Thank you, Bruce, for the introduction and good evening, everyone. So, first, I would like to thank also to the people who have joined the webinar because I know that sometimes at the end of the, of the day it could be a little bit challenging. But I want also to thank Doctor Chavia Manteca and Doctor Cecilia Villaverde as they have been available for giving this fantastic talk that you will listen now and I'm sure you will really enjoy, with these two very well recognised experts in the field, and they are also a really fantastic.

Plastic speakers as, as you will see. So thanks to both for, for being available for doing this webinar. And thanks also to the webinar bed, of course, because they are the organisers of this session.

So it's always a pleasure for us for Purina to sponsor these webinars that promote science and, and innovation. And also as a science-based company, we are especially proud of our veterinary products that are preinaro plant veterinary diets that you can see in this slide or veterinary range, which we offer a complete nutritional solution for canine and feline patients with a concise, comprehensive. An effective range of dry and wet products that are adapted to promote pets' well-being.

So if you are interested or interested or you want more information about the products, please, you can contact the bringer representative that will be in your country and we will be very happy to answer any question that you can have. And also as a science-driven pet food company, we are also very, very proud of having the PRINA Institute, which is a global professional organisation that is the voice of Purina science. So please don't hesitate to click this link because inside there you will find really very valuable scientific material and content.

So. So yeah, just click, click on it and, and you will see. And also to make you an idea of what PRINA science means, here you have some numbers that I want to share with you tonight.

So in PRIA, we have more than 500 scientists and pet care experts spread along a research and development facilities along the 5 whole continents. So we have really Huge facilities and also we have 100 years of experience in research, and we have been really pioneer in science of pet health, publishing more than 500 papers in worldwide scientific journals. So you can imagine the for that Pura is doing also to drive science and, and innovation.

So we are really proud of this too. And yeah, after this short introduction, I don't want to spend more time on this and let's enjoy this fantastic webinar on how nutrition can impact the brain. So thanks again to everyone and Doctor Manteca Dilla Verde.

We look forward really to hearing you tonight. Thank you. Jennifer, thank you for those words and explanation of Purina.

And once again to yourself and to Purina, thank you so much for the sponsorship. Folks, as I said to you tonight, we have got two different speakers, presenters, over and above. Jennifer, 3 different continents they're all sitting on.

So if we do have any technical delays or anything else, just bear with us. But, the, the trials have all run very, very smoothly. So our second speaker will be Cecilia, and she obtained a veterinary degree in 2000 and her PhD in animal nutrition in 2005 from the Autonomous University in Barcelona.

She worked as a post-doctoral researcher in feline nutrition at the University of California Davis, where she also completed a residency in small animal clinical nutrition. She's board certified in veterinary nutrition by the American College of Veterinary Nutrition and by the European College of Veterinary and Comparative Nutrition. And she has been the president of that organisation since 2016.

After working at the nutrition centre support services at UC Davis for a year, Celia is now a consultant in clinical nutrition for expert pet nutrition and also the Veterinary Information Network. She is a member of the World Small Animal Veterinary Association Global Nutrition Committee and has written several articles and book chapters on the topic of companion animal nutrition. So welcome to Cecilia.

As I said, she is going to speak 2nd. Our first speaker tonight is Javier, and he received his BVSC and PhD degree also from the Autonomous University of Barcelona, as well as an MSC in applied animal behaviour and animal welfare from the University of Edinburgh. Currently, he is a full professor of animal behaviour and animal welfare at the School of Veterinary Science in Barcelona.

One of his main fields of interest is companion animal behaviour and welfare. He has published extensively and he is a diplomat of the European College of Animal Welfare and behavioural Medicine. So we have got very well qualified people talking to us tonight.

Enough from me. Javier, welcome to the webinar vet and over to you. Well thank you very much, Bruce, and good, good afternoon, everyone.

Thank you for being here and thank you to Nestle Bria for sponsoring this webinar and thank you, of course, to the webinar vet for organising it. Now, this webinar is on how we can use nutrition to modify brain functioning and address behavioural problems in dogs and cats. This is the plan of the webinar.

We will start with a short introduction and then we will basically address 22 different issues. One is cognitive dysfunction in senior dogs and cats, and the other one is fear, anxiety-related problems in companion animals. For, for each of these two problems, I will, give a little bit of background to set the scene.

And then Cecilia will discuss how we can use nutrition to address these problems and we will finish up with a summary and a list of concluding remarks. So let's just start then with the introduction. Our dogs and cats clearly provide many benefits in terms of human health and welfare.

Sometimes though, their behaviour may become problematic. The graph you are seeing now shows the results of a study we did in Spain some time ago looking at a large number of dogs that were taken to the animal hospital at the vet school in Barcelona because they were showing behavioural problems. So you can see, roughly half of them.

