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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!