EMBRYO TRANSFER AND INVITRO FERTILIZATION

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

• Strength
• Weakness
• Opportunity
• Threat

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EMBRYO TRANSFER

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

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

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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

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Estrous Synchronization

• Prostaglandins PGF2a
• 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.

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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
  

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

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Endocrine mechanisms of oestrus cycle
(-,)
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Superovulation

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

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Superovulation

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

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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.

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Superovulation

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

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Superovulation

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

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SUPEROVULASI DG PMSG hCG - Sinkron dg PG
hCG
IB Pagi
PMSG
DONOR
PG1 11Hr PG2 9 Hr 2Hr PG3 3Hr
EST 7 HR Flushing
IB Sore
1/6 12/6 21/6
23/6 26/6 3/7
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
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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
IB Sore
1/6 12/6 15/6 24/6
26/6 29/6 6/7
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
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SUPEROVULASI DG FSH LH
FSH PAGI
FSH PAGI
IB Pagi
FSH PAGI
FSH PAGI
LH
DONOR
PG1 11Hr PG2 9 Hr PG
3Hr EST 7 Hr Flushing
IB Sore
1/6 12/6 21/6 22/6
23/6 24/6 27/6 4/7
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
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Recovery of Embryo

• flush - remove fertilized eggs from donor before
  7 days gestation
• plant embryo in the recipient animal

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Recovery of Embryo
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Recovery of Embryo
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Transfer to recipients

• Non surgical
• common
• deposit embryo in uterus through cervix with AI
  straw and gun
• Usually 70-80 success rate

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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.
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INVITRO FERTILIZATION
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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.
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INVITRO FERTILIZATION
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Transfer to recipients
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The End Thank You