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09/12/2010

Uso de Semen Sexado

Predetermination of Pre-History (6,000 B.C. to 2000 A.D.) Sex


A dream of humankind for 8,000 years
Ancient Egyptians - cattle Peruvians - llamas Bedouins - goats/sheep Mongolians - ponies

L. Cutaia, G. Brogliatti, P Baruselli y GA Bo

Tried everything they knew - but nothing XY Inc. worked

dlg

PASSADO
470 a 402 AC: DEMCRITO SUGERIU QUE O TESTCULO DIREITO PRODUZIA MACHOS E O ESQUERDO FMEAS. DVIDA PERMANECEU AT O PASSADO RECENTE

1976 CONTEDO DE DNA SPTZ


Princpio em que est baseada a sexagem de smen pela citometria de fluxo

DIFERENAS NA QUANTIDADE DE DNA ENTRE OS SPTZ X-Y NAS ESPCIES DE MAMFEROS

DIFERENAS NA QUANTIDADE DE DNA NOS SPTZ X-Y ENTRE RAAS BOVINAS

09/12/2010

XY Then Achieved:

FUTURO: Produo Prod. Mundial de Smen Congelado ~ 300.000.000 doses

1997 - First XY sexed calf

1998 - More sexed calves

Prod. Mundial de Smen Sexado ~ 7.000.000 doses (2% do conv.)

1999 - More sexed calves

1999 - First sexed calves by AI outside USA (UK)

1999 - First sexed calf with frozen semen & AI

XY Inc.

dlg

HO33342 STAINED SPERM

SX MoFlo
Separacin de espermatozoides
3. Los zoides X fluorescen con 4% ms de intensidad que los Y 4. La computadora procesa la fluorescencia detecta-da y categoriza los zoides como X, Y o dudosos 6. Las gotculas cargadas son desviadas cuando pasan frente a placas continuamente cargadas

Desde Computa Des -dora a cargador de gotas

1. Un

cristal piezoelctrico ondula y quiebra la corriente en gotculas ~ 90,000/second. 2. Un rayo laser proyecta luz azul sobre los zoides

A la Compu tdora Detecto r


5. Se aplica una carga negativa, positiva o ninguna a las gotculas 7. Las muestras son recojidas en 3 recipientes

XY, Inc.

Multiplicacin de

hembras mediante el uso


SX MoFlo

de semen sexado
Pietro S. Baruselli

Cytomation Fort Collins, CO

Departamento de Reproduo Animal, Faculdade de Medicina Veterinria e Zootecnia Universidade de So Paulo

XY Inc.

dlg

09/12/2010

Roteiro da apresentao:
Inseminao artificial com smen sexado

Uso de smen sexado para

IATF em gado de corte

1. Com deteco de cio


2. Com sincronizao da ovulao para IATF 3. Em doadoras de embrio

Hiptese experimental:
A reduo no intervalo entre a inseminao e a ovulao proporciona aumento na taxa de concepo em vacas inseminadas com smen sexado

Ejaculado

Congelamento convencional

Sexagem

Congelamento

EFEITO DO MOMENTO DA IATF


Grupo IATF 54h (n=194)
VE + NOR

eCG (400UI)

Smen sexado = 99 Smen Conv. = 95

GnRH IATF 54h


6h
D11 D11

Norgestomet (Crestar)
D0 D9

48h

Grupo IATF 60h (n=195)


VE + NOR

eCG (400UI)

Smen sexado = 99 Smen Conv. = 95

GnRH IATF 60h


12h
D11 D11

Norgestomet (Crestar)
D0 D9

48h

Figura. Efeito do momento da IA (54h ou 60h aps a retirada do implante de progestgeno) e do tipo de smen (convencional ou sexado) na taxa de concepo aps IATF em vacas Nelore (Bos indicus) no ps-parto.
Souza et al., 2007

09/12/2010

60

a
50

a,b = P < 0.05

60 50

P > 0.10

Taxa de concepo

Taxa de concepo

n = 193
40

b
n = 196

n = 195
40

n = 194
30 20 10

30

20

10

Convencional

Sexado

IATF 60h

IATF 54h

Grupos experimentais

Grupos experimentais

Taxa de concepo: convencional vs. sexado


Souza et al., 2007

Taxa de concepo: IATF 60h vs. IATF 54h


Souza et al., 2007

Research colaboration USP and Sexing Dr. Nil Campos Filho PRELIMINARY RESULTS USA EXPERIMENTS

TIME TO OVULATION IN JERSEY

EXPERIMENT 1

HEIFERS SUBMITTED FTAI

09/12/2010

OBJECTIVE
The aim of this study was to evaluate the follicular dynamics in Jersey heifers (Bos taurus) submitted the FTAI.

