Hello and welcome to this protective veterinary webinar on the microbiota, probiotics, and prebiotics. In this webinar, we're first going to look at the microbiota, what it is and how it helps us and our veterinary patients. And what happens when it becomes imbalanced, which is also known as dysbiosis.
We'll then move on to look at probiotics and prebiotics, again, what they are and how they can help our patients before finishing up looking at evidence supporting the use of probiotics and prebiotics in dogs and cats. So let's get started looking at beneficial bacteria or dysbiosis and disease. When we talk about the gastrointestinal microbeta, we are not just referring to gut bacteria.
But the entire ecosystem which exists within our gastrointestinal tract. Yes, this does include bacteria, but also fungi, protozoa, archaea, and viruses too. As it is an entire ecosystem that we are dealing with, tweaking even one component can have huge knock-on effects to the environment and other species present.
You may be familiar with the fact that reintroducing wolves to the Yellowstone National Park had huge knock-on effects. The walls predated the Eks which kept them moving more throughout the park. As a result, the vegetation in the valleys grew and beaver populations flourished, so they built more dams.
The combination of the dams and the new vegetation in the valleys meant the rivers actually changed their courses. A slight digression, but a good example of how a small change within an ecosystem can have a huge impact. Going back to the microbator, a few fun facts.
There are over 100 trillion microorganisms making up our microbiota. And if we were to line up all of the bacteria for our gastrointestinal tract next to each other, they would reach the moon and back 130 times. There are as many bacteria as there are human cells within your body.
And if you hopped on the scales, you could attribute 1 to 2 kg of your body's weight to the bacteria that live on or within it. If we think about the gastrointestinal tract, it is essentially an open tube from the mouth to the anus. It is therefore not surprising that bacteria are able to enter and colonise.
While we do have microorganisms present throughout the entire gastrointestinal tract, studies have shown that bacterial numbers and populations vary at different points along the way. In general, the number of bacteria present increases as we move distally from the stomach through the small intestines and into the colon. There are several different factors which can influence the bacterial populations throughout the gastrointestinal tract.
In the stomach, because of its oxygen content and acidity, resident bacteria numbers are limited. The bacteria in the small intestines tend to be aerobic or facultative anaerobes whose growth is limited by peristalsis. The greatest colonisation of the gastrointestinal tract occurs in the colon.
Bacteria within the colon are predominantly anaerobes. The microbial diversity is higher in the colon due to the slower flow of digester and the increased time and availability of nutrients for the microbiota to utilise. Microorganism prevalence varies between compartments of the gastrointestinal tract within an individual and between individuals.
Each vertical line on this graph represents an individual person. And each section is a different body part. The different colours represent different groups of bacteria.
We can see that there are huge variations in the types and numbers of bacteria present both at different sites within an individual and between individuals when samples are collected from the same site. Varying factors contribute to this difference, including host genetics, environment during birth, infancy, and at present. Diet, medications, stress, and so on.
Interestingly, when scientists have gone one step further and investigated the genes expressed by bacteria at different sites and in different individuals, they have found much less variation. Though this is an area which still needs a huge amount of research. There are certain bacteria which are associated with providing beneficial effects to the host, often referred to as good bacteria.
And others which tend to be more health negative, commonly described as bad bacteria. You may recognise Lactobacillus and bifidobacterium from human probiotic supplements. Or Enterococcus specum, which is the most commonly used probiotic in canine and feline products in the UK.
While there are definite trends as shown here, it has also been discovered that a healthy microbator is generally one with high diversity. So how did the bacteria get into the guts in the first place? It used to be thought that the uterine environment was completely sterile, with bacterial exposure only occurring at birth and thereafter.
However, more recently this has been brought into question. Bacterial DNA has been isolated from the placenta, an amniotic fluid from healthy term pregnancies in the absence of intrauterine infection. Furthermore, labelled enterococcus specium given to pregnant rats has been found in the meconium of their pups.
While live bacteria have not been detected to date, this does suggest there could be some degree of intrauterine colonisation. As the puppies or kittens pass through the birth canal, they are exposed to large numbers of vaginal and faecal bacteria. In fact, in people it's been shown that the delivery method can affect the baby's microbita.
Vaginal delivery results in a gut microbeta more similar to that of the vagina. Whereas babies delivered by caesarean develop a microbeta more similar to that of the mother's skin. There have even been proposed links between C-section delivery and certain diseases including asthma, diabetes, and inflammatory bowel disease.
