Symbiosis of Breeders, Gluck Equine Research Center Reaches New Heights

When Dr. Emma Adam took on the role of equine outreach veterinarian in 2018, the University of Kentucky's Department of Veterinary Science hoped that the renewed position could continue to bridge the gap between their research and diagnostic laboratories and the industry that they serve.

With five years now under her belt in the role, Adam is pleased with the strides that have been made in fostering relationships with the equine community–particularly within the Thoroughbred business in Central Kentucky–and she is optimistic that those connections will lead to further advancements in the coming years.

While Adam's role encompasses outreach for both UK's Maxwell H. Gluck Equine Research Center and the Veterinary Diagnostic Laboratory, she said the Diagnostic Lab has always been more outward facing by nature so her initial goal was to bring more external emphasis to the research center.

“Our entire community wants to be better connected with our industry stakeholders, but bandwidths can limit that reality,” Adam explained. “I think everyone recognized the need for someone who had practice and industry experience and could bring that to the Gluck Center to connect it with what we do within the building. Researchers are so busy trying to fund their projects and write papers that the time available to get out there and interact with our industry was getting less and less. We needed to take a step back and say, 'What do we need to do to remain connecting  what is happening in our industry with what is happening in our academic community and vice versa, so that we can help each other?'”

Adam's background in racing has served her well in her position, which was spearheaded by Nancy Cox, UK's Vice President for land-grant engagement and Dean of Martin-Gatton College of Agriculture, Food and Environment. A native of Newmarket, England, Adam was immersed in the racing and breeding industry from a young age as she grew up on a commercial farm and was first familiarized with the racing side of the business when she worked for champion trainer Sir Michael Stoute. As a practicing veterinarian, she worked in various racing and breeding-related positions around the globe before deciding to return to academia and earn her PhD at UK's Department of Veterinary Science. From there, she stepped into her current role.

Making connections with farm managers and veterinarians has proven to be invaluable as Adam has worked to get the word out on the Gluck Center's mission and increase awareness among owners and breeders on the resources that the research center has to offer.

“Our research community is brilliant, but reaching them is sometimes difficult,” Adam admitted. “We have a fantastic network here through the Consignors and Commercial Breeders Association, the Kentucky Thoroughbred Farm Managers Club and our veterinary community. While we'd always love for it to be bigger, we're developing a stronger network of people that know we're here to help.”

Dr. Emma Adam | photo courtesy Emma Adam

When breeders have issues arise, such as pregnancy loss, the Gluck Center's team coordinates with the Diagnostic Lab, veterinarians and farm managers to investigate those problems.

“We will go through lots of different parameters with the veterinarian and the farm manager and take some colleagues that might help with such things as examining the pastures,” Adam explained. “We'll do hay, soil and water analysis, and examine records for each mare. We'll work closely with our colleagues in the Diagnostic Lab so we can connect any dots that might be available to look at from the perspective of those unfortunate pregnancy losses.”

“We may not always arrive at an answer, but along the way we'll have often found out some other things that we can be more vigilant for,” she continued. “Those things might include checking to make sure that we're testing our pastures for things like tall fescue or trying to avoid stress when moving mares to different pastures. Everybody brings something to the table and we chew it all over and see if we currently have what we think is the best possible plan for that farm and then offer help to implement it if we can.”

Adam stressed that confidentiality and trust are important aspects of the work they do with commercial breeding farms.

“We have very strict confidentiality obligations and we take them very seriously,” she said. “All samples and veterinary and farm interactions are completely confidential. We recognize that it is vital that people can trust everything we do, not only with the quality of our science but in how we handle those interactions. We're very fortunate that people are generous with their information and we respect that they have entrusted us with it.”

Building relationships with more breeding farms throughout Central Kentucky has been an ongoing goal that Adam said she believes will lead to mutually beneficial results for both the research center and breeders.

One example of how this reciprocity has led to research findings is in the Gluck's studies on nocardioform placentitis, a cause of late-term abortions and perinatal deaths. Recent studies on this disease have been made possible in large part by the samples researchers have received from local farms. Efforts are ongoing with the goal of developing an early-warning diagnostic test for the condition and better understanding how and when mares are exposed.

