Dr.

Anil Kumar Nahak

02 ARGO/PhD/12
Department of Animal Reproduction , Gynaecology &
Obstetrics,
C.V.Sc.& A.H., OUAT, BBSR
The critical period for signaling by the conceptus to block luteolysis and allow
pregnancy to be established is called maternal recognition of pregnancy.

The developing embryo enters the uterus between 2-5 days after ovulation
depending on the species.

For the early embryo to become an established pregnancy, two major events
have to take place:
1) PGF2α synthesis and secretion must be stopped
2) Progesterone must be maintained
The conceptus must provide a timely (before luteolysis) biochemical signal
- Conceptus signals its presence to the dam - Signals enable pregnancy to
continue
- If a signal is not delivered quick enough, luteolysis will occur, progesterone
will decline,and the early embryo will die
In Sheep and Cow:
estrogen produced by preovulatory follicles
on the ovary
stimulates

expression of receptors for oxytocin in the uterine endometrium

oxytocin released from CL/posterior pituitary

stimulates

episodic PGF2α release from endometrium by enzymatic conversion of

arachidonic acid

In pregnant ewes oxytocin receptor diminishes, & luteal oxytocin is inhibited .

This reduces episodic PGF2α release ( 1.3 pulses/24 hr between day 14
and15 of pregnancy versus an average of 7.6 pulses in non pregnant ewes
during the same period.)
In pregnant ewes conceptus IFN-τ produced between days
12 and 21
supresses expression of estrogen receptors
results
Lack of up-regulation of oxytocin receptor numbers
Leads to
insensitivity of luteal oxytocin release

dampen the pulsatile release of PGF2α below critical

threshold level of 5 pulses / 24 hour.
luteolysis does not occur
 Interferons: glycoproteins that may possess antiviral action and alter
the function of target cells
In sheep these low molecular wt protein with acidic characteristic was
called as ovine trophoblastic protein-1
Later termed as - Ovine interferon-tau (oIFN τ) and in bovine
species it is termed Bovine interferon-tau (bIFN τ)
 - The trophoblast produces oIFN- τ and bIFN- τ between d 12 to d
21 as the conceptus elongates (spherical to tubular to filamentous)
 - oIFN- τ and bIFN- τ also promotes protein synthesis thought to be
critical to preattachment embryonic survival.

In pregnant cow the endometrium also produces another

substance called endometrial prostaglandin synthesis
inhibitor (EPSI) which specifically depresses PGF2α
synthesis. It appears to be a linoleic acid
In sow
The conceptus of the pig produces estradiol between d 11 and 12 after
ovulation (coincides with the elongation of the conceptus Estradiol
serves as the signal for maternal recognition of pregnancy
What happens to PGF2α ?
- PGF2α is produced by the endometrium re-routed into the uterine lumen
and metabolized, rather than being drained by the uterine veins luminal
PGF2α has little access to the circulation and can’t cause luteolysis.
- The sow must have at least two conceptuses in each uterine horn for
pregnancy to be maintained
- If there no two conceptuses, PGF2α is secreted in an endocrine manner and
luteolysis will occur, and pregnancy will be terminated.
In mare
Mechansim of action:
- The presence of the conceptus helps to prevent Luteolysis.
- The equine conceptus does produce proteins; their role in maternal
recognition is unknown
- The conceptus must migrate within the uterus between 12 to 14 times per
day during days 12, 13, and 14 of pregnancy in order to inhibit PGF2α
production.
1) This migration appears to be very important because the early
embryo does not elongate
2) Conceptus must “touch” enough receptors or secrete “proteins”
and place near (on) receptors to maintain pregnancy.
- Unlike sow, the conceptus produces estrogen(estradiol & estrol)
- Besides low/ high mol.wt. proteins ranging from 22000 to 65000 daltons
have also been isolated.
- May be these secretory protein modulate PGF2α production or it inhibits
it’s sensitivity to CL
Maternal Recognition of Pregnancy (in a nutshell)
Chemical message (Usually hormonal)
which results in maintenance of the CL.

species Embryonic Day of Day of definite

Signal maternal attachment
recognitio
n
pig Estrogen 12 18
Sheep and goat interferon 12-13 16
cow Interferon (b IFN-τ) 16-17 18-22
mare Small unknown 14-16 36-40
Peptide
Dog and cat not confirmed not 13-17
confirmed
Importance
-to identify non pregnant animals soon after mating or
insemination so that production time lost from infertility
may be reduced ,
-to certify animals for sale or insurance purposes,
-to reduce waste in breeding programs using expensive
hormonal techniques,
- to assist in the economic management of livestock.
 Techniques of pregnancy diagnosis