When the puppies or kittens start to drink milk from their mum, they pick up bacteria not only from the skin and teeth, but also from the milk itself. Mother's milk contains high numbers of bacteria, in particular vilobacterium longus subspecies Infantus, which itself has been shown to aid milk digestion. Obviously at weaning, the young are going to be exposed to more bacteria as they eat from different bowls, etc.
But the food source in itself has changed, which may feed and favour different bacterial populations within the gut, thereby altering the microbiota. Following weaning, each individual is thought to have their own bacterial blueprint, unique to themselves but stable over time. It is therefore important that the microbiota is able to be established as normal in the first few weeks to months of life.
For example, use of antibiotics during this period could have prolonged effects on an individual's microbeta. And has even been linked to increased risk of obesity and allergies. Excessive hygiene in childhood could also affect the development of the microbota, with links again to allergies and inflammatory bowel disease possible in later life.
So we've seen how the microbiot is established within the cut, but what role does it actually play within the body? It has several functions which we'll go through one by one. Firstly, it aids digestion.
Dogs and cats, like us, are unable to digest certain fibres which form part of their diet. The gut bacteria ferment these fibres into short chain fatty acids which can then be absorbed by the body. They are also involved in producing certain vitamins such as B and K vitamins and metabolising bile acids.
Only our gut bacteria can convert primary bile acids to secondary bile acids. And we need an optimal balance of these for normal body functioning. They inhibit pathogens.
They do this indirectly by competing for nutrients and binding sites on the intestinal cells, and directly by producing molecules known as bacteriocins which damage or kill the pathogens. The microrota has been shown to be important in the maintenance of the epithelial barrier. It encourages the production of mucus which lines the epithelium and ensures tight junctions form as normal.
Finally, the microbiota stimulates the immune system and is particularly important for the development of oral tolerance. The fact that we can eat foreign material and in a healthy individual, the body does not react to it. Disruption of this function can play a role in the development of dietary intolerances.
Not only is the microbiota important for the functions we have just discussed, but it is also essential for normal gut development. It has been shown that antibiotics given to rats at the end of their pregnancy resulted in wrap pups with smaller stomachs and altered intestinal palmeability. Mice, brought up in a sterile environment with no microbiota, have abnormal development of their villy.
Amazingly, Introducing gut bacteria by faecal microbial transplantation reversed this abnormality. Similar results have been shown in pigs, where piglets from sows given antibiotics during gestation had an abnormal gastrointestinal architecture. Rather than having normal finger-shaped Billy, they're flattened and irregular.
Having looked at the micromotor and what it does when healthy, we'll move on to see how it may be disrupted and what happens when it is. The term dysbiosis refers to an imbalance in the microbes on or within the body. Today we are focusing on the GI tract as there is most research in this area.
So what could cause a gastrointestinal dysposis? Any sort of GI disease can upset the balance of microbes within the gut. Many drugs can also result in a dysbiosis, in particular, antibiotics, which can kill beneficial bacteria within our intestines.
Dietary indiscretion, sudden change of diet, or periods of anorexia can all lead to changes in the microbiota. It's well known that stressful situations could upset the balance of bacteria within the guts. And finally, certain life stages may result in a dysmosis, for example, weaning or diet change with old age.
Given the many roles that our microbietter plays, it is not surprising that the clinical signs of a dysbiosis can be varied. We can see obvious gastrointestinal upset in the form of vomiting and diarrhoea. Or more subtle changes, such as weight loss, appetite change, and abdominal pain.
A dysmosis will often result in an increase in sulphur producing bacteria which we may see as increased flatulence and bulgy. Finally, symptoms may be more generalised, and the animal may just be quiet or of colour. If we think back a few slides to the function of the microbita, we will remember that the microbiota plays an important role in supporting the epithelial barrier function.
It does this in 3 ways. Firstly, by producing butyrate, the main food source for enterocytes. Secondly, by promoting mucus production, which protects the epithelial cells from pathogens and antigens.
Thirdly, by modulating the innate immune system. In dysbiosis, we see a proliferation of pathogenic bacteria. This results directly in disruption to the integrity of the mucosa.
Atrophy of the villy and increased levels of waste products which can further damage to the intestines or result in flatulence and bloat. If the mucosal barrier is disrupted, this can result not only in local inflammation. But also low grade systemic inflammation which has been linked to specific diseases such as diabetes and autoimmune conditions.
