Use of OPU-IVP and SO at CRV breeding program in the genomic era

Erik Mullaart and Sybrand Merton

Content

Introduction Genomic Selection Impact on breeding program

Selection procedures Delta Nucleus breeding program

Application of OPU-IVP and SO Insire embryo program

Holland Genetics

CRV Worldwide

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

Genomic selection

Which calf received the best genes from the parents?

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)

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

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)

Impact on breeding program; selection procedures

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

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

Renate Raisa Rianne

Inet Tot.ext Leg. Inet Tot.ext Leg. Inet Tot.ext Leg.

EBV 165 109 106 165 109 106 165 109 106

EBV +merkers

213 109 107 178 107 103 145 107 104

BV aftertest

217 110 107 173 108 105 132 108 106

InSireBV

234 110 107 176 106 103 120 107 104

Impact on breeding program; Delta nucleus breeding program

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

Application of OPU-IVP and SO

Embryo production

IVP sessions: 2000 / year 1.6 embryo/ session

SO: 300 / year 4-5 embryos/ flush

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

Holland Genetics

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)

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%

Holland Genetics

Results: Efficiency

OPU-IVP MOET

Oocytes: 8 9

Cleavage: 5,6 (70%)

Embryo: 1,6 (20%) 5 (55%)

Calf: 0,8 (10%) 3 (33%)

Insire embryo program

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

2) Newest InSire embryos (most actual combination)

Benefits to the clients

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

Thank you for your attention!