University of Juba

School of Veterinary Medicine

Department of Clinical Studies
Theriogenology note

The female reproductive organs

The female reproductive organs are composed of
 ovaries
 oviducts
 uterus
 cervix
 uteri horn
 vagina
 external genitalia
The internal genital organs (the first of four components) are supported by the broad
ligament. This ligament consists of the mesovarium, which supports the ovary; the
mesosalpinx, which supports the oviduct; and the mesometrium, which supports the uterus. In
cattle and sheep, the attachment of the broad ligament is dorsolateral in the region of the
ileum, so that the uterus is arranged like a ram's horns, with the convexity dorsal and the
ovaries located near the pelvis.
Embryology
The fetal reproductive system consists of two sexually non-differentiated gonads, two pairs of
ducts, a urogenital sinus, a genital tubercle, and vestibular folds. This system arises primarily
from two germinal ridges on the dorsal side of the abdominal cavity, and it can differentiate
into a male or a female. system. The sex of the fetus depends on inherited genes,
gonadogenesis, and the formation and maturation of accessory reproductive organs. Wolffian
and Mullerian ducts are both present in the sexually undifferentiated embryo. In the female,
the Mullerian ducts develop into a gonaductal system and the Wolffian ducts atrophy. The
opposite is true in the male. The female Mullerian ducts fuse caudally to form a uterus, a
cervix, and the anterior part of a vagina. In the male fetus, testicular androgen plays a role in
the persistence and development of the Wolffian ducts and the atrophy of the Mullerian
ducts.
Ovary

Note by Dr. Jong Anthony

 The ovary, unlike the testis, remains in the abdominal cavity. It performs both exocrine (egg
release) and endocrine (steroidogenesis) functions. At birth, a layer of follicular cells
surrounds the primary oocytes in the ovary to form the primordial follicles.
The shape and size of the ovary
The shape and size of the ovary vary both with the species and the stage of the estrous cycle.
 In cattle and sheep, the ovary is almond-shaped,
 whereas in the horse it is bean shaped owing to the presence of a definite ovulation
fossa, and indentation in the attached border of the ovary.
 The porcine ovary resembles a cluster of grapes because the protruding follicles and
corpora lutea obscure the underlying ovarian tissue.
part of the ovary
The ovary, composed of the medulla and cortex, it is surrounded by the superficial
epithelium, commonly known as germinal epithelium. The ovarian medulla consists of
(a) irregularly arranged fibroelastic connective tissue and (b) extensive nervous and
vascular systems that reach the ovary through the hilus. The arteries are arranged in a
definite spiral shape.
The ovarian cortex contains ovarian follicles and/or corpora lutea at various stages of
development of regression. The vascular pattern of the ovary changes with different
hormonal states. Variations in the architecture of the vessels allow adaptation of the blood
supply to the needs of the organ. The intraovarian distribution of blood undergoes
remarkable changes during the preovulatory period. Arterial blood flow to the ovary
varies in proportion to luteal activity. Hemodynamic changes seem to be important in
regulating corpus luteum (CL) function and lifespan. Thus, changes in blood flow precede
the decline in progesterone secretion, whereas restriction of ovarian blood flow causes
premature CL regression. At the time of luteolysis in ewes, there is a reduction in ovarian
blood flow. Blood flow to the bovine ovary is highest during the luteal phase, decreases
with luteal regression, and reaches a nadir just before ovulation. Ovarian blood flow
increases with the newly developing CL. The decline in blood flow seems to follow the
abrupt decline at the time of regression of the CL.
The Oviduct
The length and degree of coiling of the oviduct vary in farm mammals. The oviduct may
be divided into four functional segments: the fringelike fimbriae; the funnel-shaped
abdominal opening near the ovary-the infundibulum; the more distal dilated ampulla;
and, the narrow proximal portion of the oviduct connecting the oviduct with the uterine
lumen-the isthmus. The fimbriae are unattached except for one point at the upper pole of

Note by Dr. Jong Anthony

 the ovary. This ensures close approximation of the fimbriae and the ovarian surface. The
ampulla, accounting for about half of the oviductal length, merges with the constricted
section known as the isthmus. The isthmus is connected directly to the uterus; it enters the
horn in the form of a small papilla in the mare. In the sow, however, this junction is
guarded by long finger-like mucosal processes. In the cow and ewe, there is a flexure at
the uterotubal junction, especially during estrus. The thickness of the musculature
increases from the ovarian to the uterine end of the oviduct.
The Uterus
The uterus consists of two uterine horns (cornua), a body, and a cervix (neck). The
relative proportions of each, as well as the shape and arrangement of the horns, vary
according to species. In swine, the uterus is of the bicornuate type (uterus bicornis). The
horns are folded or convoluted and may be as long as 4 to 5 feet, while the body of the
uterus is short. This length is an anatomic adaptation for successful litter bearing. In
cattle, sheep, and horses, the uterus is of the bipartite type (uterus bipartitus). These
animals have a septum that separates the two horns and a prominent uterine body (the
horse has the largest). In ruminants the uterine epithelium has several caruncles. Both
sides of the uterus are attached to the pelvic and abdominal walls by the broad ligament.

Function of the Uterus

The uterus serves a number of functions. The endometrium and its fluids play a major
role in the reproductive process:
(a) sperm transport from the site of ejaculation to the site of fertilization in the oviduct.
(b) regulation of the function of the CL.
(c) initiation of implantation, pregnancy and parturition.
Sperm Transport.
At mating, the contraction of the myometrium is essential for the transport of sperm from
the site of ejaculation to the site of fertilization. Large numbers of sperm aggregate in the
endometrial glands. As sperm are transported through the uterine lumen to the oviducts,
they undergo "capacitation" in endometrial secretions.

