11

Synchronization treatments for fixedSynchronization treatments for fixed--time artificial time artificial insemination and embryo transfer in beef and dairy insemination and embryo transfer in beef and dairy

cows, factors affecting pregnancy ratescows, factors affecting pregnancy rates

Gabriel A. BGabriel A. Bóó

Instituto de ReproducciInstituto de Reproduccióón Animal Cn Animal Cóórdoba rdoba CCóórdobardoba, , ArgentinaArgentina

DONORS:Cost of hormones.Treatment protocols.Variable response.

FactorsFactors AffectingAffecting thethe WidespreadWidespread ApplicationApplication ofofEmbryo Embryo TransferTransfer Technologies in Technologies in CattleCattle

RECIPIENTS:Management.Synchronization.Estrus detection.% of recipients used.

22

33

Which one is in Heat?Which one is in Heat?

What are the alternatives?

• Synchronize estrus to decrease the daysrequired for heat detection (PGF treatments).

• Use Treatments that eliminate the need forestrus detection (Fixed-time Embryo Transfer).

44

Resincronización de CelosResincronización de Celos

Synchronization of Estrus With Prostaglandin F2α

Requires Requires trainnedtrainned Personnel for estrus detectionPersonnel for estrus detectionLow submission ratesLow submission ratesVariable interval from treatment to estrusVariable interval from treatment to estrusOnly Works in cows with a CLOnly Works in cows with a CL

LIMITATIONS OF PGF TREATMENTSLIMITATIONS OF PGF TREATMENTS

55

OvsynchOvsynch Treatments for FTETTreatments for FTET

BaruselliBaruselli et al., 2000et al., 2000

D0

PGF

7 days

ETEstrus Detection

D0

GnRH GnRHPGF

D7 D9 D16

FTET

10/70

66

Sincronização das Receptoras

G1- Ovsynch G2 - PGF2α P

Transferred 72.6% (122/168) a 45.2% (80/177) b <0.0001

Conception Rate 49.2% (60/122) 56.3% (45/80) 0.20

Pregnancy Rate 35.7% (60/168) c 25.4% (45/177) d 0.03

(a ≠ b; c ≠ d; P<0,05)

BARUSELLI et al., 2000

77

PGF

4

8.5

10

14

Day 9Day 7

mm

Day 0 Day 1-2

GnRH GnRH

Ov: 30h

P4

PGF

4

8

10

14

Day 9Day 7

mm

Day 0 Day 1-2

GnRH GnRH

No OV

P4

88

PGF

4

8

10

14

Day 9Day 7

mm

Day 0 Day 1-2

GnRH GnRH

No OV

P4

OVSYNCH+ P4 FOR EMBRYO TRANSFER

OVSYNCH+ P4 FOR EMBRYO TRANSFER

Day 0 Day 7 Day 9 Day 16

GnRH PGF GnRH

P4 ET

99

PGF

4

8

10

14

Day 9Day 7

mm

Day 0 Day 1-2

GnRH GnRH

No OV

P4

PGF

4

8

10

14

Day 9Day 7

mm

Day 0 Day 1-2

GnRH GnRH

No OV

P4

CIDR

1010

Synchronization of Recipients With Ovsynch or Ovsynch + P

82/165 (49.7)82/152 (53.9) ab152/165 (92.1) bOvsynch+P

72/165 (44.0)72/150 (48.0) b150/165 (90.9) bOvsynch

67/169 (39.6)67/108 (62.0) a108/169 (63.9) aGnRH+PGF/estrus detection

PregnancyRate (%)

Conception Rate (%)

Transferred/treated

(%)

Group

ab – P < 0.01Beal and Hinshaw, 2001

Field Trial (Ovsynch+CIDR) 980/1637 (60%)

Fixed-Time Embryo Transfer (FTET)

Progestins, Estradiol and eCG

1111

Days0 5 8 9 17

2 mg EB

Progestin

FTET

SynchronizationSynchronization ofof RecipientsRecipients

PGFPGF

1 mg EB

+ 400 IU eCG+ 400 IU eCG4.0±0.4b

6.8±0.7a

P4 ng/ml

20/93 (21.5) b

20/49 (40.8)

49/93 (52.7) b

PGFDay 8

39/95 (41.1) a

39/67 (58.2)

67/95 (70.5) a

PGFDay 5

Pregnancy Rate(%)

Conception Rate (%)

Treated/Transferred

(%)

Group

7.5±0.7 b

5.7±0.4 a

P4 ng/ml

76/156 (48.7) b

76/132 (57.6) b

132/156 (84.6) a

eCG

53/156 (33.9) a

53/127 (41.7) a

127/156 (81.4) a

Control

PregnancyRate (%)

