How can ET-praxis find his feet in the age of

genomics ?

Practitioners` Forum

Frank Becker

Key data of European cattle reproduction

• 90 millions of cattle

– Dairy cows : 23 millions

– Beef cows : 15 millions

• 22 millions of first inseminations

– Dairy : 85 % vs Beef : 15 %

– Holstein : 8.7 ; Simmenthal : 2.8 millions ;

– Charolais : 700 000 ; Limousine : 600 000

PEDIGREES

PHENOTYPE

INFORMATION ABOUT YOUNG ANIMALS UNTIL 2

YEARS AGO

GENOMIC

INFORMATIO

N

+

NEW!

CONSEQUENCES:

• genetic (breeding) progress can

be

doubled

• costs of breeding value

estimation

can be reduced by more than

90%

• young bull dams

• young AI bulls

• genomic analysis is possible immediately after fertilization

• selection is possible very early (in both sexes)

KNUT ROSCHLAU, MASTERRIND Germany

STRATEGY FOR GENOMIC SELECTION OF

EMBRYOS

• Embryos day 6 – 7

• Biopsy of 10 cells

• Whole genome amplification (over night)

• Sexing (next day)

Transfer of female embryos Freezing of male embryos

• genome analysis

• calculation of breeding values

• Selection of male embryos for transfer

(without breeding value)

Best

only!

In vivo embryo production in bovine

• Number of flushed donors 2011: 23,480

• Number of transferable embryos: 108,711

P.I.

+10 points

-10 points

Flushings and OPU/IVP sessions

1. 2. 3. 4. 5. 6.

Example: 24 calves from 6 flushings or OPU/IVP sessions – the calf with the

highest breeding value was produced only in the 6th flushing/session

(P.I. = pedigree index; gZW = genomic breeding value)

gZW

gZW

Embryo Transfers and % IVP embryos

0

5

10

15

20

25

0

20000

40000

60000

80000

100000

120000

140000

160000

180000

19

88

19

90

19

92

19

94

19

96

19

98

20

00

20

02

20

04

20

06

20

08

20

10

Embryo transfers and % IVP embryos

numb transf

% IVF

• In vivo 109,802 (56.2% frozen)

• In vitro 5,661 (39.7% frozen)

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

10000

19

96

19

97

19

98

19

99

20

00

20

01

20

02

20

03

20

04

20

05

20

06

20

07

20

08

20

09

20

10

OPU sessions

10 -13 countries

4-5 countries

Development of bovine in vitro embryo production

Source Knijn H, AETE Workshop 2012: Oocyte collection; what is new?

Use of OPU in Europe

ET teams Minimum

age of start

OPU in

pregnant

animals

Breeds

Italy

AVANTEA

6-7 month Yes HF, Brown Swiss, Simmenthal,

Chianina etc

The Netherlands

CRV

12 month Yes HF and Belgium Blue

France:

UNCEIA

15 month Yes (between

35 and

60days)

Montbeliard, HF, Charolaise,

local breeds

France

MIDATEST

15 month Yes (66% of

donors)

HF, Brown Swiss, Blonde

d’Aquitaine

France:

CREAVIA

>10 month Yes, a

selection

HF

Germany

MASTERRIND

After first

heat

8-9 month

Yes HF

All other breeds

IVP technologies used in 4 countries

2011

Activity

Description

of OPU activity

Number of

OPU

sessions

(donor cows)

Number

Oocytes Transferable

embryos

The

Netherlan

ds

OPU, twice a week, without

superovulation, CRV station (donors)

+ farm

3530 ↗ 28678 3814

(1,1)

Germany OPU, once a week, without

superovulation,

Masterrind station (donors +

recipients) >> farm

1012 4564 3217

(3,2)

Italy OPU, once or twice a week, without

superovulation, Avantea station +

farm

168 2434 423

(2,5)

France OPU, once a week, with

superovulation, 4 stations (donors,

CREAVIA, GD, MIDATEST, UNCEIA)

265 ↗ 2208 524

(2,0)

Total 4975 37984 8034

IVP results in commercial programs

• Immature COCs recovered twice weekly by OPU

• in vitro maturation TCM199/FCS/LH/FSH, with or without

cysteamine (0.1 mM).

• fertilisation with frozen-thawed gradient-separated semen and

culture for 7 d in SOFaaBSA.

Group Sessions

n

Oocytes

n

Cleavage

n (%)

Embryo production Day 7

Mor+ Blast

n (%)

Transferable

n Per

session

Relative

increase

Control 696 5454 3296 (60.4)a 1277 (23.4)a 740 1.06a

Cyst. 589 4557 2939 (64.5)b 1569 (34.4)b 1017 1.73b 63.2%

Merton et al., IETS 2006

We have to preserve embryo viability !