Note by Dr. Jong Anthony

 Luteolytic Mechanisms.
There is a local utero-ovarian cycle whereby the CL stimulates the uterus to produce a
substance that in tum destroys the CL. The uterus plays an important role in regulating the
function of the CL. Corpora lutea are maintained in a functional state for long periods
following hysterectomy of cattle, sheep, and swine. If small amounts of uterine tissue
remain in situ, luteal regression occurs and cycles are resumed after variable periods.
Following unilateral hysterectomy, corpora lutea adjacent to the excised uterine horn are
usually better maintained than those adjacent to the remaining horn. Intramuscular or
intrauterine administration of prostaglandin causes complete luteal regression in the cow
and ewe. The gravid uterine horn exerts an antiluteolytic effect at the level of the adjacent
ovary. This effect is exerted through a local utero-ovarian venoarterial pathway.
Implantation and Gestation.
The uterus is a highly specialized organ that is adapted to accept and nourish the products
of conception from the time of implantation until parturition. Uterine "differentiation" is
governed by the ovarian steroid hormones. This process must evolve to some critical
stage when the uterus is prepared to selectively accept the blastocyst. Unless such
differentiation occurs, the uterus is unsuited to permit implantation. After implantation,
the embryo depends on an adequate vascular supply within the endometrium for its
development. Throughout gestation, the physiologic properties of the endometrium and
its blood supply are important for the survival and development of the fetus. The uterus is
capable of undergoing tremendous changes in size, structure, and position to
accommodate the needs of the growing conceptus.
Parturition and Postpartum Involution.
The contractile response of the uterus remains dormant until the time of parturition, when
it plays the major role in fetal expulsion. Following parturition, the uterus almost regains
its former size and condition by a process called involution. In the sow, the uterus
continuously declines in both weight and length for 28 days after parturition; thereafter it
remains relatively unchanged during the lactation period. However, immediately after the
young are weaned, the uterus increases in both weight and length for 4 days. During the
postpartum interval, the destruction of endometrial tissue is accompanied by the presence
of large numbers of leukocytes and the reduction of the endometrial vascular bed. The
cells of the myometrium are reduced in number and size. These rapid and disproportional
changes in the uterine tissue are a possible cause of low postpartum conception rate.
Neither the presence of suckling calves nor anemia delays uterine involution. Caruncular
tissues are sloughed off and expelled from the uterus 12 days after calving.

Note by Dr. Jong Anthony

 Cervix Uteri
The cervix is a sphincter-like structure that projects caudally into the vagina. The cervix
is a fibrous organ composed predominantly of connective tissue with only small amounts
of smooth muscle tissue present. The cervix is characterized by a thick wall and
constricted lumen. Although the structure of the cervix differs in detail among farm
mammals, the cervical canal has various prominences. In ruminants these are in the form
of transverse or spirally interlocking ridges known as annularrings, which develop to
varying degrees in the different species. They are especially prominent in the cow
(usually four rings) and in the ewe, where they fit into each other to close the cervix
securely. In the sow, the rings are in a corkscrew arrangement that is adapted to the spiral
twisting of the tip of the boar's penis. Distinguishing features of the mare's cervix are the
conspicuous folds in the mucosa and the projecting folds into the vagina. The cervix is
tightly closed except during estrus, when it relaxes slightly, permitting sperm to enter the
uterus. Mucus discharged from the cervix is expelled from the vulva.
Functions of the cervix
The cervix plays several roles in the reproductive process:
(a) it facilitates sperm transport through the cervical mucus to the uterine lumen
(b) it acts as sperm reservoir
(c) the selection of viable sperm, thus preventing the transport of nonviable and defective
sperm.
Sperm Transport.
Upon ejaculation, sperm are oriented toward the internal os. As the flagellum beats and
vibrates, the sperm head is propelled forward in the channels of least resistance. The
macrorheologic and microrheologic properties of cervical mucus play a major role in
sperm migration. Sperm penetrability increases with the cleanliness of mucus, since
cellular debris and leukocytes delay sperm migration.
The cervix might act as a reservoir for sperm, thus providing the upper reproductive
tract with subsequent releases of sperm. It is also possible that sperm that are trapped in
the cervical crypts are never released, thus preventing excessive numbers of sperm from
reaching the site of fertilization. In ruminants prolonged survival of sperm in the cervix
relative to other parts of the reproductive tract suggest that the cervix acts as a sperm
reservoir. In the cervices of cattle and goats, most sperm are not randomly distributed but
are located between cervical crypts. Penetration of sperm to these sites in the cervix

