Preparing Lactating Dairy Cows for Unseen Stressors - Featuring Experts from Novonesis

Stress on lactating dairy cows isn't just about what you can see, it's about what's happening beneath the surface.

​On this episode, listeners will gain practical insights into supporting gut stability, optimizing digestion and helping cows maintain performance when stressors arise, whether they are environmental, nutritional or management related.

Listeners will also gain clarity into the evolving role of microbial feed additives in dairy nutrition, understanding how different technologies function and where they fit in today's feeding strategies.

This episode is eligible for ARPAS continuing education credits.

Transcript

SPEAKER_00 0:39

Good afternoon, everyone. My name is Betsy Whitaker and I'm a national talent manager with Nova Nestis. Thanks for joining us. And before we get started, I'd like to give a brief introduction of our speaker today, Dr. Keith Bryan. After earning three degrees and spending 20 years at Penn State University in the Department of Animal Science, Dr. Bryan transitioned to the feed out of the industry, focusing on silence documents and nutritional supplements that featured these probiotic bacteria, prebiotics, and digestive enzymes. Dr. Bryan's experience and expertise extends across a multitude of species and disciplines focused primarily on nutritional support of production, reproduction, and help for improved performance and sustainable economic returns. Dr. Bryan currently sits on the North American Text Service team for noveletis, focusing on sound inoculants and forage management along with life cycle feeding of probiotics to dairy cattle. His professional interests include the integration of microbial technologies from farm to forest, including seven soil amendments, impacting plant health, improving nutrient preservation and digestibility from forage inoculants, enhancing health and nutrient utilization of dairy animals fed probiotics daily, and the impact of these technologies on the health and well-being of a rapidly growing human population. With that, I'd like to turn it over to Dr. Keith.

