How the gut-brain connection affects the performance of horses

Diligence is the mother of good luck. –Benjamin Franklin

Article by Scott Anderson

Trainers are always looking to gain an edge in performance. At a minimum, they make sure their athletes get proper nutrition and exercise. Horses require muscle and stamina to compete, so they need to be in top physical condition. But what about their mental state? Are they jittery, distracted or disinterested? No matter how strong the horses are, their heads must be in the game to succeed.

Surprisingly, much of that mental attitude is driven by gut health, which in turn depends on the collection of microbes that live there, called the microbiota. In a horse, the microbiota is a tightly packed community of about 100 trillion microbes, composed of bacteria, archaea, fungi and protozoa. It colonizes the entire GI tract but is largely concentrated in the hindgut, where it works to ferment the prebiotic fiber in forage. The microbial fermentation of fiber into fatty acids produces 70% of the animal’s energy requirements and without it, the horse couldn’t get sufficient energy from simple forage. Intriguingly, byproducts of that fermentation can affect the brain. 

It is easy to be skeptical about this gut-brain connection, but over the last decade, research has made it clear that gut microbes have an outsized influence on mood and behavior. Microbes that improve mental state are called psychobiotics, and they may completely change the way you train and manage your horses. A horse’s health – and consequently its performance – starts in the gut.

Inflammation

When the microbiota is unbalanced by stress, diet or sickness, it is said to be dysbiotic. It loses diversity, and a handful of bacterial species compete for domination. Without the pushback of a diverse population, even beneficial bacteria can become pathogenic. Surprisingly, that can affect the brain. Multiple studies in various animal models have shown that transmitting fecal matter from one animal to another also transmits their mood. This demonstrates that a dysbiotic microbiota can reliably cause mental issues including anxiety and depression, thereby affecting performance. 

An important function of the microbiota is to fight off pathogens by outcompeting, starving or killing them. However, a dysbiotic microbiota is less diligent and may permit pathogens to damage the gut lining. A degraded gut lining can leak, allowing bacteria and toxins into the bloodstream. The heart then unwittingly pumps them to every organ in the body, including the brain. This makes the gut the primary source of infection in the body, which explains why 80% of the immune system is located around the intestines. Over time, a leaky gut can lead to chronic systemic inflammation, which weakens the blood-brain barrier and interferes with memory, cognition and mood. 

Inflammation is a major component of the gut-brain connection, but not the only one.

Neurotransmitters and hormones

Horses and humans use neurotransmitters to communicate between nerve cells. Brains and their attendant nerve bundles constitute a sophisticated network, which makes it somewhat alarming that microbes also produce neurotransmitters. Microbes use neurotransmitters to converse with each other, but also to converse with their host. The entire gut is enmeshed in nerve cells that are gathered up into the vagus nerve that travels to the brain. Microbial neurotransmitters including serotonin and dopamine thus allow certain microbes to communicate directly with the brain via the vagus nerve. We know this happens with specific bacteria, including Lactobacillus species, because when the vagus is severed, their psychobiotic effects disappear. 

As well as neurotransmitters, hormones are involved in gut-brain communications. The hypothalamus-pituitary-adrenal (HPA) axis controls the stress response in animals. The hypothalamus is located low in the brain and responds to stressors – such as a lurking predator – by producing hormones that stimulate the neighboring pituitary, which then triggers the adrenal gland to produce cortisol, the stress hormone. Cortisol acts as a threat warning and causes the horse to ramp up glucose production, supplying the energy needed to escape a predator. This is the same hormonal circuit that trainers exploit for racing.

The HPA axis produces cortisol in response to stress. Cortisol inhibits the immune system, which in combination with a leaky gut allows pathogens to enter the bloodstream. Subsequent systemic inflammation and vagal feedback lead to stereotypies.

The production of these hormones redirects energy to the heart, lungs and muscles at the expense of the immune system. From an evolutionary point of view, the tradeoff makes sense: first escape the predator and deal with infections later. After the danger has passed, cortisol causes the HPA to return to normal – the calm after the storm. 

However, continued stress disrupts that cycle, causing anxiety and diminishing the brain’s ability to store memories. This can dramatically interfere with training. Stress can also induce the release of norepinephrine, which promotes the growth of pathogenic bacteria including Campylobacter jejuni, Listeria, Helicobacter pylori and Salmonella. Prolonged high cortisol levels can increase gut leakiness, potentially leading to infection and further compounding the situation. In the long term, continued stress leads to systemic inflammation, which is a precursor to problematic behaviors.