EXPERIMENTAL DESIGN
EB + PGF

PGF

EB

FTIA

CIDR D0 D8 D9 US 24/24h D0 D8 D9 D10 D12 US 12/12h D10 D12

US 0 hours

US 24 hours

US 48 hours

US 60 hours

US 65 hours

Table - Follicular dynamics in Jersey heifers (Bos taurus) submitted the FTAI.
Animals number 26
92.3 (24/26) 66.12.2 16.02.3 15.62.3

RESULTS

Ovulation rate (%) Interval device removal/ovulation (h) > Dominant Follicle (mm) > Ovulatory Follicle (mm)

100 90 80
Percentage of ovulation

88

EXPERIMENT 2

70 60 50 40 30 20 10 0 48 60 66
Hours after device removal

6 0

6 0 72 84

Figure - Time of ovulation in Jersey heifers (Bos taurus) submitted the FTAI.

09/12/2010

OBJECTIVE AND HYPOTHESES


The objective was to evaluate the interval between

Effect of type of semen (sexed vs non-

progesterone device removal and artificial insemination on pregnancy rates in Bos taurus (Jersey) heifers fixed time inseminated with sexed or non-sexed semen.

sexed) and time of AI (54h vs 60h) on


pregnancy rates of Jersey heifers inseminated in a fixed time

We hypothesized that delay timed AI (TAI) with sexed semen increase the pregnancy rates in Bos Taurus (Jersey) heifers.

EXPERIMENTALS DESIGN
2 x 2 Fatorial: (type of semen vs. timed AI) Groups: 1) Non-sexed semen 54h - AI 12h before ovulation. 2) Non-sexed semen 60h - AI 6h before ovulation. 3) Sexed semen 54h - AI 12h before ovulation. 4) Sexed semen 60h - AI 6h before ovulation.
D0

TREATMENT
1) Group FTAI 54h
FTAI Sexed or Non-sexed EB SEMEN (1.0mg) EB + PGF (2mg) (12.5mg) PGF (12.5mg)

CIDR
D0 D8

54h
D9 D10 D12

2) Group FTAI 60h


EB + PGF (2mg) (12.5mg) PGF (12.5mg) EB (1.0mg) FTAI Sexed or Non-sexed SEMEN

CIDR
D8

60h
D9 D10 D12

P=0.001

60 50 Pregnancy rate (%)

51.2

RESULTS

40
23.7

30
n=213

20
n=207

10 0 Non-sexed Sexed

Figure 1 - Conception rate by semen type (Non-sexed and sexed) in Jersey heifers submitted to FTAI.

09/12/2010

P=0.047

60 50 Pregnancy rate (%) 40 30


n=214

P=0.001 60 42.1 33.0 Pregnancy rate (%) 50

50.5a

51.8a

40 30
20 10 0
n=101

31.4b
n=112

16.2c
n=105

n=102

20 10 0

n=206

Non-sexed FTAI 54h FTAI 54h FTAI 60h

Non-sexed FTAI 60h

Sexed FTAI 54h

Sexed FTAI 60h

Figure 2 - Conception rate by timed AI (54h vs 60h after withdrawal P4 device) in Jersey heifers submitted to FTAI.

Figure 3 - Conception rate by semen type (Non-sexed and sexed) and timed AI (54h vs 60h) in Jersey heifers submitted to FTAI.

P=0.001

60 50 Pregnancy rate (%) 40 27.3b 30 20 10


n=138 n=139 n=143

60

P=0.001 47,3 40,6


ab a a

44.2a

41.3a
Pregnancy rate (%)

50 40

47,2
ab

30,9
30 20 n=71 10 0 n=68

ab

35,2

23,9

n=64

n=74

n=71

n=72

54H

60H

54H

60H

54H

60H

755

760

762

Bull 755

Bull 760

Bulll 762

Figure 4 - Conception rate by bull (755, 760 and 762) in Jersey heifers submitted to FTAI.

Figure 5 - Conception rate by bull (755, 760 and 762) and timed AI (54 and 60h) in Jersey heifers submitted to FTAI.