Disruption to the microbita, known as dysmosis, has been linked to many diseases in people, as we see here. You may not be surprised to see the gastrointestinal diseases listed in the top row diarrhoea. Antibiotic responsive diarrhoea or small intestinal bacterial overgrowth, inflammatory bowel disease, and irritable bowel syndrome.
But dysmosis has also been linked to many other diseases, including those affecting the skin, such as eczema, the lungs, such as asthma, and even the joints when we look at arthritis and osteoporosis. We're going to look at a couple of examples just to highlight the importance of the microbiter. Firstly, we will look at the microbiota in obesity.
It is fairly well established that certain bacterial groups are more commonly found in lean people. While others are generally more common in obese people. We have already seen that bifida bacteria and lactovacili commonly feature in human probiotic supplements and are known to provide protection from pathogenic bacteria in the gut.
Bacteroides are known to be involved in the development of immune tolerance and also help to break down undigestible fibre to produce butyrate. Acamentia fed to overweight rats, was shown to stop them becoming obese and developing diabetes. And finally, Kristen Cenella seem to be particularly prevalent in those lucky people who can get away with eating whatever they like, but remaining skinny.
Unfortunately, it's very difficult to harvest, so no magic diet pills just yet. Thermacues are known to help digest the fat in the diet and extract extract the maximum energy from food ingested. Unsurprisingly, these tend to be found in higher numbers in obese people.
It has also been shown that bacterial diversity is generally lower in obesity. When assessing the richness of bacterial genes present in 61 obese women, it was found that 75% of the subjects had low bacterial gene counts. There are many studies looking at the links between microbiota and obesity.
And this is just one example. In this study, the faeces from obese or lean children were given to germ-free mice. Despite eating the same diet, the mice receiving the microbiota from obese children gained significantly more weight than the mice receiving the microbiota from lean children.
They also found that the lean mice passed more unspent energy in their stalls. Another particularly interesting area of research is that of the gut brain axis. Here are just a few examples of published studies demonstrating the gut brain axis and how our microbeta can influence various aspects of our neurological functioning.
It's been shown that transplanting faeces from a depressed person into a rat makes the rat show signs of depression, such as hiding away in dark corners and avoiding social interactions. Transplanting the faeces of a person with Parkinson's into a mouse with a genetic predisposition for the disease led to the development of symptoms in the mouse. Finally, it has been shown that 90% of the body's serotonin is made in the gut.
In dogs and cats, dysbiosis has been shown in the following diseases. In dogs with inflammatory bowel disease and acute diarrhoea, they have been shown to have decreased short chain fatty acid producing bacteria and increased Clostridium species. Disposis has also been shown in cats with chronic heopathies and acute diarrhoea, and dogs and cats with giardia duodenalis.
It's been reported that a dysposis is often associated with meningoencephalomyelitis of unknown origin or MUO, and in fact there is strong evidence that a high abundance of prevotell SAA in the gut is associated with reduced risk for developing immune mediated brain disease. Dogs with congestive heart failure have been shown to have an increased abundance of proteobacteria. And there are studies linking dysposis to obesity in dogs and epilepsy too.
Here are a couple of examples demonstrating dysbiosis in cats. This first study demonstrated that cats with IBD had reduced bacterial diversity in their faeces and an increase in dissulfo vibrio species which produced the gas hydrogen sulphide. This is likely to result in increased flatulence, wary, and possibly abdominal discomfort if they suffer from trapped winds.
In this second study, we are looking at bacterial diversity in healthy cats and those with acute and chronic diarrhoea. We can see the number of bacterial species observed on the y axis. We see that healthy cats shared the greatest diversity, but this was reduced in cats with acute diarrhoea and further still in cats with chronic diarrhoea.
More recently, researchers have taken things one step further. Rather than just looking at bacterial diversity in healthy dogs and those with chronic androopathies, they have come up with the dysbiosis index. This is a mathematical algorithm which combines changes in the levels of 8 bacterial groups into a single numerical value.
A negative dispas index represents a normal microbeta. Whereas a dis-based index of 0 or more demonstrates a dysbiosis. The dispas index could be used in future to monitor the effect of certain drugs on the microta.