ConceptionRate (%)

Transferred/treated

(%)

Group

ab – P < 0.02Moreno et al., 2005

Effect of eCG Treatment (400 iu) onPregnancy Rates in Embryo Recipients

22/70

1212

692/1542 (44.9%)692/1309 (52.7%)1309/1542 (84.9%)

Pregnancy Rate(%)

Conception Rate(%)

Transferred/Treated

(%)

Commercial FTET with Fresh and Frozen Thawed In Vivo Embryos

(Bo et al., 2005)

September 2003 - April 2004

• Total ET = 8909• Total ET evaluated = 5277• Preg. Rate (30 d) = 49.1 %, (n=2592)• Preg. Rates (60 d) = 42.2 %, (n=2225)

Results of FTET using Freshin vitro produced embryos

Luiz F. Nasser(Campo Grande, MS, Brazil)

1313

SIMPLIFICATION OF THE FTET PROTOCOL

P4

EB(2.0mg)

PGF2α+

eCG(400UI)

EB(1.0mg) FTET

24h

D0 D8 D17D9D5

Removal

Materials & Methods918 crossbred recipients

1414

Peres et al., 2006, P<0.05

DayDay ofof thethe secondsecond EB EB treatmenttreatment

0,00%

10,00%

20,00%

30,00%

40,00%

50,00%

60,00%

70,00%

80,00%

90,00%

100,00%

Transferred/Treated Conception Rate Pregnancy Rate

EB8

EB9n=460a

n=458b

n=458b

n=460a

n=350n=321

0,00%10,00%20,00%30,00%40,00%50,00%60,00%70,00%80,00%90,00%

100,00%

PGF0+eCG5 PGF0+eCG8 PGF5+eCG5

Transferred/Treated

Conception Rate

Pregnancy Rate

Peres Coelho et al., 2006, P<0.06

n=301a n=208

n=301

n=305b

n=312

n=205

n=312a

n=243

n=305

By Day of PGF & eCG Treatment

1515

Other Factors Affecting Pregnancy Rates

Peres et al., 2006

1333Total

219/409 (53.5%)Not seen in heat0.36

517/924 (55.9%)Seen in heat

P-valueConception Rates

1616

1333Total

10/14 (71.4%)353/99 (53.5%)2 0.38673/1220 (55.2%)1

CL Numbers

400/736 (54.3%)>18 mm208/369 (56.4%)>16≤18 mm 0.94128/228 (56.1%)14 to 16 mm

CL Diameter (by ultrasonography)

P ValueConception Rate

PROTOCOL WITH ECP vs EB

ECP

EB

EBECP

70h

P4

D0 D8

EB(2mg) PGF2α

D9 D10

FTETDay 17

1717

P4

EB(2.0mg)

PGF2α+

eCG(400UI)

EB(1.0mg) FTET

24h

D0 D8 D17D9D5

Removal

P4

EB(2.0mg)

PGF2α+

eCG(400UI) FTET

D0 D8 D17D9D5

RemovalECP (0.5ECP (0.5mgmg))

1818

EB EB onon DayDay 9 9 vsvs ECP ECP onon DayDay 8835/70

EB vs ECP

50.077.375ECP

44.878.775EB

ConceptionRates (%)

TransferredTreated

(%)

N

Carballo Guerrero et al., 2007

P=0.8438

1919

0.5mg VS 1.0mg ECP

P4

BE½ PGF

D0 D8 D17

FTET

9 days

½ PGFeCG

ECP (0.5mg)

Group 0.5-ECP (n=132)

P4

BE½ PGF

D0 D8 D17

FTET

9 days

½ PGFeCG

ECP (1.0mg)

Group 1.0-ECP (n=132)

Martins, C.M., SBTE (2007)

Results

0.5444.2 (46/104)

49.5 (52/105)

Conception Rate (60 d) (%)

0.5451.9 (54/104)

56.2 (59/105)

Conception Rate (30 d) (%)

0.8878.8 (104/132)

79.5(105/132)

Transferred/Treated (%)P1.0mg ECP0.5 mg ECP

2020

The QuestionThe Question……

Can we use Can we use eCGeCG in an in an OvsynchOvsynch type protocol?type protocol?