The breeding industry supports the Gluck not only in supplying samples, but on occasion, in providing much-needed financial support. When a rash of neonatal foal diarrhea broke out in the spring of 2021, the general consensus was that the cases behaved like Equine Rotavirus A, however diagnostic tests were coming back negative. Upon closer examination, UK virologists Drs. Feng Li and Dan Wang were able to perform genomic sequencing on samples to determine that a new strain of Rotavirus had emerged.

The Kentucky Thoroughbred Association, Kentucky Thoroughbred Owners and Breeders Foundation, The Grayson-Jockey Club Research Foundation, Coolmore America, Consignors and Commercial Breeders Association and the William S. Farish Fund provided gifts to develop a research plan to develop a vaccine. This summer, work is being done to test new vaccine candidates that are hoped will provide protection to foals against Equine Rotavirus B.

“That is a perfect example of how our industry is really trying to help itself by helping us,” said Adam. “We're very fortunate to have a relationship where they recognize that we're here and trying to help and where they support us in that effort because there is so little money for equine research. When it comes to things that are a bit more niche like foaling mares, it is even harder to get funds to do the kind of work that is relevant to our industry.”

Adam explained that the concentration of mares bred and foaled in Central Kentucky can oftentimes lead people to falsely believe that the area has more problems than other places. Instead, she described how the community's outstanding veterinarians and farm managers endow the region with a culture of constant vigilance and a drive to understand every problem encountered.

“Everything is so close here,” she shared. “You don't have to drive more than an hour from any of these farms to get to our world-class Veterinary Diagnostic Lab that sees probably more horses and horse samples than any lab globally. What that means is we're able to, as best as we possibly can, explore and understand what happened and how can we might prevent it in future. That is not something that a lot of other places can access so easily.”

Moving forward as a growing number of local farms have started utilizing the Gluck's resources, Adam's next goal is to develop ways to gather data each year from surveys and surveillance sampling.  These findings will be used to build profiles of how data is changing over time. The Veterinary Diagnostic Laboratory website collates some of these data, but Adam said she wishes to extend the survey to the farm level.

“We can take that data gathering further in asking our stakeholders to help us generate information on the things we deal with on a farm level–for example, pregnancy losses, red bag deliveries, foal diarrhea cases, yearling respiratory disease and so the list goes on,” she explained. “These data are absolutely essential to inform and direct our research not just at the local level here in the department, but from the perspective of being able to go out there and fight for the funding from the grant agencies to get that research done.”

Embarking on such a project is only possible with the collaboration of the entire Thoroughbred breeding industry in Central Kentucky, but Adam said she is counting on both the relationships that are newly formed and in others that have flourished over the years.

“The Department's footprint in our community continues to grow and that is a culture all of us in the University are keen to expand,” she said. “Taken together we have extraordinary natural and human resources here in Kentucky to serve and support our industry locally and worldwide.”

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Study To Help Identify Horses At Risk For Catastrophic Injury Moves Into Third Phase

The Kentucky Equine Drug Research Council, a committee of the Kentucky Horse Racing Commission, voted Friday to grant additional funding to an ongoing study at the University of Kentucky's Gluck Equine Research Center into catastrophic injury prevention.

Read more about the science behind the study here.

Dr. Allen Page, researcher at Gluck, presented the EDRC with an update on the first two phases of the three-phase project, which the council has contributed $300,000 to so far.

The goal of the research is to ultimately develop a blood test that will help officials detect horses who may be at elevated risk of a catastrophic injury. While some research has looked at biomarkers for existing injury or disease, this study is looking “upstream” at the messenger RNA responsible for sending instructions that those markers and other proteins be made. The hope is that a horse's blood sample could show warning signs of chronic or building inflammation that is not yet detectable to an observer.

In phase one, Page and fellow researcher Dr. David Horohov gathered blood samples from racetracks in four jurisdictions taken at the time of pre-race TCO2 testing, then examined the samples of horses who ultimately suffered fatal injuries alongside competitors in the same races who did not. The team looked at 21 mRNA markers and found three which were present in different levels in injured versus non-injured horses — IGF-1, MMP-2, and IL1RN.

