1. EMBRYO TRANSFER AND INVITRO FERTILIZATION ABDUL SAMIK DEPARTMENT OF VETERINARY REPRODUCTION FACULTY OF VETERINARY MEDICINE - UNAIR

2. Strength Weakness Opportunity Threat SWOT ANALYSIS

3. EMBRYO TRANSFER b provides opportunity to introduce genetic material into populations of livestock while greatly reducing risk of transmission of diseases

4. Process of removing fertilized ova from a donor female and placing them in recipient females EMBRYO TRANSFER

5. ET Process b superovulate donor cow b synchronize donor & recipients b Breed donor cow (A.I.) when in heat b Recovery of embryo from uterine of donor b Transfer of embryo into uterine of recepient cow

6. Estrous Synchronization controlling or manipulating the estrous cycle so that females express estrous at approximately the same time controlling or manipulating the estrous cycle so that females express estrous at approximately the same time b Commonly used with AI and embryo transfer b For a successful synchronization program to occur, planning is essential

7. Prostaglandins: PGF2 α Lutalyse - Natural compound 25 mg dose I.M. Estrumate - Analogue 500 ug dose I.M. Bovilene - Analogue 1 mg dose s.q. Prosolvin - Analogue 15 mg dose I.M. Equimate - Analogue used in Mare Glandin F – 25 mg dose I.M. Estrous Synchronization

8. Principle - Regress active corpus luteum i.e., Day 5-17 corpus luteum. In estrus 2-5 days after injection heifers ~50 hours; cows ~72 hours ~60-65% of herd should respond to injection. To get whole herd synchronized, give 2nd injection 11 days after 1st Cows responding to 1st injection have day 6-9 corpora lutea Unresponding cows now have day 6-11 corpora lutea Estrous Synchronization

9. Use of progestogens for Estrus synchronization Principle: Maintain the cow under the influence of progesterone until corpus luteum regresses, remove progesterone - animal respond to ¯ progesterone with estrus. 2-5 days later. Estrous Synchronization

10. Synchromate B /PRID/ CIDR system: Day 1 Inject 5 mg estradiol valerate & 3 mg norgestomet Implant Norgestomet (Progestagen) 6 mg Remove implant 9 days later. Breed 48 to 60 hours later or 54 hours later. Estradiol - Luteolytic to d 3-9 corpora lutea. Norgestomet - Inhibits early C.L. development. Estrous Synchronization

11. Endocrine mechanisms of oestrus cycle (-,+)

12. Superovulation b treatment of the female with hormones so that more eggs are ovulated than normal

13. Superovulation b alter normal level of FSH b Follicle Stimulating Hormone b Typical results are ovulation of 10 or more eggs

14. Hormone Treatments: Hormone Treatments: – eCG (PMSG) 2,000 IU from blood of mare day 40 - 100 of pregnancy. – FSH from pituitary of pig or cow. – Recombinant FSH. Superovulation

15. b FSH: given for 4 days, twice each day (starting on day 9 to 12 of cycle). Results in multiple follicles producing estrogen Results in multiple follicles producing estrogen  PGF 2 α on 3 rd day to regress CL. Removes the high progesterone allowing the preovulatory surge of GnRH/LH. Removes the high progesterone allowing the preovulatory surge of GnRH/LH. b injection of LH,or hCG at the time of heat needed for ovulation. Superovulation

16. Superovulation b recipient animals must be closely synchronized with the donor cow. b Must be at the same stage in their heat cycle

17. DONOR RESIPIEN PG1 11Hr PG2 9 Hr 2Hr PG3 3Hr EST 7 HR Flushing PMSG PG1 11 Hr PG2 3Hr EST 7 HR Transfer hCG IB Pagi IB Sore 1/6 12/6 21/6 23/6 26/6 3/7 SUPEROVULASI DG PMSG & hCG - Sinkron dg PG SUPEROVULASI DG PMSG & hCG - Sinkron dg PG DOSIS PMSG : 2000 – 3000 IU DOSIS hCG : 1500 -2000 IU 12/6 23/6 26/6 3/7

18. DONOR RESIPIEN PRID 9 Hr EST 9 Hr 2Hr PG 3Hr EST 7 Hr Flushing PMSG PRID 9 Hr 3Hr EST 7 Hr Transfer hCG IB Pagi IB Sore 1/6 12/6 15/6 24/6 26/6 29/6 6/7 SUPEROVULASI DG PMSG & hCG - Sinkron dg P4 SUPEROVULASI DG PMSG & hCG - Sinkron dg P4 DOSIS PMSG : 2000 – 3000 IU DOSIS Hcg : 1500 -2000 IU DOSIS P4 DALAM PRID = 40 mg – 1 g 17/6 26/6 29/6 6/7 CABUT PG

19. DONOR RESIPIEN PG1 11Hr PG2 9 Hr PG 3Hr EST 7 Hr Flushing FSH PAGI PG1 11 Hr PG2 3Hr EST 7 HR Transfer LH IB Pagi IB Sore 1/6 12/6 21/6 22/6 23/6 24/6 27/6 4/7 SUPEROVULASI DG FSH & LH SUPEROVULASI DG FSH & LH FSH I : 6 mg FSH II : 5 mg FSH III : 4 mg FSH IV : 3 MG TOTAL 40 mg FSH PAGI FSH PAGI FSH PAGI 12/6 23/6 27/6 4/7 FSH SORE FSH SORE FSH SORE FSH SORE

22. Recovery of Embryo b flush - remove fertilized eggs from donor before 7 days gestation b plant embryo in the recipient animal

23. Recovery of Embryo

25. Transfer to recipients b Non surgical b common b deposit embryo in uterus through cervix with AI straw and gun b Usually 70-80 % success rate

26. Surgical b incision in recip through flank b tip of uterine horn is exposed b cut open and embryo is deposited recipient is sewn shut less than 60-65% success rate.

34. Aspirate oocytes from a live donor animal ultrasound guided transvaginal probe with a needle guide. Oocytes matured, fertilized and cultured in vitro. ♦ Embryo production from animals that do not superovulate well, are injured, or dying. ♦ Procedure can be conducted every 3-5 days. Ovum Pick Up (OPU)

45. Transfer to recipients

46. The End – Thank You