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EMBRYO TRANSFER AND INVITRO FERTILIZATION

EMBRYO TRANSFER AND INVITRO FERTILIZATION. ABDUL SAMIK DEPARTMENT OF VETERINARY REPRODUCTION FACULTY OF VETERINARY MEDICINE - UNAIR. SWOT ANALYSIS. Strength Weakness Opportunity Threat. EMBRYO TRANSFER.

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EMBRYO TRANSFER AND INVITRO FERTILIZATION

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

  2. SWOT ANALYSIS • Strength • Weakness • Opportunity • Threat

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

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

  5. ET Process • superovulate donor cow • synchronize donor & recipients • Breed donor cow (A.I.) when in heat • Recovery of embryo from uterine of donor • 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 • Commonly used with AI and embryo transfer • For a successful synchronization program to occur, planning is essential

  7. Estrous Synchronization • Prostaglandins: PGF2α • 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.

  8. Estrous Synchronization • 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

  9. Estrous Synchronization 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.

  10. Estrous Synchronization • Synchromate B /PRID/ CIDR system: • Day 1 • Inject 5 mg estradiolvalerate & 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.

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

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

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

  14. Superovulation • 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.

  15. Superovulation • FSH: given for 4 days, twice each day (starting on day 9 to 12 of cycle). • Results in multiple follicles producing estrogen • PGF2 α on 3rd day to regress CL. • Removes the high progesterone allowing the preovulatory surge of GnRH/LH. • injection of LH,orhCG at the time of heat needed for ovulation.

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

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

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

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

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

  21. Recovery of Embryo

  22. Recovery of Embryo

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

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

  25. INVITRO FERTILIZATION

  26. INVITRO FERTILIZATION

  27. INVITRO FERTILIZATION

  28. INVITRO FERTILIZATION

  29. INVITRO FERTILIZATION

  30. INVITRO FERTILIZATION

  31. INVITRO FERTILIZATION

  32. INVITRO FERTILIZATION Ovum Pick Up (OPU) 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.

  33. INVITRO FERTILIZATION

  34. INVITRO FERTILIZATION

  35. INVITRO FERTILIZATION

  36. INVITRO FERTILIZATION

  37. INVITRO FERTILIZATION

  38. INVITRO FERTILIZATION

  39. INVITRO FERTILIZATION

  40. INVITRO FERTILIZATION

  41. INVITRO FERTILIZATION

  42. INVITRO FERTILIZATION

  43. Transfer to recipients

  44. The End – Thank You

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