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Use of OPU-IVP and SO at CRV breeding program in the genomic era. Erik Mullaart and Sybrand Merton. Holland Genetics. Content. Introduction Genomic Selection Impact on breeding program Selection procedures Delta Nucleus breeding program Application of OPU-IVP and SO
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Use of OPU-IVP and SO at CRV breeding program in the genomic era Erik Mullaart and Sybrand Merton
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
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
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)
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 Genomic Selection,What does it bring us?
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)
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
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
Delta nucleus • Selection of 800 calves • GS selection • After GS-selection 110 donor heifers • MOET en OPU • 25 - 30 embryos per heifer • 100 selected heifers tested • Selection of 10 -15 donor cows • ET en OPU => 2500 embryos
Embryo production • IVP sessions: 2000 / year • 1.6 embryo/ session • SO: 300 / year • 4-5 embryos/ flush
Holland Genetics Production Logistics • 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 • 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 • D -12 --- Start protocol FSH 1x • D -11 --- FSH 2x • D -10 --- FSH 2x + Prostaglandin with 5th FSH injection • D - 9 --- FSH 2x + Prostaglandin with 6th FSH injection • D - 8 --- FSH 1x • D - 7 --- Heat + AI (2x 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 • OPU twice a week (Mo, Thu) • No superstimulation • (para)veterinarian plus assistant • Pregnant heifers and first parity donor cows
Holland Genetics Working method: IVM • Maturation for 16 - 28 h, 38.5°C, 20% O2 • Volume: transport and maturation in 0.5 ml in tubes
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
Holland Genetics Working method: IVC • Medium: SOFaaBSA • Volume: wells (0.5 ml) • Incubation for 6-8 days, 38.5°C, 5% O2
Holland Genetics Working method: IVC Embryos • Day 7 - 8, fresh / frozen • Stage 4 - 9 (Morula - Hatched Blastocyst) • Grade 1 and 2 IETS
Holland Genetics Working method: Embryo transfer Recipients • Healthy, fertile recipients • Natural cycling • Day 6, 7, 8 or 9 of estrus cycle • <5% rejected recipients • 150 contracted farms
Holland Genetics Working method: Logistics OPU / IVP • OPU and SO location • Transport COC’s • Central IVP lab • Transport of fresh embryos • Recipient herds • Farmers
Holland Genetics Results: pregnancy rates • Fresh IVP embryos: • 45% - 55% • Frozen embryos: • IVP 40% - 50% • SO 50% - 60%
Results: Efficiency OPU-IVP MOET • Oocytes: 8 9 • Cleavage: 5,6 (70%) • Embryo: 1,6 (20%) 5 (55%) • Calf: 0,8 (10%) 3 (33%)
Number of InSire-embryos per year • InSire-embryo donors • Delta test heifers (IVP + SO) 2500 • Delta donor cows (IVP + SO) 500 Total from Delta program = 3000 • Marker selected Bullmothers in West-Europe. (e.g. France, Germany, Belgium, Denmark and the Netherlands 1000 • Totaal 4000 InSire-embryos
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
Segmentation InSire-embryos • Kg Milk • % Protein • Life span en health • Top exterior
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 • IVP fresh , IVP and SO frozen • 3) Available InSire embryos (in stock) • Embryo price € 325 • IVP and SO frozen
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