100 90 80
Pregnancy rate (%) 100 90 a ab

70 60 50 40
bc

61,4
Pregnancy rate (%)

80 70 60 50 40 30
n=35

52,8 39,4
e
n=71 n=72 n=66

64,9 54,5 40 57,6 51,3 42,9


n=37

cd

34,9
n=70

ed

38,9 25,8

30
20 10 0
Non-sexed Sexed

21,9
n=73

n=33 n=35

28,6 15,8
n=38 n=35

14,7
n=68

21,9
n=36

n=33

n=39

20 10

n=31

8,3
n=36

n=32

Non-sexed

Sexed

Non-sexed

Sexed

0
54H 60H 54H 60H 54H 60H 54H 60H 54H 60H 54H 60H
755 Non-sexed 755 Sexed 760 Non-sexed 760 Sexed 762 Non-sexed 762 Sexed

Bull 755

Bull 760

Bulll 762

Figure 6 - Conception rate by bull (755, 760 and 762) and semen (Non-sexed and sexed) in Jersey heifers submitted to FTAI.

Figure 7 - Conception rate by bull (755, 760 and 762), semen (Non-sexed and sexed) and time of FTAI (54 and 60h) in Jersey heifers.

09/12/2010

CONCLUSION
Jersey heifers inseminated with sexed semen showed lower pregnancy rate than non-sexed semen. However, pregnancy rates with sexed semen improved with the further increase of 4h in the interval between the P4 device withdrawal and the FTAI, confirming our initial hypothesis. The 755 bull showed lower pregnancy rate than the 760 and 762 bulls.

EXPERIMENT 3

OBJECTIVE
Rate and time to ovulation in Jersey
The aim of this study was to evaluate follicular

heifers synchronized with new and previously used progesterone

dynamics in Jersey heifers (Bos taurus) synchronized with new and previously used progesterone device in FTAI protocols.

device in FTAI protocols

EXPERIMENTAL DESIGN
EB + PGF NEW or USED CIDR D0 D8 D9 US 24/24h D0 D8 D9 D10 D10 US 12/12h D12 D12 PGF EB FTIA

RESULTS

US 0 hours

US 24 hours

US 48 hours

US 64 hours

09/12/2010

Table - Follicular dynamics in Jersey heifers (Bos taurus) synchronized with new and used device progesterone in protocols to timed AI.
Percentage of ovulation (%)

New
100 90 80 70 72 64

Used

CIDR P New Animals number Ovulation rate (%) Interval device removal/ovulation (h) > Follicle Dominant (mm) > Follicle Ovulatory (mm) 27 92.6 (25/27) 66.90.5 15.50.5 15.10.5 Used 21 100.0 (21/21) 65.5 0.7 14.3 0.5 14.2 0.5 0.98 0.24 0.18 0.33

60
50 40 30 20 10 0 48 60 65 Hours after device removal 72 84 0 0 4 8 20 12 4 0

Figure - Interval between P4 device removal and ovulation in Jersey heifers synchronized with new and previously used progesterone device in FTAI protocols

CONCLUSION
We conclude that new and used device of progesterone presented similar rate and timing

EXPERIMENT 4

of ovulation in protocols that synchronize


ovulation for FTAI in Jersey heifers .

OBJECTIVE AND HYPOTHESES


Effect of time of AI (50h vs 56h) on pregnancy rates of Jersey heifers synchronized with new and previously used progesterone device for FTAI
The objective of this study was to evaluate the effects of the progesterone device (new vs used) and the interval between progesterone device removal and artificial insemination on pregnancy rates in Bos taurus (Jersey) heifers inseminated in

fixed time with sexed semen.

We hypothesized that delay timed AI (TAI) with sexed semen increase the pregnancy rates in Bos taurus (Jersey) heifers.

09/12/2010

EXPERIMENTALS DESIGN
2 x 2 Fatorial: (Timed FTAI vs type of CIDR) Groups:

TREATMENT
Groups FTAI 50h /New or used CIDR
EB + PGF (2mg) (12.5mg) PGF (12.5mg) EB (1.0mg) FTAI

NEW OR USED CIDR

50h
D8 D9 D10 D12

1) New CIDR and FTAI 50h. 2) Used CIDR and FTAI 50h. 3) New CIDR and FTAI 56h. 4) Used CIDR and FTAI 56h.

D0

Groups FTAI 56h /New or used CIDR


EB + PGF (2mg) (12.5mg) PGF (12.5mg) EB (1.0mg) FTAI

NEW OR USED CIDR


D0 D8

56h
D9 D10 D12

P=0.18
50
45 40 Pregnancy rate (%) 35 30 25 20 15 10 5 0 50h 56h Hours after removal device
n=208 n=200

RESULTS

19.5
14.9

Figure 1 - Conception rate by timed AI (50h vs 56h after removal P4 device) in Jersey heifers submitted to FTAI.