Note by Dr. Jong Anthony

 depends on sperm viability and on the structure and, consequently, the rheologic
properties of the cervical mucus.
The Cervix During Pregnancy.
During pregnancy, a highly viscid, nonferning, thick, and turbid mucus occludes the
cervical canal, acting as an effective barrier against sperm transport and invasion of
bacteria in the uterine lumen, thus preventing uterine infections. The only other time the
cervix is open is before parturition. At this time the cervical plug liquefies and the cervix
dilates to permit the expulsion of the fetus and fetal membranes.
Vagina
The vaginal wall consists of surface epithelium, muscular coat, and serosa. The muscular
coat of the vagina is not as well developed as the outer parts of the uterus. It consists of a
thick inner circular layer and a thin outer longitudinal layer; the latter continues for some
distance into the uterus. The muscularis is well supplied with blood vessels, nerve
bundles, groups of nerve cells, and loose and dense connective tissue. The cow is unique
in possessing an anterior sphincter muscle in addition to the posterior sphincter found in
the other farm mammals.
Functions of the Vagina
The vagina has multiple functions in reproduction. It is a copulatory organ in which
semen is deposited and coagulated until sperm are transported through the
macromolecules of the cervical mucus column. The dilated bulbous vagina provides a
postcoital semen pool to supply sperm for cervical reservoirs. The rugae vaginales and the
fencelike, rhomboidshaped arrangement of the musculature allow distention of the vagina
during mating and parturition. Although the vagina contains no glands, its walls are
moistened by transudates through the vaginal epithelium (incorrectly called mucosa), by
cervical mucus, and by endometrial secretions. Following ejaculation, the seminal plasma
is not transported into the uterus; most of it is expelled or absorbed through the vaginal
walls. Some of the biochemical components of the seminal plasma, when absorbed in the
vagina, exert physiologic responses in other parts of the female reproductive tract. The
pH of the vaginal secretion is unfavorable to sperm. A complex interaction of the cervical
mucus, vaginal secretion, and seminal plasma induces a buffering system that protects
sperm until they are transported through the micelles of cervical mucus. Pathologic
conditions resulting in insufficient buffering of the seminal pool (e.g., low volume of
ejaculate, scanty amounts of thick cervical mucus, and leakage of semen) may cause rapid
immobilization of spermatozoa. The vagina serves as an excretory duct for secretions of
the cervix, endometrium, and oviduct; it also serves as the birth canal during parturition.

Note by Dr. Jong Anthony

 These functions are accomplished through various physiologic characteristics, namely,
contraction, expansion, involution, secretion, and absorption.

External Genitalia
The vestibule, the labia majora, the labia minora, the clitoris, and the vestibular glands
compose the external genitalia.
Vestibule
The junction of the vagina and vestibule is marked by the external urethral orifice and
frequently by a ridge (the vestigial hymen). In some cattle, the hymen may be so
prominent that it interferes with copulation. The vestibule of the cow extends inward for
approximately 10 cm, where the external urethral orifice opens into its ventral surface.
Gartner tubes (remnants of the Wolffian ducts) open into the vestibule posteriorly and
laterally to Gartner's ducts. The glands of Bartholin, which secrete a viscid fluid, most
actively at estrus, have a tuboalveolar structure similar to the bulbourethral glands in the
male.
Labia Majora and Labia Minora
The integument of the labia majora is richly endowed with sebaceous and tubular glands.
It contains fat deposits, elastic tissue, and a thin layer of smooth muscle. It has the same
outer surface structure as the external skin. The labia minora have a core of spongy
connective tissue.
Clitoris
The ventral commissure of the vestibule conceals the clitoris, which has the same
embryonic origin as the male penis. It is composed of erectile tissue covered by stratified
squamous epithelium, and it is well supplied with sensor nerve endings. In the cow, the
greater part of the clitoris is buried in the mucosa of the vestibule. In the mare, however,
it is well developed, and in the sow it is long and sinuous, terminating in a small point or
cone.
Puberty
Puberty is the period when a female animal becomes sexually mature and able to undergo
cyclic activities and reproduce their young ones. Or a period when a male or female
animal is able to release gametes and to manifest complete sexual behaviour sequences.

Note by Dr. Jong Anthony

 Age at Puberty
In normal breeding conditions, females of domestic species reach the age of puberty at
the following times:
 mare: 1–2 years
 cow: 7–18 months
 ewe: 6–15 months
 doe or nanny goat: 4–8 months
 sow: 6–8 months
 bitch: 6–20 months
 queen cat: 7–12 months

External factors influencing the time of onset of puberty

The time of onset of puberty is determined by the individual’s genotype, with smaller breeds
of animal tending to be slightly more precocious. However, this inherent timing is influenced
by a number of external factors.
A. Nutrition. There is good evidence that in most domestic species, the age of puberty is
closely related to body weight; therefore, it is not surprising that nutrition is an
important factor. Animals that are well fed with good growth rates reach puberty
before those that are poorly fed with slow growth rates. However, unless the animal is
severely malnourished, the onset of cyclical activity will eventually occur.
B. Season of the year. In those species which are seasonal breeders, such as the ewe,
mare and queen cat, the age at which puberty occurs will be influenced by the effect
of season of the year. For instance, a filly born early in the year, i.e. January or
February, may have her first oestrus in the May or June of the following year, i.e.
when she is 16 or 17 months old. A filly foal born late in the year, July or August,
may not have her first oestrus until she is 21 or 22 months old. The same is true of
ewes which, depending upon the time of year at which they are born, may reach
puberty as early as 6 months or as late as 18 months old.
C. Proximity of the male. Studies in sheep and pigs have shown that exposure to the
male of the species will advance the timing of the onset of puberty. This so-called

Note by Dr. Jong Anthony

 ‘ram or boar effect’ is probably mediated by pheromonal and other sensory cues
influencing hypothalamic GnRH secretion.
D. Climate. Anthropomorphic extrapolation has assumed that animals living in the
tropics reach puberty at an earlier age than those in temperate climates.
E. Disease. Any disease which can influence the growth rate, either directly or because
of interference with feeding and utilisation of nutrients, will delay the onset of puberty