Managemental method
Clinical methods Immunological
or
tests
nonreturn to estrous

Rectal Pregnancy
ultrasonography
palpation hormones associated
substances

Progesterone, Early pregnancy factor,

estrone sulphate, pregnancy specific
equine chorionic protein B
gonadotrophin
Diagnostic methods Earliest
time
Early pregnancy factor (EPF)/early conception factor (ECF) 3days
Realtime ultrasound (direct imaging) 13 days
Failure to return to oestrus and persistence of corpus luteum 21 days
Progesterone concentration in plasma and milk 21-24days
Assay of pregnancy-specific protein B (PSPB) 24 days
Palpation of the allantochorion (membrane slip) 33 days
Unilateral cornual enlargement and disparity in size, thinning of the 35 days
uterine wall, fluid-filled fluctuation of 35 days
enlarged horns
Palpation of the early fetus when the amnion loses its turgidity 45-60 days
Palpation of the carnucles/cotyledons 80 days
Hypertrophy of the middle uterine artery until presence of fremitus 85 days
Oestrone sulphate in blood or milk 105 days
Palpation of the fetus 120 days
1. cessation of oestrus :
false +ve or false –ve result may occur due to-
silent second estrous
Pathological conditions of uterus, ovary
 anestrous cases
Early embryonic mortality
Might have been sold or disposed or not reported
Gestational estrous

1. Increase in abdominal size : not reliable in pluriparous animals

2. Abdominal ballottement :possible as early as 7 months in some small breed

balloting right flank- foetal movement, womb felt
-fetal heart can be asculted in right flank between 6-7 th month
fetal electrocardiography used to study fetal heart

3. Mammary gland : - glossy, enlarged and edematous,

teats-waxy tumified appearance.
-observed after about 4 months
-best remarkable in primigravida
-
Trans rectal palpation
 Palpation of the amniotic vesicle.
- towards the end of 1st month of pregnancy, 30 -50 days of
pregnancy
- a distinct, round, turgid object.1–2 cm in diameter floating
in the allantoic fluid.
-this technique is dangerous because of the possibility of
rupture of the amniotic sac or of the embryonic heart
 Palpation of the allantochorion (membrane slip).
-best at 35 to 90 days
-possible as the intercotyledonary part of the fetal membrane
is free.
-it enables the differential diagnosis of pregnancy from
mucometra or pyometra
 Unilateral cornual enlargement.
-assymetry is due to vesicle, placental fluid, foetal
membrane and fetus which gives a fluctuating feel with a
good tone
pregnant uterus – located on pelvic cavity upto 3-4 months

6 week 12 week 16 week 19 week

 Stage of
pregnancy (d Uterine
ays of Uterine diameter Palpable Structures
position
gestation)

Uterine asymmetry/fetal
35-40 Pelvic floor Slightly enlarged
slip
Uterine asymmetry/fetal
45-50 Pelvic floor 5.0 - 6.5 cm
slip
Pelvis/abdome
60 6.5 - 7.0 cm Membrane slip
n
Small placentomes/fetus
90 Abdomen 8.0 - 10.0 cm
(10-15 cm long)

Placentomes/fetus (25-30
120 Abdomen 12 cm
com long)/fremitis
Placentomes/fetus (35-40
150 Abdomen 18 cm
cm long)/fremitis
Palpation of the early fetus.
At about 45–50 days of gestation the amniotic sac
becomes less turgid, and it is sometimes possible to palpate directly the
small developing fetus.This should be done with care.

Pregnancy Foetal length

Fetal size at various stages of
month
pregnancy in relation to the size
1 .8-1
of some commonly-known
animals 2 6
3 15
Stage of
Animal 4 28
pregnancy
2 months mouse 5 40

