Pregnancy diagnosis

Chaiwat JarassaengDepartment of surgery and

Theriogenology Vet Med KKU

Outline

1. Why?2. Rectal palpation3. Ultrasound4. Milk progesterone assay5. Bovine pregnancy-specific protein B6. Early pregnancy factor

References

1. Bearden and Fuquay 1997. Applied Animal Reproduction

2. Youngquist 1997. Current therapy in large animal theriogenology.

3. Paul M. Fricke. Methods of Diagnosis and Monitoring of Pregnancy in Dairy Cattle and their Implementation.2010

Why pregnancy diagnosis

• Ultimate goal of production• One calves one years• Pregnancy at 80 days postpartum• Early pregnancy 17-19 days post-AI• Inexpensive

Pregnancy diagnosis

1. 100 % accuracy2. No false positive3. No false negative4. Early as possible5. Viability of conceptus6. Possibly determine of sex7. Result immediately

Rectal palpation

1. Easiest, fastest, cheapest method2. 45 days post breeding (35 days is

goal??)3. positive signs of pregnancy in the cow

are fetus, cotyledons/caruncles, amniotic vesicle, and fetal membrane slip

Rectal palpation

1. You must examine the entire tract before declaring the cow open.

2. You must find one of the positive signs of pregnancy before you call a cow pregnant

3. Pregnancy examination must always be the first step in your examination. If you not sure, recheck the cow...maybe in a few minutes, maybe tomorrow or next month

Rectal palpation

First feel the uterus for asymmetry. At 35 days of pregnancy the pregnant horn will

feel slightly larger. Second, feel for fluid in the larger horn

Rectal palpation

1. Placentome (cotyledons/caruncles) >75 days

2. Fremitus : hypertrophy of middle uterine artery >90 days

• Estimation of stage of pregnancy by rectal palpation ??

Amniotic vesicleDays Gestation

Width in mm

Width in fingers

35 7 1/2 42 15 1 48 35 2 52 55 3 58 75 4 62 90 4+ 65 105 5

Fetal membrane slip

32 days thread 1 horn

45 days small string 1horn

60 days string 2 horns

>70 days large string

Diameter of Bovine uterine horn of difference stages of pregnancy

Stage in days Diameter in cm

10 Slight enlargement60 790 8120 12150 18

Average size of bovine conceptusat different ages

Ages/Days Crown/Rump length in cm

30 160 590 13120 30150 38180 56210 71245 81280 86

Placentome75 days pea size100 dime115 nickel125 quarter150 half dollar> 150 variable

Fetus

3 months rat4 months small cat5 months large cat6 months beagle dog

2 ‡µŸœ∫

3 ‡µŸœ∫

4 ‡µŸœ∫

5 ‡µŸœ∫

6 ‡µŸœ∫

FremitusAt 120 days found fremitusAt 5 months the artery fremitus.At 6 months both the artery At 7 months both arteries are large with fremitus and the fetus is ascending.

Differentials in Pregnancy Diagnosis by rectal palpation

• Adhesions • Pyometra• Segmental aplasia• Mummification • Maceration • Lymphosarcoma• Granulosa cell tumor

Rectal Palpation at 35-45 days

1. Uterine horn slight enlargement2. Slippage membrane3. Amnion size of yolk4. Corpus luteam present in ovary

Rectal Palpation at 45-50 days

1. Slightly greater difference size of uterine horn

2. Amnoin -> small hen egg3. Membranes slipped4. Corpus luteam in ovary

Rectal Palpation at 60 days

1. Slightly greater difference size of uterine horn 6.5-7.6 cm

2. Membranes slipped both ??3. Fetus bumped 4. Corpus luteam in ovary

Rectal Palpation at 90 days

1. Uterine horn well over in pelvic brim2. Fetus 10-15 cm.3. Corpus luteam in ovary

Rectal Palpation at 120 days

1. Uterine horn well over pelvic brim2. Fetus 25-30 cm.3. Placentome4. Fremitus

Rectal Palpation at 150 days

1. Uterine horn well over pelvic brim2. Fetus 35-40 cm.3. Placentome4. Fremitus 6mm-1.25 cm.

Rectal Palpation at 170-230 days

1. Fetus difficult of palpate2. Fremitus 1.25 – 1.4 cm.

Rectal Palpation at 230-280 days

1. Fetus large extend, head leg 2. Movement of the fetus

Ultrasound1. B-mode real-time, meaning produce an

acoustic image in real time. 2. The usually range from 3.5 - 7.5 MHz,

1. With greater MHz you see more detail but have less depth penetration.

2. There is more depth penetration with lower MHz, but less detail. It is always a tradeoff.

Ultrasound1. B-mode real-time, meaning produce an

acoustic image in real time. 2. The usually range from 3.5 - 7.5 MHz,

1. With greater MHz you see more detail but have less depth penetration.

2. There is more depth penetration with lower MHz, but less detail. It is always a tradeoff.

UltrasoundExternal

- small animal - 3.5 MHz, M-mode

Intrarectal• - 26-28 give about 100% accuracy and the fetal heartbeat can be seen at 21 days.•- gestational age •- Fetal sexing can be done at about 60 days of gestation.

27 days post breeding

embryonic vesicle

35 days gestation

50 days gestation

90 days gestation

Ultrasonography in Cattle

• Days 10-20 • Size 2-4 x 3-7 mm. • Round shape in 1/3 of Uterine horn

Days 13

• Days 12

Ultrasonography in Cattle

• Days 21-24 • Size 3-5 x 10 mm.• Heart 21 days• 188+- 5 times/min

Days 22

Ultrasonography in Cattle

• Days 25-30 • Size 18-20 mm.

Days 25

Days 27

Ultrasonography in Cattle

• Days 25-30 • Size 18-20 mm.

Days 30

Ultrasonography in Cattle

• Days 31-40• Crown rump length :CRL

Days 35

Ultrasonography in Cattle

• Days 31-40• Amniotic vesicle

Days 40

Days 32

Days 32

Days 37

Days 39

Days 40

Days 40

Twins Days 39

Days 40

Milk progesterone assay

Progesterone assay1. Milk progesterone concentration 21-24 days post

breeding. 2. The milk progesterone parallels blood

progesterone.3. Important to know AI or breeding dates and the

test cannot be done randomly in a herd. 4. It is based on the fact that if progesterone is low

21 days post breeding then there is no chance that the cow can be pregnant.

5. Remember that a cow must have progesterone be pregnant. Therefore the test is 100% accurate at non-pregnancy diagnosis when the progesterone is low.

Cow side test

Protein B1. Bovine pregnancy-specific protein B) (bPSB, bTP-

1) 2. Protein B is secreted by trophoblast cells

(binucleate giant cells). 3. It is a radioimmunoassy procedure, and therefore

is time consuming and relatively expensive. 4. There are false positives early in the post partum

because it is present in the post partum period.

Protein BJ Dairy Sci. 84:1884-9, 2001.

•Test sensitivity - 86%•Specificity - 4%•Positive predictive value - 49% •Negative predictive value - 23%•False-positive - 96% •False-negative - 14%

Early Pregnancy Factor•EPF-A - Uterine tube•EPF -B - Ovary •Production requires signal from fertilized ovum (ovum factor) released under prolactin presence after sperm penetration.•Appears 4-6 hours •Disappears with fetal death •Non-detectable at 20 days in milk and 30 days in serum