completed recently.
Jose Santos (22:35):
So we assigned 110 cows to one of five treatments. We used the same hepatic lipidosis induction protocol. So we had cows that were fed at Lipton for a period of five days, and then all the cows under went a period of fit restriction to supply only half of the calories that they need at that stage of gestation. And according to their maintenance needs, then we applied one of five treatments and we measure lots of things including intake, and we took blood samples and liver samples at strategic time. So liver was collected on day six and 13. I'll show you the data of day 13. And we took blood samples throughout the experiment. So the treatments were a negative control, no supplemental choline as reprotect choline, or 12.9 grams of choline ion as reprotect choline from one of two products. So a product with low cho chloride.
Jose Santos (23:33):
And within this product, we doubled the dose of choline ion to 12 5, 25 0.8 grams per day. And then we did the same, but with a product that had a high concentration of cho chloride. For the sake of time, I want to focus on the effect of colon, which are these four treatments against the control and the effect of dose of cho, which is 12.9 to 25.8 grams being fed per day. So just to show you that we cause massive lipo mobilization. This is the body weight change in nine day period, cows lost two kilograms per day. They lost body condition and by losing tissue the marker that we used was fatty acid concentration in blood. And you can see that there's massive lipo mobilization and there's increase in ketogenesis. Important point here is it didn't matter what treatment they were assigned to, they all had marked increase in fatty acid and, and blood ke.
Jose Santos (24:33):
So what I'm gonna show to you here is independent of the level of lipo mobilization, which was very similar among all treatments. So here is the dry matter intake and that energy intake of the cows and the net energy balance during the feed restriction period. So these cows were deficient in 10 mega calories per day, okay? So our treatment protocol induced negative energy balance as expected because we limit fat cows. And by doing that, we increase trio glycerol in the pad tissue. So here's the concentration on day 13 during the period of feet restriction. And we can clearly see that we have high triglycerol concentration on the hepatic tissue on a wet basis, but the concentrations become smaller, they are reduced when cows were fed colon in the diet. And you can see that the reduction is greater when they are fed a larger dose from 2012 0.9 to 25.8.
Jose Santos (25:37):
Whether we look on hepatic triglycerol and acid basis or on dry matter basis, we see exactly the same response. We see a decrease by feeding coal, and we see a greater decrease as we increase the dose of colon. When we look at glycogen, we see the opposite effect, which is a very positive effect. We see an increase in glycogen by feeding colon, and this increment is greater when we feed a larger dose of CHO such that if we look at the ratio of triglycerol to glycogen, which has been proposed as the marker of fatty liver, that ratio is much larger for cows in the control than the choline supplemented group. And the ratio becomes further smaller when the cows receive a larger dose of colon. So this data corroborate with data from anal cook the Wisconsin experiment, as well as the data from Marco z Nobi, the Florida experiment that I showed before.
Jose Santos (26:36):
So I think there's very strong information that colon has lipotropic effects. It reduces the risk of fatty liver in cows when they undergo periods of negative nutrient bowels. Now, in a subsequent experiment, we apply exactly the same treatment protocol. So we had feet restriction in those cows for a period of nine days. And then on day 19 of the experiment with those cows, with this non ionic detergent to coat lipoprotein particles such that we prevent lipoprotein lipase from hydrolyzing, very lowden triglycerides in very low density lipoprotein. So we look at the rate of accumulation of triglycerides, I'm sorry. Okay, so this is what we did. We assigned cows to one of three treatments. So we had 33 cows. They were about 230 days of gestation. They were dry cows, and we assigned them to control and then to one of two types or products containing ion are cho chloride.
Jose Santos (27:46):
And the doses of cho ion that we fed were exactly 25.8 in both products. So for the sake of this conversation here today, I wanna focus on the effect of supplementing CHO and not necessarily on the effect of product. So the idea here is to prevent hlu of triglycerides and V L D L particles in plasma or in serum such that we can look at accumulation and accumulation reflects the rate of hepatic secretion of triglycerol. And then at six hours after we doze this non ionic detergent here, we collect lymph tissue from the superficial lymphatic vessels in the back of the other.