Oestrous Cycle and Its Phases

Traditionally, the oestrous cycle is divided into a number of phases.
 Pro-oestrus. The phase immediately preceding oestrus. It is characterised by a
marked increase in activity of the reproductive system.
 There is follicular growth and regression of the corpus luteum of the previous
cycle (in polycyclic species).
 The uterus enlarges very slightly;
 the endometrium becomes congested and oedematous, and its glands show
evidence of increased secretory activity.
 The vaginal mucosa becomes hyperaemic; the number of cell layers of the
epithelium starts to increase, and the superficial layers become cornified.
 The bitch shows external evidence of pro-oestrus with vulval oedema,
hyperaemia and a sanguineous vulval discharge.
 Oestrus. The period of acceptance of the male. The onset and end of the phase are the
only accurately measurable points in the oestrous cycle, and hence are used as the
baseline for determining cycle length.
 The animal usually seeks out the male and ‘stands’ for him to mate her.
 The uterine, cervical and vaginal glands secrete increased amounts of mucus
 the vaginal epithelium and endometrium become hyperaemic and congested
 the cervix is relaxed.
 Ovulation occurs during this phase of the cycle in all domestic species with
the exception of the cow, where it occurs about 12 hours after the end of
oestrus.
Ovulation is a spontaneous process in all domestic species with the exception of the cat,
rabbit and camel, in which it is induced by the act of coitus. During pro-oestrus and oestrus
there is follicular growth in the absence of functional corpora lutea, the main ovarian
hormones produced being oestrogens. Pro-oestrus and oestrus are frequently referred to
collectively as the follicular phase of the cycle.

Note by Dr. Jong Anthony

  Metoestrus. The phase succeeding oestrus.
 The granulosa cells of the ovulated follicle give rise to lutein cells which are
responsible for the formation of the corpus luteum.
 There is a reduction in the amount of secretion from the uterine, cervical and
vaginal glands.
 Dioestrus. The period of the corpus luteum.
 The uterine glands undergo hyperplasia and hypertrophy
 the cervix becomes constricted
 the secretions of the genital tract are scant and sticky
 the vaginal mucosa becomes pale.
 The corpus luteum is fully functional during this phase, and is secreting large
amounts of progesterone.
The period of the oestrous cycle when there is a functional corpus luteum is sometimes
referred to as the luteal phase of the cycle, to differentiate it from the follicular phase.
 Anoestrus.
The prolonged period of sexual rest during which the genital system is mainly
quiescent. In this period, the following changes take place
 Follicular development is minimal
 the corpora lutea, although identifiable, have regressed and are non-functional.
 Secretions are scanty and tenacious
 the cervix is constricted
 the vaginal mucosa is pale

Endocrine Changes During Estrous Cycle

The estrous cycle is controlled by the interaction of FSH, LH, estrogen and progesterone.
These hormones are common to most domestic animals, however, their secretory patterns and
relative effects vary among the species.
 During early development, prior to sexual maturity of the bitch, very little
gonadotropic hormones are secreted and the ovaries, therefore, remain inactive.
 However, around the age of 6 months and above, the pituitary begins to secrete
higher levels of the gonadotropic hormones called follicular stimulating hormone
(FSH) and luteal hormone (LH). The rise in FSH and LH will initiate the sexual cycle
and this cyclical increase and decreases in FSH and LH in turn, control
 the cyclic ovarian changes and, as such, are responsible for the physiologic events in
the normal reproductive cycle of the female animals.

Note by Dr. Jong Anthony

  The female animal has two ovaries that will produce the ova (eggs) which are
contained within follicles that grow toward the surface of the ovary.
 When FSH and LH from the pituitary gland begin to be secreted in high quantities
during onset of sexual maturity, the ovaries and the follicles within them will begin to
grow.
 Within these follicles, a follicular fluid hormone, secreted by the ovary, called
estrogen, surrounds the ovum. This hormone is a biologic chemical that produces
physiologic and social/behavioural effects within the female that will signal a
readiness to mate.
 Two days prior to ovulation, there is a surge in the secretion of LH by the pituitary
gland preceded by rapid swelling of the follicle. This LH surge is of critical
importance because in its absence, even with the other hormonal physiologic effects
taking place, ovulation will not occur.
 Additionally, the LH surge causes the ovarian cells to switch over to secreting
progesterone hormone rather than estrogen.
 Within two days of the LH surge, the follicle reaches the surface of the ovary and
bursts, thereby releasing the ovum into a capsule that surrounds the ovary. This
process is referred to as ovulation.
 In the meantime, the ruptured follicles from which each ovum was developed will
begin to produce a rapidly dividing mass of cells called luteal bodies, which will
make up the corpus luteum.
 In addition to producing progesterone, which will maintain the pregnancy, the corpus
luteum will also produce inhibin, the hormone that will signal the pituitary gland to
decrease production of FSH and LH.
The Canine Oestrous Cycle
The normal reproductive cycle of the bitch is comprised of four stages:
 proestrus,
 estrus,
 diestrus,
 anestrus.
Proestrus Phase:
 Also known as the follicular stage because during this phase, the ovarian follicles,
each containing ova, increase in size.
 Average duration is 9 days with a range of 3-17 days.
 Swelling of the vulva, the external tissue of the vaginal opening and bloody discharge
marks the beginning of the proestrus stage.

Note by Dr. Jong Anthony

  Males are interested in the bitch but the bitch is not
 Increasing amounts of estrogen hormone, secreted by the ovarian follicles, cause the
cells of the vaginal walls to take-on a distinctive shape, a process known as
cornification.
 With vaginoscopy, the vagina mucosa glistens with rounded edges.
 Both the level of estrogen and vaginal cornification are useful indicators of proestrus.
Estrus Phase:
 Average duration is 9 days with a range of 3-21 days.
 The male and female are both interested in each other.
 The bitch will 'flag' her tail, as if allowing access to the vulva.
 NOTE: The estrus behavior results from the estrogen that peaked during proestrus
abruptly declining.
 Physiologically, estrus coincides with the predominant presence of cornified vaginal
epithelial cells and an increase in serum progesterone levels to 2ng/ml.