For example, in this study, healthy dogs were given a 2-week course of metronidazole, which we can see induced a dysbiosis. We can see here that while they were taking the metronidazole, their dysbiosis index increased above 0. While most dogs recovered post treatment, some were left with a residual dysposis 4 weeks after cessation of that metronidazol treatment.
Similarly, we could use the dysbiosis index to screen donor animals prior to faecal microbial transplantation. Or to monitor response to treatment. In this study, we can see how two dogs with chronic diarrhoea responded to FMT.
In one dog, the dysbiosis index remains negative 4 weeks after treatment, i.e., the treatment was successful and the dog no longer has a dysbiosis.
However, in the other dog, the dysbiosis resolved temporarily 1 week after treatment but has recurred by 4 weeks post FMT. This kind of monitoring could help to explain why in some patients the symptoms resolve but not in others. Moving on then to look at probiotics and prebiotics.
Probiotics are defined by the World Health organisation as live microorganisms which when administered inadequate amounts, confer a health benefit on the host. The name comes from pro meaning for, and bios meaning life. Historically, yoghurts and fermented foods were used, but nowadays there are specialised preparations available for dogs and cats.
In order for a bacteria to be registered as a probiotic, it must be approved by the European Food Safety Authority EFSA. In order to achieve this accreditation, the bacteria must be shown to be safe and not to cause infection or disease. To respond to antibiotics and not contain any genes which may promote antimicrobial resistance.
And to have beneficial effects upon the host. Until recently, there were only 2 strains of enterococcal specium which were registered for use as probiotics in dogs and cats in the EU. But this list has recently expanded.
In addition to Eococcus specium, Bacillus felaensis and Lactobacillus acidophilus have now been registered as gut flora stabilisers. A further 3 lactobacillus strains have also been registered as acid stabilisers for use in dogs. For a bacteria to be an appropriate candidate for use as a probiotic, it must have several features.
Firstly, it must not be pathogenic. Enterococcus specium has been verified as safe by the European Food Safety Authority. Healthy beagles, fed 10 times the recommended level for 21 days, had no clinical or biochemical changes detected.
The bacteria must also remain alive and viable throughout their shelf life if they are to make it as a probiotic. Specialised manufacturing conditions are required. And products should be tested at the end of manufacture and at the end of their proposed shelf life to validate their viability.
The recommended probiotic content for enterococcusheum is more than 1 times 109 colony forming units per kilogramme of product. That's more than 1 billion colony forming units per kilo. It is important to ensure that any individual probiotic product meets this specification.
Probiotics must be able to survive the low pH or acidic environment of the stomach. In order to reach the intestines where they can then have their beneficial actions. These graphs demonstrate the acid stability of enterococcus specium and Bacillus velazensis.
When kept at a low acidic pH for 2 hours, there was no reduction in colony forming units grown. Finally, if we think back to the World Health Organisation's definition of probiotics, live microorganisms which, when administered in adequate amounts, confer a health benefit on the host, we know that in order to be described as a probiotic, bacteria must be shown to have health benefits to the host. Enterococcus specium has been shown to reduce levels of pathogenic bacteria such as E.
Coli and Clostridia in the faeces of dogs and cats, and Bacillus velazensis has been shown to improve faecal consistency. So we know that probiotics have to go through rigorous testing in order to be approved by EFSA. But once approved, how can they actually help our patients?
They work in a very similar way to the beneficial bacteria already present in our guts. They compete with pathogens for nutrients and binding sites. They release toxins to kill pathogens.
They create a more acidic environment within the gut lumen which favours beneficial bacteria. And finally they stimulate the innate immune system. When searching out a probiotic either for ourselves or for our pets, we should always ensure that certain facts about a product are stated on the label.
The organism right down to the strain. The number of colonies present. And the shelf life for the product.
Moving on to look at prebiotics. Prebiotics could be considered as a lunchbox for the beneficial bacteria. They are substances which are not broken down by the animal itself but instead are fermented by the gut microbiota.
They provide a food source for bacteria and create an environment in the gastrointestinal tract which promotes growth and activity of good bacteria. Prebiotics are often provided in the form of soluble fibre. Many soluble fibres form gels when mixed with water, or have a good ability to absorb water.
As fermentability and solubility tend to go hand in hand, soluble fibres are fermented by the microbiota to produce short chain fatty acids. We will look into the benefits that these provide next. Some examples of soluble fibres include gums and mucilages.
