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Proceedings of the 16th

Italian Association of Equine Veterinarians
Congress

Carrara, Italy
January 29-31, 2010

Next SIVE Meeting:

Feb. 4-6, 2011 – Montesilvano, Pescara, Italy

Reprinted in the IVIS website with the permission of the

Italian Association of Equine Veterinarians – SIVE

http://www.ivis.org
Published in IVIS with the permission of SIVE
Breeding management of the mare with cooled
and shipped semen
Margo Macpherson
DVM, MS, Dipl ACT, Associate Professor and Service Chief,
Reproduction, College of Veterinary Medicine,
University of Florida, Gainesville Fl, USA
Catie DeLuca, DVM, MS, DACT
INTRODUCTION
Mares are seasonally polyestrous and cycle
from spring into fall (long day breeders) in the
Northern hemisphere. This pattern produces
foals that are born during favorable weather in
the spring and summer months of the year. A
January 1st birthday instituted by several breed
registries offers an incentive for early-born
foals. Consequently, the mare’s natural breed-
ing season is often altered using artificial pho-
toperiod and hormone therapy to induce early
cyclicity. The result is a breeding season that
begins in January and lasts until fall in much
of the Northern hemisphere. For most breed-
ers the ultimate goal is to produce one foal per
mare per year.
BREEDING PROGRAMS
A variety of management programs are avail-
able, each one with its own advantages and
disadvantages. Breeding mares by natural
mating has been used for centuries and until
the development of assisted reproductive tech-
niques, was the only available method of in-
seminating mares. Natural mating offers the
advantages of fresh semen and minimal labor.
Natural mating includes both pasture breed-
ing and hand-breeding. The biggest advan-
tages of pasture breeding are the availability
of fresh semen and the minimal labor in-
volved. Since the mare and stallion are housed
in the same pasture for a period of time, the
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Proceedings of the Annual Meeting of the Italian Association of Equine Veterinarians, Carrara, Italy 2010
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mare will be mated numerous times during es-
trus. Because mating is frequent, fresh semen
is likely to be present at the time of ovulation.
The frequent interaction/teasing behavior of
the mare and stallion is also proposed as ben-
eficial for uterine clearance post-mating1.
There are some distinct disadvantages to pas-
ture breeding. The risk for injury to the stal-
lion or mare is significantly higher in a pasture
breeding program. Records are frequently
poor since matings may not be observed or
recorded. Further, even with good observa-
tion, one cannot predict double ovulations or
post-mating problems, which can lead to twin
pregnancy or infertility if not managed. Tran-
srectal palpation and ultrasonography enhance
pasture breeding management, but are often
not incorporated. Evaluation of semen is gen-
erally not possible with pasture breeding sys-
tems, and stallion infertility may be missed
until late in the season. Finally, the mare own-
er is typically limited to stallions within a ge-
ographic region.
Some of these disadvantages can be overcome
using hand-breeding. Hand-breeding has
been utilized by the Thoroughbred industry
with great success. Hand-breeding is charac-
terized by controlled interaction between the
stallion and the mare. In a hand-breeding pro-
gram, the mare is typically restrained while
the stallion is brought to the mare for mount-
ing and breeding. Hand-breeding allows ob-
servation that a breeding occurred and collec-
tion of a dismount sample of semen for evalu-
ation. Risk of injury to the mare and stallion is
Published in IVIS with the permission of SIVE
decreased with hand breeding but not elimi-
nated. Restraint devices such as hobbles and
leg twitches may be employed. Mares may be
teased daily and hand-bred every other day
while in estrus; such mares are usually bred 3-
4 times per cycle. With more intense manage-
ment (transrectal palpation and ultrasonogra-
phy) and hormonal induction of ovulation, the
mare can be sent to the stallion at the appro-
priate time for a single breeding.
Many breed registries now allow the registra-
tion of foals conceived by artificial insemi-
nation (AI). Advantages of artificial insemi-
nation include the ability to inseminate multi-
ple mares with a single ejaculate, a decreased
risk of venereal disease and improved safety
for the mare and stallion. In addition, the abil-
ity to cool and ship, or freeze, semen allows
the distribution of desired genetics to distant
sites without the transport of animals. With