SPEAKER_03 2:06

Thank you, Betsy. So we're going to tackle kind of an interesting topic for me in the probiotic space. So basically, the the title is Preparing Lactating Dairy Cows for Unseen Stressors. And so when we speak to stressors, I think it's important to enumerate those and categorize them. And we all know that stress can be deleterious to performance. So these stressors impact lactating dairy cows in a variety of ways. So first and foremost, there'd be environmental stressors. That would be heat and cold stress, social stress, crowding, flies, etc. And those environmental stressors sometimes we can see, sometimes we can't see, but uh we may be able to sense them in a variety of other ways. Likewise, there are feed and water stresses. There could be pathogens in the feed, obviously, mycotoxins tend to be a big concern. Feed availability or feed restriction in certain cases, obviously, stray voltage would be an environmental stress that impacts water and water consumption. Lastly, there's the management stressors, uh, handling, transport, grouping, regrouping, and even injury or the effects of foot health, lameness, hairy heel wort, whatever the case may be, can be managed, and there's a lot of crossover between environmental feed and water and management stress. But suffice it to say that when we look at these environmental stressors or management stressors, stress generally creates systemic inflammation. And sometimes it's manifest as leaky gut. And so we're going to focus a lot of our attention on the effects of probiotic in the digestive system of the cow, but it goes well beyond that. And everybody's heard of leaky gut. Uh, it's a problem, uh, can be manifest severely in the form of hemorrhagic bowel syndrome or HBS, uh, potentially leading to death. But if you've heard me speak before, I'm a firm believer that for lactating dairy cows, a healthy rumen, a healthy lower gut, ultimately creates a healthier cow that should be more productive. And if I say that once, I may say it multiple times throughout today's discussion. So, focused on systemic inflammation and the leaky gut, it's important to realize that there are a lot of tangential axes that are impacted by the digestive system of most mammals. One of those would be the brain-gut axis influencing the immune system and the enteric nervous system. Another, obviously, is the gut-lung axis, and we know through evidence and research that feeding a science-based research-proven effective probiotic can impact lung health and respiratory disease incidents. We also know that there's a reproductive component between the reproductive tract and the digestive system of mammals, as well as a mammary access interplay with the gut. So just because we're talking about gut health and the impact of stressors on gut health, doesn't it mean that we can exclude other organs and tissue systems. And suffice it to say that there's a plethora of data in the literature that points to leaky gut or systemic inflammation resulting from any number of stressors. These could be pathogens, they could be uh nutritional, uh dietary imbalance leading to stress, uh, grouping, so on and so forth that we described on the previous page. But once we uh experience or have cows that experience systemic inflammation andor leaky gut, a lot of times that leads to dysbiosis or an imbalance in the microbiota and microbiome of the digestive tract. And we could have reduced beneficial microbial growth resulting from that. And that could lead to decreases in digestibility, decreases in nutrient absorption, there could be pathogen proliferation within that digestive tract, ultimately leading to secondary disease or diseases, which is morbidity, and we all know that morbidity leads to suboptimal performance. So if we're going to focus our attention on preparing high-producent lactating dairy cows for unseen stressors that impact the digestive system, we need to think about the types and kinds of supplements or additives that we could feed. And just to keep things simple, I've broken this down into four buckets. The first bucket is lactic acid bacteria. The second bucket are spore-forming bacteria in the form of bacilli. The third bucket is live yeast, and the fourth bucket is hydrolyzed yeast or yeast cell wall. And again, there are numerous other feed additives that could be at play in the strategies to mitigate stress, particularly digestive stress and the interplay with the other axes, but we're going to limit ourselves to these for today. Basically, globally, the first three buckets, lactic acid bacteria, bacilli, and live yeast, can be defined as live viable probiotics, or AVCO in the US defines them as direct-fed microorganisms. And so, by definition, a probiotic is a live viable microorganism that, when fed in sufficient quantities, confers a health benefit to the host. And so the focus is on health. It's not necessarily on performance, but again, when we speak to health, a healthy rumen, a healthy lower gut should lead to a healthier cow that ultimately is more per productive. Likewise, hydrolyzed yeast are non-living, and we can separate out live yeast and hydrolyzed yeast from the bacterial probiotics by splitting this figure in half right down the center vertically. By definition, direct fed microbials, according to AVCO, have been reviewed and found to be safe. And there's approximately 49 species of organisms currently on the DFM list. Now, again, I think it's important to distinguish two types of DFMs. There are DFM products, which is a dried fermentation product with a genus and species listing, and those dried products should come with a corresponding viable cell count or guarantee on the label. Conversely, that same organism, say Bacillus subtilis, could show up on the label as a dried bacillus subtilis fermentation extract, and in that case, the designation of extract simply refers to an organism that was used to produce enzymes. And similar to DFM products, where you're looking for a live, viable CFU guarantee, if enzymes are included in any feed additive, we should also look for a guarantee of enzymatic activity for those enzymes that are included. So as we take just a step back, we'll delineate the various buckets