As we have mentioned, when prebiotics are fermented by the microbiota, short chain fatty acids are produced, primarily butyrate, acetate and propionate. Butyrate is the primary energy source for colonocytes, providing about 70% of their total energy requirements. Acetate and propionate are absorbed across the gut epithelium and metabolised by the liver to other substrates.
Short chain fatty acids are really important for normal turnover and functioning of gut epithelial cells. Short chain fatty acids do not only have an effect on the gut epithelium, but also the gastrointestinal microbiota. The production of short term fatty acids lowers the intestinal pH, which supports the growth of beneficial bacteria species and prevents overgrowth of pH sensitive pathogenic bacteria.
The composition of the intestinal microbiota, as well as luminal concentrations of various short chain fatty acids are important influences for virulence factors and colonisation of some enteropathogens. A diverse microbiota is important for the production of short chain fatty acids in the first place. And a positive feedback loop occurs.
So we can see how short chain fatty acids play an important role in regulating the microbiota. Short chain fatty acids play a key role in regulating the barrier function of the intestines by supporting the multiple layers of the gut barrier. Fermentation of prebiotics by the gut bacteria produces short chain fatty acids.
These short chain fatty acids lower the intestinal pH, which favours the growth of beneficial bacteria and inhibits the growth of harmful bacteria, thereby supporting the microbial barrier. Short chain fatty acids also strengthen the chemical layer. By regulating mucus production and increasing antimicrobial peptide production.
The physical barrier of the gut is comprised of epithelial cells held together by tight junctions. Short chain fatty acids, increased tight junction formation, and as previously discussed, provide energy for normal cell turnover. Finally, we may forget that 70% of our immune system resides within our gut, and short chain fatty acids have been shown to support the gastrointestinal immune system by acting as a messenger between the microbiota and the immune system.
This is important in activating correct cellular responses and modulating cytokine production. Short chain fatty acids have also shown to have important anti-inflammatory actions, which could be relevant when we consider many intestinal diseases have an inflammatory or immune component, for example, inflammatory bowel disease. Preplex is a registered dual source prevertic combination made up of fructo oligosaccharide or hoss and acacia, or gum Arabic.
Foss has a low molecular weight and is therefore fermented quickly and can be used higher up in the gastrointestinal tract. Acacia has a more branch structure and is therefore fermented more slowly, so it can be used further down within the gut. By providing a dual source prebiotic, fermentation occurs along a greater length of the gastrointestinal tract.
And therefore can influence a larger bacterial population. Frost, fed to German shepherds with small intestinal bacterial overgrowth, has been shown to reduce aerobic and facultative anaerobic bacterial colony forming units in the duodenum and proximalduinum. These bacterial groups include pathogenic and potentially pathogenic bacteria.
Acacia, or gum Arabic, has been shown to preferentially increase levels of beneficial bacteria such as lactobacili, bifidob bacteria, and lactic acid bacteria. Whilst not increasing potentially pathogenic anaerobes. When probiotics and prebiotics are given in combination, they are referred to as symbiotics.
The effects are thought to be synergistic in maximising the benefit of bacteria. Minimising pathogens. And supporting the gastrointestinal barrier and immune function.
We are not going to go through all these studies published about probiotics. I think we'd be here for days. But probiotics, prebiotics, and symbiotics have been studied with regard to pretty much every organ system in the body.
We'll just have a look at a few examples. To continue on our themes from earlier, we'll start by looking at some examples of probiotic use in obese people. Overweight individuals receiving a 12-week course of Lactobacilluscari were found to lose significantly more weight than those receiving a placebo.
Individuals with metabolic syndrome, given bifidobacterium lacti, had significant reduction in obesity and its blood markers than those taking a placebo. And finally, a meta-analysis, ie a systematic review looking at multiple papers, concluded that symbiotics could have the ability to decrease both body weight and waist size. We will also take a look at some examples of the gut brain axis.
A double blinded, placebo-controlled, randomised study showed that depressed people's moods could be significantly improved by the administration of a mixed strain probiotic. Finally, we'll look at the use of a multi-strain probiotic by chronic and episodic migraine sufferers. Chronic migraine sufferers are defined as having more than 15 migraines per month, whereas episodic migraine sufferers get less than 15 per month.
The study found that frequency and severity was significantly reduced in both groups. In the episodic group, the frequency was reduced on average by 40% in the probiotic group compared to just 1% in the placebo group. With the severity measured on a visual analogue score reducing by 23% in the probiotic group and actually increasing in the placebo group.