5050

Day 0 7 9 10

GnRH PGF GnRH AI

CueCue--MateMate

Ovsynch + Progesterone Releasing Devices

400 IU Pregnecol

D16FTET

40/70

2121

n=292abP<0.04

0

10

20

30

40

50

60

70

Preg

nanc

y r

ate

(%)

2 3 to 4 5 to 11 eCG(-) eCG (+)

2 y old- No CIDR

a a

b

c

58.742.9 58.2 26.1

Effect of parity and Effect of parity and eCGeCG treatment treatment on pregnancy rateon pregnancy rate

59.1

d

cdP<0.032 yr old

Other Treatments Used to Increase Other Treatments Used to Increase Pregnancy Rates in RecipientsPregnancy Rates in Recipients

Gabriel A. BGabriel A. Bóó

Instituto de ReproducciInstituto de Reproduccióón Animal Cn Animal Cóórdoba rdoba CCóórdobardoba, , ArgentinaArgentina

2222

Substances Reported to Improve Substances Reported to Improve Pregnancy Rates in Cattle Following Pregnancy Rates in Cattle Following

Embryo TransferEmbryo Transfer

ProgesteroneProgesteroneHCGHCGGnRHGnRHClenbuterolClenbuterol (smooth muscle relaxant)(smooth muscle relaxant)BanamineBanamine (anti(anti--prostaglandin release)prostaglandin release)eCGeCG (PMSG)(PMSG)

The effect of Banamine (flunixin meglumine) on pregnancy rate of embryo transfer recipients

BanamineControls

65.4

65.3b

% preg.

260

1,300

No. trans.

66.2245Burry**

60.2a797Schrick*

% preg.No. trans.Study

abP<0.02 (*Schrick, 2003; **Burry, unpublished)

2323

Accesory CL and Preg. Rates.

4

6

10

14

Day 7

mm

Day 0

Induction of ovulation

2 CLsEstrus

↑P4

CL

ET

2424

Mann et al., 1999

D0 D2D7

D7

PGF2α

Estrus detectionET

7 days

G1: hCG (1.500UI); n=96

G2: LH (25mg); n=97

G3: GnRH; n=99

G4: CIDR (D7 a D20); n=95

G5: Control; n=98

2525

Resultados e DiscussãoResultados e Discussão

Gráfico 1. Concentração plasmática diária de progesterona (média + erro padrão) em novilhas Bos taurus x Bos indicus tratadas no dia 7 do ciclo estral.

0,00

2,00

4,00

6,00

8,00

10,00

12,00

6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23

Dias do Ciclo Estral

Prog

este

rona

(ng/

mL)

Controle

GnRH

hCG

LH

CIDRTratamentos

2626

Tratamento

05

101520253035404550

Taxa

de

Con

cepç

ão (%

)

ControleGnRHhCGLHCIDR-B

a

bb

ab ab

MARQUES et al., 2002

%

n=100 heifers/group

30

Días0 5 8 9 17

2 mg EB

P4

FTETPGFPGF

1 mg EB

+ 400 IU eCG+ 400 IU eCG

TreatmentsTreatmentsGnRH

hCG (1500 UI)

Control

2727

Pregnancy Rates

0

20

40

60

80

100

CONTROL GnRH hCG

n=78 n=83 n=80

62,8% 53,8%54,2%

Tríbulo et al., 2005

Transferred

(%) Conception

Rate (%) Pregnancy Rate

(%)

With GnRH 87.3 (103/118) 51.5 (53/103) 44.9 (53/118)

No GnRH 88.4 (107/121) 49.5 (53/107) 43.8 (53/121)

Nasser et al., SBTE 2005

Effect of GnRH at the time of Embryo Transfer on Pregnancy Rates

2828

Effect of CIDR on Pregnancy RateEffect of CIDR on Pregnancy RateEffect of CIDR on Pregnancy RateControlControl CIDRCIDR--PP CIDR+PCIDR+P

Overall Overall 115/186 115/186 79/151 79/151 103/166 103/166

PregPreg 62% 62% 52% 52% (62%) (62%) Em. Code 1Em. Code 1-- 71/110 71/110 52/100 52/100 52/94 52/94 PregPreg 65% 65% 52% 52% 55%55%Code Code 2&32&3-- 41/72 41/72 25/49 25/49 51/71 51/71 PregPreg 57%57% 51%51% 72%72%MorulaMorula 40/7440/74 30/6230/62 46/7046/70PregPreg 54%54% 48%48% 66%66%

Bousquet and Twagiramungu, unpublished

50/70

CIDR at Embryo Transfer

0102030405060708090

100

Conception Rates

CIDR (n=93) No CIDR (n=69)

P>0.24 Purcell et al., 2005

2929

Mann et al., 1999

D 0 D 13 D 20 D 25

CIDRED

D 0 D 16 D 21 D 28 D 30 D 31

CIDR

GnRHUS

PGF

GnRH

FTAI

3030

Centro Córdoba

291/113b

(76.6%)113/202 b(55.9%)

