Equine Umbilical Cord Blood: What You Need to Know
Scott Hopper, DVM, MS, Dipl. ACVS
People have been collecting and storing umbilical cord blood from their children for 20 years. Recently there has been increased publicity and encouragement for horse owners and breeders to collect umbilical cord blood from their foals. Owners and breeders need to understand the facts concerning umbilical cord blood as a source of mesenchymal stem cells (MSC) and the potential use of these stem cells in horses.
Why Store Umbilical Cord Blood?
Cord blood allows us to harvest the most immature and possibly a more potent stem cell with a better potential to differentiate into the tissue we desire. An example of this was recently reported where the University of Copenhagen evaluated the chondrogenic potential (ability to produce cartilage) in MSC derived from both bone marrow and umbilical cord blood. Research showed that MSC’s from both sources were able to produce hyaline (joint) like cartilage but MSC from umbilical cord blood had an increased cartilage producing potential based upon certain testing parameters.
The culture and expansion of MSC from umbilical cord blood allows the horse owner to have almost immediate access to a therapeutic dose of MSC instead of waiting for 3 weeks while the MSC are being processed. Current thought is that injuries are better treated sooner than later and that multiple doses may be needed. Having stored cord blood also avoids the risks and expense associated with bone marrow collection.
Recently in the human literature it has been shown that induced pluirpotent (ability to differentiate into many different cell types) stem cells (iPS) can be derived from using umbilical cord blood as a source. This is important because iPS behave like embryonic stem cells which have the ability to differentiate to more types of cells than the typical mesenchymal stem cell. The potential of these cells is much greater. Human cord blood derived MSC can be used on siblings for hematopoetic disorders such as leukemia. Currently umbilical cord blood derived stem cells have been used to treat 80 different diseases in people.It is important to know what the probability is of a having a viable stem cell product when it is needed. The University of California-Davis recently published that they were able to collect, store and expand MSC to therapeutic doses in 80% of the cases. If there was greater than 21,000 nucleated cells per microliter harvested from the cord blood, then 91% of the cases yielded an adequate number of MSC. This is the best result harvesting MSC from cord blood reported to date.
Why is Cord Blood Processing Important?
A key factor that contributes to having a viable stem cell product is the ability of a lab to process, store, culture and expand the MSC to a therapeutic dose in the same facility without the need for shipping to a second facility. Many stem cell laboratories have storage capabilities but do not culture and expand the MSC into therapeutic doses. The Rood & Riddle Stem Cell Laboratory is able to provide complete cord blood stem cell services, and also employs the same technique used by U.C.-Davis that yields the highest success rate for obtaining viable stem cell dose from cord blood.
A common question is how long stem cells can be stored. Current data in humans indicate that cells have the same composition after 15 years of storage that they did at the time of storage. Although no long term data is available for horses, we employ the same technique used for human stem cell storage and expect to have similar results.
Umbilical Cord Blood Collection
Collection of umbilical cord blood is a relatively easy process that can be performed by an owner, breeder or night watch man. Collection kits are available from Rood & Riddle or possibly from your regular veterinarian. Below are the simple steps to collection.
1. Once foal is delivered apply an umbilical clamp approximately 4 inches from the foal’s umbilicus.
2. A second clamp or cable tie can be placed 2 inches below the umbilical clamp and the cord cut if needed.
3. Clean the umbilical cord. If no gross contamination of the cord wipe with alcohol before prior to blood collection. If contaminated, then clean with surgical scrub and wipe clean with alcohol prior to collection.
4. Using the blood bag provided, insert the needle into the umbilical vein and “milk” the blood into the bag. The goal is to collect approximately 250 mls of cord blood. If cord blood cannot be collected for some reason then an 8-10 inch section of umbilical cord can be collected for submission. Two cable ties can be placed approximately 8-10 inches apart and the cord collected. The cord should be rinsed to remove any gross debris and placed in a ziplock bag.
5. Place the blood bag or ziplock bag containing cord tissue in a refrigerator or in the provided shipping container (ice packs included). Sample should be shipped or brought to the Rood and Riddle Stem Cell lab the following morning.
Cost of Umbilical Cord Blood Collection and Storage
There are a variety of plans available for umbilical cord blood collection and storage ranging from $695 to $1,395. If you are interested in collecting your foal’s cord blood please contact Rood and Riddle for a detailed plan and price list or for any additional information you need.