Were aggressive animals and then we had around 10% of dogs with elimination related problems and between 5 and 10% of dogs that were showing excessive barking or destructive behaviour or fear related problems. These behavioural problems can be seen across all ages in, in dogs and, and to them, we have to add the cognitive dysfunction syndrome, we will talk later on in the webinar which is found in geriatric dogs. Now, if you look at this graph and the sort of behavioural problems we encounter in dogs, I think it becomes clear.

That behavioural problems have a negative effect on the quality of the human animal bone. And it is not only the quality of this bone which is affected by behavioural problems. They also have a negative effect on the welfare of the animal itself.

This graph comes from a paper which was published in the journal Animal Welfare and in the paper, the authors were ranking different welfare problems in companion dogs based on a variety of, of criteria. And as you can see, undesirable behaviours or problem behaviours rank among the top 3 welfare problems in companion dogs. So in, in summary, in dogs, and this is applicable to cats as well, behavioural problems are both common and important.

And they are important for two different reasons because they have a negative effect on the quality of the human animal bone and because they have a negative serious effect on the welfare of the animal itself. Now, how can we address these behavioural problems? Well, behaviour comes from the brain and brain functioning and therefore behaviour may be modified through different strategies.

Nutrition is one of them. And as I said before, Cecilia will be looking at how we can use nutritional expertise to address behavioural problems in dogs and cats. Now, we will use as an example as two case studies, the cognitive dysfunction syndrome in geriatric dogs and then later on, fear or anxiety-related problems.

So, let's look at cognitive dysfunction in geriatric companion animals. You are all familiar with this problem, the cognitive dysfunction syndrome, you know that it is a set of symptoms that are a consequence of a progressive loss of cognitive capacity in senior dogs. And it has been well established that cognitive dysfunction syndrome has an important effect, a negative effect on the welfare of the doctor.

The reasons are several, but one of them is that because Dogs, show a loss of, memory and, and learning capabilities. The ability to control their environment is, seriously affected and this, becomes stressful for, for the animal. Now, cognitive dysfunction syndrome is a highly prevalent condition.

We have here an example of one of the very many studies looking at the epidemiology of cognitive dysfunction syndrome. This in particular was done in Spain and as you can see, more than 20% of dogs older than 9 years show cognitive dysfunction syndrome. Now, this condition has several risk factors, but by and large, age is the main risk factor for cognitive dysfunction syndrome.

And age is important in two different ways. First, as dogs grow older, they have a higher risk of developing the problem. So for example, here, roughly 30% of dogs, from 11 to 12 years old had the condition whereas this percentage went up to almost.

70% in dogs from 15 to 16 years old. But the second consequence of age is that as dogs grow older, they are more likely to show more than one symptom of cognitive dysfunction syndrome. So, a higher prevalence overall and a higher likelihood of dogs with the condition showing more than one symptom.

And talking about symptoms, this is a summary of what can we expect in dogs with cognitive dysfunction syndrome. The set of symptoms has been summarised using this acronym, dogs with cognitive dysfunction syndrome may show disorientation, so they may be unable to find its way back home or around objects. They show changes in the social interactions.

Normally they, they show less social interactions, but in some cases they may be fearful or, or, or even aggressive. Changes in the sleep wake, cycle, more active at night, night, vocalisations. Changes in house training issues so they stop responding to commands, unable to learn new commands and, and, and may show how soiling problems.

Changes in activity are commonly, reduction in exploration and, and play, but in some cases, we may see an increase in activity mainly because of repetitive behaviours. And finally, and more recently, it has been shown that dogs with cognitive dysfunction syndrome show a higher risk of anxiety. Now, what are the mechanisms underlying cognitive dysfunction syndrome?

Well, there are many, but I would like to emphasise two of them which I think are highly relevant when it comes to treatment. One mechanism is neuron loss. We know that geriatric dogs and cats lost neurons across the brain mainly in the hippocampus and the prefrontal cortex.

We know as well that this loss of neurons is caused mainly by a reduction in neurogenesis, meaning the formation of new neurons, particularly in the hippocampus. And this reduced neurogenesis is correlated with learning and memory deficits. So we have neuron loss.

And another well, studied mechanism is related to mitochondria dysfunction. We know that age, mitochondria are less efficient at producing energy and they produce more free, radicals which cause oxidative processes in the brain. Now, how can we address cognitive dysfunction syndrome in dogs?

Well, Cecilia is gonna talk about nutrition, nutrition is extremely important. When addressing CDS cognitive dysfunction syndrome, I think it's important to realise that nutrition has to be part of a broader strategy that includes other things as well. And among these things, I believe that changes in the environment and in the general husbandry of the animals are very important.

Now, this slide shows a very interesting study looking at how physical activity may be beneficial to stop this loss in neurogenesis and cognitive capabilities in senior animals. Of course, we're talking About, geriatric dogs, so physical activity has to be tailored to the health status and to the age of the dog. But, but considering that, providing opportunities for play, walking, and physical activity in general has been proved to be, an effective strategy.

