PREGNANCY DIAGNOSIS OF DAIRY
COWS AND HEIFERS FOR AI TRANEE’S
The Four Positive
Signs of pregnancy
–Fetal membrane slip
–Amniotic vesicle
–Placentomes
–Fetus
By Feyisa girma
AT THE END OF TRAINING THE TRANEE’S SHOULD
ABLE TO PERFORM/UNDERSTAND
Different Methods of pregnancy diagnosis in cows/heifers and its role
Physiological/pathological
diagnosis
condition
confusing
with
pd/Differential
Purpose and Economic significances of early pregnancy diagnosis
As Pregnancy diagnosis is an important tool to measure the success of
reproductive management of a cattle herd.
Estimation of gestation stage ,physiological and pathological condition of
RT
Identify positive and negative sign of pregnancy and golden rule of rectal
palpation
Changes/stages of pregnancy and structure developed during pregnancy
etc………….
Introduction
The accurate and early pregnancy diagnosis is essential for
 the maintenance of high levels of reproductive efficiency by early
identification of fertility problems and
 decreasing the interval between AI services and calving.
The purpose for examining cows for pregnancy
is not to detect those that are pregnant, but to
detect those that are not pregnant
Why ……
The three Methods of Pd diagnosis
1.Direct /clinical methods of pregnancy diagnosis
 trans rectal palpation
Rectal palpation/Trans rectal palpation of the uterus for pregnancy diagnosis in
cattle was first described in the 1800’s (Cowie, 1948) and is the oldest and most
widely used method of pregnancy diagnosis up to now.
 ultrasonography.
2.BIOCHEMICAL TEST/LAB/INDIRECT METHOD
Biological changes in milk, blood, urine, cervical mucus
 early conception factor (ECF)/Early pregnancy factor(EPF) test
 early Pregnancy-associated Protein Interferon-τ
 hormonal measurement protocols (progesterone assay)
 Pregnancy associated Glycoprotein(PAG)
 ESTRONE SULFATE
3.Visual/ Manegmental method
 Development of udder
 non-return to heat/estrus
 History taking
Why it is far from perfection ?
 Body size increment
B/C: PD is verified on the bases of physiological & morphological changes of a
fetus and dam that are manifested by internal changes on (ovaries, uterus, uterine
arteries, cotyledons & fetal development) but not only external indicators changes
on (udder, abdomen, vulva, body weight gain, & fetal movement)
The general objective of understanding rectal early pregnancy
identification is
 To
measure the success of AI service efficiency
 To control fertility problems
 For reproductive performance
 For heat synchronization
Development of the Early Bovine Embryo
Event
Day
Estrus
0
Ovulation
1
Fertilization
1
First cell division
2
8-cell stage
3
Migration to uterus
Maternal recognition of
pregnancy
Attachment to uterus
Adhesion to uterus
Placentation
Birth
5-6
15-17
19
21-22
25
285
Figure below shows that the ovium is fertilized at the ampulla of the
oviduct, and the fertilized Egg develops moving down the oviduct. About
4-5 days after the fertilization, the embryo Will enter into the uterus.
Fetus & placenta
physiology of cattle Pregnancy
Gestation/:- The Periods of pregnancy/ development in an infant`s life are divided into
three part ( Zygote, embryo and fetus.) usually from conception to parturition.
Gestation period of domestic animals depends on the species and breeds. An average
gestation length in cattle is 279 days (275-285days).
Pregnancy /The physiological condition of a female organism holding offspring
in the uterus.
It starts from union of ovum & sperm for zygote formation and that ends up
with abortion ,still birth or parturition.
Zygote: - is the fertilized egg to day 12.
Embryo: -
is the developing organism from day 13 to 45.
It is age that begins the formation of parts & organs.

Fetus: -
This is the stage from day 46 to parturition
what are the Advantage of/Economical significances of
accurate & Early pregnancy diagnosis/detection? Or why ?
Improve reproductive performance.
Earlier the pregnancy diagnosis performed, the more profitable is the
return.