Acupuncture and Broodmares
Kathleen Paasch, DVM
Acupuncture has been used to treat the ailments of humans and animals for thousands of years. In China, texts detailing acupuncture points and indications have been found dating back to 1399AD. In fact, until fairly recently, acupuncture, a part of Traditional Chinese Medicine (TCM), was the predominant health care system in China. The United States has been relatively slow to embrace the benefits of acupuncture but is coming around quickly. Many people now are familiar with the idea or have even been treated themselves. Animal owners, in particular, are very enthusiastic about this treatment.
The approach taken with acupuncture is a little different from the Western approach to medicine that is familiar to us. Where Western medicine seeks to control and treat disease, TCM seeks to restore balance to all systems of the body and allow the body to heal itself. An important point is that TCM and Western approaches are not mutually exclusive and actually serve to complement each other. In our practice, both approaches are often used to achieve the desired outcome in our patients. Acupuncture can be used for many types of ailments including: body soreness, gastrointestinal disturbances, allergies, headaches and infertility problems. While there are many scientific studies stating that acupuncture treatments have positive effects, we are not entirely sure why this is so. Traditional Chinese Medicine believes acupuncture improves the flow of qi or energy which then helps the body to heal itself. From a Western perspective, we know that when needles are placed in specific locations (acupuncture points), there is an increase in circulation (of both blood and lymph) and a release of endorphins (natural pain-killers). It is also likely that other chemicals, hormones and neurotransmittors are released with needle placement influencing our internal regulatory system. Applications in horses are not so different from those in people. For example, acupuncture can be used in horses to treat arthritis, COPD (heaves) and mild colics. One of the most striking and useful applications of acupuncture in equine practice has been with broodmares.
Much of the time things go routinely with broodmares. Days lengthen, mares start to cycle, they are bred, ovulate and become pregnant. However, for those mares that have problems getting in foal, acupuncture can help them to cycle normally and conceive. The causes of subfertility are numerous and include poor breeding conformation, number of previous foalings, uterine infections, and chronic pain. Any of these can cause mares to cycle abnormally and/or have a poor uterine environment in which to sustain an embryo or fetus. In recent years, in addition to more well-known treatments (e.g., intrauterine lavages, implants and oxytocin), veterinarians have started to employ acupuncture.
Acupuncture can help broodmares in different ways depending on their problems and the points used. For example, a mare in constant pain from chronic arthritis will circulate cortisol (a stress hormone) which has a negative effect on fertility. Acupuncture can help to lessen the pain leading to more normal cortisol levels. A less painful horse will likely be more active in the field benefiting the entire mare not just her reproductive tract. Another common problem is the mare that has a poorly toned uterus or difficulty clearing uterine fluid. Acupuncture is beneficial to these mares in helping to improve smooth muscle tone thereby enabling the uterus to clear fluid. Acupuncture can have a beneficial effect on hormone levels as well. Mares that are slow to begin cycling may be ‘restarted’ by a series of acupuncture treatments.
Mares that are treated are usually seen between one and three times depending on the initial complaint and response achieved. Treatment may involve placement of needles at specific points with electrostimulation or may consist of injections of Vitamin B12 at the same points. Most mares tolerate the procedure very well, likely due in part to the release of endorphins that result from acupuncture. An acupuncture treatment generally takes less than half an hour.
The uses of acupuncture are many and varied. It can significantly improve chances of conception in broodmares. So, if your trusted reproductive veterinarian suggests that acupuncture may be helpful, don’t think that he or she has lost his or her mind. Instead, know that acupuncture is a treatment that has been around benefiting both horses and people for thousands of years.
Colt or Filly on the Way? --- The Basics of Fetal Sexing
Tom Riddle, DVM
The determination of the sex of a mare’s fetus, or fetal sexing, as it is commonly called, is an extremely accurate procedure performed by rectal ultrasound examination of the pregnancy. With an experienced practitioner, the correct determination is made in well over 99% of cases. The procedure is difficult to perform, not because it is challenging to interpret the ultrasound image, but rather because it is difficult to place the ultrasound probe in the position necessary to view the required image. Most mares tolerate the procedure very well, but in some cases it does require a slightly deeper placement of the ultrasound probe than in the routine pregnancy ultrasound.
In my practice there are two windows of opportunity for fetal sexing, the first between 58 and 76 days of gestation, and the second between 110 and 130 days. In some occasions the sex may be determined a few days earlier or later, but if one attempts to sex a fetus which is too young, the anatomical structures needed for the determination may not be mature enough. If one attempts the procedure too late, the fetus may be dropped down too far in the abdomen to reach the needed structures. I prefer the earlier window (58 – 76 days) because the fetus is almost always in a location that permits the necessary view of the anatomy. With the second window, approximately 15% of the time the fetus cannot be viewed properly and a repeat examination at a later date is needed. Because a second examination may be needed, it is best to check mares early in the second window to allow time for a second examination.