 With vaginoscopy, the vulva and vaginal epithelium appear to wrinkle because the
decreased estrogen results in water loss of the cells.
 Ovulation usually occurs 2 days following this increase in progesterone
Diestrus Phase:
 Average duration is 2 months approximately 6 days after ovulation. The cornified
vaginal epithelial cells will revert to a non-cornified state. This condition marks the
beginning of diestrus
 This stage ends when progesterone levels fall to less than 1 ng/ml just prior to
whelping in the pregnant bitch or approximately 2 months after ovulation in the non-
pregnant bitch.
Anestrus Phase:
 This is a time of mandatory endometrial repair.
 The endometrium is being 'repaired' after the progesterone effects during diestrus for
the preceding 60 days.
 True anestrus lasts 90 -150 days’ post whelping, or post diestrus.
 Though Duration of anestrus is quite variable among bitches and may be governed by
both genetic and environmental variables.
 Interestrous (anestrus + diestrus) lasts 150 - 210 days after the last estrus.
 Fertility is low if at least 90 day anestrus (or a 150 interestrus interval) is not attained.
This is because the uterus has not repaired enough to maintain pregnancy.

Note by Dr. Jong Anthony

  NOTE: Anestrus is not the same as interestrous. Anestrus is a variable time after
diestrus while Interestrous is (diestrus + anestrus) averages 5 - 7 months.

Anestrus events:
 The male shows no sexual interest in the female.
 The female shows no sexual interest in the male.
 The vulva appears normal. It is not swollen or edematous.
 The vaginal cytology has very few cells and they are non cornified.
 The vaginal wall is very thin and appears pale on vaginal speculum examination.
 Progesterone is at baseline concentrations Even spayed bitches run basal levels of
progesterone. This baseline progesterone is probably of adrenal origin.
 Prolactin secretion by the pituitary may promote anestrus, because prolactin inhibitors
can be used to terminate anestrus (i.e. induce estrus).
 The beginning of proestrual bleeding marks the end of this stage.

The Feline Oestrous Cycle

There are some peculiarities of feline estrous cycle which makes their reproductive pattern
unique and distinct from that of other domestic species.
 Like the mare, the queen is seasonally polyestrous and responsive to photoperiod,
thus referred to as long day breeders (i.e. requires 12hrs or more of light to maintain
normal cyclicity)
 Although it has been noted that long haired breeds tend to be more seasonal than the
short haired breeds.
 Like the bitch, the queen is unusual in maintaining sexual receptivity for a period of
some days after ovulation, while the corpus luteum is forming.
 Most importantly from a clinical standpoint, like the rabbit and ferret the queen is an
induced ovulator, requiring a copulatory stimulus or exogenous hormones for
ovulation and corpus luteum formation.
 Puberty in the cat usually occurs at 9 to 10 months of age although may occur as
early as 4 months or late as 2 years.
 However, because cats are seasonal breeders and the season in which the kitten was
born influences the age at which puberty occurs.
Proestrus Phase:
 Proestrus, the period preceding estrus, lasts 1 to 2 days. This phase is often
unobserved and may be seen in only 16% of estrous cycles.

Note by Dr. Jong Anthony

  During this time, the female is attractive to but do not accept the male.
 Behavioral changes may begin to be seen during proestrus in which-
 The queen may rub against objects, vocalize and assume a lordotic posture
sometimes referred to as a “dragster posture”.
 She will place her front quarters on the ground, elevate her hind quarters and lift her
tail to one side. When the dorsal caudal area is stroked, she will tread with her hind
legs.
Estrus Phase:
 Estrus is defined as the period of sexual receptivity. Estrus lasts 3 to 16 days (average
of 7) and then subsides for 3 to 14 days (average of 9 days).
 Behavioral changes are more pronounced in estrus than in proestrus; however, no
conspicuous changes in the appearance or size of the external genitalia are evident.
 The period following estrus is affected by ovulation whether induced by copulation
or exogenous hormones.
Interestrus Phase:
 Interestrus is the period between successive estrus periods if ovulation does not
occur.
 If the queen is not bred, she will cycle into estrus on an average of every 2 to 3
weeks.
Diestrus Phase:
 If the queen ovulates, corpora lutea are formed and secrete progesterone. Note that
elevated progesterone levels are the hallmark of diestrus.
 If the queen is not pregnant, diestrus is also termed covert pseudo-pregnancy and
lasts for 35 to 40 days.

Anoestrus Phase:
Anestrus is the seasonal period when the cat does not cycle.

Note by Dr. Jong Anthony

Folliculogenesis, Egg maturation and Ovulation
Folliculogenesis
In the primordial follicle reserve, formed during fetal life or soon after birth, some primordial
follicles begin to grow continuously throughout life or at least until the reserve is exhausted.
When any follicle is released from this reserve, it continues to grow until ovulation or until
the follicle degenerates, which is the case with the majority of follicles. The largest follicle is
responsible for most oestrogen secretion by the ovary at estrus. Estrogen secretion by the
largest follicle decreases rapidly at the time of the LH peak.
Cattle ovulate a single follicle that can be identified by its size about 3 days before the onset
of estrus when there are one or two large follicles on ovaries. In sheep, one or two large
follicles secrete more estrogens and bind more gonadotropins to granulosa cells than smaller
follicles. In sows, recruitment of the ovulatory follicles into the ovulatory population
continues during the follicular phase. Thus, the development of smaller follicles may be
promoted rather than inhibited by larger "dominant" follicles. The final follicular growth in
ewes, cows, and sows ranges between 12 and 34 days; the total duration of follicular growth
is longer than 20 days and presumably about six months.
The growth of the follicle up to the stage of antrum formation is not strictly gonadotropin
dependent. In hypophysectomized females, the formation of preantral follicles continues at a
more or less normal rate. On the other hand, antrum formation and final growth are entirely
FSH/LH dependent.