Jose Santos (28:30):
Okay, so this is what we found. This is the concentration of triglycerol in serum over time, over a period of 12 hours. And you can see that the accumulation is great greater in the dash lines than in the continuous line. So much so that we have a treatment by time interaction. The dash lines represent the cows that received 25.8 grams of room protected choline. So we, when we analyze the area under the curve, we can clearly see the cows supplemented with room protected choline have a greater accumulation of trie glycerol, meaning that they have a greater secretion of pathetic v ltl, which is the vehicle to export trie glycerides from the liver. And we think this is part of the mechanism for reduction in fat livering cows, as it is the mechanism for reduction of fat liver when choline is supplemented to monogastric animals.
Jose Santos (29:25):
So again, cows seem to behave very similar to monogastric species. And coal clearly shows an increased ability to export trie glycerin, which would reduce fatty livering cows. Now, when we evaluate the lymph composition in those cows, so these are the cows in green that receive ribbon protected colon. The cows in red are those that were controlled. We have 30 cows that we sample lymph that we were able to collect lean fluid as we go from blue to brown or red. The concentration of that analyte increase in the lymphatic fluid. And here are the analytes that were differentially concentrated in the hepatic in the lymphatic fluid of cows. So you can see that they included triglycerides. They included triglycerides with different fatty acid composition. They include amino acids such as isoleucine, isestine, alanine, lanine, but they also include the choline.
Jose Santos (30:34):
So we can see that the concentration of choline in lymphatic fluid increase as we go from right to left. And as I go from right to left, I go from the control cows that are clustered here to the cow supplementary with REM and protect coal. So we believe that by supplying coal in the diet, as REM protect Cho, this choline is being absorbed and it's been incorporated into the cells, but also is being transferred either to the bloodstream by the portal system, but also being transferred to the lymphatic system either through recirculation or by being taken up by the ALS in intestine. Now, I made the case that choline is lipotropic, so this should reflect into benefits to calcium lactation. So lemme show you data from a meta-analysis in which we look at the effects of dose of Cho that was supplementing the diet of transition cows on subsequent postpartum lactation performance in health.
Jose Santos (31:41):
So to be part of the meta-analysis, the experiment had to have a control treatment in which cows received zero grams of choline ion, and they had to have a treatment that cows were fed room protected cho starting pre partum. And here you can see the amounts of cho ion that were fed to cows starting pre part. So all the way from 5.6 grams per day to as much as 25.6 grams per day of choline ion. You can see it also the most experiments, most treatment means that we collect for the meta-analysis or in this range here of nine to 13 grams, because most experiments use 12.9 grams feeding rate for cows per day. Okay? So this is one of the response that we evaluate is energy corrected milk. And the way you read this figure, this is a forest plot. Each dot represents a comparison in the metaanalysis.
Jose Santos (32:39):
The references of the experiments are listed here on the left. When the point estimate falls to the left of the central line, it tells you that the response was reduced by the intervention. And the comparison here is always co rum protect choline against the control, which is zero grams of supplement of room and protective co. When the dot falls to the right, it means that the response was increased. The gray square represents the weight of that comparison, the meta, which is listed here, and this is the 95% confidence limit, which is also listed here. And you can see that most point estimates are to the right of the central line. In fact, this is the overall response from the mixed model, and it shows an increment in energy, correct milk of point 38 standard deviations with repre, which represents two point 18 kilograms in this dataset.
Jose Santos (33:36):
So, but on one important aspect of this is that there is a lot of external validity to this data because most points fall to the right of the line, meaning that in different experiments, different farms, different management, different diets, coal increased the response, in other words, energy, correct milk, but it also showed that there is heterogeneity in the level of response. Okay? So the effect of choline is dependent on other factors, okay? And that's what we explored here, one of which is the dose of choline fat. So here are the control treatments, this is the response to level of feeding. This is the overall effect, which is the central line. And you can see as the dose increase from zero to 25.6 grams per day, there was an increase in milk yield and there was an increase in energy, correct? Milk yield, which is this linear effect of colon in the diet of dose choline.