3 months rat 6 52

4 months small cat 7 70

5 months large cat 8 80

6 months beagle dog 9 90

 Palpation of caruncles/cotyledons:
- placentomes are formed during 60-70 days but after about 80 days they
can be identified as discrete structures in the midline
-felt like a sackful of small potatoes (8-10cm)
` -but once the uterus has sunk into the abdomen between 5-7
months, it becomes impossible to palpate them
 Palpation of the cervix:
-as pregnancy advances there is tension on cervix, becomes less mobile,
pulled forward and dowmward over the pelvic brim
 Hypertrophy of the middle uterine artery and development of fremitus
-The artery runs in the broad ligament, along a tortuous course, passing
downwards, forwards and towards the midline over the pelvic brim close to
the junction between pubis and ilium.
-at pregnancy it is pulled more cranially and located5-10cm cranial to shaft of
uterus. light pressure detects a characteristic feeling like whirr or thrill or
fremitus, heavy pressure detects a pulse wave
-at 60-75 days , the diameter is 0.15 to 0.3 cm.
first recognised at 80-120 days of pregnancy
 Palpation of the late fetus.
From 7th month to termination along with cotyledons, fremitus , fetal parts
are also palpable.
 Detection of pregnancy in cow by transrectal palpation

Uterus gravid 70 days uterus gravid 90 days

Uterus gravid 110 days pregnancy approaching term

 In the 1980s, real time utrasonography was developed for use in domestic
animals. An ultrasound machine resembles a radar device.
 It employs doppler principle
 A probe is inserted through the rectum and positioned above the uterus.
 This probe generates pulses of ultrasound that are transmitted to adjacent
tissues.
 These pulses are then reflected back to the probe from different tissue
surfaces.
 The reflected pulses to the probe produce an electrical signal that is
processed by a scan converter and displayed on a video monitor.
 On the video, the intensity of the ultrasound pulses returned to the probe
is converted to different shades of gray, as compared to black and white.
 Structures that contain fluid (such as the fluid-filled placenta) absorb most
of the ultrasound pulses and the result is a black image on the video
screen.
 On the other hand, more dense structures (such as an embryo) are more
ecogenic (i.e., have greater reflectivity) and result in a light gray or white
image on the screen.
-Ultrasonic depth analyser (A mode) is used to detect pregnancy as early as 40 days.
-More advanced ultrasound technology uses B (brightness) mode to display echoes
-for early pregnancies a 7.5 MHz linear transducer is required,
-for late pregnancies a 3.5 MHz transducer is preferable.

Advantages of the use of ultrasound for pregnancy diagnosis are

1) high reliability of the results
2) pregnancy diagnosis may be conducted relatively early after
insemination (i.e., as early as 25 days after insemination).
3) also estimates fetal age upto 140 days of age
4) used to determine sex of the uterus by assessing migration of
genital tubercle. In male- towards umbilicus, in female- towards tail
Disadvantages - 1) Ultrasound machines are expensive and
2) takes more time compared to rectal palpation
3) requires considerable experience & good equipment
a) Identification of early pregnancy factor/early conception
factor.
b) Assay of bovine pregnancy-specific protein B (bPSP-Bs),
also known as bovine pregnancy- associated
glycoproteins (bPAGs)
c) Progesterone concentration in plasma and milk.
d) Oestrone sulphate in milk.
a) early pregnancy factor/early conception factor.
 it is a immunosupressive glycoprotein.
 Commercially available test kits use the ‘dip-stick’ principle and detect early
conception factor (ECF) in serum and milk from as early as 3 days after
artificial insemination
 more accurate results obtained after 7 to 8 days.
b) bovine pregnancy specific protein- B (bPSP-Bs)
 identified in the maternal serum of cows from 24 days of gestation
 secreted by the binucleate cells of the trophoblastic ectoderm
 measured by radioimmunoassay(RIA)
 false positives can occur after embryonic or fetal death due to its long
biological half-life (3 days)
 Can be used to identify twins due to the correlation of its plasma
concentration with fetal numbers.
c) Progesterone concentration in plasma and milk

 Since the CL persists as a result of the pregnancy, if a blood sample is taken

at about 21 days after the previous oestrus, progesterone levels remain
elevated (Robertson &Sarda,1971)
 Since progesterone is soluble in milk fat, after crossing mammary gland, it
appears in milk (Heap,1969)

 20ml milk collected at afternoon (high fat content) into glass/plastic bottle,
then a tablet of potassium dichromate & mercuric chloride as preservative
is added.
 Concentration assayed using radioimmunoassay.
 A no. of ‘cow side’ tests based on enzyme immunoassays are also
developed
 advantage - diagnosed as early as 21 days after insemination.
d) oestrone sulphate in milk.
- one of the major oestrogens in the milk of pregnant/ lactating cows.
- during gestation the concentration increases gradually so that after
day 105 it is present in the milk of all pregnant animals,
- whereas in non-pregnant individuals it is low or undetectable;
- the source of the hormone is the fetoplacental unit.
- the precise date of sampling is not required unlike prosesterone
estination method.
- However, it has limited applications because of the lateness of the
time that a positive diagnosis is obtained.
A.CLINICAL METHODS:

1.Rectal palpation - earliest & most accurate method .

