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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.

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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Potential Biomarker Found For Ascending Placentitis

Ascending placentitis is an infection of a mare's placenta that commonly causes late-term abortions; it can have a devastating economic impact on the breeding industry.

A new study has helped to identify which mares might be at risk of developing this infection by focusing on one variant of an inflammation-related protein, reports The Horse. The study was led by Dr. Yatta Linhares-Boakari, who found that mares that developed ascending placentitis had more messenger RNA (mRNA) of serum amyloid A1 (SAA1) in their placental tissues than those mares who didn't. Yatta Linhares-Boakari says her research might allow scientists to determine the difference between placentitis and other causes of inflammation.

The study team bred 10 pony mares and introduced Streptococcus equi spp zooepidemicus into the cervixes of half of them to induce ascending placentitis. Three days later, the researchers took blood samples, sedated and euthanized all of the mares used in the study to investigate the tissues of the mares and the fetuses.

The scientists found that mares that had placentitis had significantly higher SAA values. They also discovered that haptoglobin (Hp, another protein associated with inflammation) values in the fetuses' blood was significantly higher in the placentitis group.

Additionally, the team found distinct patterns of proteins in the tissues of infected mares and a never-before-seen mRNA trend: Both SAA and Hp were found at increased levels in the mares with ascending placentitis. SAA1 and mRNA were found in greater numbers in the placental membranes of infected mares, as well.

The team concludes that their findings — specifically the elevation of SAA1 in blood – may eventually be able to assist in pinpointing ascending placentitis in its early stages when it might be treated more easily.

Quest Continues For A Blood Test To Identify Horses At Risk For Injury

A multi-part study by the University of Kentucky to help identify at-risk horses before racing will soon move into its third phase. Gluck Equine Research Center's Dr. Allen Page provided an update on the ongoing research to the Kentucky Equine Drug Research Council last week.

The EDRC was key in funding the first two phases of the study, which has yielded promising results. In the first phase, Page and fellow researcher Dr. David Horohov took blood samples from horses suffering fatal injuries in races and compared them with blood samples from competitors in the same races who did not suffer injuries. The researchers looked for messenger RNA (mRNA) markers that were present in the injured horses but not the uninjured ones. Messenger RNA is responsible for carrying instructions from genes leading to the production of proteins. The number of these instructions, or gene copies, change if the body is increasing or decreasing inflammation somewhere.

Initially, the research identified 21 markers and determined that three of them were present in 88 percent of horses suffering injuries. The tests developed by the research had a 76 percent sensitivity rate, meaning they could practically identify an at-risk horse 76 percent of the time. Page said he wants to get that sensitivity rate higher, as it's important to minimize false positives that would keep healthy horses out of races.

The study's second phase is in progress now and will sequence RNA from the phase one samples, which will help researchers look at a total of 22,000 markers. So far, about 150 markers are promising enough to merit additional research to see if they could be combined with the results from the first phase findings for more accurate and sensitive testing.

Page is now preparing to launch the third phase of the study. In this phase, researchers want to take pre-race blood samples (rather than post-race samples) to validate that the markers could be used effectively as predictors in a practical application. The team will collect blood samples taken at the time of furosemide administration or pre-race TCO2 testing and bank those samples. If a horse is fatally injured in a race, the team will go back and analyze that pre-race sample to see whether it showed the same indicators they've seen in post-race analysis.

In this phase, the relative infrequency of fatal racing breakdowns means the team will have to cast their net wide.

“Obviously, the rate of catastrophic injury is quite low,” said Page. “So if we want to look at pre-race samples, it requires a large number of samples be collected.”

Page projects the team will need to collect 10,000 blood samples, expecting that will result in about 12 or 13 samples related to fatal orthopedic breakdowns (sudden deaths or accidental deaths will not be included). Those samples will then be stored for future research, so the process will not have to be repeated or funded again for subsequent projects.

“We're really happy and encouraged by the results we've got so far and fully anticipate that by using these pre-race samples, we'll be able to validate what we've done and potentially come up with a commercially viable and useful test that we can use in the racing industry to help further decrease the catastrophic injury rates,” said Page.

The sample collection for the project's third phase will take about a year to complete.

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