Pregnancy diagnosis

(Cyesiognosis)
Important for managing profitability of dairy herds
Early diagnosis of pregnancy helps in timely therapy of nonpregnant animals

R Required to certify female animals for sale

male animals for sale
To age the conceptus

To determine the viability of the fetus

2. PREGNANCY MAINTAINING HORMONE

2. PREGNANCY MAINTAINING HORMONE

P4 produced by the CL is secreted into the

blood and subsequently into milk.
In a normally cycling cow, levels of P4 are
relatively low in both blood and milk around
estrus.
The role of CL in maintenance of pregnancy
varies among species.

3. GESTATION LENGTH OF COW

3. GESTATION LENGTH OF COW
Gestation - intrauterine development of
the fetus.
Gestation length - fertile service to
parturition.
Duration of gestation is genetically
determined.
Gestation length of cow - 276 to 295
days.

Attributes of the Ideal Pregnancy Test

For successful integration into a reproductive
management system, an ideal early pregnancy test
for dairy cattle would be
1) sensitive (i.e., correctly identify pregnant
animals)
2) specific (i.e., correctly identify nonpregnant
animals),
3) inexpensive,
4) simple to conduct under field conditions, and
5) able to determine pregnancy status at the time
the test is performed.
Most direct and indirect methods for pregnancy
diagnosis exhibit one or more of these attributes,
but none currently available or under development
exhibit all of them. A final attribute of an ideal test

Methods of
pregnancy
diagnosis

Visual
Clinical
Laboratory
Return to estrus
Rectal palpation
1.Hormones
increase in abdominal size
Abdominal ballotment P4, E2, eCG, relaxinIncrease in udder size (4 mon
Radiography
2.Pregnancy assoc proteins
Increased fetal movements (6
Ultrassonography
bPAG, ePF
Tail cocking camel
(14-15 day of mating)

Return to Estrus as a Diagnostic

Indicator of Pregnancy Status

Pregnancy diagnosis

Direct

Transrectal palpation
Ultrasonography
Radiography
Laparoscopy

Indirect

marker molecules

Chemical tests
Immunodiagnostics
Identification of

Pregnancy diagnosis
Recto-genital palpation
Cows: Increase in uterine size & softening
Amniotic vesicle 35-45 days (1.5 cm)
(65-70 days not palpable)
Fetal membrane slip 35-90 days
Placentomes 80 days to term
Fetus 65-70 day onward
Middle uterine artery 90 day onward
Ovaries not palpable beyond 4-5 months
Vaginal discharge beyond 5 months

Table 1. Rectal examination.

Gestation
(Month)

Genital tract changes

First

Fluctuation of the uterus and fully developed CL in one ovary.

Second

Enlargement and dorsal bulging of pregnant horn; membrane

slip.

Third

Descent of uterus commences, pregnant horn retractable,

palpable fetus and placentomes and fremitus in some cases.

Fourth

Pregnant horn not retractable and situated on abdominal

floor. Fetus may be out of reach. Fremitus present and
enlarged middle uterine artery.

Fifth

Pregnant horn on abdominal floor, fetus may be out of reach,

fremitus distinct from gravid side.

Sixth

Fetus out of reach, placentomes quite variable in diameter

and very distinct fremitus.

Seventh to
term

Cotyledons, fremitus and fetal parts are palpable.

Table 2. Calf fetal size at various stages of

pregnancy.
Stage of pregnancy

Calf fetal size in relation to

the size of commonly
known adult animals

2 months

mouse

3 months

rat

4 months

small cat

5 months

large cat

6 months

beagle dog

Figure 1. Dorsal view of

of where
pregnant and non-pregnant
during rectal
reproductive tract cow.

Figure 2. Side view

arm is located
palpation.

Differential diagnosis of pregnancy include:

The urinary bladder,
The rumen and kidney,
Pyometra, fetal mummification, fetal

Maceration,
mucometra, hydrometra, and tumor.

Figure 3. Pyometra in cow

Figure 4. Fetal mummification in cow.

Figure 5. Fetal Maceration in cow.