And the other thing is environmental enrichment. We know and we, we learn that with lab animals and then this knowledge has been applied to other animals including dogs. We know that providing an a rich stimulating environment may increase the beneficial effects of physical exercise of both physical exercise and environmental enrichment.

Have additive effects on the health of the, of the brain of geriatric animals. So, a broad strategy including changes in husbandry, changes in the environment, and of course, changes in, nutrition. And nutrition, as I said before, is the main topic of this webinar and now I hand it over to Cecilia, who will talk about how we can use nutrition in dogs with cognitive dysfunction syndrome.

OK. I hope everybody can hear me OK. So I'm gonna be talking about what are the nutritional strategies that we can do to manage dogs with cognitive dysfunction syndrome or CVS.

Of course, the first goal is always to, provide nutrition. So when we feed our dog or a cat, independently of their health status, we feed them to provide a complete and balanced diet, for their species and their life stage. And that means that we provide all the nutrients and all the ingredients that they need to, to function and to live really.

So that's the first thing we need to do when we feed our dog with CDS. It's very important that the diet that we choose to feed is also fed in, in specific amounts in order to maintain a stable weight and an ideal body condition score. Of course, dogs that are too thin is not desirable because that would, signal malnutrition, so they wouldn't have enough nutrients and energy to do their body functions.

But the opposite is also concerning. Obesity can promote, a low grade chronic inflammation and could contribute to oxidative stress and free radical production, which Chavi already explained, can play a role in CVS, pathogenesis, let's say that. However, there are also some specific nutritional modifications that we can make to try to attenuate the clinical signs, of course, always in conjunction with, behavioural and medical treatment, and I'm gonna focus on two groups here.

So we'll be talking about antioxidants and also MCTs or medium change triglycerides. Remember when Chavi was talking about two of the reasons why CDS develops, one of them would be the sedative stress. And the other will be that the mitochondria would be less efficient with energy.

So these two things would affect those things. So antioxidants can deal with oxidative stress, MCTs will provide us a source of energy. So starting with antioxidants, just a definition of what we're talking about here, and antioxidants is a substance that can inhibit oxidation and or minimises oxidative damage.

So antioxidants can be endogenous, which mostly are enzymes but also exogenous, which are nutrients. And they act at different levels and synergistically. So we have antioxidants that will prevent the initiation of the oxidation chain, so you will have free radical scavengers in this case, for example, or metal binders, things like that.

You also have antioxidants that will stop the chain reaction once it has started, so stopping the process and then a lot of these antioxidants are redox substances, so go from oxidised to reduced and vice versa. So they need to be recycled back to their active form, and those substances that are in charge of recycling are also part of the antioxidant machinery of the body. So what happens, we get these free radicals or reactive oxygen species, reactive nitrogen oxide species.

These are created by the normal functioning of the body, when, when we obtain energy, when we make functions, these are created normally and we need to deal with them, but they can also be increased in some situations like ageing, but also contamination or usually light, for example. When the production of these free radicals overloads the antioxidant systems of the body, we get into the oxidative stress. So it's with this oxidative stress that, that The provision of antioxidants in the diet could potentially help these patients.

So it's important to know that the brain as a tissue is specifically sensitive to oxidative stress because it requires a lot of oxygen and a lot of these free radicals or reactive oxygen species. It has a high content in polyunsaturated fatty acids in their cell membranes and if you remember, of, oxidation can affect everything. It can affect protein, it can affect DNA, but it can also affect fat and specifically polyunsaturated fatty acids because those double bonds make them weaker and more, more vulnerable to oxidation.

And finally, the brain has a lower antioxidant capacity compared to other tissues. So what do we have? We have a couple of studies that show that provision of additional antioxidants in the diet could potentially help with CDS.

There's this one very beautiful study, where they used for the 8 beagle dogs which were between 8 and 12 years old. So they were not really sick dogs, but, they were ageing dogs and they had some dysfunction in their, in their learning capabilities and things like that. So what they did is, They had 4 treatments in here according both to diet and environmental enrichment, and that meant we had either a controlled diet or a test diet, and I'll talk about those a little bit later, and the dogs could be in an enriched environment or in a non-enriched environment.

So enrichment in this case consisted in being housed with kennel mates rather than being in isolation. They were exercised twice a week, and they had toys available all the time, and these toys were rotated weekly, and they got different toys every week. What about the diet?

The diet had a cocktail of nutrients, so they had antioxidants in there. So vitamin E, is a very important lipid-phase antioxidant, and it's a chain-breaking antioxidant. So once the lipid oxidation starts, vitamin E will sacrifice itself and stop the reaction.

Vitamin C is an antioxidant in itself, but in a water soluble capacity. But it's also a vitamin E recycler. So, you know, you have the alphatocopherol.

When it, when it stops the reaction, it will become an alphatocopherol radical, and this radical will be converted back to alphatocopherol or vitamin C, which in, in, in its turn will be recycled by the antioxidant systems in the body like superoxide. Thismahase will also be involved in this path peroxidase. There's a lot of them.