Management.
Feeding.
Sale
Monitor development of pregnancy.
Confirm absence of twins.
Monitor early embryonic death.
• Thus, any method for early pregnancy diagnosis must be integrated as a
component of the overall reproductive mang’t strategy
The goal of any method used to do the PD is to
determine the pregnancy status with
 100% accuracy
 No false positive
 No false negative
 Determine the pregnancy as early as possible.
 The ability to age the concept us.
 Be able to determine the viability of the concept us.
 Possible determine the sex of the concept us.
 Have the result immediately.
clinical method
Ultrasound
Techniques of using ultrasound:- a probe is passed over the cow’s abdominal
wall or into the rectum to transmit two-dimensional images to a monitor that
can be viewed by a technician. It is the easiest method of pd checking.
Organs of the reproductive tract, as well as a developing fetus, can be viewed
using ultrasound technology.
Trans-rectal ultra sonography after stages of
pregnancy (day28/ 30 or later).
At day 30 it is possible to observe the foetal
heart beats.
 Detection of amniotic vesicle, fluid, foetus,
and foetal heartbeat.
Sex foetuses from day 55 - 75 of pregnancy
2.RECTAL PALPATION
Pregnancy is diagnosed by per-rectal examination of the animal and the
anatomical changes in the reproductive organs like ovaries, uterus, uterine
artery and palpation of foetus is taken as the indicator of pregnancy
 Pregnancy diagnosis by palpation is an important tool
to measure the success of reproductive management
of a cattle herd
 Skilled practitioner can confirm pregnancy at 30-40
days of gestation.
 Detection of uterine tone and corpus leutum at 18-24
days after service gives suggestive idea about
pregnancy.
Cont…
 The corpus luteum on the ovary and no tone in the uterus 21
days post-breeding indicates high progesterone and the cow
may be pregnant
 Re-examination at 40-50 days with typical changes of gravid
uterine horn assures up to 100% accuracy of conception
N.B: Rectal palpation helps not only for identification of nonpregnant from pregnant but it helps also for estimation of the
gestation stage, Characterization of the physiological and
pathological status of the uterus and/or the ovaries and other
relevant events that is associated with ovulation or infertility
The Golden Rules of Rectal Pregnancy
Examination/Palpation
1.
Examine the entire tract before declaring the cow is open.
2.Pregnancy exam. must always be the first step (If you are not sure, recheck. This maybe in a few minutes, maybe tomorrow or after a
while.
3. Take a time and be patient
4. Find one of the positive signs of pregnancy before you call a cow is
pregnant or not.
Example:-The only positive signs of pregnancy in the
cow are fetus, amniotic vesicle, fetal membrane slip
Techniques of pregnancy diagnosis by
rectal palpation
1. The first step is to locate the cervix.
Usually the cervix lies on the midline of the floor of the pelvic cavity, but may
be displaced laterally by a full bladder or a short broad ligament
2. Retract back the cervix to
pelvic cavity
 In principles if there is some thing inside uterus, it does not retract
back in respective to its content.
3. Find the bifurcation of the uterus
 to examine left and right horn easily
.
Cont.…
4. Gentle palpate and Find the positive sign of pregnancy
first feel for uterine asymmetry and tone.
Second, feel for fluid in the larger horn.
At 35 days of pregnancy the pregnant horn will feel slightly larger and non-tonic
1st month of gravid uterus is about 1.5 times non-gravid uterus with 80-100 ml of
fluid.
The fluid has a smooth velvety feel because the uterine wall is thin during pregnancy.
The fluid almost feels like a balloon that is not totally filled with water.
 You must systematically feel the uterus for
the amniotic vesicle,
the fetal membrane slip,
the fetus and
the Placentomes
5. If cow/heifer is open palpate the ovary
Open uterus
 The "open" uterine horns are coiled on the front edge of the pelvis or,
in older cows, may hang slightly into the abdominal cavity.
 The entire tract may be held in the hand at this stage.