In the first time period (58 – 76 days) the sex is determined by identifying the location of a structure called the genital tubercle, which is present in both colts and fillies. In colts the tubercle will develop into the penis, and in the filly the tubercle becomes the clitoris. In the second window (110 – 130 days) the anatomical structures have matured and the determination is made by identifying the penis in the colt and the udder in the filly.
Fetal sexing is performed for a variety of reasons, ranging from appraising the value of the pregnant mare to just plain curiosity. A partial list of reasons for fetal sexing includes:
- Valuation of mare prior to selling
- A factor in deciding who the mare should be bred to the following year (some clients want a foal of a particular sex by a certain stallion)
- A client may ship mares carrying only colts (or fillies) to foal in another state or country; for example, the Japanese frequently will ship only mares carrying colts to Japan
- Agents may have an order for mares carrying only colts or only fillies (many European agents look for well bred mares carrying fillies)
- A factor in deciding whether a mare will be sold
Probably the most common reason for requesting fetal sexing is to provide prospective buyers at the broodmare sales with the sex of the mare’s fetus. Whether the mare is carrying a colt or a filly may have a significant effect on her value. Some buyers may prefer a filly by a sire known for siring good female runners or a filly from an impressive female family, while many buyers prefer a colt because colts on average bring more money when selling as weanlings or yearlings. An analysis of the past ten Keeneland September Yearling Sales shows colts averaging 25.7 % more than fillies (thanks to Mark Taylor of Taylor Made Farm for providing this information). Providing prospective buyers with the sex of the foal the mare is carrying gives the buyer one more piece of information to factor into the price he /she is willing to pay for the mare.
In these challenging economic times spending money wisely is more important than ever. The decision to fetal sex a mare should be made based on the mare owner’s circumstances and the intended use of the information. In many cases the relatively small investment in the cost of fetal sexing ($135 in our practice) can generate a significant return for the owner.
Tricide®-Treatment for Uterine Infections
Michelle M LeBlanc DVM, Dip ACT
Standard treatment for uterine infections is uterine irrigation and either intra-uterine or systemic antibiotics. However, not all infections respond to these therapies. Treatment failure may be due to continual contamination of the uterus because of anatomical abnormalities in the reproductive tract, degradation of antibiotic in uterine exudates, or biofilm production by the micro-organism. While most anatomical defects can be surgically corrected and exudates can be removed by uterine irrigation, biofilms (thin surface layer produced by microorganisms) are difficult to treat. Bacterial biofilms consist of a community of different bacterial species, surrounded by an extracellular matrix, that co-exist in a symbiotic relationship. Such biofilms are found throughout the human body, e.g. the oral cavity, the skin, the intestines and the vagina. In most cases, the inhabitants of this community are considered as normal flora and serve as a protective mechanism to prevent the colonization of frank and opportunistic pathogens. If the balance of this biofilm community is upset or disrupted, pathogens may colonize, proliferate and cause disease.
An important aspect of biofilms is that the bacteria within them are much more resistant to antibiotics. Research has demonstrated that bacteria in biofilms are up to a 1000 times more resistant to antibiotics than the same bacteria grown in broth. Antibiotics kill rapidly dividing bacteria and eliminate those that are more susceptible to antibiotics. In doing so, antibiotics tend to select for the more resistant bacteria. In addition, bacteria in biofilms are usually not actively dividing and therefore do not succumb to the effects of antibiotics.
Certain bacteria are known for producing biofilm. These include Pseudomonas aeruginosa, Staphylococcus epidermis, Escherichia coli, Enterobacter cloacae and a number of yeast and fungi. These organisms more commonly cause endometritis in older, barren mares that have anatomical defects than in young, fertile mares, although uterine defenses can be broached in the latter resulting in chronic infection. Infections by these organisms can be difficult to treat, are often resistant to a 3 to 5 day course of antibiotics, and may result in a population of bacteria colonizing the uterus that is highly resistant to the drug initially used for treatment. Work in other species and in the mare has shown that buffered chelating agents (Tris-EDTA, Tricide®) may potentiate the actions of antimicrobials, dissolve exudate, and break up biofilm.