Similarly, in the group of chronic migraine sufferers, we can see that they reduced on average from having about 22 attacks per month at the start of the study down to 12 attacks per month by the end of the study. I think it's fair to say that these changes would be fairly life changing for these people. Likewise, the migraine intensity was also significantly improved in the probiotic group, but not the placebo group.
We shall finish up this webinar by looking at some of the evidence available supporting the use of probiotics and prebiotics in dogs and cats. Here are some examples of studies looking into the use of probiotics in various diseases in dogs and cats. As expected, we see various presentations of GI disease.
There also have been studies looking at probiotic use with regards to immune function, skin and renal disease, and even canine behaviour. Research is ongoing and current topics of interest include probiotics in the management of obesity and urinary tract infections. Let's start by looking at acute diarrhoea in dogs.
We know that this is often self-limiting. Uncommonly occurs following scavenging, sudden diet change, stress, or use of other medications such as antibiotics. It could also be due to anatomical abnormalities or systemic disease, though diarrhoea due to these causes may be less likely to self-resolve.
Despite the fact that most cases of acute diarrhoea would likely resolve with no intervention, a study in 2010 found that 71% of dogs presenting a first opinion general practise with diarrhoea were dispensed antibiotics. This figure has reduced in a more recent study to 49.7%, so it does look like things may be going in the right direction.
However, even in the latter study, only 3.2% of cases had faecal analysis carried out. So it would be difficult to correlate antibiotic use with confirmed bacterial infection.
We could do a whole webinar on this topic, so I'll stop there and just leave it for you to think about. Are antibiotics truly indicated in acute diarrhoea? We should also bear in mind that their use could even cause adverse effects to the patient.
If we return to the dysbiosis index, which we covered earlier in the webinar, we can see in two separate studies that 4 and 6 weeks after cessation of a 2-week course of metronidazole given to healthy dogs, some dogs still had an ongoing dysmosis, which appears to have been induced by the metronidazole. Given the important function of a stable microbiota, we want to avoid inducing a disposis where at all possible. Aside from possible adverse effects to the individual animal, there are wider implications of antibiotic use from a public health stance.
Antibiotics are a common cause of dysbiosis due to their non-selective bacteriostatic or bactericidal nature. As such, the microbiota's protective function is compromised, and host colonisation resistance is reduced. This creates the opportunity for pathogens to proliferate and fill the void.
Simultaneously, antibiotic administration can exert a selection pressure promoting the development of antibiotic resistant genes or ARGs in both pathogenic and commensal bacteria of the microbiota. Many studies have documented ARG transfer between mentals and pathogens, meaning that the microbita is an important reservoir for development of an antibiotic resistant bacteria. Examples of ARGs include genes for extended spectrum betylectomma enzymes, allowing bacteria to be resistant to penicillins and some cephalosporins, or plasmid-mediated quinone resistance genes which confer a resistance to pluroquinones.
There also appears to be a zoonotic risk with farm workers carrying higher levels of ARGs compared to non-exposed controls, and a higher carriage of extended spectrum betylactomma producing E. Coli detected in veterinary staff when assessed over a six week period. Therefore, it becomes even more important to employ a responsible antibiotic use either when there is a true clinical indication or in line with the current research.
For example, a recent study reported that a 7 day course of amoxicillin clapulanic acid predisposed dogs with non-complicated acute diarrhoea to the establishment of amoxicillin resistant E. Coli in their faeces, which persisted for as long as 3 weeks post treatment. In the graph shown here, we can see a significant increase in the percentage of resistant E.
Coli at 6 and 30 days in the antibiotic group shown on the right compared to the placebo shown on the left. Furthermore, antibiotic use in these patients did not provide a clinical benefit with regards to the speed of resolution of diarrhoea compared to those given placebo. If we know that acute diarrhoea would often self resolve, what are we trying to achieve with our management protocols?
I guess the main aim would be to accelerate the resolution of the diarrhoea. Primarily for the animal's benefit, but also the owner's benefit, to reduce concern and the need for repeated cleaning at home. We should not just be using a product in the hope that it will do this, but in the knowledge that it has been proven to do so.
We will start by looking at a study which demonstrated just this. It looked at the efficacy of an oral probiotic prebiotic, clay-based paste in dogs with acute diarrhoea. 148 dogs were recruited onto the study that were presented at one of 14 veterinary practises in the UK and Ireland for acute diarrhoea.