This, treatment also had mitochondrial cofactors. In this case, two of them, it had L cardittin, which is a substance that's involved in mitochondrial lipid metabolism. So it's basically a shadow substance that will ensure that fatty acids can go from the cytosol to the inside of the mitochondria to be oxidised for energy.

So it's very important to ensure energy provision to the brain cells. And the other one would be alpha lipoic acid, which is a co-factor of the mitochondrial enzymes that are in charge of the respiration to obtain the ATP. So again, very important in mitochondrial function, especially in their function related to energy.

So the goal of the researchers providing this cocktail of substances was to minimise the damage created by the free radicals, prevent further free radical degeneration, and also stabilisation of membrane function. In the mitochondria. So, What they did is they took all of the dogs and they made them do several tasks that would investigate different aspects of, of cognition, and they performed them at baseline to separate the, the dogs into, into the four groups, and then after 1 year and then after 2 years, they repeated the same tasks and basically what they found is that the supplemented dogs had improvements in spatial attention, learning abilities, visual discrimination and reverse learning.

And these improvements were more marked, even better when combined with the environmental enrichment. So in this case, using such a diet would result in a more functional bed, a pet with less stress and less anxiety because it would be able to perform better and deal with life, especially if combined with behavioural modification or enrichment. And there's another study, that also supplemented with antioxidants.

In this case, they used a nutrient blend of vitamins E and C, similar to the previous study, but they also added B vitamins, fish oil, and arginine. B vitamins are involved in the energy metabolism, so it's, the rationale for including them was to see if they could help get more energy out of nutrients. Fish oil is a source of omega 3 fatty acids which can reduce inflammation, and arginine is the precursor of nitrous oxide, which is a very important second messenger.

This was a shorter study, for about 6 months, and they had, senior dogs between 9 and 12 years old, separate into two groups according to, to their cognition tests. And similarly to the previous study, the supplemental group did better, not in all of the tasks, but in several of them. The same group of researchers, did a very similar study, in cats, .

And they also found similar positive results. So even though we're talking about dogs right now, I just wanted to mention that. So that's all I have to say about antioxidants.

So, we have a couple of studies. I, I wish we had more, but right now the ones we have are very encouraging. The second nutrient that I wanted to talk about would be the medium chain triglycerides, which provide medium chain fatty acids and in nature, you can find them in some vegetable oils like coconut oil, mostly, but also palm oil.

So just as a reminder, fatty acids can be classified according to their chain length. They're basically a very long carbon chain, and you can see that medium chain fatty acids are those that have a carbon chain length of between 8 and 12 fatty acids. So you'll see caprylic would be the 8 carbon chain fatty acid, Capric, and loic.

You'll see that the study that I'm gonna show you that was done by the Purina researchers, it's mostly using caprylic or 8. And this is important because sometimes, we, we might try to mimic the results using coconut oil or some commercial products, but they might have a different composition in MCTs. So what's interesting about MCT is, is that compared to other fatty acids like longer chain fatty acids, they are more ketogenic, and I'll show you a little bit what I mean by that.

They basically increase hepatic ketone body synthesis. Just as a reminder. This, the, the pathway to synthesise, ketone bodies, you will start with two acetylcho A molecules and after a series of reactions you would get your three ketone bodies ace acetate, acetone, and beta hydroxybrate and we'll be talking mostly about the latter, but other fats tend to be more, either for storage or they would be.

Used in beta oxidation or complete oxidation of the molecule rather than ketone body synthesis. So ketone body synthesis was very interesting for the researchers because it could be an alternate source of energy for the ageing brain. So the brain usually relies a lot on glucose as a source of energy and doesn't really have the capability to oxidise, oxidise or use fatty acids in the beta oxidation, the long process.

However, they can use ketone bodies. So the liver can make those ketone bodies, provide them to the blood, and the brain can use them. They would be converted to acetylcho A which would be incorporated in the citric acid cycle and that will provide ATPs to the brain cells.

So that's the hypothesis that came from that, that ketone bodies, could be an alternative source of energy. And this is inspired a little bit by what's happening in people with fasting, right? When also animals, when we fast and we don't have an access to energy, we will metabolise our own fat, and we will use ketone bodies and the brain will switch from glucose to ketone bodies to survive starvation.

In the human patients with Alzheimer's, where the dog is used as a model, using medium chain triglycerides, and studying, ketone bodies has been done for a while now with very encouraging results. And I'm gonna show you what we have on dogs right now. So the idea would be that we provide these MCTs in the diet, they get absorbed mostly in the portal vein.

You get your medium chain fatty acids. They will go to the liver, be converted to ketone bodies, which will go to the bloodstream, cross the blood brain barrier, and then provide energy to the brain, and up to 60% of the energy of the brain could be provided by this ketone. So the study that the Purina group did, they recruited 24 age dogs between 8 and 12 years old.