Jose Santos (34:36):
But there was also an interaction between dose of CHO and concentration of metabolized methionine as a percent of the MP of the diet postpartum, which was something that we explored. So let me show you what this interaction here look like. So this is the metabolized methionine as percentage of the metabolized protein of the diet. So we had experiments, we put all this data into the National Research Council software of 2001. So the predictions are all based on the NRC 2001, which probably predicts a little less methionine supply than other models such as the C N C P S. And you can see if we look at zero grams of choline ion compared with 12 grams of choline ion supplemented, we can see that indicts that are low in mee, there is a substantial increase in milk and in diets that are high metin, there is also an increase in milk kill.
Jose Santos (35:43):
So in both cases we see an increase in performance, but the in the increment becomes smaller in as we increase the supply of metabolize onion. If we look at energy, correct, milk is the same type of response we see here. A 2.4 kilogram increase with 12 grams. If we go to the maximal level, we see a four kilogram increase in milk. If we get to diets that are very high in methionine 2.4, 2.5, the increments smaller, but still present, okay? One to two kilograms more. So, the point I'm trying to make here is that although their biochemistry has an interplay, okay, obviously this is showing that both that meth, that the response of colon is dependent on methy, but there seems to be an additive effect that even in the presence of high methy, we still capture a response in milk yield and fat crack milk yield.
Jose Santos (36:47):
Now, lemme show you the effects on health of cows. An important aspect is not all the, we had 21 experiments reported for the meta-analysis, but only 11 or 12 of them report disease incidents in cows. Ok? So the dataset is not as robust for disease as it is for milk. And this is probably why for many of the disease we were unable to detect any impact of calling. But for the risk of retained placenta and the risk of mastitis, we saw a reduction with supplementing colon compared to control. And the reason why I'm showing you here, cho as a category, is because almost every experiment of reported disease fed 12.9 grams of cho ion as we're in protected calling. So here I make the case that there is a lot of external validity for the production response.
Jose Santos (37:46):
When cows receive calling, they receive by increasing milk, yield and energy, correct milk, okay? But they also respond by reducing the risk of some disease postpartum. Now, in two experiments that we conducted here at the University of Florida, cows were fed room protected coal starting at 255 days of gestation and stopping at exactly 21 days postpartum. So they were fed during the transition period. And you can clearly see that supplementing coal, which is the close symbols here, increase milk production. But more importantly, the increment was extended well above the period of supplementation up to 40 weeks in the experiment of Marcus Nobi, and up to, to 25 weeks in the experiment by one bti. So in both cases, we saw a two kilogram increase in milk kill for a period of 25 to 40 weeks, although cows were supplemented for only three weeks during only six weeks during the transition period.
Jose Santos (38:52):
And I think that probably part of this extended benefit comes back to the fact that during the transition period, we have the smallest concentrations of colon or phosphite choline in plasma cows, and probably by supplying Cho, we increase the supply of substrate for fossil lipid synthesis. And this is right at the time in which proliferation of memory cells are upregulated. So this is the transition period here, and this is activity of CHO kinase, that enzyme that phosphorylates choline into phosphocoline. And this enzyme is highly active in the memory aphelion, such that when there is proliferation of tissue, the enzyme is upregulate. So not only the gene is upregulated, the protein is, and the activity of that protein is upregulate. And this enzyme here is a marker for cell proliferation, in particular during breast cancer in human. So it is possible that by supplying more choline, we increase the abundance of phosphorylcholine that supplies the memory gland.
Jose Santos (40:01):
We substrate for synthes of phospholipids, and this upregulate the activity of colon kinase because we provide more colon. And this may stimulate memory cell proliferation that results in this extended benefit for lactation performance. Lastly, I wanna touch on this concept that probably every cow benefits from choline, and the example that I'm gonna use is body condition score. So we went back to our data in two experiments that has about 230 cows. And we asked the question, does body condition score a cows when they entered the closeup period or the closeup pin affect how they respond to colon? So this is the distribution of body condition score of cows in the control treatment in Marco Nobi experiment and in the colon treatment. And the same for one Bois experiment. So you can see that we have cows that have low body condition or moderate body condition.