P=0.28
50 50 45 45 40

P=0.28

Pregnancy rate (%)

40 Pregnancy rate (%) 35 30 25 20 15 10 5 0 n=206 n=202

35 30 25 20 15 10 5 0 New CIDR - FTAI New CIDR - FTAI Used CIDR - FTAI Used CIDR - FTAI 50h 56h 50h 56h n=107 n=99 n=101 n=101 13.1 18.2 20.8 16.8

18.8 15.5

New CIDR

Used CIDR

Figure 2 - Conception rate by progesterone device (new vs 1x used) in Jersey heifers submitted to FTAI.

Figure 3 - Conception rate by bull (755, 760 and 762) in Jersey heifers submitted to FTAI.

10

09/12/2010

50
50 45 40 Pregnancy rate (%) 35 30 25 20 15 10 5 0
8.5
n=47 n=25 n=19 n=82 n=72

45 40
34.2 28.6 24 19.1
n=79

34.2

Pregnancy rate (%)

35 30 25 20 15 10 5 0 n=47 n=82 n=72 n=29 12.2 8.5 n=79 8.3 10.3

10.5

12.2 8.3

n=7

10.3
n=21 n=29

10 7.1
n=10 n=17

492

503

504

753

760

761

762

763

766

767

773

492

753

760

761

762

Figure 4 - Conception rate by bull in Jersey heifers submitted to FTAI.

Figure 5 - Conception rate by bull in Jersey heifers submitted to FTAI.

50 45

40
Pregnancy rate (%) 35 30 25 20 15 10 5 0 FTAI 50h FTAI 54h
n=208 n=105

Experiment 2 Experiment 4
31,4

19,5 14,9 16,2


n=102 n=200

Efeito do touro na taxa de prenhez com semen sexado

FTAI 56h

FTAI 60h

Hours after removal device

Figure 6 - Conception rate by timed AI (50h, 54h, 56h and 60h after withdrawal P4 device) in Jersey heifers submitted to FTAI.

Danish A.I. field data with sexed semen


80 70 60
Pregnancy rate (%)

Sexed Unsexed

50 40 30 20 10 0 Bull 1 Bull 2 Bull 3 Bull 1 Bull 2 Bull 3 Bull 1 Bull 2 Bull 3

Holstein

Red Breed

Jersey

Figure: Conception rate by bull within breed


Borchersen and Peacock, 2009 (Theriogenology, v. 71, p. 5963)

Taxa de prenhez de vacas Nelore inseminadas com smen sexado conforme o touro (n=3.200 IA), Fazenda Campo Verde e Paraso, Viana-ES, 2006/2007

11

09/12/2010

Danish A.I. field data with sexed semen

EXPERIMENT 5
Holstein (-12,6%) Red Breed (-7,3%) Jersey (-5,2%)

Efeito de raa ?

Borchersen and Peacock, 2009 (Theriogenology, v. 71, p. 5963)

OBJECTIVE
Rate and time to ovulation in Jersey
The aim of this study was to evaluate the follicular dynamics in Jersey heifers (Bos taurus) in normal estrous cycle.

heifers in normal estrous cycle

ULTRA-SONOGRAPHY EXAMS
ESTROUS US US US US

D0 AM

D0 PM

D1 AM

D1 PM

D2 AM

RESULTS

US D0 AM

US D0 PM

US D1 AM

12

09/12/2010

Table - Follicular dynamics in Jersey heifers (Bos taurus) in normal estrous cycle.
Animals number Ovulation Rate until 48h (%) Interval ovulation: Onset heat (h) End heat (h) > Follicle Ovulatory (mm) 31.2 0.9 16.3 1.2 14.1 0.3 43 86.1 (37/43)
Percentage of ovulation

50 45 40 35 30 25 20 15 10 5 0 18 to 24 24 to 30 30 to 36 >36 Hours after onset heat

32,4

29,7

27,0

10,8
n=04

n=12

n=11

n=10

Figure - Heat onset to ovulation interval in Jersey heifers (Bos taurus ) identified by Heat Watch system.