They were assessed on their cognitive function at baseline and they were randomised into two groups to make sure that there were no differences at baseline on their cognitive function, and they also make sure that the groups were balanced on gender. They had a test diet which had 5.5% as fed of a combination of MCT, so mainly caprily, the C8, fatty acid, and some Capri, which is the C10.

And that was compared to the standard diet which used animal fat instead of MCTs. That diet was fed for 8 months and then cognitive function was reassessed using several tests. Here you can see them, landmark discrimination, learning ability, visual spatial function, and attention were all assessed by different very sophisticated tests.

What they found as a result, they did take blood samples of these dogs and they found that, beta hydroxypy rate was higher in test dogs than in controls. They, of course, did not measure, brain values, but we assumed that the higher in the blood, the higher they would cross the blood brain barrier. Test dogs perform better in almost all tests, and those really, really encouraging great results.

And the more difficult the tests, the more improvement with the supplemented diet. So, again, very encouraging for those refractory cases or those harder to treat cases. And this is one example of the landmark task that assesses discrimination and memory, and then part of the analysis was have the dogs do the task and then measure the number of errors.

So you can see in this graph that the supplemented dogs, Made much fewer errors compared to the control dogs, which again can help them in their day to day life, like they'll be able to know where the food is, where, where they should be, what's the time to go to work, things like that. So very meaningful impact on their quality of life of both dog and owner. So also improve spatial memory, executive functions, and attention.

So more interest in the environment and more interactions with their loved ones. And that's all I have to say for now about CDS, and I'm gonna have, give this back to Chavi to talk a little bit about fear and anxiety-related problems. OK, thank you.

Thank you, Cecilia. OK. So our second topic tonight is, fear or anxiety-related problems.

I am not implying that fear and anxiety are the same. They are not discussing, both concepts in depth is well beyond the, the scope of this, of this webinar. But the, the important thing now is that we do have many behavioural problems in dogs and cats.

Which are somehow related to fear or to anxiety. We have suppression anxiety, mainly in dogs. We have noise phobias, mainly in dogs as well, and we have a variety of social fears and some forms of aggressive behaviour in both dogs and cats, among others.

Now, these, problems are important, because the, they have an effect on, on the welfare of the animals and they are, they are highly prevalent and, and complex in nature and in the underlying mechanisms. So let me illustrate that complexity by looking first at separation anxiety. This picture on the left, .

Shows a dog with separation anxiety. It is clear that one dog like this, may, may have, may, may have an impact on the quality of the human animal bone. It is a highly prevalent condition.

We have some studies done in the United Kingdom suggesting that more than 40% of the canine population may develop separation anxiety. And it has been ranked by itself as one of the main welfare, problems in companion dogs. It is not only because of the anxiety these animals suffer when they are alone, but also because, work done at Bristol University has, shown and Lincoln University shown that, these, dogs develop what we call a neck.

Cognitive bias. So it's, it's quite a serious, a serious problem. And noise phobias are quite common as well.

Some studies suggesting a prevalence between 20 and 30% of the whole canine population and they may show when dogs are exposed to, fireworks or thunderstorms and mainly, so noise phobias cause stress and They may even endanger the the life of the, of the dog. Now, these problems have to be addressed once again, considering the, the complexity of the symptoms and of the underlying mechanisms. And just, let me illustrate this complexity in the particular case of separation anxiety.

Now, separation anxiety. Is a condition, itself, but it may show up in, in a variety of, of symptoms as this study, as this study shows. A significant proportion of dogs with separation anxiety may bark excessively.

Some of us may, show, elimination-related problems or destructive, behaviour, then we have, . Symptoms which are less common but, but still important such as vomiting, self trauma and, and of course we, we think there are dogs with separation anxiety that suffer the welfare problems caused by separation anxiety and yet do not show any of these symptoms and, and therefore may be overlooked by, by their ownness. So why do dogs develop separation anxiety?

Well, it's a rather fascinating problem really with, with a variety of causes. We, we think that in many cases separation anxiety is related to attachment problems, attachment between the dog and the owner. And for many years we thought that attachment problems mean basically hyper-attachment, so meaning an excessive attachment between dogs and owners.

We know now that in some cases attachment-related problems may not mean hyper-attachment, but what we call inappropriate attachment. Which is the consequence of owners which are inconsistent in their relationship with, with the dog. So they, they may be supportive at times and basically ignore the dog in, in other occasions and this inconsistency may lead to separation anxiety as well.

And finally, we have cases where separation anxiety is not Related to attachment at all and we think that in those cases it may be the consequence of what we call condition fear. So the dog goes through a frightening experience when being alone and from then on, the dog will associate being alone with that negative experience and that will result in separation anxiety. So how do we address separation anxiety and for that matter, other behavioural problem related to, to fear or to anxiety.