Dogs were randomised to receive either a placebo or the anti-diarrheal paste alongside a commercially highly digestible diet. The paste and the placebo were identical in product appearance and packaging. Neither the vets nor the owners knew which one they were administering.
The owners were taught to score the consistency of every faecal motion and did so for the duration of the study. A scale ranging from 1 to 6, very watery was used. The study found that the dogs given the anti-diarrheal probiotic paste recovered significantly faster than those given the placebo.
1.6 times faster to be precise. We can see that about 50% of the animals in the anti-diarrhea probiotic paste group had recovered within 24 hours.
Whereas it took about 48 hours for 50% of the dogs to recover in the placebo group. An interesting extra finding in this study was that the group given the anti-diarrhea probiotic paste was significantly less likely to require additional medical intervention such as antibiotics, intravenous fluid therapy, or other gastrointestinal medications. 15% of dogs in the placebo group had to be removed from the study for additional medical intervention.
Whereas only 4% of dogs in the anti-diarrhea probiotic pace group had to leave the study for this reason. This could be beneficial clinically for the animal, not needing to be hospitalised for intravenous fluid therapy. And from a broader perspective of antimicrobial resistance by potentially reducing the usage of antibiotics.
Interestingly, if we return to this graph, which excludes patients which were removed from the study for additional medical intervention, we know that 4% of dogs taking the anti-diarrheal probiotic past required additional medical intervention. So 96% recovered with the specific anti-diarrheal probiotic paste and a highly digestible diet within 5 days. Even in the placebo group, 85% of patients recovered within 7 days with a highly digestible diet alone.
So have another think about last year's study which found that nearly 50% of dogs presenting with diarrhoea were prescribed antibiotics. Did they all really need them? I'll leave that for you to think about.
The next study we should look at assessed the efficacy of giving a probiotic prebiotic supplement on the incidence of diarrhoea in a dog shelter. 773 dogs were enrolled in the study. Following a mission to a rehoming shelter, they were randomised to receive either the probiotic prebiotic combination or a placebo.
The capsules were identical and were just labelled A or B. None of the staff knew which one was which. The staff monitored the dog's faecal scores during their stay, using a faecal scoring chart.
First, they looked at the number of dogs that developed diarrhoea during their 1st 14 days at the kennels. 75 dogs in the probiotic prebiotic group and 102 dogs in the placebo group developed at least 1 episode of diarrhoea during this time. As there were slightly less dogs overall in the placebo group, when we work these out as percentages, it equates to a 33% reduction in the incidence of diarrhoea in the dogs receiving the probiotic prebiotic combination.
Secondly, they looked at how many dogs had diarrhoea for more than 48 hours. This was particularly relevant because when dogs at this shelter had had diarrhoea for 48 hours, the protocol was that they should be seen by a vet. Again, there was a significant reduction in the probiotic prebiotic group 18 versus 30.
And when calculated as a percentage of the total number of dogs in each group, it worked out as a 44% reduction. Moving on now to look at some studies assessing the use of probiotics in feline diarrhoea. We're going to look in particular at the use of a powdered formulation containing probiotics and prebiotics alongside of other ingredients to support gut health in cats with tri-trichomonas foetus.
Trichomonas foetus is a protozoa which causes recurrent large bowel diarrhoea. And at times faecal incontinence in young purebred cats. It requires faecal PCR for diagnosis, so it may go undetected with routine faecal cultures.
Renidazole is known to be the most effective treatment. However, this is not licenced for use in cats and can have some nasty side effects. Also, it is not uncommon for symptoms to recur despite treatment with rannidazole.
This was a prospective double-blinded, placebo-controlled pilot study that was based out of Edinburgh vet school that involved cases from first opinion practises across the UK. 26 cats were recruited and as expected all were young pedigree cats. They were started on the recommended protocol of rannidazole at 10 to 30 milligrammes per kilogramme, once daily for 14 days.
At the same time, they were also started on a probiotic prebiotic digestive support powder or a placebo powder for a total of 4 weeks. The results showed that both groups improved significantly during the first two weeks. There was an improvement in their faecal score, and they also maintained or gained weight.
In the cats receiving the probiotic prebiotic digestive support powder, they were much less likely to have a recurrence of clinical signs. Only 2 out of 13 cats relapsed in any way. On the other hand, in the placebo group, 8 out of 13 cats relapsed.