IGF-1 is known to play a role in bone development and repair, and it was increased in injured horses, suggesting chronic inflammation was present. Matrix metalloproteinase-2, or MMP-2, is thought to assist with tissue repair and fracture remodeling and was also elevated in injured horses. Interestingly, IL1RN is more commonly known as IRAP–a anti-inflammatory material derived from a horse's own blood and given therapeutically by veterinarians to reduce inflammation and aid in healing an injured horse. IRAP was decreased in fatally injured horses, suggesting the body's natural anti-inflammatory process had been disrupted for some reason. Horses with higher levels of IRAP were actually seven times less likely to suffer fatal injuries.

In phase two, Page reported that the team used the university's super computer network to analyze terabytes of data, looking for other significant markers. The data analysis looked at 22,000 different mRNA markers and was able to uncover three new ones which seemed to change significantly depending on a horse's injury status. He could not reveal those markers, since he is hopeful the findings will be published in a peer-reviewed journal, and such publications require key information not be divulged beforehand. He was able to say that two of those mRNA markers were encoded from different chromosomes but appear to play very similar roles in the horse's body. The third has been described in research in humans and horses, but previously the genetic marker's purpose had been unclear.

Page said a blood test looking at all six mRNA markers – the three from phase one of the project, and the three new ones – is now correct 80 percent of the time when trying to identify a horse that will have an injury. Considering that research from pathologists indicates about 90 percent of fatal musculoskeletal breakdowns show signs of chronic damage on necropsy, Page thinks that's a pretty good success rate for the blood test.

“I'm starting to become part of the camp that thinks we can identify a large portion of the horses that may be at risk for a catastrophic injury, but we may not be able to identify all of them,” said Page. “We're certainly making some headway and I'd argue that it's substantially better than anything out there that we have now to detect horses at risk for injury.”

Phase three of the project will involve 15,000 new samples pulled from horses in Southern California in an attempt to validate the work identifying the six mRNA markers. Page does not think the results should be significantly impacted by the fact they'll be coming from a different racing circuit than the phase one samples, and that the possibility the team could end up sampling the same horses multiple times will give them even better opportunities to see how inflammation may change in an individual over time.

When asked about the practical applications of the test on Friday, Page said that currently it takes about 24 to 48 hours to get results, so its use as a screening tool pre-race would need to take that into account. With time and improved testing methods however, it is possible a racetrack could someday analyze the samples more rapidly on-site.

The EDRC approved $40,000 to help fund the project's third and final phase. Industry stakeholders, including Keeneland, The Stronach Group, and the New York Racing Association have already committed to contributing to the project.

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New Injury Prevention Study To Begin Using Horses In Southern California

Southern California will participate in a groundbreaking study aimed at identifying and avoiding potential injuries in racehorses by analyzing messenger RNA (mRNA) markers in blood samples. The research project is being conducted by the University of Kentucky's Gluck Equine Research Center.

The goal is to develop mRNA technology into a reliable, non-invasive tool that will empower owners and trainers by providing an additional method for improving the welfare and longevity of their racehorses. Participation in this study is a means by which owners and trainers can contribute towards this important goal.

The project involves the collection of an additional blood sample at the time of TCO2 testing. These samples will then be used to validate previously compiled data to identify specific markers for horses at risk for a catastrophic injury. The study will be overseen by Dr. Tim Grande at Santa Anita. Sample collection is expected to begin in mid-February.

Since samples are collected quickly (5 seconds) and during a time when the horse is already being stuck with a needle, participation in this study entails minimal disruption for the trainer and minimal risk to the horse. If you do not want your horse to participate in this study, please make that known at the time of TCO2 sample collection.

Click here for more information

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A ‘Horse-on-a-Chip’? The Future Of Equine Drug Research Could Look Very Different

The research process for drug toxicology in horses has always been long, slow, and expensive. Too often, when veterinarians want to more about the way a drug behaves in horses, they find themselves relying on limited data collected from a small number of horses. That's because there is a lot of expense and regulation associated with using live animals for research of any kind, even a simple drug administration study aimed at determining how quickly horses' bodies metabolize a therapeutic substance. It's also expensive for universities to maintain horse research herds of significant size year after year, awaiting their use in a short study.

A research group at the Gluck Equine Research Center is hopeful they have a solution that will make it quicker and easier for scientists to understand how drugs behave in horses, and it sounds like something out of a sci-fi drama: microscopic equine organ systems.