Short-chain fatty acids

When microbes consume proteins and fiber, they break them down into their constituent molecules, such as amino acids, fatty acids and sugars. These are the metabolites of the microbes. As well as neurotransmitters and hormones, the gut-brain conversation is mediated by metabolites like butyrate  – an important short-chain fatty acid that plays multiple roles in the body. 

In the gut, butyrate serves as a preferred nutrient for the cell lining. It encourages the differentiation of stem cells to replenish gut cells that are routinely sloughed off or damaged. It plays an important role in the production of mucus – an essential part of gut protection – which coats the gut from mouth to anus. In the muscles, butyrate boosts the growth of skeletal muscle, which is crucial to athletic performance, as well as for inducing the production of glucose  – the primary muscle fuel. One-quarter of systemic glucose is driven by butyrate. In its gut-brain role, butyrate passes through the blood-brain barrier, where it nourishes and enhances the growth of new brain cells. 

These factors make butyrate a star player in the gut-brain connection. They also highlight the benefits of prebiotic fiber, especially when high-energy, low-fiber feeds are provided.

Starting a microbiota

We’ve explored the major pathways of the gut-brain connection: inflammation, neurotransmitters, hormones and fatty acids. Some of these pathways are at odds with each other. How does such a complicated system come together?

As mentioned, the microbiota is an animal’s first line of defense against pathogens, attacking and killing them, often before the immune system is even aware of them. That means a healthy microbiota is an essential part of the immune system. However, the immune system is designed to attack foreign cells, which includes bacteria. For the microbiota to survive, the immune system must therefore learn to accept beneficial microbes. This lesson in tolerance needs to take place early in the foal’s development, or its immune system may forever fight its microbiota.

There are multiple ways nature ensures that foals get a good start on a microbiota that can peacefully coexist with the immune system. The first contribution to a protective microbiota comes from vaginal secretions that coat the foal during birth. After birth, microbes are included in the mare’s milk. These microbes are specially curated from the mare’s gut and transported to the milk glands by the lymphatic system. The mare’s milk also includes immune factors including immunoglobulins that help the foal to distinguish between microbial friends and foes. An additional way to enhance the microbiota is through coprophagia, the consumption of manure. Far from an aberration, foals eat their mother’s manure to buttress their microbiota. 

Microbes affect the growth and shape of neurons in various brain sites as the foal develops, a remarkable illustration of the importance of a healthy early gut microbiota. 

The cooperation between the immune system and the microbiota is inevitably complex. Certain commensal bacteria, including Clostridiales and Verrucomicrobia, may be able to pacify the immune system, thus inhibiting inflammation. This is a case where microbes manage the immune system, not the other way around. These convoluted immune-microbial interactions affect the mental state – and consequently the behavior – of the horse, starting at birth.

Stereotypies

A 2020 study of 185 performance horses conducted by French researchers Léa Lansade and Núria Mach found that the microbiota, via the gut-brain connection, is more important to performance than genetics. They found that microbial differences contributed significantly to behavioral traits, both good and bad. A diversified and resilient microbiota can help horses better handle stressors including stalling, training and trailering. A weakened or dysbiotic microbiota contributes to bad behaviors (stereotypies) and poor performance. 

The horses in this study were all carefully managed performance horses, yet the rates of stereotypies were surprisingly high. A kind of anxiety called hypervigilance was observed in three-quarters of the horses, and almost half displayed aggressive behavior like kicking or biting. The study found that oral stereotypies like biting and cribbing were positively correlated with Acinetobacter and Solibacillus bacteria and negatively correlated with Cellulosilyticum and Terrisporobacter. Aggressive behavior was positively correlated with Pseudomonas and negatively correlated with Anaeroplasma. 

Some of these behaviors can be corrected by certain Lactobacillus and Bacteroides species, making them psychobiotics. That these personality traits are correlated to gut microbes is truly remarkable. 

Intriguingly, the breed of a horse has very little impact on the makeup of its microbiota. Instead, the main contributor to the composition of the microbiota is diet. Feeding and supplements are thus key drivers of the horse’s mental state and performance. 

The gut-brain connection and training

How might the gut-brain connection affect your training practices? Here are some of the unexpected areas where the gut affects the brain and vice-versa:

High-energy feed. Horses evolved to subside on low-energy, high-fiber forage and thus have the appropriate gut microbes to deal with it. A high-energy diet is absorbed quickly in the gut and can lead to a bloom in lactic acid-producing bacteria that can negatively impact the colonic microbiota. High-energy feeds are designed to improve athletic output, but over time, too much grain can make a horse antisocial, anxious and easily spooked. This can damage performance  – the very thing it is trying to enhance. Supplementary prebiotics may help to rebalance the microbiota on a high-starch regimen.