EXPERIMENT 6

Timing of insemination with sexed or nonsexed semen on pregnancy rates of Jersey heifers identified in estrus by radiotelemetric estrus detection system

TREATMENT
2 x 4 Fatorial: (type of semen vs timed AI)
Groups:
a) Sexed semen 1) AI 0 to 6 hours after onset heat 2) AI 6 to 12 hours after onset heat 3) AI 12 to 18 hours after onset heat 4) AI 18 to 24 hours after onset heat b) Non-sexed semen

EXPERIMENTALS DESIGN
AI Sexed or Non-sexed SEMEN Heat Watch + PGF
Heat detection for Heat Watch

US

1)
2) 3) 4)

AI 0 to 6 hours after onset heat


AI 6 to 12 hours after onset heat
D0 D7 D37

AI 12 to 18 hours after onset heat AI 18 to 24 hours after onset heat

13

09/12/2010

P=0.001
100 90 80

RESULTS

70 Pregancy rate (%) 60 50 40 30 20 10 0

a 64,2 b 49,5

n= 371 n= 382

Sexed

Non-sexed

Figure 1 - Conception rate by semen type (Non-sexed and sexed) in Jersey heifers identified in estrus by Heat Watch system.

Sexed
80
70

Non-sexed

70
60

66,7 54,4 45,8

65,5

Pregnancy rate (%)

60 50 40
n=60 46,9 47,2

58 53,6

TIMING OF INSEMINATION (Onset heat)

n=91 n=89 n=97

n=77 n=72

n=84 n=57

n=58

30
n=64

20 10 0
0 to 5 5 to 10 10 to 15 15 to 20 20 to 25

Interval from first standing event to AI (h)

Figure 4 - Timing of insemination with sexed or non-sexed semen on pregnancy rates of Jersey heifers identified in estrus by Heat Watch system.

755 BULL
Sexed
80 70 60 Pregancy rate (%) 48,2 50 40 30
n=85 n=78 n=111 n=104 n=99 n=100 n=84 n=88

Non-sexed
Sexed
68,1

Non-sexed

62,8 48,7

60,6 49,5 52,4

64,7

80 68,4 70 60 Pregancy rate (%) 50 40 30 20 10 0


n=19 n=18 n=30 n=31 n=28 n=27 n=22 n=22

61,1

63,6 56,7 51,6 39,3 55,6 40,9

20 10 0 0 to 6 6 to 12 12 to 18 18 to 24 Interval from first standing event to AI (h)

0 to 6

6 to 12

12 to 18

18 to 24

Interval from first standing event to AI (h)

Figure 5 - Timing of insemination with sexed or non-sexed semen on pregnancy rates of Jersey heifers identified in estrus by Heat Watch system.

Figure 6 - Timing of insemination with sexed or non-sexed semen on pregnancy rates of Jersey heifers identified in estrus by Heat Watch system.

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09/12/2010

760 BULL
Sexed 80 70 60 Pregancy rate (%) 50 40 30 20 10 0 0 to 6 6 to 12 12 to 18 18 to 24 Interval from first standing event to AI (h)
n=40 n=38 n=51 n=42 n=36 n=43 n=41 n=42

762 BULL
Sexed
65,9 69,1 80 70 60 Pregancy rate (%) 59,1 51,6 44,1 27,6
n=22 n=25 n=29 n=31 n=35 n=30 n=19 n=24

Non-sexed 74,4

Non-sexed
70,1 58,3 48,5 36,8

73,8 65,8 56,9 42,5 58,3

50 40 30 20 10 0

0 to 6

6 to 12

12 to 18

18 to 24

Interval from first standing event to AI (h)

Figure 7 - Timing of insemination with sexed or non-sexed semen on pregnancy rates of Jersey heifers identified in estrus by Heat Watch system.

Figure 8 - Timing of insemination with sexed or non-sexed semen on pregnancy rates of Jersey heifers identified in estrus by Heat Watch system.

Sexed
70 60,1 60 48,8 Pregancy rate (%) 50 40 30 20 10 0 > -8 -8 to 0
n=86 n=70 n=110 n=120

Non-sexed
66,7 63,7 51,5 63,3

48,2

48,5

TIMING OF INSEMINATION (End heat)

n=113 n=103 n=66

n=60

0 to 8

>8

Interval from end heat to AI (h)

Figure 9 - Timing of insemination with sexed or non-sexed semen on pregnancy rates of Jersey heifers identified in estrus by Heat Watch system.

Sexed
70 59,8 60 Pregancy rate (%) 50 40 30 20 10 0 0 to 10 10 to 20 20 to 30
n=91

Non-sexed
64 ab 50,7 a 56,1 ab 45,8 66,7 67,2

a 57,8

64,5

NUMBER OF MOUNT

b 38,5

n=92 n=90

n=76 n=77

n=75 n=50

n=63 n=59

n=58

30 to 40

> 40

Number of mount during heat

Figure 10 - Conception rate by number of mount in Jersey heifers identified in estrus by Heat Watch system.