It's no longer science fiction. Dr. Carrie Shaffer said researchers aren't reconstructing full-size organs, but rather are using defined layers of cells that mirror what you'd find in an equine kidney, liver, lung, or intestine. The cells come from tissue-specific stem cells collected from a Thoroughbred foal that had to be euthanized due to an unrelated structural deformity. Stem cells have the ability to become any kind of differentiated cell upon command, so the researchers are able to direct the cells to form a particular organ tissue.

“We can prove, using a variety of different methods, that our equine microscopic organ systems are stem-cell derived and have the same characteristics and architecture as the corresponding tissue in the horse.”

These microscopic organ systems are grown in clear, plastic microfluidic chips that are about the size of a AA battery. In human medicine, similar microfluidic chips have been developed to mimic the human liver, lung, intestine, kidney, and blood/brain barrier and are used to study various aspects of cell biology and tissue responses to therapeutics.

The metabolism of a drug isn't dependent on the full-size physical structure of an equine liver or kidney, according to Shaffer – it's how the cells of those organs interact with drugs they encounter as the substance passes through an animal's bloodstream and into the organ tissue. Shaffer is able to grow specific liver cells in one channel of the microfluidic chip while creating artificial blood vessels and blood-like fluid flow on the opposite channel of the chip. This simulates a continuous blood supply interfaced with the mucous membranes that are normally found in the body. The blood flow can go in only one direction, which also mimics the horse's body, where veins and arteries carry blood through an organ in only one direction at a time.

“In the case of the lung chip and the intestine chip, we can also introduce relevant biomechanical forces that simulate complex biological processes,” she said. “We can introduce physical stresses into the chip that mimic breathing and lung inflation, or recreate defined patterns of stretch across the intestine chip that simulate the wave-like pattern of nutrients and waste products moving along the equine intestinal system.”

These forces have been shown to direct gene expression in the cells, which create small, but critical, changes that make the microfluidic chips behave more like the cells found in a live animal.

Previous iterations of this technology didn't include biomechanical forces like stretch, so the tissue wasn't as true to that in a horse's body. Additionally, previous tissue culture systems did not allow for directional fluid flow, but rather exposed a single type of liver or kidney cell to static fluid containing a drug at a fixed concentration. That's not how real kidneys and livers actually work, said Shaffer – the organs contain multiple cell types that are exposed to blood flowing at a relatively high rate. Therapeutics within the bloodstream pass through various organ systems within seconds, and carry metabolized drug away from one organ system for delivery to another.

“Under normal drug testing conditions, we are able to analyze a blood sample from a horse after a drug is administered, but we cannot tell in that blood sample where the drug metabolism occurred,” she said. “We don't know whether the drug was liver-metabolized, intestinal-metabolized, or metabolized in the lung. Our horse-on-a-chip microfluidic technology allows us to isolate exactly where drug metabolism occurs within the horse.”

Some drugs metabolize at different rates in different organs, and organs probably take turns at metabolizing a drug but there's currently no way to know in what order metabolism occurs for a given therapeutic. That information could be useful because some drugs linger longer in the body than expected, and scientists often don't know where the hold-up is.

Shaffer said her lab has performed only a handful of studies with the technology because it's so new. So far, the team has pulsed a drug through an equine lung-chip and a liver-chip for sample collection from the apparatus at defined times post-administration to see how much of the drug had been metabolized by specific tissues in a set timeframe.

The team is still validating these emerging  methods and drafting papers for peer-reviewed journals describing the process they've used to create this technology. Shaffer said they're still a few months away from using the organ chips en masse for huge studies – and they need to expand to include tissues from other breeds – but she thinks the microfluidic chips could be useful for pre-clinical analysis of new therapeutic drugs.

“The big sell with our horse-on-a-chip technology is that it's going to significantly reduce animal use for studies – reduce euthanasia, reduce the need for research herds,” she said. “We can now perform the majority of upstream pre-clinical analyses  in the lab using our technology that recreates the dynamic environment within the horse. Before, we'd study the effects of a new drug using expensive and limited research herds. Now, we can perform critical toxicity and safety studies before the candidate drug is ever injected into a horse.

“The key to our technology is that we don't need to euthanize additional horses.  We can go back to our cryobank of Thoroughbred tissue and enrich for tissue-specific stem cells to essentially grow equine microfluidic organ-chips indefinitely. My research team has developed several innovative methods that allow us to keep using and expanding these diverse equine tissues indefinitely.”

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