- the same equipment ,dress, mode of procedure is used as for
examination in cow.
exception- Restraint is more essential in mares.
(nose twitch to control & make them stand quietly &
prevent them from kicking the operator)

It is possible to detect pregnancy in a high percentage of mares at any

time from the 30th day onwards, although during the 30-60 day
period accurate detection is easier in primigravida & non-foaling
mares than those have conceived at the foaling heat.
 At the 30th day
-the gravid cornu (more often the right )is turgid & sausage-like
it contrasts with the flaccid uterine body & non-gravid horn.
 By the 45th day
-the embryonic mass forms a soft distension towards the apex of
the cornu, oval rather than spherical in outline,
- measuring about 6cm. from side to side.
 By the 60th day
-the localized corneal distension attends a breadth of about 8cm
( size of a large orange.)
- is still relatively soft to the touch & gentle pinching of the
uterine wall may reveal the foetal membranes slipping through
the fingers.
 At 90th day
-distension involves the uterine body as well as a cornu.
- width is about 12.5 cm. & it lies somewhat transverse with its
dorsal surface at the same level as the pelvic brim.
 At 110 days
-the outline of the distended uterus resembles a rugby football &
the organ sunk a little below the pelvic brim.
- By gentle tapping the dorsal surface with finger it may be
possible to feel the foetus as it touches the uterine wall.
 90 to 120 day
- optimum time for early detection of pregnancy by this method.
-chance of confusion between a distended uterus & inflated condition
of the large colon,
but with a little experience the difference in tone which these organs
present become readily distinguished.
 By the 150th day
-appreciable distension of the uterine body tends to be lost owing to the
uterus having sunk downwards & forward into the abdomen & the
fluids having gravitated with it .
 From 200days onwards
-the size of the foetus is such that parts of it like limbs , head, ribs can
generally be palpated without difficulty.
2.BIOLOGICL METHOD:-
A. Blood serum for PMSG hormone :
-PMSG is first found in blood between 37th to 42nd days of pregnancy &
reaches its maximum between 50th & 80th day.
-The presence of this hormone is demonstrated by the stimulation of the
ovarian activity in immature rat or mice.
Method -30-40 ml of blood is allowed to coagulate & the serum is kept for 24 hour.
The serum is stored at 4 c .& to 4 /5 mice of 3 ½ week old 0.25ml of serum is
injected twice daily for two days. Some 24-30 hours after the last injection the
ovaries & uteri of the mice are examined post-mortem.
In negative case –the uterus is small & underdeveloped ,
the horns being little thicker than ordinary sewing thread;
the ovaries are small & inactive each being uniform pale pink in colour.
In positive case- uterus is generally turgid & swollen to five times its normal size.It
may be somewhat ballooned with fluid.The ovary show variety of changes,
depending on the concentration of the hormone & degree of resposivenessof the
mouse.In all case ovaries are swollen but in weaker + ve sera a crop of ripe
follicles in both or one ovary & in strong +ve a crop of hemorrhagic follicles
apper as dark –red or black spots numbering from 2-3 to as many as 7-8 in each
ovary. There is marked swelling in vulva & vagina.
 B. URINE TESTS:-
The pregnant mare generally excretes estrogen in her urine in appreciable
amount at about 75th day of gestation ; it may be delayed until the 85th-90th day.
THE MOUSE TEST:- ovariectomized mice are used & a positive reaction comprise s of
the changes in the vaginal epithelium which characterized by estrus.
--For this young female mice are spayed , & they are ready for the purpose of the test
2-3 week after operation .
--the urine samples are taken from the mare in her normal resting state. Prior to
use the urine it is treated to overcome its toxicity to mice .
--Sulpho-salicylic acid (1g/25ml) is added until all reaction between it & the urine
ceases. The urine is allowed to stand for 30 minutes .Its pH is neutralized at 7.0 -
7.5 & filtered by using Whatman No. 2 paper. Filtrate is diluted twice by adding
distilled water .In this form it is ready to use .
--To four mice of equal size & age 0.5 ml of prepared urine is injected subcutaneously
twice daily for 3 days. After 1-2 days of last injection the test is “read”. Using a wire
loop, smears are from the anterior vagina of each mouse & these are stained by
usual Geimsa technique.
--Under the influence of estrone ,the normally small, round ,nucleated epithelial cells
are converted into large , angulated ,cornified, non-nucleated cells,& it is on the
