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

Strength Weakness Opportunity Threat SWOT ANALYSIS

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

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

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

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

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

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

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

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

Endocrine mechanisms of oestrus cycle (-,+)

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

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

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

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

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

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 : IU 12/6 23/6 26/6 3/7

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 : IU DOSIS P4 DALAM PRID = 40 mg – 1 g 17/6 26/6 29/6 6/7 CABUT PG

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

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

Recovery of Embryo

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

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.

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)

Transfer to recipients

The End – Thank You