Mare
Uterine tone 16-50 days
Amniotic vesicle 25-28 days bulge (60 days football size)
Fetus palpable by day 90 (difficult 5-7 months)
Uterine descent by day 100 Ovaries descent 3-5 months
beyond 5 months ovaries not palpable
Radiography
Sheep and goat 70 days onwards
Bitch 6 week onward
Good for counting fetal numbers
Spalding sign: Overlapping of fetal cranial
bones in dead fetuses

Ultrasonography
Uses sound frequencies beyond
20000 Hz (1 MHz to 12 MHz)
Real time A(Amplitude mode)
B(brightness) mode and M (motion)
mode
Probes Linear array, sector or
curvilinear (Transrectal,
transabdominal and transvaginal
probes)
Trans-rectal probes for large
animals 5-10 MHz frequency
Trans-abdominal probes for small
animals 1.0 to 4.5 MHz frequency
Coupling gel neccessary

graphy technique and sonograms showing anechoic fluid and echogenic fetus du
y in cattle

Ultrasonographic appearance of different structures

Fetal heart beat
18-20 days
Fetal heart beat
25-30 days
Cotyledons
35-40 days cow 40-50 day sheep
(Trans-abd)
Fetal sex
57-60 day cow
60-70 day mare
--------------------------------------------------------------------------------------------------Transrectal probe 5.0 7.7 MHz
Cow, buffalo, mare, camel
Transrectal (Prostatic) probe 5.0 7.5 MHz
Sheep/Goat before
day 40
Trans-abdominal probe 2.5 3.75 MHz Sheep/Goat- beyond day
40
Bitch, cat
Doppler ultrasound
Sow

Abdominal ballotment
Cows beyond 7 months
Fetal sacs sometimes palpable in the bitch
at 30 days
Rectal abdominal method sheep(rod method)
Digital manipulation goat
Palpation of fetus 4months goat 50 days bitch

Laboratory tests
Assay of hormones
Progesterone 18-24 days post-breeding 98-100%
accurate for non-pregnant regularly cycling cows
75-80% accurate for pregnant cows.
Useless for the bitch
Estrogens
Mare

Maternal estrogens high after 60 days. Conjugated urinary

estrogens high after 150 days

Cow/buffalo

Maternal estrogens high after100 days.

Goats

Maternal estrogens high after 50 days.

Sows

Rise at 20 days peak at 25-30 days, decline at 45 days

and rise again at 70 80 days to term

Bitch

Slightly elevated at implantation and high for rest of

gestation

Figure10.Progesterone concentrations in
milk
during the reproductive cycles.

Figure 11. Progesterone concentrations

in milk during pregnancy.

Chemical tests to detect urinary estrogens

cows barium chloride testUrine + BaCl2heat a white ppt if
positive.
mares mucin test ( dark staining cells on vaginal smears)
cubonis test15 ml urine + 3 ml HClheat(10 min and cool)
add 18 ml benzolcollect benzol layer and add 10 ml H 2SO4 and
heat(5min) a green fluorescence in a positive case
eCG

Currently Elisa, RIA have replaced older tests

Older tests (biologic tests)
Ascheim Zondek test Inject pregnant mare serum to rats
Freidmans test (Rabbit test)
Toad test
Relaxin Pregnant bitches placenta secrete relaxin
from 20-30 days of gestation
Assay of pregnancy specific proteins
bPAG (PSPB) placenta specific 29-30 days post breeding RIA needed
EPF (RIT) 24-48 h of fertilization and these molecules disappear
within
48 h of fetal death
Vaginal biopsy vaginal epithelial cells decrease from 20 layers at estrus
to 3-4 layers at 18-22 days of pregnancy in sows and sheep.

4.2.2.2. Pregnancy Associated Antigens

Antigens specific to pregnancy reported in
maternal tissues of varies species including
cattle.
Table 3. Immunological methods of pregnancy diagnosis in the cow.

Test

Principle

Stage of

Sample Technique Earliest

Gestation
P4
EPF

Predicts luteal activity

day

implantation Serum/

EIA

Serum

RIT

4 day

Serum/

RIA/EIA

72

HI

Latter

Pre

suppressive factor.

implantation

Estron

Determines feto-placental

Post

sulphate

function

implantation milk

Pregnancy

Identifies antigens specific Post

associated

to pregnancy

antigens

21

milk
Detects immuno-

implantation

RIA/

Serum

stage

4.3.2 Prostaglandin Induced Milk Flow Test

(PGIMF Test)
This method is based on the observation of
milk ejection by IV administration of 256 g of
PGF2.
The tests can be performed in lactating
cows, 18-22 days after AI.
If CL is present, the alveolar milk flow starts
within 51-121 seconds and if not functional
the cow would be considered not pregnant.
The accuracy of positive results was 72.3%.
The test has the advantage of being
inexpensive, rapid and easy to interpret.
With such a test, non pregnant cows can be