-------178/380(46.8%)

2006

229/309 b

(74.1%)65/144 b

(45.1%) -------164/326

(50.3%)2005

174/279 a(62.4%)

25/130 a(19.2%)

25/50(50.0%)

50/130 a(38.4%)

149/279(53.4%)

2004

Pregnancy RateConceptionRate

ED rate

OverallResynchronizationFTAIYear

Persistent Follicle PGF2α

2

6

10

14

mm

Day 0

P4

PGF2α↑CL↑FP

60/70

3131

Wehrman et al 1997

CIDR-B®

BE (2mg) +P4 (50mg)

D8 D17

CIDR-B®

BE (2mg)P4 (50mg

D0 D14 D22

Group CIDR 8 days

Group CIDR 14 days

D15

BE(1mg) TETF

D9

BE(1mg) TETF

D0

PGF2α

PGF2α

D5

3232

Groups N ∅ DF (mm) CL (cm2) [P4 ] ( ng/ml)

G1 (CIDR 8 days

PGF on D8)

70 7.85 ± 0.36b 1.96 ± 0.06b 2.30 ± 0.25b

G2 (CIDR 14 days

+ PGFD0 & D5) 71 11.11 ± 0.31a 2.32 ± 0.07a 3.78 ± 0.25a

G3 (CIDR 14 days

+ PGF D5) 67 10.61 ± 0.37a 2.36 ± 0.08a 3.78 ± 0.30a

G4 (CIDR 14 days

+ PGF D0 & D14) 70 10.60 ± 0.32a 2.16 ± 0.07a,b 3.12 ± 0.28a, b

a, b (p < 0,05)

MANTOVANI et al., 2005

3333

Groups N Transferred/treated (%)

Conception Rates (%)

Preg. Rates(%)

G1 (CIDR 8 days

PGF on D8) 70 51.4 (36/70)b 59.1(21/36)a 30.0 (21/70)a

G2 (CIDR 14 days

+ PGFD0 & D5) 71 77.4 (55/71)a 38.9 (21/55)b 29.6 (21/71)a

G3 (CIDR 14 days

+ PGF D5) 67 74.6 (50/67)a 37.1 (19/50)b 28.4 (19/67)a

G4 (CIDR 14 days

+ PGF D0 & D14) 70 68.6 (48/70)a, b 50.0(24/48)a, b 34.3 (24/70)a

a, b (p < 0,05) MANTOVANI et al., 2005

Use of Embryo Transfer to Increase Pregnancy Rates in Heat Stressed and Repeat Breeder Holstein Cows

66/70

3434

0%

10%

20%

30%

40%

50%

60%

1 2 3 4 5 6 7 8 9 10 11 12

IA TE

aa a

a a

aa

a aa

aa

b b bb

bb

b bb b

b

b

Rodrigues et al SBTE (2007)

Conception Rates at AI (n= 18.568) or embryo transfer (n= 4.871) in Holstein cows.

AI ET

3535

28.50%

41.90%

0%

5%

10%

15%

20%

25%

30%

35%

40%

45%

IA TE

TAX

A D

E C

ON

CE

PÇ

ÃO

IA TE

b

a

Rodrigues et al SBTE (2007)

Conception Rates at AI (n= 18.568) or embryo transfer (n= 4.871) in holsteincows.

AI ET

AI ET

0%

10%

20%

30%

40%

50%

1 2 3 4 5 6 7 8 9 10 11 12

Mês

Taxa

de

Conc

epçã

o

IA TE* * * * * * * * * * * *

0%

10%

20%

30%

40%

50%

1 2 3 4 5 6 7 8 9 10 11 12

Mês

Taxa

de

Conc

epçã

o

IA TE* * * * * * * * * * * *

REPEAT BREEDER (≥4 SERVICES)

(AI vs Embryo Transfer)

Rodrigues et al SBTE (2007)

5.693 AI and 3.858 Embryo Transfers

3636

CONCEPTION RATES 2000 – 2005 CONCEPTION RATES 2000 – 2005

05

101520253035404550

1ª = 3.4712ª = 3.6383ª = 2.451>3ª= 3.183TE = 4.594

Summary• Estrus synchronization and management are critical

factors in a successful embryo transfer programs

• Treatments that synchronize ovulation improve the efficiency of synchronization programs by:

–eliminating the need for estrus detection

–improving the efficiency of embryo transfer programs by increasing the proportion of recipients transferred

3737

Sometimes collecting embryos is not so easy !!!

Neither to transfer them!!!

3838

But Nature is so great that sometimes we end up with some calves on the ground!!!

Thank You!