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Transported Semen for the Small Breeder
Part 1 The Stallion Side This article may be downloaded for personal use only. For information on reprint rates please send me e-mail at oogiem@desertweyr.com Artwork copyright 1992 Jodi Abrams and used with permission.
Our View
This article will give one breeders view of the pitfalls and benefits of starting a transported semen program. It is not meant to be an extensive article about all aspects of transporting semen. This article assumes the reader is familiar with the basic concepts of AI and has some information on the specific requirements for transporting semen.
Why Bother?
We are a small breeder of Crabbet, Mayensboro and old Kellogg Arabians. We greeted the news that transported semen would be allowed for purebreds with cries of relief since it would enable us to breed our own mares to CMK stallions far away from us and we could offer our stallions to mare owners who could not send their mares to our farm. We can promote our horses to mare owners who could not ship their mares to us. For those of us breeding CMK horses transported semen offers a way to spread our limited gene pool across the country.The down side of transported semen is the expense, both of the initial testing and equipment purchases and of the ongoing permit fees. The stallion owner contemplating offering this service should be aware of the costs and challenges before committing to shipping semen. The mare owner too must be aware of the additional costs of using transported semen. More intensive veterinary management of estrus cycles and more frequent palpations are all part of good management for success with transported semen. As a mare owner do not expect to spend less just because you do not have to ship your mare. It may cost you more but you may be able to breed to a previously inaccessible stallion.
What's Involved?
When we initially decided to pursue transported semen we contacted a number of breeding farms using this technology and discussed their experiences. We based our program on what had worked for other breeders setting up their own transported semen programs. Our contacts included very large warmblood farms and one breeder who produces hunter ponies. In addition to talking with breeders we discussed using transported semen with our veterinarian. Our discussions with our veterinarian identified a series of procedures we would have to do to verify the ability of our stallions' semen to survive transport. These tests are described below:First we had to be sure our stallions would serve an artificial vagina (AV). Since some stallions won't this can be a serious block to transporting semen. It is not very successful when you try to collect the semen after it is already in a mare! Fortunately our young stallion had not been used for breeding yet. We started him on a mare first then collected him using the same mare as a jump mount. He seems to be readily learning the ropes and we are preparing a collection dummy for use next year. Our senior stallion had never been collected before but we were successful with him on the first try.
The Missouri Model Artificial Vagina
Second we must verify the semen's ability to survive the cooling process in extender. There are a number of formulas for extender and each is slightly different. Each individual stallion must be tested to determine which type works for him.
Third we must find an antibiotic which does not kill the sperm. Again, each stallion may show differing sensitivity to different antibiotics. The antibiotic protects the sperm from infection during the transportation process.
The "Horse Kitchen"
Testing is a trial and error process until you get the right "recipe". It is recommended that at the start of each breeding season a 24 hour semen evaluation test be performed to be sure that the collection procedures are still working and that there have been no changes in the semen and its ability to survive transport. According to our veterinarian the sensitivity of semen to extender and antibiotic may change over time.Once you have verified that a particular stallion can be collected and extended you must be able to store and ship the semen. Hamilton Thorn is the leading supplier of equine semen shipping containers. Although less expensive insulated box systems are available the quality of the temperature control and survivability of the container were major questions. We decided not to risk a possibly inferior container and instead went exclusively with the Hamilton Thorn Equitainer. The Equitainer is shown in figure 1. This was the only AI equipment we purchased this year since our veterinarian had the required AV, filters, thermometers, phase contrast microscope and other paraphernalia required to collect and test semen. Our veterinarian uses a Missouri model AV. This model is depicted in figure 2. The temperature and fullness of this model can be adjusted to suit each stallion. We found that the stallions like an AV that is slightly warmer than normal horse body temperature. Based on the prior experience of our veterinarian we decided to try Kenney prepackaged extender and ticarcillan antibiotic for our stallions first.