So again, we have to use a combination of, of strategies, different types of strategies and changes in the environment of paramount importance again, in particular, providing the dog or the cat with what we call a safe area, the area where the animal. I feel secure with important resources and has been shown to be extremely important to ameliorate the signs of separation anxiety and other fear or anxiety-related problems. Then we can apply desensitation pro programmes whereby the animal is gradually exposed to the stimulus which may cause fear so that the animal learns to stop responding to that stimulus with, with fear.

We can use pheromones and pharmacological treatment and of course, once again, we have nutritional strategies which are helpful to address these problems and this is what Cecilia is going to discuss now. So over to you again, Cecilia. OK, so we do have some dietary strategies that we could do for anxiety and fear.

There's less evidence definitely compared to CDS where both the antioxidants and the MCT, even though there's not a lot of studies, the studies are very solid and, and. And very promising fairly. For for anxiety, there's definitely less data.

One of the molecules that has received the most attention though is alpha quesozepine, which many, if you look at the ingredient list of the diets, sometimes it comes as a milk hydrolyzate or something like that. The, the history of this molecule is that, cow's milk has been considered a common food for a long time, especially in babies. So it, it, it was believed that when babies ingest the milk and then digested, there are some biopeptides, produced after this digestion that could have some sort of calming effect on the brain.

In babies compared to adults, trypsin digestion is more prevalent than Pepsin digestion, which is what we do in adults. So alpha cazozepine is one of those biopeptides obtained by the tripsy digestion of cow's milk. .

And the idea is to test if this could have a calming effect on, on dogs and cats. So we would have alphaS1 casein, which is a very important protein that's present in the milk. We would use trypsin to hydrolyzat to break that down and then we would obtain the alpha quesozepine.

This is a decapeptide, so there's 5, 10 amino acids. In this molecule and it binds GABAB receptors, so it has an anxiolytic activity. It is considered an ultraceutical, at least in Europe, so it's not a drug medication.

It is an ultraceutical, so you don't need prescription to buy it. That said, it does act a little bit like the benzodiazepines, for example, it's basically the same route. We have one study, this was done in France.

It was a multicentric randomised double blind placebo control, so very high quality study in that sense. It had 34 cats that were recruited by certified behaviourists, veterinarians, and those cats had social phobias, so mostly fear of strangers, things like that. So they tested the alphaozepine, the brand name is on the Paper title you can see it's called Zulken.

I never know how to pronounce that. And they gave this at 15 milligrammes per kilo compared to the placebo capsules. They gave that once a day orally.

And all of the cats underwent the behaviour modification that Chavi was talking about. So the cats were treated holistically like with multi-modal approach, but they received this retraceutical or just a placebo. .

This was multicentric, so there were different veterinarians involved in the checkups, but the appointments were Initially a recruitment and then after 4 weeks and then at 56 days. There were two phone calls in between to check that things were going well at days 14 and 42. So what they found is that they did not find differences in aggression in those cats that were showing aggressive behaviour, but they did see an improvement in the relationship with humans and also an improvement in fear-related behaviour.

So for example, the fear of strangers had a significant, significant reduction, those cats that were treated with the alfoxozepine, compared to the cats that were treated with the placebo. All of them. Treated with behavioural modification.

There's another study also in cats, with 2021 cats, also pretty small number of cats, and in this case, instead of testing the, the medication directly or the nutraceutical directly, they were fed a test diet. So this is diet in the market that has alphacazozeine but also has increased tryptophan, which is a serotonin precursor, which could also have a calming effect. And they compare that to a controlled diet and they fed that diet for 8 weeks.

In this case, they tried to measure some more objective measurements of improvement because, you know, doing behavioural asking owners and asking vets can be a bit subjective, so they measured cortisol both in the serum and in the urine. And even though they saw a decrease in the cortisol in the urine after 8 weeks, they did not see an effect of serum cortisol following a stressful event, and that stressful event was a vet visit and a blood draw. So in this case, with the neuroceutical included in the diet, very short-term study, they did not see really a positive effect on stress or anxiety.

However, in this study, they did not measure behavioural changes, so those improvements might have been missed if they happened. I also think that with the diet, the dosage is not so precise. So when, when you give the nutraceutical directly, you can give 15 milligrammes per kilo.

However, the dose of the diet will depend on the energy needs of the cat, so the dosages can, can definitely vary a bit from your goal dose. They did actually a follow-up of this study, 24 cats that were fearful and they were separated into mildly fearful versus markedly fearful, and then they gave them this diet compared to a controlled diet, and they were assessed again very short term, 2 and 4 weeks, and it was the same commercial diet used in the previous study. So in this case they did assess behaviour.

So they did a home test. So they went to the, they put a human in the cat's home. They also assessed response to a human in an empty test room, so an unfamiliar space, and then they also assessed the response to an unfamiliar human, so an unfamiliar person rather than unfamiliar room.

So they were assess the baseline and then at the rechecks. In this case, they did not see an effect on the response to an unfamiliar human, but there was a mild improvement on anxiety due to placing the cat on an unfamiliar location. So all of these tests are done in cats, especially because cats are, get really stressed out, especially when they go to the vet.