Jose Santos (41:01):
We have fat cows, cows are over condition, okay? So the question we ask, do these cows that are low body condition respond as well as this cows that have high body condition to supplemental colon during the transition period? And I'll show you the distribution of the body condition score. You can see that we have a wide spectrum of body condition all the way from less than three to more than four in both treatments. And there's about 115 cows in each group here. And this is the milk killed and the the energy correct milk killed for those cows. And the way you look at this figure is that you can clearly see that as I move from the blue line to the orange line, there is an increase in milk and there is an increase in energy, correct milk of about two kilograms per day. And this increase is independent on body condition score.
Jose Santos (41:55):
So let me remove the dots, which represents every cow. So you can see here no interaction between supplementing room and protect CHO and the body condition score of the cow when she entered the closeup pin. So skinny cows respond to CHO with more milk and energy, correct milk fat cows respond to CHO with more milk and more fat cracked milk. So just to finalize, I wanna present some potential mechanisms for improved production health when cows receive cho, obviously. The first one is this classic mechanism that CHO provides substrate for since the phosphate colon, which is important to synthesize very low density lipoprotein particles that export triglycerol from the liver as it's been shown in Usman ad's recent work, and this probably explains the reduction in fat livering cows that I showed you very consistently from three experiments by supplying choline to choline deficient diets, we enhance the synthesis of chylomicrons, which is illustrated here in laboratory animals based on the concentration of polyprotein B 48, which is a marker for chylomicrons.
Jose Santos (43:11):
And chylomicrons are lipoproteins that facilitate the transport of absorb lipids in the gastrointestinal tract to be delivered to other tissues. So this would make the case that by supplying choline to animals that are deficient, enhances the ability to transport, absorb nutrients from the gastrointestinal tract to be delivered to other tissues. And in fact, when Marco nobi supplemented cho to cows and those them with fat, he showed that cho supplemented cows had increased concentrations of trias glycerol in plasma compared to control, suggesting that probably absorption and transport of lipids enhancing the gastrointestinal tract of those cows. We are very aware of the fact that cows are predisposed to this disruption in gut structure that results in leaky gut and efficiency of phosphocoline is a risk factor for leaky gut. Fost choline is important for Ethel cell integrity because it's part of synthesis of cell membrane.
Jose Santos (44:17):
And the structure of this gap junctions is enhanced when we have intact enterocytes. So when animals are deficient in phospholipids, it's possible that leaky gut that is, that might be happening during transition period, may be exacerbate. Also, phosphorylcholine has anti-inflammatory effects. It attenuates the pro-inflammatory response, which probably enhances the integrity of the gastrointestinal tract and improves liver health by attenuating a pro-inflammatory response that might be induced by accumulation of lipids. But phosphorylcholine has some very, or choline in general, has some other very cool effects. And one of them is this experiment here in which they bred mice to have microbes in the gastrointestinal tract that are unable to destroy dietary colon in into these three methylamine oxides. So they had the wild animals that had microbes that destroy colon. Therefore, the supply of colon to the offsprings, okay, to the offspring of those dams was limited.
Jose Santos (45:30):
Whereas in mice that they bred to have this stringent microfluid that is, doesn't express the enzymes that degrade colon in trimethyl amine, they would have increased supply of colon to the dams as well as to the offspring. And what they observed was that by supply more of these methyl donors, there was increased DNA methylation. So methylation of stones in the DNA were enhanced. And by methylating, you suppress opening of chromatin. So there is less gene expression. And some of the genes that were suppressed, they were involved with reduction in apo reduction, metabolic disease, and, and changes in behavior, the animals, so less anxiety. So by providing more calling through the less action of this degrade and enzymes and the gastrointestinal tract, they changed the phenotype of the offspring of those animals, which is a very cool response.
Jose Santos (46:35):
And lastly, I wanna leave you with this concept that there is this enzyme choline kinase that phos correlates choline into phosphocoline. And this provides substrate for the novo synthesis of phosphorylcholine in our cells. But this is probably very important to mammary of arteri cells that are replicating during the onset of lactation, and perhaps by supplying colon, we influence the activity of this enzyme in addition to providing more substrate. And this enhances cell proliferation, maintains cell membrane integrity. And this may explain the fact that cals keep responding in with additional milk yield and energy, correct milk well above, well beyond the period of coal supplementation. So I just wanna conclude that I think it's unquestionable that fits the criteria of required nutrient in as it does for other species. It reduces triglycerol infiltration and delivered by enhancing export of triglycerol as lipoprotein, it benefits lactation and reduces the risk of disease.
