“Reoccurring period of sexual receptivity which is associated with the ripening in the ovaries of

one or more Graafian follicles and culminates in the shedding of one or more ova.”

Phases of Estrus Cycle

Traditionally, estrus cycle is divided into a number of phases.
1. Proestrus
- It is the phase immediately preceding estrus.
- It is characterized by a marked increase in activity of reproductive system.
- It is the period of rapid follicular growth under gonadotropin stimulation and
regression of corpus luteum of previous cycle.
Hallmarks of reproductive organs:
- The uterus enlarges very slightly; the endometrium becomes congested and
edematous, and its glands show evidence of increased secretory activity.
- The vaginal mucosa becomes hyperemic
2. Estrus
- It is the period of sexual receptivity during which mating & ovulation occurs.
- The animal usually seeks out the male and “stands” for him to mount her.
- Duration of estrus in influenced by breeding age, specie variation and ambient
temperature; cow in hot environment has shorter period (about 10-12 hrs) than
a cow in a cold climate (18 hrs).
Hallmarks of reproductive organs:
- The uterine, cervical and vaginal glands secrete increased amounts of mucus.
- The vaginal epithelium and endometrium become hyperemic and congested.
- The cervix is relaxed.

Follicular Phase: Pro-estrus and estrus are frequently referred to collectively as the follicular
phase of the cycle. During pro-estrus and estrus there is follicular growth in the absence of
functional CL, the main ovarian hormones produced being estrogens

3. Metestrus
- It is a transitional period between ovulation and full development of CL.
- The granulosa cells of the ovulated follicle give rise to lutein cells which are
responsible for the formation of the corpus luteum.
Hallmarks of reproductive organs:
- There is reduction in the amount of secretion from the uterine, cervical and
vaginal glands.
4. Diestrus
- It is the period of corpus luteum.
- The reproductive organs are under dominant influence of progesterone.
Hallmarks of reproductive organs:
- The uterine glands undergo hyperplasia and hypertrophy.
- The cervix becomes constricted.
- The secretions of genital tract are scant and sticky.
- The vaginal mucosa becomes pale.
Luteal phase: The period of estrus cycle when there is a functional corpus luteum is
sometimes referred to as the luteal phase of the cycle.

5. Anestrus
- It is the prolonged period of sexual rest during which the genital system is
mainly quiescent.
- Follicular development is minimal; the corpora lutea, although identifiable,
have regressed and are non-functional.
Hallmarks of reproductive organs:
- Secretions are scanty and tenacious; cervix is constricted and the vaginal
mucosa is pale.
Other facts:
- Anestrus is the normal physiological state of reproductive function in;
prepubertal, aged animals, pregnant animals and postpartum period.
- It also occurs as pathological condition caused by a variety of factors include:
Nutritional deficiencies, environmental influence causing endocrine
imbalance, ovarian disorders, infectious diseases causing early
embryonic death and abortion.

A Bird’s Eye View on Estrus Cycle

OVARIAN PICTURE
Proestrus
- Developing follicles on ovary
- Small regressed corpus luteum
- Slight tonicity in uterus
- Cervix is slightly dilated
- Pink to red swollen vulva
Estrus
- Fully developed follicle(s) near to ovulate
- Marked tonicity in uterus
- Cervix is dilated; stringy transparent mucus arises from vulva.
- Other estrus signs must be under consideration
Metestrus
- Corpus hemorrhagicum on ovary
- Ovulation depression (fossa) may find on ovary
- Small follicles; of first wave of follicles
- Uterus becomes flaccid
- Cervix is tightly closed
Diestrus
- Fully developed corpus luteum
- Developing follicles may find on ovary
- Uterus becomes flaccid
- Cervix is tightly closed

Signs of Heat
Primary sign:
- Stand to be mounted by other herd mate (standing heat) and attempt to mount
on other cows
 Secondary signs:
- Nervousness and restlessness; reduction in the milk production
- Roughened tail head
- Dirty streaks and marks on lower hips
- Vulva is swollen; due to which wrinkles disappears
- Frequent urination and clear transparent stringy mucus discharge from vulva.
Internal Changes:
- Dilatation of cervix
- Marked tonicity in uterus
- Fully matured (Graafian) follicle(s) on ovary
- Hyperemic vagina
- Increased secretory activity of uterine, cervical and vaginal glands.

Duration of phases of estrus cycle in domestic animals

Phase/stage Cow Ewe Mare Doe

Estrus cycle (d) 21 17 22 21
Proestrus (d) 3-4 2 -3 2 -3 2 -3
Estrus (hrs) 12 - 18 24 -36 4-8 (d) 30-40
Metestrus (d) 3-4 2-3 2- 3 2- 3
Diestrus (d) 10 - 14 10 -12 10 -12 13 -15
Ovulation time 10-12 hrs after Late estrus 1-2 days before 1 -2 hrs after end
end of estrus end of estrus of estrus

PREGNANCY DIAGNOSIS
Techniques of pregnancy diagnosis are divided into three categories:
(i) Return to the estrus
(ii) Direct method; also know as clinical method
(iii) Indirect method; also called immunological method
Direct Method