So having such a diet will be really helpful. And the results, even though some of them positive are not super strong at this point. This type of diets are also very interesting in cats with idiopathic cystitis, where we believe stress is a very important part of the pathogenesis of that problem.

So if, if these results end up panning out, it would be great to, to have another tool in our, in our belt to manage stressful cats with urinary disease or not. What about dogs? We have one study that I found, they had 38 dogs with anxiety-related complaints that were recruited again by certified veterinary behaviourists.

This was also multi-center randomised and blinded. And they, in this case they did not use a placebo. They actually compared the alfoxozepine with medication selegiline.

Which is an anti-anxiety medication and they assess, they did, they have a test, standardised test to assess emotional disorder. So that's how they checked the effect of the alpha cazozepine after 56 days. And what they actually found is that both treatments resulted in improvements of the scores.

So using medication compared to using the nutraceutical. Both was positive according to this measure, which is very interesting, because it seems that, that we again, we can have this tool to treat anxiety and stress in dogs without having to go to the medication route, at least initially or even in combination if need be. So I'm gonna switch to Chay right now.

OK, so, thank you very much again, Cecilia, and So, we thought that we, we couldn't finish this webinar on nutrition and behaviour without, mentioning even if briefly, the link between serotonin and aggressive behaviour. Now, it has been said that there is a, a clear cut, nice, simple link between serotonin activity in the brain and Aggressive, behaviour in the sense that, when serotonin activity goes down, aggressive behaviour is more, likely to happen. Well, this clear cut relationship doesn't seem to be, to be true.

There is a link between serotonin activity and aggression, but this Link doesn't apply to all dogs which are aggressive or all animals which are aggressive, but only those showing out of context aggression and aggression without warning signals, which is what we mean here by impulsive aggression. So in these dogs with out of context or impulsive aggression, there is a reduction in the activity of serotonin in, in the brain. And as serotonin is produced from tryptophan.

And there is the hypothesis that increasing the levels of tryptophan in the diet and reducing protein and therefore other amino acids which may compete with tryptophan for access through the blood-brain barrier. This may have a, a positive effect by reducing some forms of aggression. And this hypothesis has been supported by some studies.

The effect doesn't seem to be, dramatic, but, we believe there is scope for further research on the effect of, changes in cryptophan. And, and, and dietary protein that effect on aggressive behaviour on some forms of aggressive of aggressive behaviour. OK, so that's very, very briefly, but, we thought we had to touch upon this at the, at the end of the webinar.

And now, for the last time, I will, hand it over to Cecilia, which, who will finish the webinar by going through the main concluding remarks. So, to summarise, behavioural disorders in dogs and cats are complex and their management should be a multi-modal and definitely tailored to each individual and each. Taking of course into account the situation of each caretaker, for to maximise success.

Dietary management, besides providing a complete and balanced diet, to a stable body weight, and an ideal body condition score can help manage some of these conditions. Antioxidants and potentially other nutrients like mitochondrial cofactors, omega 3 fatty acids, and the vitamins can help improve signs associated with cognitive decline, ageing, . Potentially related to, to reducing the oxidative stress caused by this age mitochondria.

However, we do need those response studies to identify the best combination and dosages of antioxidants and other brain protective substances. At this point, what we have really is product testing. We have a diet or a supplement, we test that and we see the effects.

But it'd be lovely to have those response studies and, and more studies on the individual nutrients and see how they interact with each other. This is something that we still need. The use of a diet enriched with medium chain triglycerides, specifically C8, shows long-term positive effects on HK9 cognition, and we think that this is mostly due to the provision of an alternate source of energy to the brain to glucose, which would be the ketone bodies, and this is the diet that Purina has the neurocare, .

There are no studies on supplementation, meaning that we don't have, again, these dose response studies like using an MCT supplement. So I, I do, if, if we want to try this strategy, it's best to use the diet as is, again, because the combination that they use in this diet might not be easy to find outside, but hopefully we'll get more research done and supplementation of MCTs and also those responses. Alphacaozepine shows, I would say promising results in the management of anxiety in dogs and cats.

In cats, for the data that we have, it seems that it's much positive when we use it separately than in the diet. However, we need more studies with a higher number of cats, and dogs and more objective outcome measurements. They do have a similar mechanism of action.

Than benzodiazepine, so I would potentially avoid using both in case we run into issues. So choose one or the other at this point. The inclusion of tryptophan, an amino acid, which is a serotonin precursor, plus protein reduction to facilitate this, this, this blood brain barrier cross, can have some positive effects on canine aggression, but right now we don't have a lot of data and we definitely need more studies to give a strong recommendation in that sense.

And I think I would like to thank you all for listening, thank you, the webinarve for hosting and Purina for sponsoring and we're happy to answer any questions that you have. Cecilia and Javier, that was absolutely insightful. And I can hear that both of you are really, really passionate about about your subject.