To prepare for the testing process we had to mix and package the extender. We package the extender in Whirl-Pak bags. These sterile dairy sample bags are relatively inexpensive and easy to use. We mixed the antibiotic Ticarcillan in half of the extender and packaged the other half plain. We package extender in roughly 100 ml aloquats and place the packages in our freeze. It took us approximately 30 minutes to prepare and package the extender the first time. During testing we ran out of extender and had to purchase and prepare more. Our second liter only took five minutes to prepare and another 15 to package. Extender can be stored for about a year in a deep freezer.
A Typical Test
Vet arrives and we start boiling water to fill the AV and start a plain and an antibiotic aloquat of extender thawing. We used regular tap water to fill the AV although some texts recommend distilled water.The AV is prepared for use with an in-line dairy filter over the end , then two whirl Pak bags to collect the semen. We use double bags to be sure we don't have a problem due to bag failure. It is hard to separate dirt from semen and dirty semen won't ship well! The filter is to remove the gel fraction. Semen doesn't ship well unless the gel fraction is removed. We fill the AV and get the internal temperature to 130 degrees F. This is a bit higher than is generally recommended but we've found that it cools off to the correct temperature by the time we get it lubricated and the jump mare ready.
The extender is slowly thawed and brought to a temperature of just over 100 degrees F. Because we do not have an incubator we use several pots and a warm water bath. We use a heating pad on medium to warm the sterile cup and two ballast bags used to wrap the semen packets. It is not precise but has been OK so far. Careful attention to checking the temperature before placing the ballast around the packaged semen is necessary when using this type of system. The whole process would be much simpler with an incubator but they are very expensive. One note, don't plan on using a microwave to thaw extender, since antibiotics do not survive microwaves very well. The microscope is set up, and several slides are washed and prepared for use. The slides are also placed on the heating pad to warm. A phase contrast microscope is best for viewing sperm. Other types do not provide the clear view necessary for evaluating motility and checking for abnormal sperm. From the time the vet arrives until we are ready to go out and get the animals usually takes just over an hour.
When we are ready to go there is a jumble of activity. The vet takes the AV and using a palpation sleeve places a small amount of sterile non-spermicidal lubricant in the AV. I was amazed at just how little is used, approximately 1 tablespoon or less. The end of the sleeve is folded down over the AV to keep dust and dirt out.
The Collection Process
For safety during the collection process we use three people; the vet, who is doing the actual collection, and two animal handlers. Everyone dons appropriate protective clothing (gloves, helmet and either boots or running shoes depending on position) and proceeds into the fray. All people wear helmets. I have witnessed people hit on the head by an overanxious stallion as he mounts so I require all people in the pen to have appropriate head protection.We are fortunate in having an older mare who with a bit of persuasion (hobble on one leg and lip chain or twitch) can be made to stand for any stallion even when she is not in heat. One animal handler gets the mare, places her in the hobble, adjusts the lip chain or twitch if she is not in heat, wraps her tail and washes her if necessary. We only wash the mare if she is very dirty and only with plain warm water. We use neoprene tail wraps because they are easier to put on and can be easily washed and disinfected.
The other animal handler gets the stallion in his breeding halter and brings him to the designated breeding area. He is allowed to tease the mare until he drops and his cleanliness is ascertained. We only wash the stallion if he is very dirty and use only plain water. This is because current research suggests that extensive washing with soap can lead to a larger population of harmful bacteria and cause more infections than the few normal ones left by a less vigorous washing. We are also using a special halter for breeding and the stallion is only allowed to show libido when he is wearing this halter. For safety both animal handlers also have a whip.
If we have washed either the mare or the stallion we wait until they are dry before starting the collection process.
Collection takes from 10 minutes to over an hour depending on the stallion. Our youngster does not always know what to do and tends to fumble around a lot. He hasn't quite gotten the sequence down correctly and still tends to mount, tease and then get an erection. Patient work is helping and his last two collections went more smoothly and in the proper sequence. Our older stallion is ready and collected in only a few minutes.