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Sexed
70 60 Pregancy rate (%) 50 40 30 20 10 0
n=55

Non-sexed
66,7 ab a 50,9 67,6 a 62 ab ab 46,2

b 54,1

a 57,1

DURATION OF HEAT

b 34,6
n=61 n=84 N=81 n=177 n=151 n=52 n=71

0 to 6

6 to 12

12 to 18

18 to 24

Duration of estrus (h)

Figure 11 - Conception rate by duration of estrus in Jersey heifers identified in estrus by radiotelemetric estrus detection system .

CONCLUSION
Our initial hypothesis that AI using sexed semen performed around the end of the estrus

Acknowledgements
Sexing Technologies and Dalhart Jersey Ranch

increase
confirmed.

the

pregnancy

rates

was

not

Next studies using more animals should be conduced to confirm this preliminary results.

INSEMINACION ARTIFICIAL A TIEMPO FIJO PROFUNDA CON SEMEN SEXADO EN VAQUILLONAS HOLANDO
G. M. Brogliatti, G. Domnguez, M. G. Lusenhofft, J. Perkins, G. A. Bo, Cutaia L.
1Centro de Inseminacion Artificial La Argentina Chica, Vdo Tto; 2 Actividad Privada Venado Tuerto; 3 Adeco SA; 4 IRAC Instituto de Reproduccin Animal de Cordoba,

Introduccion
Poder elegir el sexo en los mamferos ha sido uno de los objetivos ms buscados durante mucho tiempo por los cientficos y ganaderos del mundo.

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09/12/2010

Limitantes del Semen Sexado


Semen Sexado = Baja Preez
Baja concentracion? Espermatozoides activados? Lugar de deposicion? Horario a la ovulacion? Plasma seminal?

Vagina, Cervix, Utero, Oviducto para llegar a la fertilizacion?

Test Termo-Resistencia
Porcentaje de espermatozoides mtiles progresivos

Trabajos previos
Lugar de Inseminacion Seidel y Col. CSU (1999)
Horn 27/49 68% Body 25/50 60% 85/158 53% 70/163 40%

Hora de incubacin(hs) Semen sexado Semen no-sexado

0 (%) 23,14,9

3 (%) 3,7 3,2

Brogliatti y col Goyaike (2001)


Cuerno 16/46 Cuerpo 14/54 34,8% 25,4%

43,74,9 38,7 10,2


Brogliatti y col 2005

Trabajos previos
Horario de la IATF Veneranda y Col
52 Horas 18/50 60 Horas 20/50 54 Horas 72/185 36 % 40 % 37 %

Objetivo
Comparar los porcentajes de preez con semen sexado (2 M de esp.)
Depositados en Cuerpo (tradicional) vs Cuerno (profundo) 52 h vs 58 h posterior al retiro del dispositivo intravaginal.

Capitaine
1241 servicios en 17 establecimientos en vaquillonas, 37 % (34-40 %)

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Materiales y Metodos
Vaquillonas Holando Argentino entre 15 y 18 m
condicin corporal 3,05 + 0,22. evaluadas por ecografa para determinar estructuras ovricas y tamao del tero inicio

Materiales y Metodos
Un solo inseminador con una jeringa tradicional para IA en el cuerpo y la jeringa de TE para las inseminaciones en el cuerno uterino (vainas de ET, SBS Cryotec sa, Arg.). Diagnstico de preez ecografia transrectal 30 d Analizados por regresion logistica y al combinar los datos por Chi Cuadrado (Infostat).

Al momento de la IA, ecografa para determinar el tamao y el lugar del folculo preovulatorio.

Protocolo utilizado para la IATF


Cuerpo y Cuerno
PG

Resultados I
52 hs
Cuerno Cuerpo 21/43 49 % 17/45 38 % 38/88 43 %

58 hs
15/24 63 % 21/51 41 % 36/75 48 %

Total
36/67 53 %a 38/96 39% b

Semen sexado IATF 52 h


0
B.E

PG

IATF 52
9
B.E

CIDR (8 d)
8 10

Cuerpo y Cuerno IATF 58

PG

PG

Semen sexado CIDR (8 d) IATF 58 h 0 8


B.E

9
B.E

10

Total

Resultados II (IETS 2010)


52 hs
Cuerno
Cuerpo

58 hs
32/65 49,2 % 35/104 33,6 % 67/169 39 %

Total
64/155 41 % 70/202 34 % Vaq. 357

Total

32/90 35,5 % 35/98 35,7 % 67/188 35 %

Emprego do smen sexado em doadoras superovuladas e inseminadas em tempo fixo (SOTF)

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Ejaculado

Grupo smen convencional (n=10)