Jose Santos (47:41):
How important criteria that makes a nutrient required. The response is influenced by the supply of metabolized methionine and by the dose of colon fed, but still present even in high methionine diets. The benefits are observed regardless of body condition scar. And I think we need to be sensitive to this. It's not just the fat cow respond, the skin cow also responds to that, okay? So it's not, it should not be target only to over condition cows. It should be target at this point to the entire population and the mechanism that justify improved productive performance. It may be related to increased nutrient intake. I didn't show you this data, but there's some data to substantiate that perhaps improved nutrient absorption and integrity of the gastrointestinal tract. Obviously, there is less fatty liver, there is reduction in clinical disease, and perhaps we have this other non-classical mechanisms such as reduced inflammatory response and improved memory cell proliferation,
Scott Sorrell (48:43):
Mcclay, that's gonna be last call. Dr. Santos covered a lot of ground in many research studies. In your mind, what is the biggest takeaways for nutritionists and dairy producers from his talk?
Speaker 3 (48:53):
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Dr. Clay Zimmerman (49:18):
The pro, you know, probably the biggest thing was, you know, he spent quite a bit of time, you know, discussing the, the most recent meta-analysis that that was done out of his lab published back in 2020. You know, showing, showing the consistency of response to rumor protected colon across all these transition cal studies. And and also the fact that you, you get a response to rumor protected colon no matter what the production level of the herd. It's a very consistent response that really ranged from about, you know, 45 pounds of milk in in the control cows at the low end and up to a hundred pounds per cow in these control treatments. So, so nice responses across a very wide range production levels. He discussed, you know, some potential mechanisms of, you know, how, how choline may work in the transition cow.
Dr. Clay Zimmerman (50:23):
Some of these, we know some are still theories at this point. We know, you know, we, we clearly know that it, it has a positive effect on, on liver health that, and that really has lots of positive effects for this, for this fresh cow. But, but you know, he, he discussed some other potential modes of action that that quite frankly we're studying, trying to learn, learn more about. We at, you know, as, as we're running continue to run research studies with reassure, we learned something new every time. So he, he was, he was alluding to some, some different potential modes of action that are, that are there with colon and the transition cow. And really one, one of the last things he talked about was based off of actually another, another small meta-analysis that they did looking at whether or not the response to room protected colon is related to body condition score of these of these dry cows. Because, you know, there's, there certainly has been a thought in the industry that, that maybe river protected colon is only for fat cows, and that's not true. There is they looked at, they looked at data from different studies, it was over 300 cows total, and there was absolutely no correlation between pre partum body condition score and the res and the production response to rumor protected colon.
Scott Sorrell (52:05):
Yeah, so,
Dr. Clay Zimmerman (52:06):
So really shooting down that, that dog by, that's, that's been out there in some circles.
Scott Sorrell (52:11):
Yeah. So Clay, one of the things I found interesting in, in the talk was the discussion around body condition score and the feeding of rumor protective. Could you kind of touch on that for a moment?
Dr. Clay Zimmerman (52:24):
Yeah. So they Jose published a paper, I think it was back in 2021. Now at looking at is, is there, is there a relationship between pre partum body condition score and the response to rumor protected colon? Cuz there, there's been a dogma out there that only fat cows respond to, to rumor protected colon supplementation. And that was absolutely not true. There was absolutely no, there was no correlation between pre-party, pre partum body condition score and the response to rumor protected colons. So whether they were thinner cows or heavier cows, they all responded equally with, with increased milk, milk and energy corrected milk yield.
Scott Sorrell (53:15):
Hmm, great summary, clay. Remember, this is just one in a series of four podcasts around the new revelations in transition cow nutrition. Be sure to tune in for the rest of the series, and if you haven't listened to the first one, go back and listen to the first one by Dr. Mike Van Amberg. The next podcast will take us into a deeper dive into some of the newest transition Cow nutrition research coming out of Michigan State University with Dr. Barry Bradford. So we'll look forward to seeing you there. To all of our loyal listeners, thank you again for coming along for more than 60 episodes and sticking with us as we explore more topics. 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 3 (53:57):
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