When the stallion mounts, the vet rushes in and deftly deflects his penis into the AV. This looks easy but it is a very special skill! So far we have been lucky and once we get the stallion in position with an erection we do not have any problems getting him to serve the AV. The vet keeps one hand under the stallion's penis to feel when he ejaculates. At the conclusion the vet rushes to the house to prepare the semen while we take the horses back to their corrals.
Processing Semen
A small sample of the raw semen is placed on a slide for viewing. Then the semen volume is calculated and the amount of extender required is calculated. One half is placed in extender with antibiotic while one half is placed in plain extender. The warm extender is mixed into the semen rather than vice versa to reduce shock when the sperm are placed in it. The temperature of the extender is critical and must be exactly 100 degrees F. Samples of the semen in both plain and antibiotic extender are placed on waiting slides while the rest in placed in a Whirl-Pak bag. The Whirl-Pak bags are carefully closed without any air and wrapped in the warm ballast. The ballast must also be at 100 degreesF. The two bags are placed in the cup, put in the isothermalizer and packed into the waiting Equitainer with coolant cans according to directions.
The Hamilton-Thorn Equitainer (r)
The container is closed and we can all take a deep breath. Then we evaluate the raw, extended and extended with antibiotic samples. Waiting to do this until after the semen is packed ensures that what is in the Equitainer is in the best condition. Even slight delays and exposure to light are detrimental to all sperm so this is an important point.
The slides are evaluated for motility and the three samples are compared. This gives us a rough indication whether we got a good collection but is not a conclusive test for transportability. These same tests are done at 24 and again at 36-48 hours. The 24 hour test is the one which determines whether the stallion can be shipped. The last test indicates the range in which successful insemination can take place. These later tests must be done on samples that have been warmed to 100°ree;F. Checking them while they are still cool can result in false negative results.
After the semen is packed and the samples have been viewed comes the cleanup. The AV is washed with plain water and then rinsed with alcohol until it is clean. It is left to air dry. We did not use disposable AV liners this year which means you must be very sure the AV is clean and disinfected between each use.We plan to use disposable AV liners next year to reduce our cleanup time. The Whirl-Pak bags, sampling tubes, slides etc., are all thrown away. We have found that the clean up takes about 1 hour.
Test and Test Again
This testing process continues with changes in extender and antibiotic until a protocol is defined that works for the particular stallion.Our first test of our younger stallion resulted in all dead sperm. Since we were not sure of our collection procedure we tested the stallion again. We got good motility at 24 hours and performed a third test to confirm our results. The extra practice was useful in proving that we could duplicate our success. We did not want to say we could ship semen and have it all arrive dead! According to our veterinarian three tests to get a working protocol is doing well. If we had had problems with the extender or antibiotic we could have been looking at 10-12 tests before we could verify success. An initial test failure could have been the result of a flaw in the collection or packaging procedures, temperature fluctuations or, since it was one of the stallions' first breedings, simple immaturity of the sperm. Our older stallion was collected and the collection thrown away. About an hour later he was collected again and this collection was packaged and prepared for testing. His semen tested fine on the first test so we repeated it to verify our results. Only after the completion of these tests did we feel we could advertise our ability to offer transported semen.
Hamilton Thorn suggests packaging a sample of semen from every ejaculate and keeping it on the farm in another Equitainer. Then if the mare owner receives "dead" sperm the retained sample can be tested. If the on farm sample is OK then the problem is most likely on the mare end or in shipping. If the sample is also dead then the problem is on the stallion end. This requires an additional Equitainer for the retained sample. We have only one and did not do this. It might be a valuable option for the larger farm.
To avoid the problems with a jump mare, even one who is cooperative, we decided to build a breeding dummy and train the stallions to mount it. We used some old four by fours and other scrap wood and padded the phantom with a foam mattress and covered it in naugahide. This is an inexpensive way to go since commercial breeding phantoms can run as high as $2,500.00.