 Slight pressure by the middle finger will separate the horns of the
uterus.
 Manual rectal palpation of the cervix, uterus, ovaries and supporting
structures have a central role in determining pregnancy and status
of reproductive organs.
 So inseminators should develop accuracy skill of this technology.
 on palpation of uterus, Early pregnancy indicators are tone,
consistency, and asymmetry of two horns. So attention has to be
given for these indicators
palpation of uterus to identify +P from -P
the content
tone
Size and symmetry of the two uterine horns
consistency and
uterine position
structures developed during pregnancy that helps to
identity pregnancy such as:1. Amniotic vesicle(30-45 days)
2. fetal membrane slip
3. Placentomes
4. freemitus
Position of pregnant uterus:  age of the animals &
stage of pregnancy (for details see changes during pregnancy).
Uterine asymmetry: - Due to accumulation of fluids within the
pregnant uterine horn, one of the initial signs of pregnancy is a difference in
size of uterine horns
Uterine asymmetry is the simplest (easiest) indicator of pregnancy for
inexperienced personnel.
.
Embryonic Vesicle /Amniotic vesicle
The embryonic vesicle forms around the fertilized ovum after it
has moved into the uterine horn.
The
embryonic
vesicle
causes
an enlarged
in the
a thin
membrane
filled
with fluid
and the area
embryo;
thehorn
embryonic
vesicle serves to protect the embryo and nourishes it, until it
attaches and becomes a fetus.
pregnancy in cattle can be terminated by manual rupture of the
amniotic vesicle (Ball and Carroll, 1963),hence care should be taken
in the free portion of uterus at 30-45 days of pregnancy is helpful
diagnosis because of its spherical structure as feeling like a softshelled hen's egg
35 day Amniotic Vesicle
Feeling Amniotic Vesicle
Damage to the Amniotic Vesicle
Abortion
55 day Twins
Membrane slip
along the greater curvature within the uterus is best performed from 35-90
days of gestation in determining early pregnancy status.
The technique is carried out by gently pinking up & pinching either horn
and felling the fetal membrane slip b/n the thumb and fingers.
avoided rough pinching of the uterus particularly over the amniotic vesicle,
This technique is especially valuable in differential diagnosis of pregnancy
from uterine distention with pyometra and or muco-metra
Slip Membranes
Slip Membranes 50 days
Adulatory vibration of middle uterine artery(Fremitus )
This is 1st recognized about 80-120days of gestation.
The artery lays over the dorsal part of the shaft of the ileum into
pelvic cavity
 In heifers may be noted as early as 60-75 days of pregnancy,
while
in older cows at 90 days.
 Especially about 5-6 months of pregnancy
Uterine Artery
Placentomes
Placentomes are the structures formed by the union of maternal caraculs
and fetal cotyledons by which the placenta is attached to the uterus.
 They can be felt as ovoid thickened structure floating when balloting
through the uterus from 80-120days of pregnancy
Cotyledons – soft, button-like nodules on the fetal membrane that attach to
the caruncles lining the uterus during fetal development.
Caruncles – flattened, oval, raised prominences that line the wall of the
uterus and serve as connecting points for the fetal membrane.
After connecting to the cotyledons, the caruncles serve as a nutrient and
waste exchange site between the fetus and its mother
Caruncle & Cotyledon
Fetus
 After 90 days of gestation the fetus can be palpated.

At about 120 days the head is about the size of a lemon and enlarged
fetus fills some portion of the abdominal cavity and is easier to feel than the
3-month of age.

Between5-6 months of pregnancy in cows with deeper abdominal cavity it
becomes difficult to palpate the fetus.

Starting from 6 months onwards parts of the fetus can be palpable but, in
heifers and thin abdominal walled cows it can be palpated earlier.
 An important signs between 5th to 6th months are the thought cervix lies on
the pelvic brim and Placentomes.
 From 7-9 months parts of the fetus are easily felt.