Egg Maturation
The maturation of oocytes comprises two stages:
a) a period of growth

Note by Dr. Jong Anthony

b) a period of final nuclear and cytoplasmic preparation prerequisite to fertilization and
normal development.
Oocyte Growth.
When a primordial follicle is released from the reserve, the oocyte and its follicle begin to
grow. Oocyte growth is almost complete at the time of antrum formation. Through cellular
processes, the inner cumulus cells actively cooperate to achieve oocyte growth, as they
establish close contact with the oocyte cell membrane. During the formation of the external
membrane of the oocyte (zona pellucida), the cumulus cell processes are strengthened.

The maturation of oocytes is independent of:

a) the nature of follicular stimulation
b) the diameter of the follicle from which oocytes have been removed
c) the source of the follicular fluid or its filtrate from diverse follicles and different females.
Ovulation
Ovulation is a phase of the female oestrus cycle that involves the release of an egg (ovum)
from one of the ovaries
Preovulatory follicles undergo three major changes during the ovulatory process:
a) cytoplasmic and nuclear maturation of the oocyte
b) disruption of cumulus cell cohesiveness among the cells of the granulosa layer
c) thinning and rupture of the external follicular wall After the ovulatory surge of
gonadotropins, blood flow increases to all classes of follicles. The follicle destined to ovulate,
however, receives the largest volume of blood in absolute terms (as measured in ml/min).
Site of Ovulation
The mammalian ovary is normally arranged so that ovulation can occur at any point on its
surface, except at the hilus. However, ovulation in mares always occurs in a limited ovarian
area called the ovulation fossa. The ovary of horses begins its development in the usual way,
and the germinal epithelium covers the whole ovary. In cattle, sheep, and horses’ ovulation
occurs at random with respect to which ovary contains the previous corpus luteum. However,
in some mammals’ ovulation consistently alternates between the ovaries. In the ewe, the site
of ovulation is independent of the location of the corpus luteum of the previous ovarian cycle,

Note by Dr. Jong Anthony

and the duration of the oestrous cycle is unaffected by the relative locations of the corpus
luteum.
There is no difference between left and right ovaries in size or occurrence of ovulation in
horses. The frequency of multiple ovulations for ponies is about 10%. The ovulatory season
appears to be shorter. with ponies ovulating during the fall in the younger age groups (under
5 years) less than in the older groups, indicating a shorter breeding season for young mares.
Ovarian activity appears to decrease after 15 years. The onset of puberty occurs at 12 to 15
months of age. In ponies, there is a decrease in the number of large follicles during late
estrus.

Cellular Events
Several tissue layers separate the oocyte from the outside of the follicle. These are the
surface epithelium. the collagen-rich tunica albuginea. the theca externa, the thin basement
lumina separating the capillary network from the membrana granulosa, and the membrane
granulosa itself. Before ovulation, all tissue layers are broken down. Moreover, the necessary
increase in follicular elasticity during preovulatory growth is associated with changes in
granulosa and theca cell relationships. Such changes are also prerequisite to further corpus
luteum organization. As the enlarging follicle begins to protrude from the surface of the
ovary, the vascularity of the follicular surface increases except at its centre, which seems
devoid of blood vessels. This avascular area is the future point of rupture
Oocyte. Only the cumulus cells anchored in the zona pellucida remain, surrounding the
oocyte and forming the corona radiata, Cumulus cell dissociation frees the oocyte from the
granulosa layer, and meiosis resumes about 3 hours after the gonadotropin surge.
Cumulus cells actively secrete glycoproteins, which form a viscous mass enclosing the
oocyte and its corona. After follicular rupture, the viscous mass spreads at the ovarian surface
to facilitate the "pick up" of the oocytes by the fimbriae. Ultrasonography is used extensively
to recover preovulatory eggs from the ovarian follicle to be used in vitro fertilization.
Granulosa Cells. The granulosa layer is completely dissociated only at the follicular apex
and finally disappears. About 2 hours before ovulation, granulosa cell growth processes
penetrate through the lamina basalis, preparing the invasion of theca cells and blood vessels
into the granulosa after ovulation in the developing corpus luteum. This process is associated
with the production of the early pregnancy factor (EPF).

Note by Dr. Jong Anthony

Theca Cells. The follicular volume rapidly increases in the few hours preceding ovulation
without any increment of follicular fluid pressure, owing to the increased elasticity of the
follicle. This results from a looser cohesion of the theca externa cells owing to the invasive
oedema of this layer and to collagen fibre dissociation, which begins 4 hours after coitus.
Apex Changes. The rupture of the follicle involves interaction between the ovarian
epithelium and the underlying follicular wall. The wall of the follicle apex becomes
exceedingly thin in an area called the "stigma." The stigma thins out, bulges on the surface of
the ovary, and becomes completely avascular. At ovulation the bulging stigma ruptures at the
apex, releasing some of the follicular fluid and the viscous glycoprotein mass embedding the
oocyte.