Changing feed regimens quickly. When you change feed, certain microbes will benefit and others will suffer. If you do this too quickly, the microbiota can become unbalanced or dysbiotic. Introducing new feeds slowly helps to prevent overgrowth and allows a balanced collection of microbes to acclimate to a new regimen. 

Stress. Training, trailering and racing all contribute to stress in the horse. A balanced microbiota is resilient and can tolerate moderate amounts of stress. However, excessive stress can lead, via the HPA axis, to a leaky gut. Over time, it can result in systemic inflammation, stereotypies and poor performance.

Overuse of antibiotics. Antibiotics are lifesavers but are not without side effects. Oral antibiotics can kill beneficial gut microbes. This can lead to diarrhea, adversely affecting performance. The effects of antibiotics on the microbiota can last for weeks and may contribute to depression and anxiety. 

Exercise and training. Exercise has a beneficial effect on the gut microbiota, up to a point. But too much exercise can promote gut permeability and inflammation, partly due to a lack of blood flow to the gut and consequent leakiness of the intestinal lining. Thus, overtraining can lead to depression and reduced performance.

Knowing how training affects the gut and how the gut affects the brain can improve outcomes. With a proper diet, including sufficient prebiotic fiber to optimize microbiota health, a poor doer can be turned into a model athlete. 

References

Mach, Núria, Alice Ruet, Allison Clark, David Bars-Cortina, Yuliaxis Ramayo-Caldas, Elisa Crisci, Samuel Pennarun, et al. “Priming for Welfare: Gut Microbiota Is Associated with Equitation Conditions and Behavior in Horse Athletes.” Scientific Reports 10, no. 1 (May 20, 2020): 8311.

Bulmer, Louise S., Jo-Anne Murray, Neil M. Burns, Anna Garber, Francoise Wemelsfelder, Neil R. McEwan, and Peter M. Hastie. “High-Starch Diets Alter Equine Faecal Microbiota and Increase Behavioural Reactivity.” Scientific Reports 9, no. 1 (December 9, 2019): 18621. https://doi.org/10.1038/s41598-019-54039-8.

Lindenberg, F., L. Krych, W. Kot, J. Fielden, H. Frøkiær, G. van Galen, D. S. Nielsen, and A. K. Hansen. “Development of the Equine Gut Microbiota.” Scientific Reports 9, no. 1 (October 8, 2019): 14427.

Lindenberg, F., L. Krych, J. Fielden, W. Kot, H. Frøkiær, G. van Galen, D. S. Nielsen, and A. K. Hansen. “Expression of Immune Regulatory Genes Correlate with the Abundance of Specific Clostridiales and Verrucomicrobia Species in the Equine Ileum and Cecum.” Scientific Reports 9, no. 1 (September 3, 2019): 12674. 

Daniels, S. P., J. Leng, J. R. Swann, and C. J. Proudman. “Bugs and Drugs: A Systems Biology Approach to Characterising the Effect of Moxidectin on the Horse’s Faecal Microbiome.” Animal Microbiome 2, no. 1 (October 14, 2020): 38.

Probiotics – The key to a well-balanced equine gut

Article by Kerrie Kavanagh

It is no surprise that the health maintenance of the racehorse is a top priority for trainers. And probiotics can be used as a treatment modality to manipulate the gut microbiome to improve or maintain health. Equine studies to date have shown that probiotic strains can offer an advantageous approach to minimising disturbances in the gut microbial populations, repair these deficiencies—should they occur—and re-establish the protective role of the healthy gut microbiome. Other probiotic-associated health benefits include reducing diet-related diseases such as colic and laminitis, preventing diarrhoea, conferring host resistance to helminth infection, improving stress-related behavioural traits (e.g., locomotion) and even promote the development of an effective gut-brain communication pathway. 

Probiotics have been used by humans for more than 5,000 years with their development closely linked to that of dairy products and fermented foods. Today, probiotics are seen as an excellent non-pharmaceutical way to improve the health of both humans and animals, and there are a plethora of products to choose from. But what exactly is a probiotic, and how do they work? Why would your horse need one? What types of probiotics are available for horses? These are all questions that horse trainers ask frequently, which we will attempt to answer here. 

The Equine Gut Microbiome

Probiotics and the equine microbiome can benefit from a valuable symbiotic relationship; probiotics are seen as a restorative treatment modality for the gut, to re-establish the bacterial populations there and also to re-establish the protective role that the health gut microbiome confers to the host. But when we discuss the equine microbiome, what are we really talking about? 