105 day Fetus
Fetal Size in Relation to Common
Animals/Estimation of stages of Pregnancy
There is a rule of thumb that is quite useful in estimating fetal
age based on the size of the fetus in relationship to the size of
some well known animals.
Stage of Pregnancy Size of fetus
2 Months
Mouse
3 Months
Rat
4 Months
Small Cat
5 Months
Large Cat
6 Months
Beagle Dog
Estimation of Stage of Gestation
Why do you need to estimate the stage of pregnancy?
There may be lost records and you need to predict dry off dates of the herd.
You may need to confirm AI dates, or sell/feed preparation/milk day
 AI date may not match what you feel.
You may be asked to estimate the stage of gestation and parturition dates for beef
herds
Parameters /Based on
–Size of A.V.
–Size of placentomes
–Size of middle uterine artery
–Size of fetus
–Fetal crown-to-nose length
Pregnancy size from day 30 to >day 210
In 30-day pregnancy, the uterus will be
filled with fluid and feel slightly thinner.
One horn will be enlarged a
little more than the other.
This enlargement in the horn is the
embryonic vesicle.
The spherical vesicle is nearly ¾” in
diameter and is filled with fluid.
In most cases, on the side of the uterus
(uterine horn) that the vesicle is found,
a corpus luteum on the ovary will also
be found
30 days:
Slight asymmetry of horn
Presence of CL
Chrio-allontoic membrane
Amniotic vesicle pea size
Uterus in pelvic cavity
Rectal Palpation
• 30 to 45 days
 Foetus is 1.5 to 2.5 cm long.
 Cervix & uterus are usually in the pelvic
canal
 One horn is slightly enlarged and fluidfilled
35-45 days:
Uterus in pelvic cavity
Thinning of uterine wall
 Membrane slip technique used to detect
Presence of CL
pregnancy
Chrioallontoic membrane
Amniotic vesicle size is
 Proceed with caution because damage
yolk of hen (0.7 cm).
to membrane can cause pregnancy to
be terminated.
45 day:
Uterus in pelvic cavity
Thinning of uterine wall
Presence of CL
Chorioallantoic membrane
Amniotic vesicle size small egg of
hen
Rectal Palpation
• 45 to 60 days (60 days is A in
figure)





Foetus is 3 to 7 cm long.
Cervix is typically in the
pelvic cavity.
Pregnant uterine horn
begins to fill with fluid
with little fluid in nonpregnant horn.
Amniotic cavity is about
the size of a hen egg.
Use membrane slip
technique. Caution: too
much pressure can
damage membranes
60 day:
Uterus in pelvic cavity
Presence of CL
Chorioallontoic m/m
Amniotic vesicle 9-10 cm
Fetus size 2.5 inch
Fetal membrane slip test
is positives
60-Day Pregnancy
The uterus has now enlarged
until one horn is approximately
the size of a banana and
measures 8” to 10” long.
The weight of the fetus and
other contents has pulled the
uterus over the pelvic ridge into
the body cavity The fetus
measures 2 ½” in length and
the embryonic vesicle is still
prominent
60 day:
Uterus in pelvic cavity
Presence of CL
Chorioallontoic m/m
Amniotic vesicle 9-10 cm
Fetus size 2.5 inch
Fetal membrane slip test is
positives
Rectal Palpationa
• 60 to 90 days (90 days is B in
figure)
Foetus is 8 to 30 cm long.
Foetus about size of a small rat.
Difficult to move hand
completely around the pregnant
uterus.
90-Day Pregnancy
The uterus in this stage is considerably
larger because of increased fluid and
fetal growth.
The fetus is now nearly 6 ½” long and
is located
on the floor of
the body cavity
90 day:
Uterine horn 3 inches in
diameter
Placentome (1-1.5 cm) pea
size
Presence of CL
Chorioallontoic membrane
Fetus 6.5 inch (rat size
Rectal Palpation
• 90 to 120 days (120 days is C in figure)
 Foetus is 35 to 50cm long
 Fetus is the size of a small cat.