frozen semen, international transport is pos-
sible, allowing the import and export of de-
sirable genetics overseas. Semen evaluation
is possible with AI and should be a routine
part of semen handling both at the time of se-
men collection and prior to insemination of
the mare.
Mares inseminated with cooled semen are
typically managed with teasing, transrectal
palpation and ultrasonography and adminis-
tration of an ovulation induction agent. Preg-
nancy rates using cooled-shipped semen are
about 5-10% lower per cycle than they are
with fresh semen from a fertile stallion. Se-
men from some stallions does not withstand
the stress of cooling, and fertility can be dra-
matically reduced. Prior to the breeding sea-
son, semen from a stallion should be mixed
with several types of extenders and antibiotics
to determine longevity of semen under cool-
ing conditions.
Breeding with frozen semen offers the advan-
tage of semen availability at all times. The
major disadvantages of breeding with frozen
semen are increases in labor and cost, and a
decrease in pregnancy rates compared to fresh
semen. As with cooled semen, some stallion’s
semen does not tolerate the freezing and thaw-
ing process and can result in a dramatic re-
duction in fertility.
131
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BEFORE THE BREEDING SEASON
BEGINS
Prior to the onset of the breeding season, it is
wise to determine which management system
is desired or most appropriate for each farm.
Mare reproductive history, cost and available
labor are important factors to consider.
A brief but accurate history should be taken
on each mare. Initial information includes the
mare’s age and number of foals she has pro-
duced in her lifetime and the time of her last
foal. The management plan and type of semen
used in the past are important, and the number
of cycles required to obtain pregnancy should
be recorded as well. Any significant reproduc-
tive issues (dystocia, abortion, repeated at-
tempts to breed without success, etc.) are also
important points to consider. A breeding
soundness exam should be considered for old-
er mares, problem mares and new purchases.
Information from culture, cytology and en-
dometrial biopsy can provide important prog-
nostic information prior to breeding the mare.
EXAMINATION OF THE MARE
AND PREDICTING OVULATION
The estrous cycle of the mare is approximately
21 days (range 18-21) and can be divided into
a period of estrus (4-7 days) and diestrus (14-
15 days). The mare ovulates approximately 24
hours prior to the end of estrus. Given the long
period of receptivity in the mare, a thorough ex-
amination must be performed in order to deter-
mine the optimal time for breeding.
Initial examination of the mare should include
evaluation of body condition, perineal confor-
mation and examination of the vulva for the
presence of discharge. A thorough examina-
tion of the reproductive structures is per-
formed by transrectal palpation and ultra-
sonography. Uterine and cervical tone, uterine
edema score, the presence and size of ovarian
follicles, presence/absence of a corpus luteum
(CL) and presence/absence of uterine fluid are
evaluated and recorded. Any pathology, in-
cluding size and location of uterine cysts,
should be noted in the mare record. This will
Published in IVIS with the permission of SIVE
be invaluable at the time of pregnancy diagno-
sis. A speculum exam may also be performed
to further assess cervical relaxation. Teasing
to a stallion should also be performed, when
possible. Some mares will tease to a gelding
or another mare when a stallion is not avail-
able, however this is not as reliable. Teasing
should be performed every day while a mare
is in estrus, as subtle changes will be noted on
a day to day basis.
Based on the findings of the reproductive ex-
amination, pharmacologic agents may be used
to increase the predictability of ovulation. The
goal of most programs is to breed each mare
one time per cycle and have most mares preg-
nant within three cycles. This not only limits
cost and labor but also allows a popular stal-
lion to accommodate more mares per season.
Hormonal control helps achieve this goal.
In the presence of a mature corpus luteum (i.e.
in diestrus), prostaglandin (PGF) may be giv-
en to cause luteolysis and allow the mare to
come into estrus. Prostaglandin is generally
thought to be only effective between days 5
and 16 of the estrus cycle (ovulation = day 0).
If prostaglandin is given outside of this win-
dow, it does not reliably induce luteolysis or
shorten the time to ovulation. The average
mare ovulates approximately 7-10 days after
prostaglandin treatment, however the range is