Rectal palpation Ultrasonography Visual observation

Mammogenesis Fetal ballottement

Indirect Method

Hormones Preg. Associated factors

Progesterone Estrone sulphate PMSG Early preg. factor Preg. associated

protein

RECTAL PALPATION
Rectal palpation is the fastest, cheapest, easiest and more accurate method of pregnancy
diagnosis.
Anatomical Features for rectal palpation of reproductive tract
- The reproductive tract of bovine is suspended with the dorsal body wall with
the help of broad ligament.
- The posterior part of the tract is located in the pelvic cavity (vagina, cervix and
some part of uterus) whilst the anterior part of tract hangs over the pelvic brim
(horn uteri, oviduct and ovaries) and suspended in the abdominal cavity.
- The reproductive tract is located immediately below the rectum.
- The rectal wall of the cow is so strong that chances of tearing are minimal.
- The rectum of the cow allows to grasp the reproductive tract structures
Indications for rectal palpation
- Pregnancy diagnosis
- Artificial insamination
- Embryo transferring and flushing
- As diagnostic aid in uterine and other genital organs disorders.
- To determine the suitability of the animal for breeding
- To inspect the status of genital organs; ovarian picture
- As clinical diagnosis of cystic ovarian disorder

Findings in Rectal Palpation

Cervix:
It is a landmark structure in bovine for palpator. It is recognized as a firm, rope like structure
(cervical rings; give a typical feeling) lying on the mid line of the pelvic floor.
The size of the cervix varies with the age, stage of the reproductive cycle and state of the
reproductive tract (either normal or abnormal).
In normal non-pregnant adult animals, it ranges from 7 to 10cm in length and 3 to 4 cm in
diameter. Increased in age, number of parturition and any infection in reproductive tract results
in larger size of the cervix.
Uterus:
It can be examined by following the cervix forward. To do so, the cervix is grasped in hand so
that fingers lie over the tract and thumb underneath it. The external bifurcation where two horns
of uterus separate each other can be palpated. Both uterine horns should be roughly of the
same size. This does not happen in late pregnancy and early postpartum period. They vary
greatly in size depending on the reproductive state of the animal.
Ovaries:
Ovaries are located on each the side of the uterus at variable distance. Once located, ovaries
can be grasped with fingers and then move your fingers along the entire surface of ovary and
palpate the structure (if any) on the ovary by using thumb. Fully developed follicles gives
feelings of a blister like fluid filled structure but CL is liver like which is protruded from the
surface of the ovary. Just after ovulation, it is possible that you may find ovulation fossa on the
ovary.

Note: oviducts are not found by rectal palpation unless some pathological condition develops.

Size of Fetus
Age of fetus Size (resemblance)
2 month Mouse
3 month Rat
4 month Small cat
5 month Large cat
6 month Beagle dog
7 month to onward Parts of the fetus may be palpated

Fetal Membrane Slip Test

Time to perform Result
32 days Small thread in 1 horn
45 days Small strings in 1 horn
60 days Strings in both horns
70 days Large strings in both horns

Size of Placentomes
Gestation Size of placentome (cm) Resemblance
75 days 1 – 1.5 Pea size
100 days 1.5 – 2.5 Dime
115 days 2.5 – 3.0 Nickle
125 days 3.0 – 4.0 Quarter
150 days 4.0 – 5.0 Half dollar
180 days 5.0 – 6.0 Silver dollar

Palpable structure during different stages of pregnancy

Stage (day) Ut. Position Ut. Diameter (cm) Palpable structures
35 – 40 Pelvic floor Slightly enlarged Uterine asymmetry, +ve fetal
membrane slip test
45 -50 Pelvic floor 5.0 – 6.5 Same as above
60 Pelvis / abd 6.5 – 7.0 Same as above
90 Abdomen 8.0 – 10.0 Small placentomes, fetus
120 Abdomen 12.0 Samll placentomes, fetus, freemitis
150 Abdomen 18.0 Fetus, freemitis

Estimation of Stages of Pregnancy

30 days
- Asymmetry of uterine horn (slightly)
- Presence of corpus luteum (CL) on ovary
- +ve membrane slip test
- Amniotic vesicle – pea size
- Uterus – in pelvic cavity
35-40 days
- Uterus is at the floor of the pelvic cavity
- Thinning of uterine wall
- +ve membrane slip test
- Presence of corpus luteum on ovary
- Amniotic vesicle – Hen egg yolk size
45 -60 days
- Gravid horn somewhat enlarged
- Fetus attached to the uterine wall
- Amniotic vesicle – small egg of hen.
60-90 days
- Uterine horn is about 3 inches in diameter
- Placentomes – (1-1.5 cm) pea size
- Fetus is about 6 ½ inches long = rat like
- Uterus located on the floor of pelvic brim
120 days
- Fetus is about 10-12 inches long = Small cat like
- Freemitis is a hallmark feature
- Uterus is descended down in the abdomen
- Placentomes - 1.5-2.5 cm
150 days
- Fetus; large cat like
- Palpation of uterus is difficult due to descending in abdomen
- Freemitis is a hallmark feature
- Placentomes - Quarter size
180 days
- Fetus size; beagle dog
- Freemitis is palpable
- Placentomes – (5.0-6.0 cm) Silver dollar
- Uterus is descended in the abdominal cavity
210-270 days
- Freemitis is a hallmark feature
- Fetus parts are also palpated
- Fetus ascend again in the pelvic cavity

Methods of pregnancy diagnosis in the cow

Methods Earliest time
Early pregnancy factor (EPF)/early conception factor (ECF) 3 days
Realtime ultrasound (direct imaging) 13 days
Failure to return to estrus and persistence of corpus luteum 21 days
Progesterone conc. in plasma and milk 21-24 days
Assay of pregnancy specific protein B (PSPB) 24 days
Palpation of the atlantochorion (membrane slip test) 33 days
Unilateral corneal enlargement, thinning of uterine wall and fluid-filled 35 days
fluctuation of enlarged horns
Palpation of early fetus when the amnion loses its turgidity 45-60 days
Palpation of the caruncle/cotyledons (placentomes) 80 days
Presence of freemitis 85 days
Oestrone sulphate in the blood or milk 105 days
Palpation of the fetus 120 days
Source: Arthur’s Veterinary Reproduction; Page 87