Buffered chelators such as first generation Tris-EDTA (ethylene-diamine tetra-acetic acid (3.5 M)-tromethamine 50mM;) and third generation Tricide® (8mM disodium EDTA dehydrate and 20 mM 2-amino-2-hydroxymethyl-1,2-propanediol) potentiate the actions of antimicrobials. Tricide® has been developed by scientists and veterinarians at the University of Georgia College of Veterinary Medicine. Tricide® appears to be more stable than Tris-EDTA because of its buffering agent. Chelating agents such as Tris-EDTA and Tricide® enhance the bactericidal effects of antimicrobials in dogs with refractory otitis, pyoderma, osteomyelitis, multiple fistulas, and cystitis. In the 1970s, isolates of Pseudomonas collected from the uterus of mares with chronic endometritis were killed when exposed to Tris-EDTA solution. However, the mechanism of action was not known. Since that time others have shown that addition of Tris-EDTA to gentamicin in vitro improved killing of Pseudomonas aeruginosa by 1000 fold more than treatment with only gentamicin. A recent study showed that Tricide®, a third generation buffered chelating agent, increased in vitro activity of antifungal drugs against common fungal pathogens isolated from eyes of horses with mycotic keratitis. The mechanism of action of buffered chelating agents is not completely understood but it is speculated that the chelating agent (EDTA) binds to calcium and/or magnesium on the outer membrane of bacteria, thereby altering the integrity and permeability of the cell wall. Basically, it punches holes in the cell wall and allowing substances to flow into the cell.
We include Tricide® in our treatment for chronic bacterial or yeast endometritis when one of the organisms listed above is isolated from the uterus or if the organism isolated is resistant to antibiotics that can be infused into the uterus. In order to kill the bacteria or yeast, buffered chelating agents must come in direct contact with the bacterial cell wall so the volume of solution needed for infusion must be large enough to cover the entire endometrial surface. Doses ranging from 250 to 500 ml are recommended. The chelating agent binds to the bacteria within minutes resulting in cell death and accumulation of debris so the uterus should be lavaged within 12 hours to remove these by-products. Our current recommended therapy for gram negative bacteria and yeast repeatedly isolated from the uterus of a mare with endometritis is to infuse 250 to 500 ml of Tricide® or Tris-EDTA into the uterus on Day 1, lavage the solution out within 24 hours and examine the efflux. If the efflux is cloudy or has mucus strains the chelating agent is infused into the uterus again on Day 2. Antibiotics are then begun on Day 3 following uterine irrigation and continued daily for a minimum of 5 days. The mare is short cycled and re-evaluated. As many of these infections are long standing, treatment over 2 to 3 cycles may be needed. As most infections are due to overgrowth of a bacteria that colonizes the vagina, anatomical defects most be corrected if the infection is to resolve especially if the defect interferes with physical clearance.
Education for Veterinarians and Horse Owners Part of the World Games Experience
Rood & Riddle Equine Hospital’s role as the Official Veterinary Partner of the Games will not be limited to providing veterinary support during the competition but will also include hosting educational forums for veterinarians and horse owners. Rood & Riddle, Alltech and the American Association of Equine Practitioners, have joined forces to sponsor a sport horse symposium for veterinarians and another for horse owners, to be held in conjunction with the 2010 Alltech FEI World Equestrian Games which opens September 25 at the Kentucky Horse Park, in Lexington, Kentucky.
The Veterinary Sport Horse Program will be held from September 22-24, 2010, at the Marriot Griffin Gate Resort in Lexington, Kentucky. An international roster of speakers will present in-depth, current information on orthopedic problems, diagnostic imaging, equine podiatry, lameness versus neurological disease, upper and respiratory disease, muscle disease, and nutrition with a focus on the veterinary care as it applies to the equine athlete. Featured speakers include orthopedic surgeon Dr. Alan Ruggles, and internal medicine specialist Dr. Steve Reed from Rood & Riddle, and noted experts Dr. Kent Allen, Dr. Wayne McIlwraith, and Dr. Jean-Marie Denoix. Complete program and registration information can be found on the AAEP website.
Rood & Riddle’s Dr. Chris Newton and farrier Rodney King NZCEF, CJF, AWCF are among the speakers for the one-day horse owner workshop held September 24, 2010 at the Embassy Suites in Lexington. Titled “A Winning Edge, Promoting Peak Performance in Equine Athletes”, the workshop will be conducted by veterinarians and nutritionists to provide horse owners, trainers, managers, and riders with valuable information for managing injuries and maintaining peak performance in the sport and performance horse. Interested horsemen and women can register through the AAEP website.
In addition to the two seminars, Rood & Riddle will continue to offer educational presentations throughout the 16 days of the Games. The Rood & Riddle pavilion inside the Alltech Village will have a theater section where presentations will be offered 6 times each day. These presentations will focus primarily on equine health but will also include guest speakers presenting their equine connection including WEG riders, a Hall of Fame Jockey, and representatives from Normandy, France, host of the 2014 FEI World Equestrian Games.