large (as early as 2 days or as long as 10+
days) and depends on the size of the largest
follicle at the time of PGF treatment. Because
administration of PGF shortens the duration
of diestrus, it is often called “short cycling”
the mare.
In the absence of a CL and in the presence of
a large dominant follicle (> 35 mm in diame-
ter) accompanied by other evidence of estrus
(relaxed cervix, uterine edema), semen should
be ordered or an appointment made at the
breeding shed. Once a booking is confirmed,
ovulation can be hormonally induced with hu-
man chorionic gonadotropin (hCG) or
GnRH agonists (deslorelin, buserelin). hCG
has LH-like activity and acts directly on the
ovary to induce ovulation in an approximately
36 hours in 90% of mares. GnRH agonists
(deslorelin, buserelin) act directly on the pitu-
itary to cause the release of LH and induction
132
Proceedings of the Annual Meeting of the Italian Association of Equine Veterinarians, Carrara, Italy 2010
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of ovulation approximately 40 hours after ad-
ministration. Response to hCG (or deslore-
lin) requires that the mare be in estrus with
a follicle 35 mm (30 mm) or greater in di-
ameter with uterine edema and a relaxed
cervix. It is unwise to administer an ovulatory
agent prior to confirming stallion availability.
While some light horse mares with 35 mm fol-
licles will routinely ovulate in response to
hCG or deslorelin, large breed mares (Draft or
Warmblood mares) may need to develop a
larger follicle (45-50 mm) before induction of
ovulation. These large mares can ovulate a
larger (50 mm) follicle, and may not respond
to hCG if it is given too early. Conversely,
smaller breeds, such as ponies, may ovulate
smaller follicles and respond to hCG stimula-
tion earlier.
BREEDING WITH COOLED SEMEN
Ultimately, the preceding steps all serve to ac-
curately predict the optimal time for breeding.
Ideally, breeding should occur shortly before
ovulation, however due to the life-span of sper-
matozoa and the oocyte, a window of opportu-
nity exists for post-ovulation breeding as well.
Conservatively, it can be assumed that maxi-
mal fertility for most stallions can be achieved
using fresh or cooled-shipped semen within
24-48 hours before ovulation or within 6-8
hours after ovulation. Cooled-shipped semen
is typically 24 hours old at the time of arrival.
Ovulation should therefore occur within 24
hours after insemination. This can be achieved
by monitoring the mare until she has a large
(>35 mm) dominant follicle and uterine ede-
ma, and administering an ovulation-inducing
agent at the time that semen is collected and
shipped. This type of management will ensure
that most mares ovulate within approximately
12 hours following insemination. In many cas-
es, two doses of semen are shipped. When a
second dose is available, it can be inseminat-
ed 24 hours after the first dose. Some practi-
tioners prefer to inseminate the mare with
both doses when the semen arrives, feeling
that the mare provides better storage for se-
men than a cooling system.
 Published in IVIS with the permission of SIVE
BREEDING WITH FROZEN SEMEN
If breeding with frozen semen is desired, the
semen should be ordered early and stored in
an appropriate storage container on-site. The
advantage of breeding with frozen semen is
the ready availability of the semen, rather than
relying on the stallion’s availability for semen
when the mare is ready to breed.
Due to the decreased lifespan of frozen-
thawed semen (12 hours) and the limited lifes-
pan of the oocyte once it has ovulated (6-8
hours), frozen-thawed semen should be placed
in the uterus within 12 hours before to 6-8
hours after ovulation.
Timed insemination protocol using two
doses of frozen-thawed semen
Two timed inseminations at 24 and 40-42
hours following the administration of an ovu-
latory agent is a convenient method for breed-
ing mares with frozen-thawed semen if multi-
ple breeding doses are available. Using this
method, the mare only needs to be examined
2-3 times after administration of an ovulatory
agent. One dose of semen will be in the uterus
within 12 hours prior to ovulation, and one
dose of semen will be in the uterus within 6
hours post-ovulation (Figure 1).
Under ideal circumstances, one dose of semen
is inseminated pre-ovulation and a second
dose is inseminated post-ovulation. Pregnancy
rates using good quality frozen-thawed semen
were comparable to those using cooled semen
in one study.
Insemination protocol using one dose
of frozen-thawed semen
In some cases, frozen semen from a horse is
very limited. When only a single dose of frozen
semen is available, standard protocol is to
