1. Use of OPU-IVP and SO at CRV breeding program in the genomic era
Erik Mullaart and Sybrand Merton

2. Impact on breeding program
Holland Genetics
Content
Introduction
Genomic Selection
Impact on breeding program
Selection procedures
Delta Nucleus breeding program
Application of OPU-IVP and SO
Insire embryo program

3. CRV Worldwide

4. Genetic products of CRV
7 million units of dairy and beef semen
CRV exports semen to over 50 countries
Test program:
310 Holstein bulls
60 Red Holstein bulls
35 Fleckvieh bulls
35 Jersey bulls

5. Genomic selection

6. Which calf received the best genes from the parents?

7. Marker selection
In early days: selection on individual genes (direct or via markers)
Now: selection based on many markers spread across the whole genome, i.e. selection on all genetic material
=GENOMIC SELECTION
(or Genomewide Selection)

8. Genomic Selection, What does it bring us?
Reliable breeding values at young age
higher success rate to get top bulls
lower costs for test proofs (60,000 markers for ~ € 200)
shorter generation-interval
Toelichting geven

9. Effects of Genomic Selection
Increased genetic improvement (30-40%)
Higher genetic level of test sires and embryo’s
Wider portfolio of commercial bulls & higher level
More outcross; less inbreeding
Availability of packs of elite young sires (InSire)

10. Impact on breeding program; selection procedures

11. Use of GS in selection procedures; method implemented by CRV
Step 1:
Genotype proven bulls with reliable
EBVs (reference sires)
Find markers with an effect on EBVs
Calculate marker effect for each trait
Step 2:
Genotype selection candidates
Calculate GS Breeding Values (GBVs) with the marker effects
Select the animals with the highest GBVs

12. 3 full sisters on Delta. Pedigree ( Oman x Jocko Besne)

13. Impact on breeding program; Delta nucleus breeding program

14. 16 years of ‘Delta nucleus’
Test heifers 4381
Selected donors 561
Before Genomic Selection
large number of test heifers per year
emphasis on independent test
With Genomic Selection.
limited number of test heifers
optimal use of reproductive possibilities

15. After GS-selection 110 donor heifers
Delta nucleus
Selection of 800 calves
GS selection
After GS-selection 110 donor heifers
MOET en OPU
embryos per heifer
100 selected heifers tested
Selection of donor cows
ET en OPU
=> 2500 embryos

16. Application of OPU-IVP and SO

17. Embryo production IVP sessions: 2000 / year SO: 300 / year
1.6 embryo/ session
SO: 300 / year
4-5 embryos/ flush

18. Production Logistics 3 x SO at 13-17 months of age: 12 embryos
Holland Genetics
3 x SO at months of age: 12 embryos
Inseminate at 17 months
17 x OPU at months: 20 embryos
Calving at 26 months
32 embryos

19. Future Production Logistics
Holland Genetics
2 x SO at months of age: 8 embryos
10 x OPU at months: 12 embryos
Inseminate at 18 months
17 x OPU at months: 20 embryos
Calving at 27 months
40 embryos

20. Working method: SO D -12 --- Start protocol FSH 1x D -11 --- FSH 2x
D FSH 2x + Prostaglandin with 5th FSH injection
D FSH 2x + Prostaglandin with 6th FSH injection
D FSH 1x
D Heat + AI (2x 8-12 hr / hr)
D Flush + Prostaglandin + Progesterone application
D Remove PRID
D Heat
D D start SO again (D - 12)

21. Working method: OPU-IVP
Day -1 Ovum Pick Up (OPU)
In vitro maturation (IVM)
Day 0 In vitro fertilization (IVF)
Day 1 Start in vitro culture (IVC)
Day 4 Cleavage evaluation
Day 6 Prognoses embryo production
Day 7 Embryo evaluation (fresh, frozen)
Day 8 Final embryo evaluation (frozen)

22. Working method: OPU OPU twice a week (Mo, Thu) No superstimulation
Holland Genetics
Working method: OPU
OPU twice a week
(Mo, Thu)
No superstimulation
(para)veterinarian plus assistant
Pregnant heifers and first parity donor cows

23. Working method: IVM Maturation for 16 - 28 h, 38.5°C, 20% O2
Holland Genetics
Working method: IVM
Maturation for h, 38.5°C, 20% O2
Volume: transport and maturation in 0.5 ml in tubes

24. Working method: IVF Semen: Frozen-thawed
Holland Genetics
Working method: IVF
Semen:
Frozen-thawed
Separation semen and extender by gradient centrifugation (90% / 45%)
Volume: wells (0.5 ml),
Incubation for 24 h, 38.5 °C, 20% O2

25. Working method: IVC Medium: SOFaaBSA Volume: wells (0.5 ml)
Holland Genetics
Working method: IVC
Medium: SOFaaBSA
Volume: wells (0.5 ml)
Incubation for 6-8 days, 38.5°C, 5% O2

26. Working method: IVC Embryos Day 7 - 8, fresh / frozen Stage 4 - 9
Holland Genetics
Working method: IVC
Embryos
Day 7 - 8, fresh / frozen
Stage 4 - 9
(Morula - Hatched Blastocyst)
Grade 1 and 2 IETS

27. Working method: Embryo transfer
Holland Genetics
Recipients
Healthy, fertile recipients
Natural cycling
Day 6, 7, 8 or 9 of estrus cycle
<5% rejected recipients
150 contracted farms

28. Working method: Logistics OPU / IVP
Holland Genetics
Working method: Logistics OPU / IVP
OPU and SO location
Transport COC’s
Central IVP lab
Transport of fresh embryos
Recipient herds
Farmers

29. Results: pregnancy rates
Holland Genetics
Results: pregnancy rates
Fresh IVP embryos:
45% - 55%
Frozen embryos:
IVP 40% - 50%
SO 50% - 60%

30. Results: Efficiency Oocytes: 8 9 Cleavage: 5,6 (70%)
OPU-IVP MOET
Oocytes: 8 9
Cleavage: 5,6 (70%)
Embryo: 1,6 (20%) 5 (55%)
Calf: 0,8 (10%) 3 (33%)

31. Insire embryo program

32. Number of InSire-embryos per year
InSire-embryo donors
Delta test heifers (IVP + SO)
Delta donor cows (IVP + SO) Total from Delta program = 3000
Marker selected Bullmothers in West-Europe. (e.g. France, Germany, Belgium, Denmark and the Netherlands
Totaal 4000 InSire-embryos

33. Costs InSire-embryos InSire-embryos with option € 325,-
InSire-embryos with segment option € 275,-
CRV claims /buy 15-20% of the calves born
male € 1000
female € 3000

34. Segmentation InSire-embryos
Kg Milk
% Protein
Life span en health
Top exterior

35. InSire-embryos offered by Internet
1) Random embryo selection (most actual combination)
Choice of segment ( 4 )
Embryo price € 275
IVP fresh , IVP and SO frozen
2) Newest InSire embryos (most actual combination)
Choice of combination
Embryo price € 325
3) Available InSire embryos (in stock)
IVP and SO frozen

37. 2) Newest InSire embryos (most actual combination)

38. Benefits to the clients

39. Win-Win situation
Client InSire embryos:
- Top class calves
for a relative low price
CRV:
- increased genetics of bulls and bull dams against
an acceptable costs

40. Thank you for your attention!