presence of the latter that a positive diagnosis is based.
3.CHEMICAL TEST:-
--Cuboni (1937) first developed a chemical test for the detection of estrogenic
hormone in mare pregnancy urine.
--A mixture of 15 ml of urine & 3ml of conc. HCl is heated in a water- bath at boiling
point for 10minutes . the mixture is cooled, poured into a separating funnel, 18ml of
benzole added & shaken.
--The benzole layer is collected, poured into 10ml of conc. sulphuric acid & heated in a
water-bath at 80˚Cfor 5min;utes.the mixture is then cooled again.
--In the pregnant mare a green fluorescence is developed. A positive reaction first
appears at about 120th day of the gestational period.
Phenolsulphonic Acid Test:
by Mayer (1944) & is based on the kober colorimetric test for estrogen.
 The hydrolysis of the conjugated estrogen is accomplished by slowly adding 3ml of
conc. HCl to 51ml of mare’s urine in a pyrex test tube & heating in a boiling water-
bath for 10 minutes. following hydrolysis the urine is cooled to 15˚C .in an ice
water-bath to precipitate urinary pigments & substances formed during hydrolysis
which mask the final red colour. the precipitated substances are removed by
filtration & amber coloured filtrate collected in a separating funnel. the filtrate in
the funnel is extracted twice with 20ml & once with 10ml of peroxide free ether to
remove the estrogen.
 the combined ether extract are then shaken in the separating funnel with 100ml
portions of 2%sodium carbonate solution until the Carbonate washes are
colourless .
The Carbonate washes are followed by a final wash with 20ml of distilled water. The
ether extract at this stage is nearly colorless but in many instances may contain a
purpulish –red pigment .
The washed ether is now extracted with three15ml portion of N/10 NaOH ,in which
the esrogens are soluble but the ether soluble contents are insoluble. The NaOH
extracted is washed once with 15ml of distill water .
The water wash & three NaOH extract are combined & 25% sulphuric acid added
until the solution is acid to congo-red paper .
The combined acidified N/10 NaOH extracts &water wash are then extracted with
ether to remove estrogen as previously described for the filtrate of the urinary
hydrolysate.
The three ether extracts of the acidified NaOH solutions are poured into a clean
,dry 215 ml Erlenmeyer flask, preventing however, the insulation of water adhering
to the separating funnel. After evaporation of ether in a water- bath 3ml of specially
prepared phenolsulphonic acid reagent are added & the contents of the heated for
5minutes in a boiling water bath .The flask is immediately plunged into anice water-
bath & cooled. To the coolest mixture ,3ml of a 5%H SO solution are added ,the
flask again heated for a3minute period ,cooled & diluted to10ml by the addition of
4ml of water or 5 %H SO .
 The formation of a pink to cherry -red colour ,the intensity of the colour
depending upon the the stage of pregnancy when the urine was collected,
constitute a positive test for pregnancy.
 The non-pregnant state is indicated when the residue of a urine
extract does not form the red colour with the reagent.
Mucin Test:
 Attention has already been directed to the characteristic physical
changes which occur in the vaginal mucous of mare during pregnancy
.Kuroaswa(1931) has described microscopical examination of this mucous
as a means of pregnancy diagnosis & the method has been developed in
the country by Miller (1938) .
 A little of the tenacious mucous is obtained from the region of the
external os by using a vaginal speculum & a swab & several smears are
made on glass slides.After fixing with alcohol & allowing to dry the smear
is stained for 20minutes with Delafield’s haemotoxilin & then washed .The
affinity of the mucous for stain is more in pregnant than non-pregnant
mare.
 In positive case the smear will appear dark blue colour.
 ULTRASONOGRAPHY METHOD:
 most common and reliable method
 For pregnancy diagnosis, real time linear array B-mode portable ultrasound
machines that are fitted with a rectal (linear array) scanner or probe are
used.
 the rectum of the mare is emptied of feces, and the lubricated probe is
introduced into the rectum with the gloved arm to visualize the ovaries,
uterus and the new developing embryo within.
 enables detection of pregnancy as early as 11 days post-ovulation. Mostly
prefered between 12 and 15 days post-ovulation, when the embryonic
vesicle is larger and easier to find.
 every inch of the uterine horns and body followed to find the early
embryo (which can be as small as a quarter of an inch in diameter) and to
rule out the presence of twins for early twin reduction.