Retirada do CRESTAR

PGF2
BE (2mg)

GnRH 12h FSH CRESTAR IA IA

Colheita de Embrio

D0

D4

D5

D6

D7 T

D8 M

D8 T

D9 M

D15 M

Grupo smen sexado (n=10)

Congelamento convencional

Sexagem
BE (2mg)

Retirada do CRESTAR

PGF2 GnRH 12h FSH CRESTAR IA IA

Colheita de Embrio

Congelamento

D0

D4

D5

D6

D7 T

D8 M

D8 T

D9 M

D15 M

Experimental design
1st Replicate
Smen convencional (n=5)
(35d)

2nd Replicate
Smen sexado (n=5)

Tabela 1 - Produo de embries em vacas Nelore (Bos indicus) superestimuladas e inseminadas em tempo fixo com smen sexado e convencional, Poos Caldas, MG, 2007.
Smen Convencional (n=10)
Total estruturas (n) Emb. transferveis (n) Emb. congelveis (n) 9.90 0.78 6.80 0.66 5.90 0.71 1.50 0.48 1.60 0.37 68.7

Smen Sexado (n=10)


8.40 1.40 4.20 0.74 3.50 0.65 3.70 0.88 0.50 0.16 50.0

P Trat Rep Trat vs Rep

0.28 0.03 0.03 0.01 0.04 0.01

0.81 0.88 0.43 0.82 0.54 0.68

0.71 0.88 0.99 0.46 0.78 0.54

Smen sexado (n=5)

Smen convencional (n=5)

Emb. no fertilizados (n) Emb. degenerados (n) Taxa de emb. viveis (%)*

Taxa de emb. congelveis (%)**


Group P24/LH48 Group P24/LH60

59.6

41.7

0.02

0.32

0.73

*Taxa de emb. viveis = (emb. viveis/total de estruturas)*100

The same sires were used in all replicates

**Taxa de emb. congelveis = (emb. congelveis/total de estruturas)*100

Martins et al., 2007

Group Conventional ans sexed semen


DIB insert BE (D0)

DIB removal Flushing

Experimental design
1st Replicate Group Conv 12/24h (n=4)
D15 AM (60d)

PGF2 LH 12h FSH (133mg


Folltr.)

2nd Replicate Group Sexed 12/24h (n=4) Group Sexed 18/30h (n=4) Group Conv 12/24h (n=4)

(60d)

3rd Replicate Group Sexed 18/30h (n=4) Group Conv 12/24h (n=4) Group Sexed 12/24h (n=4)

AI

AI

DIB vaginal device


D-0 D4 D5 D6 D7 PM D8 AM

12h
D8 PM

24h
D9 AM

Group sexed semen


DIB inset BE (D0)

DIB removal

Group Sexed 12/24h (n=4)


Group Sexed 18/30h (n=4)

PGF2 LH 12h FSH (133mg


Folltr.)

Flushing

AI

AI

DIB vaginal device


D0 D4 D5 D6 D7 PM D8 AM

18h

30h
D8 PM D9 AM

D15 AM

The same sires were used in all replicates

Group Convencional semen (AI 12/24h after LH) Group sexed semen (AI 12/24h after LH) Group sexed semen (AI 18/30h after LH)

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8 7 6 5 4 3 2 1 0
Mdico Veterinrio: Claudiney de Melo Martins Fazenda: Santa Brbara, Botenhos - MG

7.25 a 5.17 3.25

Transferable embryos

Conv 12/24h

Sexed 12/24h

Sexed 18/30h

Figure - Effect of type of semen (conventional vs. sexed) and timed artificial insemination after LH (12 and 24 vs. 18 and 30) in Nelore cows (Bos indicus) superstimulated, Poos de Caldas, MG, 2008. Martins et al., 2008

Group Conventional ans sexed semen


DIB insert BE (D0)

DIB removal Flushing

PGF2 LH 24h FSH (200mg


Folltr.)

AI

AI

DIB vaginal device


D-0 D4 D5 D6 D7 PM D8 AM

12h
D8 PM

24h

D9 AM

D15 AM

Group sexed semen


DIB inset BE (D0)

DIB removal

PGF2
LH 24h FSH (200mg
Folltr.)