The gut microbiota/microbiome can be categorised by anatomical location such as the oral microbiota/microbiome in the mouth and the intestinal microbiota/microbiome in the intestines, etc. Therefore, the gut microbiome pertains to the microbiota in the gastrointestinal tract. This population of microorganisms (bacteria, fungi, viruses, protozoa) is referred to as the ‘microbiota’ of the gut, while the term ‘gut microbiome’ refers to the genetic material associated with these microorganisms. The microbiome can be defined as the sum of the microbes and their genomic elements in a particular environment. If we look at the definition of the microbiome having the propensity to an equation, then any equation must be balanced; to maintain that balance is key. If the microbial community exists in an environment in a balanced state, then any upset or disturbance to the microbial populations will cause the balance to shift (known as dysbiosis). To maintain the balance, we need to firstly understand the way the microorganisms exist within their community (i.e. their microorganism-to-microorganism interactions and also microorganism-to-environment interactions) and secondly, their functioning role. If we can understand their (microorganism) position and role, then we can maintain the balance or re-establish the balance if a shift occurs.  

The human intestinal microbiome is now recognised as an organ and likewise, the equine intestinal microbiome is deemed an ‘organ’ of the body and is vital for the breakdown of complex food and subsequent release of energy, protection against the pathogenic bacterial colonisation and in regulating the immune system and metabolic functions. There has been much debate regarding the content of the healthy equine microbiome, and even to deduce what ‘healthy’ or ‘normal’ is requires a level of understanding of the microbiota associated with healthy horses. This question has been posed by many researchers and frankly has yet to be answered with certainty. There are many reasons why the ‘normal’ microbiota keeps eluding us; and this can be attributed to the many reasons as to why the gut microbiota (of a healthy horse) can be affected (see Figure 1). It is thought that the diversity of the human gut microbiota and the general assembly of microbial communities within the gut (with the dominant phyla being classed as belonging to Firmicutes and Bacteroidetes) is a shared hypothesis across most species (i.e., humans and animals share a similar gut microbiome structure). Firmicutes and Bacteroidetes have been shown to constitute the main dominant phyla in equine, bovine, canine and feline gut microbiome studies indicating the cruciality of the role they play in the maintenance of a healthy microbial ecology in the gastrointestinal tract. Several studies do agree that dominant phyla of the equine gut microbiota are obligate anaerobes: the gram-positive Firmicutes and the gram-negative Bacteroidetes; other phyla are identified as Proteobacteria, Verrucomicrobia, Actinobacteria, Euryarchaeota, Fibrobacteres and Spirochaetes. Ninety-five percent of the  Firmicutes phyla contains the Clostridia genus in addition to genera related to gut health such as Lachnospiraceae, Faecalibacterium and Ruminococcaceae. The other main dominant phyla, Bacteroidetes, on the other hand contains a large variety of the genus. 

Role of the Equine Gut Microbiota

The role of the gut intestinal microbiota serves to protect and prevent disease. The gut microbiota has several purposes: prevention of pathogen colonisation by competing for nutrients, enrichment and maintenance of the intestinal barrier—their ability to renew gut epithelial cells and repair damage to the mucosal barrier, the breakdown of food and releasing energy and nutrients, such as synthesising vitamins D and K and also conserving and restoration of the immune system by the formation of antimicrobial metabolites and blocking access to the binding sites of the mucosal wall. The gut microbiota is also thought to play some role of influencing the neuro-active pathways that affect behaviour. It is not surprising to see that gut disorders and gastrointestinal diseases can arise when gut dysbiosis occurs. The role of the gut microbiota may have even more importance than is realised and may have a role to play with developing illness or disease later in life.

The microbial colonisation of the intestinal tract begins at birth. The foal begins its colonisation through contact with the microbiota of the mare’s vaginal and skin surfaces plus the surrounding environments to which the foal is exposed and reaches a relatively stable population by approximately 60 days in age. It is perhaps a fight for dominance to achieve establishment in the gut among the bacterial populations that sees the foal’s microbiota as being more diverse and quick to change when compared to that of the older horse. The subsequent colonisation of the intestinal tract will reflect the foal’s diet, changing environment, introduction to other animals, ageing and health.

Figure 1: Factors that can lead to gut dysbiosis

What exactly is a probiotic?

The word ‘probiotic’ is of Greek origin meaning ‘for life’ and the WHO/FAO have defined probiotics as ‘live microorganisms which when administered in adequate amounts confer a health benefit on the host’.  People have long believed that exposure to non-pathogenic microorganisms can benefit the health of humans and animals. The thinking behind this is that daily consumption of sufficient numbers of ‘good’ microorganisms (either bacteria or fungi) can maintain a healthy population of microorganisms in the gut and benefit overall health.  