 Allows monitor normal growth and health of the embryo. Additionally, if
early embryo loss is detected, one can arrange to rebreed the mare before
the end of the breeding season.
 Estrus (heat) in the mare lasts for an average of 5 to 7 days, and ovulation usually
occurs towards the end of behavioral estrus. The egg within the follicle is released
into the Fallopian tube at ovulation, where fertilization occurs if the mare has been
bred at the appropriate time. The fertilized oocyte or embryo reaches the uterus
around 6 to 7 days after ovulation, but at that time is still too small to be seen on
ultrasound examination.
 A peculiar fact about pregnancy in the mare is that the early embryo travels all
around, covering every inch within the uterus, from days 7 to 16. This is a very
important event in what we term maternal recognition of pregnancy; basically, the
embryo is asserting its presence by preventing prostaglandin release from the
endometrium (uterine lining), which would induce the mare to come back into heat,
and thus the pregnancy would be lost. This is why when we perform early
pregnancy diagnosis (before day 16) the embryo can be located anywhere in the
uterus and thus it is very important to be thorough and systematic with the
ultrasound examination.
 At around day 16, the embryo becomes too large for such movement and gets
lodged at the base of one of the uterine horns, where development progresses.
Linear array B-mode real-time ultrasound scanners are the most commonly used for
examination of the reproductive tract of mares.
 An ultrasound machine has two important parts: A transducer or probe and a
processing computer with a monitor.
 The transducer is the instrument that touches the part of the body to be imaged, in
this case inside the rectum to visualize the reproductive tract of the mare.
 Piezoelectric crystals arranged in a linear mode within the transducer vibrate
emitting sounds in a frequency above that heard by the human ear.
 These ultrasounds travel through different body tissue densities at different speeds
and then bounce right back to the transducer, producing an image in different
shades of gray, from black to white, is processed and seen in the monitor.
 Ultrasounds travel best through fluids giving rise to a pitch-black image on the
monitor.
 The early equine embryo appears like a small vesicle or bubble filled with fluid (yolk
sac stage), and therefore on our ultrasound screen we see a black circumference
within the lumen of the uterus.
 After day 21 of the pregnancy, the embryo proper can be seen within a fluid-filled
vesicle. Structures within the fetus appear in different shades of gray and the bones
appear bright white.
Management method
 failure to return to oestrus at 18–22 days after service or artificial
inseminations
 However, the detection of oestrus can be difficult, and it is timeconsuming
even the back pressure or riding test, which is generally accepted to be the
most reliable (Reed, 1969), is inconsistent
 False +ve result due to reluctance to show signs/ anoestrous / ovarian cysts
Clinical methods
Transrectal palpation
0–21 days of gestation
-the bifurcation of the cornua becomes less distinct and the uterus becomes
slightly enlarged, with soft walls
-middle uterine artery increases to approximately 5 mm in diameter towards
the third week
21–30 days of gestation.
-The bifurcation of the cornua is less distinct, and the cervix and uterine
walls are flaccid and thin.
-The middle uterine artery is 5–8 mm in diameter and more easily
identified
31–60 days of gestation.
- The cervix feels like a soft-walled tubular structure
- the uterus is ill defined and thin-walled.
-Fremitus can be first identified at 35–37 days (Meredith, 1976), the pulse
pattern can be compared with that of the external iliac artery
60 days to term.
-The middle uterine artery is greater in diameter than the external iliac and
it has strong fremitus; it now crosses the external iliac artery more dorsally
than before. --Only towards the end of gestation is it possible to palpate
piglets at the level of the cornual bifurcation.
Transrectal B-mode (5 MHz transducer) ultrasound image of a pregnant
sow 23 days after service. embryos surrounded by amniotic fluid (black)
Estimation of plasma progesterone
-values in the plasma ≥7.5 ng/ml were indicative of
pregnancy
Plasma oestrogen assay
-oestrone sulphate not detected in the blood of non-
pregnant cyclic sows but in pregnant animals it was
detectable from day 20 of gestation
-the optimum time for diagnosis, when maximum
concentrations of oestrone sulphate are present, is at
about 24–28 days.
To be continued…

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