breed in the post-ovulatory period. An ovula-
tion induction agent is administered to the mare
with the development of a > 35 mm follicle and
good estrus edema. Typically, the mare is ex-
0 24
Figure 1
hCG or deslorelin
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Proceedings of the Annual Meeting of the Italian Association of Equine Veterinarians, Carrara, Italy 2010
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amined by transrectal palpation and ultrasound
every 6 hours beginning approximately 24
hours following administration of an ovulatory
agent until ovulation is detected. This type of
management ensures that semen will be placed
in the uterus within 6 - 8 hours following ovu-
lation. This method is significantly more labor-
intensive for the veterinarian and can incur
much more expense for the mare owner.
SEMEN QUALITY
Regardless of the type of semen used, evalua-
tion of the semen is an important part of the
service provided by a reproductive veterinari-
an. Even in hand-breeding situations, a veteri-
narian often will evaluate semen collected
from the stallion’s penis as he dismounts the
mare. A simple examination of a drop of se-
men on most portable light microscopes pro-
vides good evidence that the stallion ejaculated
motile sperm. Cooled–shipped semen should
be evaluated prior to packaging for shipment.
The veterinarian managing the mare for insem-
ination should also evaluate the semen for
motility, morphologically normal sperm and
adequate numbers.
A standard dose of fresh semen used on farm is
250 to 500 million progressively motile sper-
matozoa. A standard dose of cooled-shipped
semen is at least 1 billion progressively motile
spermatozoa at the time of shipment, with at
least 500 million progressively motile sperma-
tozoa at the time of insemination. Accepted
standards for frozen-thawed semen are 1 billion
to 800 million total sperm/dose, with at least
30% motility after thawing.
POST-BREEDING MANAGEMENT
After insemination, the mare should be moni-
tored for ovulation on a daily to every other day
basis. If ovulation does not occur, the mare may
36 42
AI OV AI
Published in IVIS with the permission of SIVE
need to be re-inseminated to ensure viable
sperm within the oviduct at the time of ovula-
tion. Mares also should be evaluated for
pathologic changes, such as the presence of
anovulatory hemorrhagic follicles and in-
creased uterine fluid or other evidence of per-
sistent post-mating endometritis. All mares
experience a physiologic endometritis after
breeding, however it should resolve complete-
ly within 12-24 hours. Some mares may have
delayed uterine clearance or pathologic levels
of inflammation which should be addressed
quickly to prevent infertility. Mares with a his-
tory of post-mating endometritis will require
more intense management (uterine lavage and
tocolytics, among other treatments) than nor-
mal mares.
PREGNANCY DIAGNOSIS
Early pregnancy diagnosis should be a routine
part of breeding management. This not only
allows timely management of subsequent cy-
cles if the mare fails to become pregnant, but
also allows for management of twin pregnan-
cies while the vesicles are still in the mobility
phase. Ideally, the first pregnancy exam should
take place at day 14-16 post-ovulation. When a
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Proceedings of the Annual Meeting of the Italian Association of Equine Veterinarians, Carrara, Italy 2010
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double ovulation is observed, the mare should
be examined at 14 days post-ovulation. At the
time of pregnancy exam, careful attention
should be paid to the tone of the cervix and
uterus as well as the echogenicity of the CL and
the size and location of the vesicle.
Subsequent examinations are recommended
to ensure continued pregnancy. Timing of
these exams varies based on client and clini-
cian preference, however an exam between 25
and 30 days is often recommended as the fetal
heartbeat can be well visualized, yet the peri-
od of endometrial cup formation has not yet
commenced. A “Fall check” may be routine
on some farms in order to identify open mares
prior to the initiation of pregnant mare vaccine
and nutrition programs.2,3
REFERENCE LIST
1. Madill S, Troedsson MHT, Santschi EM, Malone
ED. Dose-response effect of intramuscular oxy-
tocin treatment on myometrial contraction of repro-
ductively normal mares during estrus. Theriogenol-
ogy 2002; 58(2-4):479-481.
2. Equine breeding management and artificial insemi-
nation. 1 ed. Philadelphia: W.B. Saunders. 2000.
3. Blanchard TL, Varner DD, Schumacher J, Love CC,
Brinsko S, Rigby S. Manual of Equine Reproduc-
tion. second ed. St. Louis: Mosby. 2003.