Probiotics are used to manipulate the bacterial populations of the gut in order to re-establish the delicate microbial balance there which, in turn, confers health benefits on the host. As the benefits associated with some of the ‘good’ bacteria within the gut became known, these were referred to as probiotic bacteria. 

How do probiotics work?

There are 4 main mechanisms by which probiotics are thought to exert their effects.

1. By inhibiting pathogen colonisation in the gut through the production of antimicrobial metabolites or by competitive exclusion; in other words, they prevent the ‘bad’ bacteria from growing in the gut.

2. By protecting or re-stabilising the commensal gut microbiota, probiotics can be a means to re-establish the balance of the gut microbial populations.

3. By protecting the intestinal epithelial barrier, they maintain the health of the intestinal wall.

4. By inducing an immune response, probiotics can boost the immune response and help prevent disease.

If we consider the definition of a probiotic as ‘live non-pathogenic microorganisms that, when administered in adequate amounts, confer a health benefit on the host’, then this reference to ‘adequate amounts’ must be emphasised, and the dose administered is critical to ensure that the probiotic has the desired effect. For horses, we must consider the route through the digestive tract that the probiotic strains must travel to arrive at their destination is a distance over 15 metres long. It is a race for survival! The gastrointestinal system has many obstacles along the passage such as the acidic stomach environment and the dangers of exposure to bile and digestive enzymes, in which they must survive. The initial dose of ‘live’ probiotic strains is therefore crucial to ensure survival in the gut. Prebiotics are ingredients such as carbohydrates and fibre, which promote the growth of these probiotic bacterial/yeast strains in the gut. Prebiotics are essentially the food for the probiotic strains and can help form a symbiotic relationship with the probiotic to improve the overall health status of the horse. 

Why would you need to give your horse a probiotic?

Gut dysbiosis is a fluctuation or disturbance in the population of microorganisms of the gut, which may be linked to a wide range of diseases in horses. Gut dysbiosis can be caused by many factors ranging from dietary changes, antibiotics, disease, intense exercise and training, age, worms, environment, travel, or even minor stress events—resulting in major consequences such as colic. Dysbiosis is generally associated with a reduction in microbial species diversity. 

Diet is one of the major factors contributing to gut dysbiosis. Unlike the ruminant cattle and sheep that use foregut fermentation, horses are hindgut fermenters. The large intestine is the main area where fermentation occurs. The horse utilises the microbial enzymes of the hindgut microbial population in the colon and caecum to break down the plant fibres (cellulose fermentation) sourced mainly from grasses and hay. The horse itself does not possess the hydrolytic enzymes that are required to break the bonds of the complex structures of the plant carbohydrates (in the form of celluloses, hemicelluloses, pectins) and starch; so therefore, it strongly relies on the microbiota present to provide those critical enzymes required for digestion. The main phyla Firmicutes and Bacteroidetes possess enzymes capable of breaking down the complex carbohydrates (such as starch and cellulose).

Research has shown that forage-based diets (grasses and hay) promote the most stable gut microbiomes, but ultimately the equine athlete requires far more energy than a forage-based diet can supply. Supplementing the diet with concentrates containing starch such as grain, corn, barley and oats can affect the number and type of bacteria in the gut. Optimising diet composition is so important as carbohydrate overload—as seen with high-starch diets (>1g/kg body weight per meal)—can change the populations of bacteria in the gut, alter pH, upset digestion and the gut environment, and ultimately result in diseases such as colitis, colic and laminitis. The correct diet is essential for maintaining the delicate balance of bacterial populations. Probiotics can be used to either replace the bacteria missing in the gut and/or can help maintain the delicate microbial balance even in the face of adversity such as abrupt dietary changes, antibiotic treatment and stress.

What types of probiotics are available for horses?

There are several probiotic products on the market, and most are in powder or liquid form. There are two main categories of probiotics: generic and autogenous. Generic probiotics are off-the-shelf products that contain specific strains of bacterial or yeast, singularly or in combination. The Lactobacillus and Bifidobacterium families, Enterococci and yeasts such as Saccharomyces cerevisae and boulardii are the most common equine probiotic strains. Advantages of generic probiotics are that they are widely available, easy to administer, and they may be beneficial to horse health (if the strains are alive in sufficient numbers). Autogenous probiotics are specifically formulated using bacteria obtained from the horse’s own faecal sample and, as such, are uniquely adapted to that individual animal. These host-adapted bacteria are more likely to survive in the gut than non-adapted generic strains and can quickly replenish absent or low levels of bacteria unique to the individual horse, thus maintaining health.