Egg "Pick-Up"
The ovary, attached to the back of the broad ligament, lies free in the peritoneal cavity. The
oviduct curls over the ovary to facilitate egg "pickup" by the mucosal folds of the fimbriae.
At the time of ovulation, the ovum, together with the surrounding cells in the gelatinous
mass, protrudes at the ovarian surface and is swept into the ostium of the oviduct by the'
action of the motile kinocilia of the fimbriae.

Note by Dr. Jong Anthony

Reception of Eggs (Ova Pickup)
The viscid mass of cumulus oophorus that contains oocyte and corona cells adheres to the
stigma and remains attached to it unless it is removed by the action of the kinocilia of the
fimbriae. Ovum transport through the ostium itself and the first few millimetres of the
ampulla is affected by the action of the cilia.
The physiologic mechanism by which freshly ovulated eggs are picked up into the
oviducts depends, on four main factors:
1. The structural characteristics of the fimbriae of the infundibulum and its relationship
to the surface of the ovary at the time of ovulation.
2. The pattern of release of the cumulus oophorus and its contained egg from the follicle
at the time of ovulation.
3. The biophysical properties of the follicular fluids and the fluids that comprise the
matrix of the cumulus oophorus.
4. The coordinated contraction of the fimbriae and the utero-ovarian ligaments.
At the time of ovulation, the fimbriae are engorged with blood and are brought into close
contact with the surface of the ovary by the muscular activity of the mesotubarium. The ovary
is moved slowly to and from and around its longitudinal axis by contractions of the
ligamentum ovarii proprium. The ovary is located inside the ovarian bursa to which the
ampulla of the oviduct and part of the fimbriae are attached. The ovary can move readily
from this location to the surface of the fimbriae, which is positioned at the open portion of the
ovarian bursa. This movement is controlled by both the ligamentum ovarii proprium and the
mesovarium, which hold the ovary and oviduct in position. The contractile activities of the
fimbriae, oviduct, and ligaments are partly coordinated by hormonal mechanisms involving
the estrogen/progesterone ration. Egg reception is most efficient about the time of estrus, but
it occurs to some degree throughout the cycle.

Egg Transport in The Oviduct

The transport time of ova in the oviduct varies with the species. In cattle, sheep, and swine,
the transport time ranges from 72 to 90 hours. Unfertilized ova are retained in the oviduct of
the mare for several months. It is critical that fertilized eggs reach the uterus at an appropriate
progestational stage of the oestrous cycle. The rate of egg transport is faster from the
infundibulum to the ampullary-isthmic junction than through the isthmic portion. This delay
in ovum transport appears to be required for subsequent implantation of the embryo. The time

Note by Dr. Jong Anthony

of entry of the ovum into the uterus is relatively precise compared with the movement of the
ovum past the ampullary-isthmic junction into the isthmus.
The transport of the egg through the oviduct is regulated by four primary forces:
1. The frequency, force, and programming of contraction of oviductal musculature and
related ligaments, as influenced by endocrine, pharmacologic, and neural
mechanisms.
2. The direction and rate of currents and counter-currents of luminal fluids as affected by
the rate and direction of the beat of kinocilia lining the mucosal folds
3. The secretory activity of non-ciliated cells in the oviductal epithelium as influenced
by the estrogen-progesterone ratio
4. The hydrodynamics and rheologic properties of luminal fluids at the critical times that
ova are being transported

Travel of Ova in Oviduct of farm animals

SPECIES Time in Oviduct (HOURS)

Cattle 90
Sheep 72
Horse 98
Pig 50
Cat 148
Dog 168

Fertilization

Fertilization is a biological process by which a male haploid reproductive cell and a female
haploid reproductive cell interact and unite to form a diploid zygote from which an organism
is developed. The process of fusion of sperm with egg (ovum) to produce a zygote.

Note by Dr. Jong Anthony

Fertilization in most animals is similar to that in humans. In other references it refers to the
amalgamation of haploid male and female gametes (sperm and oocyte) in a series of well-
coordinated molecular events, which eventually leads to the formation of diploid zygote.
Fertilization begins with the sperm's approach to the egg and ends with the fusion of egg and
sperm pronuclei.

The process of fertilization involves a series of specific interactions between spermatozoa and
the oocyte. Spermatozoa acquire maturity during epididymal transit. However, the
maturational changes that occur in the epididymis do not render spermatozoa completely
fertile. For maximum fertility to be achieved, spermatozoa must reside in the female
reproductive tract for a minimum period of time. During the time in the female reproductive
tract, some spermatozoa will undergo changes that allow them to become fertile. These
changes are referred to as spermatozoa capacitation. The site for capacitation varies among
species. In species, where spermatozoa are deposited in the cranial Vagina, capacitation may
begin as sperm ascend and pass through the cervix. In species where semen is deposited into
the mid-cervix (sow) or caudal cervix, (mare) and immediately enters the uterus, capacitation
is probably initiated within the uterus and completed in the isthmus of the oviduct. All
spermatozoa are not capacitated at the same rate. Instead they are capacitated over a
relatively long period of time; (several hours) and this reflects individual sperm differences as
well as location within the tract.