Flushing

9 8 7 6 5 4 3 2 1 0

8.92

a
5.92 4.41

b
Transferable embryos

Conv 12/24h

Sexed 12/24h

Sexed 18/30h

AI

AI

DIB vaginal device


D0 D4 D5 D6 D7 PM D8 AM

18h

30h
D8 PM D9 AM

D15 AM

Figure - Effect of type of semen (conventional vs. sexed) and timed artificial insemination after LH (12 and 24 vs. 18 and 30) in Holstein heifers (Bos taurus) superstimulated, Poos de Caldas, MG, 2008. Martins et al., 2008

Grupo smen convencional (n=10)

Retirada do CRESTAR

PGF2
BE (2mg)

GnRH 12h FSH CRESTAR IA IA

Colheita de Embrio

EVALUACIN DE SEMEN SEXADO EN LA PRODUCCIN DE EMBRIONES Y TRANSFERENCIAS EMBRIONARIAS EN FRESCO

D0

D4

D5

D6

D7 T

D8 M

D8 T

D9 M

D15 M

Grupo smen sexado (n=10)

Retirada do CRESTAR

PGF2
BE (2mg)

GnRH 12h FSH CRESTAR IA IA

Colheita de Embrio

D0

D4

D5

D6

D7 T

D8 M

D8 T

D9 M

D15 M

20

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INTRODUCCION OBJETIVOS

MATERIALES Y METODOS

PROTOCOLO DONANTES CON DISPOSITIVO Dia 0 8.00 h. Colocacion Disp.+p4 + 17bEstradiol Dia 5 8.00 h. 3cc Folltropin V 18.00 h. 3cc Folltropin V Dia 6 8.00 h. 2cc Folltropin V 18.00 h. 2cc Folltropin V Dia 7 8.00 h. 1.5cc Folltropin V+ 25mg dinoprost 18.00 h. 1.5cc Folltropin V+ Ret. Disposit. Dia 8 8.00 h. 1-0.5cc Folltropin V 18.00 h. 1-0.5cc Folltropin V Dia 9 Control de celo + IA Dia 16 Lavaje y Transf

PROTOCOLO DONANTES SIN DISPOSITIVO


Dia -11 Dia 0 Dia 1-5 Dia 13 Dia 14 8.00 h. 18.00 h. 8.00 h. 18.00 h. 18.00 h. 18.00 h. Pg f2a Pg f2a Deteccion de celo 3cc Folltropin V 3cc Folltropin V 2cc Folltropin V 2cc Folltropin V

PROTOCOLO RECEPTORAS CON Y SIN DISPOSITIVO

Dia15
Dia 16 Dia 17

8.00 h.
18.00 h. 8.00 h. 18.00 h.

1.5cc Folltropin V+ 25mg dinoprost


1.5cc Folltropin V+ 25mg dinoprost 1-0.5cc Folltropin V 1-0.5cc Folltropin V Control de celo + IA

Dia 24

Lavaje y Transf

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PRODUCCION DE EMBRIONES MEDIAS +- EE EN VACAS ANGUS SUPEROVULADAS E INSEMINADAS CON SEMEN SEXADO O CONVENCIONAL

PRODUCCION DE EMBRIONES MEDIAS +- EE EN VACAS ANGUS SUPEROVULADAS E INSEMINADAS CON SEMEN SEXADO O CONVENCIONAL
SEMEN SEXADO Toro Rp N de donantes inseminadas Ovocitos/ embriones totales Embriones fertilizados Embriones transferibles % Embrionestransferibles /ovocitos embriones totales Transferencias en fresco Preadas % Preez en fresco RP 251 29 13+2,5a 9+1,6 7,3+1ab 60a 32 19 59.4a RP 253 5 17.3+4,9a 11,7+2,9 10,2+2,4b 51.6a 20 13 65a RP 913 10 20.3+7,5 a 3,4+1,2 2,8+1a 15.6b 10 3 30a TOTAL 44 15.51+2,3 a 8,4+1,3a 6,94+1a 41.69a 62 35 56.5a SEMEN NO SEXADO 27 10.5+ 1,5a 7,6+1,2a 6,83+ 1,16a 64.43b 65 25 38.5b

NO SEXADO Tratamientos Superovulatorios Ovocitos/embriones totales Embriones Fertilizados Emb Transferibles/Vaca 29 10.5+ 1,55 a 7,62+ 1,25 a 6,83+ 1,16 a

SEXADO 34 14,21+ 2,22 a 9,72+ 1,38 a 8,07+ 1,02 a

% EMB. / ovocitos embriones totales Transferencias en fresco Preadas(Transf. En fresco) % Preez en fresco

64.4a 65 25 38.5a

52.8b 52 32 61.5b

En cada fila valores con diferentes letras difieren significativamente (p 0.05)

En cada fila valores con diferentes letras difieren significativamente (p0.05)

Emprego do smen sexado em IVF

Cometarios

Comentarios Generales

Muchas Gracias

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