 Start to feel the placentomes.
120-Day Pregnancy
The fetus in this stage is 10” to 12”
long and is still on the floor of the
body cavity.
The head of the fetus is nearly the
size of a lemon and may be the first
portion of the fetus that the palpator
touches
Because the fetus is larger in this
stage, it is normally easier to locate.
Each cotyledon is 1” to 1 ½” in
length and the uterine artery has
increased in size (¼” in diameter
120 day:
Uterus descending in pelvic brim
Placentome (1.5-2.5 cm)
Presence of CL
Chorioallontoic membrane
Fetus 10-12 inch (small cat size)
Presence of Fremitus (uterine
artery)
Rectal Palpation
• 120 to 150 days (150 days is D in figure)
 Cervix is almost completely over the pelvic
brim.
 Uterine body and horns are not easily
palpated.
 Placentomes present
 Fetus is the size of a large cat.
150-Day Pregnancy
The main change from this stage until
birth is in the increase in the size of the
fetus.
At 150 days, the fetus is the size of a
large cat (approximately 16” long).
The uterine artery is ¼” to 3/8” in
diameter and each cotyledon is 2” to 2
½” in diameter.
Palpation of the fetus may still be
difficult because of its low position in
the body cavity.
150 day:
Abdominal descending of uterus
Palpation is difficult because fetus is in
abdominal cavity
Placentomes (2.5-4 cm)
Presence of CL
Fremitis (pulse feel in uterine artery)
Fetus is large cat sized
180-Day Pregnancy
At this stage, the fetus is still deep in
the body cavity.
The uterine artery is 3/8” to ½” in
diameter and the cotyledons are
larger.
From 180 days until birth, the fetus
can be made to move by grasping its
feet, legs, or nose
180 day:
Abdominal descending of uterus
Placentome (4-5 cm) equal to large
coin
Presence of CL
Fremitus
Fetus is small dog sized
Rectal Palpation
• 180 to 210 days (150 days
is E in figure)
 Pregnancy easy to detect.
Foetal head is at or near pelvic cavity and can be
palpated.
 Fetus about the size of a beagle dog.
 Placentomes
 There is a very strong buzz to the uterine artery.
210 day:
Fetus easily felt due to increased size
Placentome
Fremitus
Rectal Palpation
• > 210 days to term
 Pregnancy is easy to detect.
 Foetal calf quite often located in pelvic cavity and its
head is easy to feel.
 From seven months until calving, the fetus may be
easily felt because of its increasing size
 Very strong buzz to the uterine artery when digital
pressure is applied to it.
 Bounce foetus like a basketball.
Uterine position/diameter and structures during
pregnancy. (source: P.J. Hansen)
Stage of pregnancy
(days of gestation)
Uterine diameter
Uterine position
35-40
Pelvic floor
Slightly enlarged
Uterine asymmetry/fetal
slip
45-50
Pelvic floor
5.0 - 6.5 cm
Uterine asymmetry/fetal
slip
60
Pelvis/abdomen
6.5 - 7.0 cm
Membrane slip
90 Small
Abdomen
8.0 - 10.0 cm
Placentomes /fetus (1015 cm long) 120
120
Abdomen
12 cm
Placentomes/fetus (2530 cm long)/fremitis
150
Abdomen
18 cm
Placentomes/fetus (3540 cm long)/fremitis
Palpable Structures
Pregnancy stages
For easy pregnancy determination,
pregnancy can be divided in to three
stages:
Early (1-3months of pregnancy)
 Mid (4-6 months of pregnancy) and
 Late pregnancy (7-9 months of pregnancy)
Early pregnancy indicators (1-3 months)
 asymmetry of the uterine horns
Different Size of Horns
 decrease in the tone uterus or
flaccid uterus of the pregnant horn
 Appreciation of an amniotic vesicle.
