Use of OPU-IVP and SO at CRV breeding

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

Content Holland Genetics Introduction g Genomic Selection g Impact on breeding program g 4 Selection procedures 4 Delta Nucleus breeding program Application of OPU-IVP and SO g Insire embryo program g

CRV Worldwide

Genetic products of CRV g g g 7 million units of dairy and beef semen CRV exports semen to over 50 countries Test program: 4310 Holstein bulls 460 Red Holstein bulls 435 Fleckvieh bulls 435 Jersey bulls

Genomic selection

Which calf received the best genes from the parents?

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

Genomic Selection, What does it bring us? g Reliable breeding values at young age 4 higher success rate to get top bulls 4 lower costs for test proofs (60, 000 markers for ~ € 200) 4 shorter generation-interval

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

Impact on breeding program; selection procedures

Use of GS in selection procedures; method implemented by CRV g Step 1: 4 Genotype proven bulls with reliable EBVs (reference sires) 4 Find markers with an effect on EBVs 4 Calculate marker effect for each trait g Step 2: 4 Genotype selection candidates 4 Calculate GS Breeding Values (GBVs) with the marker effects 4 Select the animals with the highest GBVs

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

Impact on breeding program; Delta nucleus breeding program

16 years of ‘Delta nucleus’ g g Test heifers 4381 Selected donors 561 Before Genomic Selection 4 4 large number of test heifers per year 4 emphasis on independent test 8 With Genomic Selection. 4 limited number of test heifers 4 optimal use of reproductive possibilities

Delta nucleus g Selection of 800 calves h g GS selection After GS-selection 110 donor heifers h h MOET en OPU 25 - 30 embryos per heifer => 2500 embryos g 100 selected heifers tested g Selection of 10 -15 donor cows h ET en OPU

Application of OPU-IVP and SO

Embryo production g IVP sessions: 2000 / year 41. 6 embryo/ session g SO: 300 / year 44 -5 embryos/ flush

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

Holland Genetics Future Production Logistics g g g 2 x SO at 13 -15 months of age: 8 embryos 10 x OPU at 16 -17 months: 12 embryos Inseminate at 18 months 17 x OPU at 19 -21 months: 20 embryos Calving at 27 months --------40 embryos

Working method: SO g g g g g D -12 --- Start protocol FSH 1 x D -11 --- FSH 2 x D -10 --- FSH 2 x + Prostaglandin with 5 th FSH injection D - 9 --- FSH 2 x + Prostaglandin with 6 th FSH injection D - 8 --- FSH 1 x D - 7 --- Heat + AI (2 x 8 -12 hr / 16 -24 hr) D - 0 --- Flush + Prostaglandin + Progesterone application D +12 --- Remove PRID D +14 --- Heat D + 23 - D + 28 --- start SO again (D - 12)

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)

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

Holland Genetics Working method: IVM g Maturation for 16 - 28 h, 38. 5°C, 20% O 2 g Volume: transport and maturation in 0. 5 ml in tubes

Holland Genetics Working method: IVF g Semen: Frozen-thawed Separation semen and extender by gradient centrifugation (90% / 45%) g Volume: wells (0. 5 ml), g Incubation for 24 h, 38. 5 °C, 20% O 2

Holland Genetics Working method: IVC g Medium: SOFaa. BSA g Volume: wells (0. 5 ml) g Incubation for 6 -8 days, 38. 5°C, 5% O 2

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

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

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

Holland Genetics Results: pregnancy rates g Fresh IVP embryos: 445% - 55% g Frozen embryos: 4 IVP 40% - 50% 4 SO 50% - 60%

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

Insire embryo program

Number of In. Sire-embryos per year g In. Sire-embryo donors 4 Delta test heifers (IVP + SO) 2500 4 Delta donor cows (IVP + SO) 500 Total from Delta program = 3000 4 Marker selected Bullmothers in West-Europe. (e. g. France, Germany, Belgium, Denmark and the Netherlands g Totaal 4000 In. Sire-embryos 1000

Costs In. Sire-embryos g In. Sire-embryos with option € 325, - g In. Sire-embryos with segment option € 275, - g CRV claims /buy 15 -20% of the calves born 4 male € 1000 4 female € 3000

Segmentation In. Sire-embryos g Kg Milk g % Protein g Life span en health g Top exterior

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

2) Newest In. Sire embryos (most actual combination)

Benefits to the clients

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

Thank you for your attention!