Coming to the Games?
Tour Rood & Riddle and Horse Farms During Your Stay!
Rood & Riddle Equine Hospital will open its doors to tourists during the Alltech FEI World Equestrian Games, September 25-October 10. Individual tours will run at 10am and 12pm Monday through Friday. A third tour will be included in a bus tour package from area Bluegrass tour companies which will bring visitors to several area attractions including Rood & Riddle. All of these tours will be listed on HorseCapitalTours.com. Horse Capital Tours is a tourism initiative supported by the Kentucky Thoroughbred Association to promote the local Thoroughbred industry.
The Rood & Riddle Equine Hospital Tour provides a one hour educational experience and inside view of equine veterinary medicine. Tour guests will walk through the hospital and view surgery, diagnostic imaging systems and the podiatry center, and learn about advances in equine veterinary medicine and patient care. Tours are limited to 16 persons per tour, and costs $5 per person. All proceeds from the tours will be donated to the Kentucky Equine Humane Center and the Kentucky Horse Park Foundation. All tickets and reservations will be taken by HorseCaptialTours.com.
Around the Practice
Congratulations are in order for veterinary podiatrist, Dr. Raul Bras, who passed his American Farriers Association exam for Certified Journeyman Farrier (CJF) certification in early April. Dr. Bras becomes the third veterinary podiatrist at Rood & Riddle to hold CJF status, joining fellow associate Dr. Vern Dryden, and podiatry intern, Dr. Dan Carter into this exclusive company. Currently, there are only 5 veterinarians nationwide with CJF certification.
The Rood & Riddle logo has a fresh new look after a graphics makeover early this year. In the same way that our facility and technology have been updated periodically, it was time to update our logo. So, with a few minor changes in position, color and font, we have a new, modern look to take us into the next decade. Transition to the new logo begins this month!
The new Client Center was activated on RoodandRiddle.com in early March. Clients are now able to access their account balances and pay their bill online by e-check or credit/debit cards. To sign up for account access on the web, clients with an existing account must first request a pin number and complete the sign up form available in the Client Center. Once your account is verified, the billing department will provide instructions for initiating an account.
Geoffrey Rosenberger, an employee of Lakland Farm in Versailles, Kentucky, recently presented Dr. Riddle with a gift of 50 veterinary books from the 1920s and older and a preserved equine skull to use for educational purposes. Geoffrey, a life long horseman, works closely with Rood & Riddle veterinarians when veterinary care is needed for the horses in his care and is well respected for his horsemanship. Rood & Riddle is truly thankful for his generous gift and pleased to have this wonderful historical collection of veterinary books.
Another special thank you goes to Dr. Val Nicholson who presented Dr. Chris Newton with a portrait of Dr. Chris Newton with a patient. The artist painted the watercolor from a photograph from “Equine ER: Stories from a Year in the Life of an Equine Veterinary Hospital.” Author Leslie Guttman was also on hand for the presentation.
Equine ER: Stories from a Year in the Life of an Equine Veterinary Hospital was chosen by WEKU-FM, one of the largest public radio stations in Kentucky, as its April book club selection. The book written by journalist Leslie Guttman, chronicles stories gathered at Rood & Riddle where Guttman spent one year as an embedded reporter. The book club honor culminated with a discussion at Joseph-Beth Booksellers in Lexington, on April 13. Drs. Tom Riddle, Bonnie Barr, Blair Cornman, Vern Dryden, Bruce Howard, Peter Morresey, Kathleen Paasch, Steve Reed and Bryan Waldridge were in attendance and answered club members’ questions ranging in topics from veterinary medicine technology, ethics, life balance and personal career stories to issues facing Kentucky’s equine industry. “It was a very interesting discussion, and we are pleased that the book has led to increased awareness of the profession and the equine industry,” said Dr. Tom Riddle. “It was our pleasure to answer their questions.”
Dr. Hannah Wellman recently launched the Equine Education Blog, a web forum that reports on educational articles, presentations and research developments from Rood & Riddle Equine Hospital, including short interviews with our veterinarians and staff. The information posted on the blog does not replace direct consultation regarding diagnosis, treatment or therapy of horse health conditions with your veterinarian. It is Dr. Wellman's goal to provide accurate educational information that will empower horse owners with veterinary information written exclusively by veterinarians. It also aims to give insight into the many services and specialties at Rood and Riddle Equine Hospital. Please visit the Equine Education Blog We welcome your feedback!
Are you a Fan of Rood & Riddle? Find us on Facebook to keep up with our weekly activities and try your hand at “What’s Your Diagnosis!”