So thank you for your time tonight and once again to Purina, thank you so much for your sponsorship of this evening. I'm sure Jennifer from Purina is going to be coming back on with us shortly as well. So we do have some questions that have come through.

. The first one we have is which oil is richer in omega 3 fatty acids and which ones are richer in omega 6 fatty acids. OK, I'm taking that one. So, omega 6 fatty acids are rich in vegetable oils.

So sunflower, but the, the non-modified sunflower, you know, there's a sunflower oil that it's modified for better cooking, not that one. Corn oil is very rich in omega 6. And in omega 3s, we have some vegetable oils that are rich in the precursor like canola or flaxseed.

However, the omega 3s that we like more, which are EPA and DHA, are especially rich in fish oils like salmon oil, manhan oil. There's also some very interesting oils in the market now like algae oil or krill oil which are especially rich in DHA, and And for example, algae oil is technically vegetarian, so a lot of vegetarian people use that one as a source of omega 3s, and they are also interesting because of sustainability and also because of contamination. The problem with fish oil is that especially if they come from larger predatory fish, it can have bio accumulation of heavy metals, dioxins and things like that.

So companies need to do very good quality control, but if you use something like algae, that would be less of a problem because they're lowering the traffic chain. But that would be it. Vegetable oils, mostly omega 6s, marine or fish oils, omega 3s, that would be a short answer.

OK, great. Caroline, when this question came in from Caroline when you were talking about coconut oil and she says, is there a difference in the stage of it, so cold pressed or not cold pressed, and that would be better or worse for the brain. OK, that's a good question.

I don't know that I know the answer. That said, the, the main difference between the cold pressing or the hot pressing and, for example, regarding olive oil, so as a good Spanish person, I eat a lot of olive oil, and we have the regular olive oil and you got the extra version which is called bred. It's not for the fatty acid, it's for the other substances.

So in, in extra virgin olive oil, you got all the other phytonutrients that are there that won't be affected. However, the fatty acids, the composition is gonna be the same. And coconut oil, they are medium change fatty acids and also saturated, so very resistant to oxidation.

So I, I, in theory, I wouldn't think that that would have an effect really on their, on their efficacy. OK. We've got a number of questions that have come through about which oil and how much oil and everything else.

I think I'm gonna pull Jennifer in here and say, my answer is, let's rather use a Purina food then start adding oils in an imbalance to the diet, Jennifer. Mhm. Yeah, thanks, rice.

Yes, so what I will say is that in this case, the diets that we have, we have two diets with entities. One is Purinaroplan Neurocare, and the other one is Purina proplant OtiH that both are only for dogs. I want to make this clear.

And what we are putting there is the MCTs, but they are purified MCTs and as Cecilia said before, not all the medium chain fatty acids have the same effect. So mainly the fatty acids that we put inside were C8 and C10, so they are purified. I think that, for example, coconut oil, it's mainly rich in C12.

It also has C8 and C10, but Then what we did is to extract this C8 and C10 from this coconut oil. Maybe if you want to achieve the proper dose of C8 and C10 with coconut oil, maybe you have to put, I don't know how many millilitres, but maybe it's a lot and will not be feasible to add it to the diet. Maybe it's CC and also can also.

The problem with adding oils on top is that they're very high calorie, so it's, it's. As an example, each teaspoon of oil is about 5 millilitres, so that's about 40 kilocalories just for a teaspoon. And, and then you would probably need to add a lot and that would unbalance the diet.

So you have to add a lot of calories from unbalanced items, they would eat less of the main diet and you could result in nutrient deficiencies. That's why even with omega 3s, like when you want to enrich omega 3s for arthritis or in this case for MCTs, using the diet that already have them. It's much more efficient, and, and you don't have these nutritional risks of balancing the diet by adding oils.

I, I sometimes try to use the oils if for whatever reason I cannot place the dog on such a diet, and then I'll do my calculations, but it can, it can get very high calorie really fast, and we want to prevent overweight and dietary imbalances. Yeah, and especially in the older animals when they've got cognitive dysfunction and that sort of thing, you, you're encouraging them to eat but the extra weight is going to play a bad effect on their joints and everything else. Yeah, those are dogs with usually comorbidities, so you need to take all of those into account when you're doing your plan.

Yeah. Yeah, excellent, excellent. Folks, we've come to the end of our time tonight, to our speakers, Cecilia and Javier, thank you so much for your time.

Thank you for your insight, and, I'm sure we are going to see more of you on the webinar vet. Jennifer, to yourself and Purina, thanks for coming on tonight. Thank you so much for the sponsorship.

Guys, I cannot emphasise how important it is that we support the companies that support us. So let's get out there and, and support Purina in their efforts with this. Just one last thing from my side.

When we log off tonight, there will be a questionnaire that pops up. Please take some time to answer it. Give us some feedback.

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Good night to you and to Rich, my controller in the background. Thank you for all your help. From myself, Bruce Stevenson, it's goodnight everybody.