Capacitation

In the oviduct the motility patterns of spermatozoa become hyperactive. The motility pattern
changes from a progressive, linear motility in which they swim in a relatively straight line
(like an Olympic swimmer), into a frenzied, dancing motion that is not linear and is localized
in a small area (like dancers in a disco) In general, hyperactive motility occurs in the ampulla
of the oviduct and is believed to be brought about by specific molecules produced by the
epithelium there. Hyperactive motility is believed to facilitate sperm-oocyte contact.
Spermatozoa are known to contain specific proteins on their plasma membrane surfaces
overlying the acrosome that bind specifically to zona pellucida proteins. These zona-binding

Note by Dr. Jong Anthony

proteins on the plasma membrane must be exposed during the capacitation process before
binding to the zona pellucida can occur.

zona Binding

The zona pellucida of the oocyte consists of three glycoproteins. These glycoproteins have
been named zona proteins 1, 2 and 3 (ZP1, ZP2 and ZP3). Zona proteins 1 and 2 are
structural proteins providing the structural integrity of the zona. Zona protein 3 is much like a
receptor for a hormone. It binds to proteins on the spermatozoal membrane. The binding of
spermatozoa to the zona pellucida is believed to require between 10,000 and 50,000 ZP3
molecules. The sperm plasma membrane contains two zona binding sites. The first binding
site, referred to as the primary zona binding region is responsible for adherence of
spermatozoa to the zona pellucida. The second binding site on the spermatozoal plasma
membrane is believed to be acrosome T-reaction promoting ligand. When binding occurs
between this region and the ZP3 molecule, a signal transduction occurs. This is much like a
typical hormone receptor binding complex. Binding initiates the acrosomal reaction.

Acrosomal reaction

Acrosomal reaction is an orderly fusion of the spermatozoal plasma membrane and the outer
acrosomal membrane. The purpose of the acrosomal reaction is twofold.

 First, the-reaction enables spermatozoa to penetrate the zona pellucida.

 Second, it exposes the equatorial-segment so that it can later fuse with the plasma
membrane of the oocyte.

The acrosomal reaction begins when the plasma membrane of the spermatozoon forms
multiple ‘fusion’sites with the outer acrosomal membrane. When the two membranes fuse,
many small vesicles are formed and this process is called vesiculation. After vesiculation has
occurred, the acrosomal contents are dispersed; and the sperm nucleus is left with the inner
acrosomal membrane surrounding it. Vesiculation characterizes the acrosomal reaction and
morphologically distinguishes it from a damaged acrosome. Damage to the acrosome

Note by Dr. Jong Anthony

membrane and plasma membrane is irreversible. Damage to these membrane is brought about
by changes in osmotic pressure, sudden cooling, sudden heating or marked changes in pH.
Damage to the membranes causes premature loss of acrosomal contents and such sperm
cannot accomplish fertilization.

Penetration of Zone Pellucida

The penetration of the zona pellucida by a spermatozoon is believed to be a rapid process and
probably takes no more than a few minutes. Following attachment to the zona pellucida, the
acrosome reaction, allows the release of a variety of enzymes. Acrosin is one enzyme that is
released from spermatozoa during “the acrosomal reaction. It hydrolyzes zona proteins as,
well as enhances the sperm’s ability to bind to the zona. In the inactive form, acrosin is
known as proacrosin which has” a strong affinity for the zona. Thus, proacrosin aids in
binding the spermatozoon to the zona as the acrosomal reaction proceeds. As proacrosin is
converted to acrosin, the sperm begins to penetrate and make its way through the zona
pellucida. The mechanical force generated by the flagellar action of the tail may be sufficient
to push the sperm through the zona. It is important to note that the acrosomal reaction allows
the spermatozoon to digest the zona through which it can pass. This small regional
dissolution leaves the zona predominately intact.

When the spermatozoon completely penetrates the zona and reaches the perivitelline space
(the space between the zona and the oocyte plasma membrane), it settles into a bed of
microvilli formed from the oocyte plasma membrane. The plasma membrane of the oocyte
fuses with the membrane of the equatorial segment and the fertilizing spermatozoon is
engulfed. The actual fusion of the oocyte plasma membrane with the equatorial segment is
believed to be brought about by a so called fusion protein located on this portion of the
membrane. Prior to the acrosome reaction, this fusion protein is inactive. After vesiculation
and release of the acrosomal contents, the fusion protein is activated, enabling the sperm
membrane to fuse or bind with the oocyte membrane.

After membrane fusion, the oocyte undergoes a series of changes that prepare it for early
embryogenesis. The most easily recognizable is the cortical reaction, during the first and

Note by Dr. Jong Anthony

second meiotic divisions of oogenesis, small, dense granules called cortical granules move to
the periphery of the oocyte cytoplasm. The contents of the cortical granules consist of
mucopolysaccharides., proteases, plasminogen activator, acid phosphatase and peroxidase.
After membrane fusion between the oocyte and spermatozoon, the cortical granules undergo
exocytosis and their contents are released into the perivitelline space as shown above. Exo
cytosis of the cortical granules results in the zona block, a process whereby the zona pellucida
undergoes biochemical changes so that further sperm cannot penetrate it. There by preventing
polyspermy (fertilization of an oocyte by more than one spermatozoon which results in
embryo death)

Note by Dr. Jong Anthony

Types of fertilization

Internal Fertilization

In sexual reproduction, the male inserts the semen into the female reproductive tract to fuse
with the egg. If the fusion takes place within the female parent, it is called internal
fertilization. In humans and most animals like cats, lions, pigs, dogs, hens, etc., the fusion of
gametes takes place internally. In this type, a zygote is formed within the mother and gets its
nourishment from her.

External Fertilization

When the fusion of sperm and egg takes place outside the female reproductive system, it is
called external fertilization. Only a minority of organisms exhibit this type of gamete fusion.
For example, fish, frogs, etc. Here the female parent deposits her eggs in the external
environment and later, the male parent ejects his sperm over them, and then the fusion of the
gametes takes place in the external environment.

Most external fertilization happens during the process of spawning where one or several
females release their eggs and the male(s) release sperm in the same area, at the same time.

Note by Dr. Jong Anthony