and what is actually in those buckets. So again, the first bucket is lactic acid bacteria. These are non-modal colonizing symbiotic organisms. The caveat among the lactic acid bacteria relative to, say, the bacilli is that they're non-sporulating, so they're not resistant to heat. So generally, temperatures above 40 degrees C or 100 degrees Fahrenheit can be detrimental to these organisms, and certainly temperatures of feeds that pass through pelleting or extrusion are going to kill lactic acid bacteria. These bacteria want to be where it's comfortable. They want to be warm, they want to be fed, and they're not modal, so they have to be carried around with the digesta. Like most other probiotic bacteria, they can elicit durable changes in the microbiome and/or the microbiota within the digestive tract of animals. When we look specifically for important outcomes from feeding lactic acid bacteria as probiotics, it generally focuses on intestinal health. Whether it's vilus height and cryptopth or tight junction protein integrity that basically connects the intestinal epithelial cells together. It could be mucus production or competitive exclusion. And we'll talk about some of these later on. When we look at spore-forming bacteria in the form of bacilli, converse to lactic acid bacteria, they are modal. They can move on their own as individual organisms or as sheets or layers, or similar to what you might envision for a swarm of bees. They are sporulating, and because of this, they're resistant to high temperature, they're resistant to extremes in pH and acids, and they don't care where they are. They're designed to survive. If you walk out through the woods and you see a rotting or decaying log or pile of leaves, there's probably bacilli among those leaves or on that log that are helping to break it down into its substrate components. Interestingly enough, bacilli form biofilms. They produce effective concentrations of enzymes and they attack potentially pathogenic bacteria through two primary modes, whether it's bacteriocins or quorum quenching molecules. And those quum quenching molecules simply are the ability to disrupt the communication among pathogenic organisms to elicit a disease response. As we move on to the third bucket, which is yeast, there's a variety of yeast definitions used by AVCO. You can see yeast culture is different than active dry yeast because active dry yeast has to have 15 billion live yeast cells per gram, whereas yeast culture, there's no guarantee as far as live organisms. Yeast extract is generally the contents of a yeast cell, excluding the outer layers, and hydrolyzed yeast would be just some fragmented portion, either mechanically or enzymatically, disruption of that cell to break it into its component parts of cell wall versus cell content. For the sake of today's discussion, we're going to focus solely on active dry yeast. And when we do that, there are specialized fungi as opposed to mold that are capable of scavenging oxygen. And when we put those into the diets of lactating dairy cows, they scavenge oxygen, making the rumen more anaerobic, and those fibrolytic bacteria prefer an anaerobic environment and lead to improvements in NDF or fiber digestibility. They increase rumen pH, they produce vitamin B12, and can stimulate microbial protein and VFA production. The last bucket, hydrolyzed yeast, a lot of you are probably already familiar with this. It's a significant source of manan oligocyccharides or MOSS, as well as beta-glucins. It's capable of immunomodulation, and it has also been shown to non-specifically bind mycotoxins and pathogenic bacteria. But by definition, it is a prebiotic. It is a source of food for the naturally occurring bacteria in the rumen and the lower digestive tract, as well as any probiotic bacteria that may be added to the diet of these high-producing cows. So today we're going to focus on biome balancer CS. It's a neutral end product that's supported by novinesis. And so when we look at the four buckets relative to biome balancer CS, we have lactic acid bacteria in biome balancer CS in the form of LA51 and PF24. Those of you who are familiar with bovamine dairy know that this is what's contained in the bovamine dairy product. And we guarantee that at 2.27 billion CFU per daily dose. Likewise, the second bucket, the spore-forming bacteria, we now have three strains of bacilli. Bacillus subtilis and bacillus lichiniformis correspond to a commercially available product known as bovacillus. And what we've done is we've added the bacillus subtilis king strain, and I'll show you why we did that here in a few minutes. Those bacilli are guaranteed at 13 billion CFUs per daily feeding. And then we have live yeast, strain Y 1242. It's included at 50 billion CFUs per daily dose. There is no hydrolyzed yeast or yeast cell wall in this product. When we look at the primary modes of action of probiotics, it can be compartmentalized into three main categories. One is the probiotics interaction with feedstuffs, two, the probiotics interactions with other microbes in the digestive tract, and then lastly, the probiotics interactions with the host specifically. When we look at interactions with feedstuffs, bioconversion is simply taking something like a simple sugar and making an acid, which most probiotic organisms are capable of doing. When we look at the interactions with other microbes, there are probably some favorable interactions with desirable microbes, but there are also some very favorable interactions with undesirable microbes. So direct antagonism, particularly relative to pathogenic bacteria. So I'll show you some examples of diffusion plate assay and multiple pathogen inhibition by probiotic bacteria that can be found in biome balancer CS. The last of the three primary modes of action deals with increased absorptive capacity, barrier, and immune function. And through extensive research, we've been able to show repeatedly that when dairy animals are fed an effective probiotic, we see an increase in absorptive capacity manifest as increased villi height andor crypt depth, increasing the surface area of those finger-like projections in the small intestine. So, what exactly do the lactic acid