• In vivo derived embryos : high Pregnancy rates achieved under experimental and farm conditions

Author Biopsied

embryos

Biopsy procedure Freezing Pregnancy rate

Gonzales

et al.,

2008

56 day 7 in vivo

derived embryos

(E)

Home-made microblade

fixed to a

micromanipulator +

holding pipette

G1 (24), G2 (7) and

G3 (23) embryos

frozen in 1.5 M EG2,

direct transfer

On D 35 and D 90 :

G1 : 62.5 54.2 %

G2 : 57.1 57.1 %

G3 : 52.2 47.8 %

Lacaze et

al., 2009

671 day 7 in vivo

derived G1

embryos (F)

Home-made microblade

fixed to a

micromanipulator +

holding pipette

Fresh (284) 59 %

1.5 M EG

+ 40 % FCS1 (53)

or 0.1 M sucrose

(340)

50 %

%

Accuracy (%) of genomic breeding values (gZW) in comparision to the accuracy of pedigree breeding values (P.I.)

procuction related traits functional traits

RZG = total merit index

RZM = milk production index

RZS = somatic cell count

RZE = exterior index

RZN = functional herd life

RZR = female reproduction

KV = calving ease & stillbirth

(paternal and maternal)

Reents et al., 2011)

WORLD MILK PRODUCTION (x 1,000 t) AND

DEVELOPMENT OF WORLD POPULATION (x

10,000)

World milk production in milion tones

Source: FAOSTAT

BREEDING SYSTEM FOR PRODUCTION OF AI BULLS

(HOLSTEINS)

PURCHASED

EMBRYOS

POPULATION OF FEMALE CALVES

GENOMIC SELECTION

NUCLEUS 50 – 60 TOP HEIFERS (BULL DAMS)

2 FLUSHINGS OPU/IVP (UP TO 4TH MONTH PREGN.) (5 EMBRYOS/FLUSH) (3 – 4 EMBRYOS/OPU SESSION)

START WITH 10 MONTH

20 – 60 EMBRYOS PER DONOR

RECIPIENTS

TRANSFER (FRESH)

30 – 40 %

FREEZING 60 – 70%

CONTAINER

CALVES

GENOMIC SELECTION

FEMALES

FARMS

BEST HEIFERS!

BULL CALVES

SLAUGHTERHOUSE AI STATION

BEST BULLS!

PURCHASED BULLS

AI STATION, GERMANY

GENOMIC SELECTION TRANSFER

KNUT ROSCHLAU, MASTERRIND

Germany

Continent Flushes Transfer.

Embryos

Number of Transferred Embryos

Fresh Frozen Total & Percentage

Africa 1.438 9.401 4.056 2.469 6.525 1.14 %

Asia 15.444 124.362 24.026 51.697 75.723 13.23 %

Europe 23.480 108.712 41.040 69.381 110.421 19.29 %

N. America 54.837 362.781 109.197 139.418 248.615 43.33 %

S. America 12.174 68.187 36.953 26.054 63.007 11.01 %

Oceania 10.755 59.419 32.921 35.130 68.051 11.52 %

2011 totals 118.128 732.862 248.193 324.149 572.342 100.00

%

2010 totals 104.651 732.000 243.885 291.279 590.561

Per Cent

Change 12.88 % 0,12 % 1.77 % 11.28 % -3.09 %

Bovine In vivo Derived Embryo Activity in 2011

Continent

s

Transferrabl

e Embryos

Number of Transferred Embryos

Fresh Frozen Total & Percentage

Africa 0 0 0 0 0.00 %

Asia 62.418 4.086 6.699 10.785 2.88 %

Europe

(8 cts) 8.034 8.034 3.419 11.453 3.06 %

N.

America 48.474 17.850 2.930 20.780 5.56 %

S. America 325.349 307.278 15.879 323.157 86,44 %

Oceania 9.196 6.679 1.015 7.694 2.06 %

Total 453.471 343.927 29.942 373.869 100.00 %

2010

Totals 450.549 315.715 23.970 339.685

Per Cent

Change 0.65 % 8.94 % 24.91 % 10.06 %

Table: Bovine In vitro Produced Embryos in 2011

Impact of Embryo genotyping on:

• early breeding evaluation • sexing • Preservation of genetic diversity ? • Handling

AI business:

Recipient-management: • Fewer recipients needed

• Strong selection

• Pregnancy management

Thank you for your attention!

Acknowledgement : K. Roschlau, C. Ponsart,

H. Knin, S. Merton

S. Lacase, P. Vos