Permits and Fees
A major expense is permits. When we first started the fees were: The International Arabian Horse Association requires a transported semen permit which costs $75.00 and is good for the life of the stallion. In addition each mare owner who wishes to produce a registered Half-Arabian or Anglo-Arabian must pay a $250.00 permit fee. The Arabian Horse Registry required a permit fee of $200.00 per year for the stallion.Since we started with our program the fees have changed considerably. The current fee schedule is as follows:
International Arabian Horse Association requires a transported semen permit which costs $150.00 per mare and $100.00 per stallion and is good for the life of the horse. It covers cost of bloodtyping. If the bloodtype is on file check with IAHA regarding any reduced fees.
The Arabian Horse Registry now has a one time fee of $350.00 for a Transported-Stored Semen permit for the stallion. It is good until ownership of the stallion changes. In addition each foal to be registered must include a Transported-Stored Semen Service Certificate with the registration application. These certificates are purchased by the stallion owner at a cost of $35.00 each.
These changes will increase the use of this technology.
We, like most farms that offer transported semen, charge mare owners a per shipment fee. The fee ranges from $200 to $260 per shipment and includes collection and shipment costs. Large warmblood farms who do nothing but transported semen can place all their stallions on a regular collection schedule. They have the investment in numerous Equitainers and can depend on shipping all the aloquats of collected semen. This mass production results in much lower costs to the mare owner. The small farm cannot do this. We have to charge for each mare separately because there may only be 2 or 3 mares bred each season using transported semen and the probability of them being ready at the same time is very small. We offer reduced fees when the mare owner has two or more mares who can be cycled at the same time using hormones and therefore share a single shipment. It is unfortunate that the expenses and permits required to take advantage of the new rules make it very expensive for the small breeder to get set up. Our calculations indicate that the costs to breed via transported semen were about the same as the cost for on-farm breeding for the mare owner breeding purebred Arabians. The lack of mare care and transportation charges was made up for by additional veterinary procedures and semen shipping.
In addition to the direct costs for testing and equipment we spent more time during the preparation and collection process than we do for a normal live cover. A typical live cover requires two people and about one hour of time. A typical test or collection requires three people and over two hours of time. We expect that this time factor will decrease as both we and the stallions get more experience.
Mare Owner Responsibilities
The mare owner must also be aware that they are responsible for determining the time to breed when using transported semen. This may entail additional veterinary fees for ultrasound evaluations and palpations to determine the proper time to order semen. The average for Arabians is three shipments per pregnancy. The warmblood farms we spoke with get an average of 1.2 to 1.5 shipments per pregnancy. The discrepancy is probably due to the newness of the technology for Arabian breeders. Education about artificial insemination procedures will do much to reduce the confusion regarding who is responsible for setting the time to breed. This should improve the shipments per pregnancy ration considerably. We made it a farm requirement that each mare owner must purchase the mare owners' handbook sold by Hamilton Thorn. This concise handbook provides all the information required by the mare owner to successfully use transported semen. Our approach is similar to one large warmblood farm in that we include the cost of the handbook in the booking fee.
Conclusion
It can be seen that providing transported semen is both time and procedure intensive. The additional veterinary fees for the collection etc. can be expected for the first year or two. Our veterinarian has suggested that we attend an AI technicians class at one of the vet colleges so we can do our own collections. This new procedure will become less expensive as more people start using it and breeders become proficient in the collection and other procedures required for success. After many discussions with other horse breeders (not Arabians) we re-did our breeding contracts. We are very up-front about the expenses the mare owner will have to pay to use transported semen and our contract includes sample rates for the various items including collection and shipment.CMK horses are a valuable resource that we hold for future generations. Transported semen is one tool that can help us preserve and maintain this unique genetic treasure. We had hoped that transported semen would be easy, quick and inexpensive. We found out that it is a bit more difficult than regular natural cover, takes time to learn and is as expensive as on-farm breeding. In spite of the expense and difficulties I would encourage breeders to try transported semen and would welcome the opportunity to discuss our experiences with other breeders. In addition to our horses we also tested our donkey jack and have information which may be of use to donkey and mule breeders.