bacteria do, and just as importantly, what are they in Biome Balancer? Well, we have a unique combination of a lactic acid producing bacteria in the form of Lactobacillus animalis or LA51, and a lactic acid utilizing bacteria. In the form of PF24, which is that propriani strain listed toward the bottom of this slide. So the lactate producer improves gastrointestinal stability, enhances immune responsiveness while producing lactic acid that can ultimately be consumed by the PF-24 as it converts lactate to propionate, a significant source of energy for ruminants. And again, it's two strains at 2.27 billion per daily dose. Just looking at the two strains by themselves, we have a considerable amount of research that's been conducted looking at the efficiency of energy corrected milk production. So we calculate energy corrected milk divided by kilograms or pounds of dry matter intake, and that gives us the efficiency of energy corrected milk production. So among the seven studies conducted from 2004 to 2016, you can see variable responses, but on average, we typically see a 2-3% improvement in energy corrected milk production efficiency. Likewise, we've looked at apparent digestibility of a variety of nutrients or feed components, and generally this is done as a before and after. So we'll evaluate apparent digestibility before a pen of cows is on the product and then while they're on the product. And you can see the p-values listed and the number of positive results out of the total of 16 herds over the three-week period. And so there's a significant improvement in protein digestibility, starch digestibility, as well as a significant reduction in fecal starch. All of these variables point to an improvement in overall energetic and feed efficiency. One of the in vitro methods that we use pretty regularly at Novanessus is a transepithelial electrical resistance assay. Now I need to describe the components. So basically, there's an inner well and an outer well. And that represents the apical or the gut lumen side versus the basolateral or the body proper side. You can see that there's electrodes placed in each of the wells. We pass an electrical charge. Obviously, if this cell layer is intact, there's going to be very little electricity that passes through one from one electrode to the other. That is high electrical resistance. As this layer breaks down, more electricity can pass from one electrode to the other, so we have lower electrical resistance, and that's depicted on this graph out here to the right, where high electrical resistance due to an intact monolayer of cells rises above 100%, whereas if we break that down somehow, we get a lower electrical resistance. So we can do a variety of things in these wells. We can put probiotics in by themselves, we can put pathogens in by themselves, we can put the probiotic and the pathogen in together and see what these electrical resistance responses look like over time. So again, we grow those cells for about three weeks to confluence. Again, we can put the probiotic in by itself, the pathogen in by itself, or we can include these fluorescing molecules. And ultimately, we're going to hopefully see very few, if any, of those fluorescing molecules translocate into the outer well, indicative of a cell monolayer that has maintained its integrity, or conversely, if it starts to break down, as we would experience in a leaky gut scenario, you can see some of these fluorescing molecules translocate into the outer well. So the next couple slides I'm going to show you will have both the electrical resistant component to it as well as the fluorescing molecule component to it. So when we put Lactobacillus animalis, LA51 against Salmonella typhemurium, the figure looks something like this. So we set the baseline control at 100% indicated by the black circles. We can put LA51 in by itself, the gray squares, and we have higher electrical resistance, meaning we've actually improved the integrity of that monolayer. We can add a high dose of salmonella by itself, which is the dark green squares, and you can see that the electrical resistance diminishes, meaning there's more breakdown of that monolayer. And then when we put in some LA51, in this case, let's call it a medium dose of LA51, there's a very slight improvement in electrical resistance because we want to get back up to either control levels or probiotic alone levels. And you can see over here that there's quite a bit of translocation of those fluorescing molecules when the pathogen is present by itself. But when we add the pathogen and the probiotic together, we reduce that translocation, meaning we maintain some integrity of that monolayer. When we take a medium dose of Salamenella against a medium dose of LA51, the line shifts upward, and the gap between the pathogen by itself and the pathogen probiotic combination widens. And we're approaching control levels with our pathogen-probiotic combination. And then lastly, when we kind of win the battle, if you will, because remember, I have a medium dose of LA51 against a low dose of salmonella, again, both lines shift upward, and the gap between the two gets wider, and we're a lot closer to our control level. So we're seeing the benefit in vitro in this tier assay of putting LA51 in this system when we have Salmonella Typhemurian. Alternately, we're here from the field that Salmonella Dublin is rearing its ugly heads, so we ran the same thing with LA51 and Salmonella Dublin. So the control line again is set to 100%. When we put LA51 in by itself, we have greater electrical resistance. The red line, when we put Salmonella Dublin in by itself, it wreaks havoc on that monolayer, and a lot of electricity gets through, hence lower electrical resistance. And when we put LA51 and Salmonella Dublin in together, we can see a significant improvement back toward control levels. In terms of fluorescing molecules, you can see control and LA51 allow a very, very low percentage of fluorescing molecules through that monolayer. The pathogen by itself is considerably higher percentage of fluorescing molecules that pass through, and we get a lot closer back to control level when we put the probiotic and the pathogen in together. As we shift