 Fetal membrane slip
 fluid contents in the pregnant horn (later both horns)
 A palpable corpus luteum on the ovary on the same side as the
pregnant horn
32 day pregnancy
60 day fetus
70 day Conceptus
2.5 Month Conceptus
Diagnosis in mid (4-6months)
• Thought cervix located on the pelvic brim
•
• the uterus can not be retracted
• Place tomes, and sometimes the fetus, are palpable
• Pulsation of middle uterine artery (Fremitus) 80-120days
5 months, CL on ovary on other side
Late stage 7-9 month
 Fetus located in pelvic or abdominal cavity
 Limbs ,heads of fetus palpable structure
During PD simple indicators of pregnancy at different
stages for inexperienced inseminators
Stages of
pregnancy
uterine position
palpable structures
Early
(1-3 months )
In heifers; pelvic
floor
In older; partially
in abdomen
uterine asymmetry,
Mid
(4-6 months)
abdomen
Placentomes & fremitis
Late
(7-9Months)
Abdomen
fremitis and fetus (in
advanced gestation fetal
extremities & head are
within pelvic cavity)
Differential diagnoses
ENDOMETRITIS, METRITIS
UTERINE DISTENTION (PYOMETRA, MUCO-METRA,
MUMMIFICATION),
UTERINE ABSCESS
OLD CHRONIC INFECTION OF THE UTERUS
TUMOR...ETC
 BLADDER
OVARIES
LARGE UTERUS
THE RUMEN.
Fetal Mummification & Maceration
Mummification - is a condition in which the fetus dies and the fluid &
soft tissues are reabsorbed.
fetal membranes wrapped tightly around the fetus & cervix of the
uterus closed.
Maceration - a condition in which fetus become mashed up (only
some bones & rotten tissue are found) in uterus.
 in such cases the cow does not come into heat until the fetus or
remaining parts of it have been expelled.
Handling
uncomplicated cases could be expelled manually
injection of estilbestrol or estradiol can help for expelling
Mummified fetus inside the uterus
Mummified fetus inside uterus
Macerated fetus ( bonny remnant of fetus )
Macerated fetus ( note bones of macerated fetus )
summary :Pregnancy Signs in Rectal Palpation:
Asymmetry of uterine horn
Position of uterus
Presence of fetal membrane - allento-chorion (upto 30-90 days
these are detected), amniotic vesicle (as early as 35-40 days
these are detected), placentomes (formed by fusion of crunkles
and cotyledons. After 90 day detectable).
Presence of conceptus/fetus.
Presence of CL on ovary
Pregnancy diagnosis by palpation is an important tool to measure the
success of reproductive management of a cattle herd.
Comparison of early pregnancy
diagnosis techniques
Pregnancy Diagnosis Technique
Rectal Palpation
Early Testing
Time
♦
Diagnosis
Pregnancy
Accurately
Diagnosis NonPregnancy
Accurately
♦♦♦
♦♦♦♦
Ultrasound
♦♦
♦♦♦♦
♦♦♦♦
Milk Progesterone
♦♦♦
♦♦
♦♦♦
♦♦♦♦
♦
♦
ECF
Positive sign of pregnancy at rectal palpation
Stage of Membrane Amniotic
pregnancy
slip
vesicle
30 days
+
+
45 days
+
+
60 days
+
+
+
+
+
75 days
90 days
105 days
4 months
5 months
Fremitus A.uterine media
Foetus Placentomes
+
+
+
+
6 months
7 months
+
+
+
+
+
+
+
+
Ipsilateral
+
+
+
+
+
Contralateral
+
+
+
Generally,
Positive pregnancy at different stages can be suggested based
on a combination of some or all of pregnancy signs.
 In heifers or young cows 2 to 3 months pregnant, the uterus
often lies in the pelvic cavity and about over 95% of the fetus
can be palpable.
By 5-6 the uterus is well-forwarded and down warded in the
abdominal cavity the fetus cannot be palpated, so that in some
cases only cervix can be palpated, however at this stage about
40-70 % of the fetus can be palpable.
By 6-7 months of the fetus becomes large enough and nearly in
all cows it can be palpated.
10Q