The following studies were carried out using a different strain of enterococcus specium, so we need to take care extrapolating the results. However, the results are interesting and still highlight how probiotics can be beneficial. In this study, 217 cats in a rehoming shelter were randomised to receive either probiotics or a placebo for a four-week period.
Following admission, they had 4 weeks to settle in. Then they were split into two groups to receive either the probiotic or a placebo. They then had a washout period of one week before switching to receive the opposite.
Their faeces were scored on a daily basis by individuals who did not know which treatment each group was receiving. As we see here, the incidence of cats experiencing diarrhoea for more than two days reduced significantly from 20.7% in the placebo group to 7.4% in the ehesium group.
Moving away from gastrointestinal disease, we'll now look at a study which assesses the use of probiotics in dogs with atopic dermatitis. Puppies were bred from two beagles with severe atopic dermatitis over two subsequent pregnancies. Lactobacillus rhamgnosis GG was administered during the second pregnancy and to perhaps from that litter from 3 weeks to 6 months of age.
All puppies were sensitised to house dust mites, and they then had serum IgE measured every 6 weeks. And then after 6 months had intradermal allergy testing carried out and were exposed to the allergen. The results found that there was a significant reduction in serum IgE levels in the probiotic group.
Whilst there was a milder reaction to intradermal testing, there was no significant difference in clinical signs between the two letters. A follow-up study carried out after 3 years demonstrated that there were long-term clinical and immunological benefits to having taken the probiotics. Here is another study looking at how administration of enterococcus specium can stimulate immune function in young dogs.
14 puppies were fed either a controlled diet or a diet supplemented with enterococcal specium. They were given their normal vaccination course at 9 and 12 weeks. Then had various samples collected in order to assess antibody levels in the faeces and plasma.
Here we can see that both faecal and total IGA was significantly increased in the enterococcus specum group. And if we look specifically at antibodies for distemper, one component of the normal vaccination protocol, Again we can see that these are elevated in the Ephesian group. At this point in time, it's not known whether this would have had a clinical benefit on these puppies.
Would they surmount a greater response in the face of disease? Are they at a lower risk of other infections? An interesting finding, they are likely to require more research into this area.
The penultimate study that we will cover was looking at the use of enterococcus specium in cats with herpes virus. We know that herpes virus causes recurrent upper respiratory tract and ocular symptoms, namely conjunctivitis, which often recrudece at times of stress. The cats were given either eheium or a placebo during the study period.
They were moved between group and individual housing and were neutered during the study period. These kinds of events may typically trigger recurrence of the clinical signs. Clinical signs were monitored and samples were collected regularly.
They found that cats fed the placebo had decreased faecal microbial diversity, i.e., they developed a dysbiosis during these stressful times.
But microbiota diversity was maintained in those supplemented with their ehesium. Ehesium supplemented cats had less days with conjunctivitis than placebo cats following neutering. And if all observation points were taken into account, there was a significant reduction in conjunctivitis detection in the ehesium group compared to the placebo.
Despite the clinical differences, there was no detectable difference in the antibody responses between both groups. Finally, a slightly comical study to finish up with. Yes, people actually took the time to study canine flatulence.
Adult dogs were fed a standard diet for 14 days. At which point baseline rectal gas samples were collected. Following this, enterococcus specium was added to the diet for an additional 14 days, and sample collections were repeated.
Since flatulence varies between dogs, each dog acted as its own control, though admittedly there was no specific control or placebo group. Number of emissions and hydrogen sulphide gas content were measured via a perforated tube localised near each dog's anus and attached to a monitoring pump fitted with a sensor that recorded measurements every 4 seconds for a period of 4 hours. As you can see here, the mean number of emissions was significantly lower following 14 days of enterococcus specium supplementation.
Likewise, the hydrogen sulphide concentrations were also significantly lower following EPCM supplementation. As we saw earlier, there have been human studies looking at probiotic use for diseases affecting pretty much every organ system in the body. While the vetamin research is by no means at this level yet, research is ongoing, and this demonstrates the potential that there may be for probiotic use in the future.
So in summary, we have looked at the microbiota, dysbiosis, probiotics and prebiotics, and then finished up looking at evidence to support their use. I hope you found this webinar useful. Please do check out the Protex and veterinary website for further information.
Thank you for listening. If you have any questions, please feel free to contact the veterary technical team at Protexin using the following email address, [email protected].