back to biome balancer and the spore-forming bacilli, there are three strains of bacilli in this product. Bacillus lichiniformis 809 is known for robust pathogen mitigation, and we added just recently the bacillus subtilis king strain and basically increased the bacillus feeding level by about 50% above previous. The interesting thing about the bacilli is that they're very robust, and we have done extensive recovery of live viable bacillus spores of any of the three strains I mentioned on the previous slide in a variety of dry applications. As you can see from the slide, the premixes, mash feeds, pellets, range cubes, mineral mixes, and complete feeds, and those even tubs, whether they're heated tubs or you know chemically hardened blocks. And the viability of those bacillus organisms is quite high. We've also tracked germination and replication of these bacilli. The four figures on the current graph simply show that we get growth of those bacilli in rumen fluid, in the ileal content of the small intestine, in the cecum and colon as well. The other noticeable benefit of live viable bacillus as a probiotic is their ability to produce enzymes. And so when we put either of the two strains, uh 809 or 810, on a purely starch substrate, they will produce amylase in order to survive. If we put them on a pure fat substrate, they will produce lipase in order to survive, and so on for pure protein substrate or pure fiber substrate. So they're very adept at shifting their enzyme production in order to ensure their survival regardless of the nutrient substrate that's available. Specific to the addition of the king strain in this product is the fact that it's an excellent producer of protease. And so that's depicted on this slide. When we talk about health, we have to focus our attention on pathogens, and I'm a firm believer that we never know what pathogen is going to present, the extent to with it what it's going to present, or the immune status or physiological state of the cow to combat that pathogen at a particular level. So we use diffusion plate assays to help get a handle on what might be happening in vivo in terms of pathogen mitigation. So we know that Clostridium perfringens is mitigated or inhibited by most bacillus-based probiotics. And initially we focused all our attention on Clostridium perfringens type A. But based on the figure on this slide, we now know that our bacillus-based probiotics are capable of inhibiting all five types of clostridium perfingens. We also use the diffusion plate assay, and you can see the scoring system versus the visual inhibition rings in that assay system. In the table, we've broken out salmonella, E. coli, clostridium, and enterococci, and you see the relative inhibition of the king strain versus two other Bacillus subtilis organisms in our extensive library. And without fail, the king strain is antipathogenic for the vast majority of organisms and classes that we investigated here relative to the other two bacilli. When we deep dive this just a little bit to look at a couple different strains of Salmonella, particularly Salmonella typemurium, you can see that the king strain is superior to those other two bacillus subtilus organisms in terms of Salmonella inhibition. We did the same thing for E. coli and arrived at the exact same conclusion that for the enterotoxigenic E. coli listed, the king strain was superior to the other two bacillus subtilus strains in terms of its antipathogen activity. We continue to investigate a variety of pathogens relative to the probiotic of interest. In this case, we looked at truperella, which has been implicated in foot rot in cattle, as well as Fusobacterium necroform, which has been implicated in liver abscesses, primarily in feedback cattle, but also in cald dairy cow when harvested. Lastly, as far as buckets go, we've got the live yeast bucket component of biome balancer. So it's a unique strain of Sacchoryces cerevisiae with functional attributes that have been optimized for ruminants in terms of robust oxygen scavenging, production of vitamin B12, and improved fiber digestibility. And we include 50 billion CFUs of live yeast per daily dose. To view the benefits and outcomes a little bit differently for the yeast strain that is in biomebalancer, you can see the direct actions of utilizing lactic acid or lactate. The yeast compete with other bacteria for sugars in order to survive. They're very good at oxygen scavenging and produce vitamin B12. And so, from a benefit standpoint, the two that are most important in my estimation are the stabilization or actual increase of rumen pH and the increased fiber digestion resulting from feeding live yeast. In terms of confirmation, we can look at the meta-analysis across a variety of yeasts and a variety of trials that was published back in 2008, and you can see a significant increase in rumen pH, VFA production, a reduction in lactic acid, organic matter digestibility, dry matter intake, milk yield are all significantly improved with the addition of live yeast versus an untreated control trend or tendency for milk fat content to increase as a result of increased organic matter and mainly NDF digestibility with no significant difference in milk protein content. So, in summary, I think we can all agree that there are a variety of stressors that cows may experience throughout the day or throughout the year, and they fall into three broad categories, but we never really know exactly when some of these stressors are going to impact a pen of cows or individual cows within a pen. We all know that these stressors can lead to systemic inflammation andor leaky gut, and biome balancer CS is just one tool in your toolbox to help mitigate this systemic inflammation and leaky gut scenario, and hopefully return cows that are stressed back to having a healthy rumen, a healthy lower gut, ultimately becoming a healthier cow that should be more productive. And with that, I appreciate everyone's time. Any closing comments for the group, Betsy?

SPEAKER_01 41:27

No, thank you so much for your participation today, and uh thank you, Dr. Keith, for providing us with that information. And uh we look forward to speaking with all of you more out in the field.