Practitioners` Forum How can ET-praxis find his feet in the age of genomics? 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
INFORMATION ABOUT YOUNG ANIMALS UNTIL 2 YEARS AGO PEDIGREES genomic analysis is possible immediately after fertilization selection is possible very early (in both sexes) young bull dams young AI bulls + NEW! PHENOTYPE GENOMIC INFORMATIO N CONSEQUENCES: genetic (breeding) progress can be doubled costs of breeding value estimation can be reduced by more than 90% 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 (without breeding value) Freezing of male embryos genome analysis calculation of breeding values Best only! Selection of male embryos for transfer
In vivo embryo production in bovine Number of flushed donors 2011: 23,480 Number of transferable embryos: 108,711
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) +10 points gzw P.I. gzw -10 points 1. 2. 3. 4. 5. 6. Flushings and OPU/IVP sessions
1988 1990 1992 1994 1996 1998 2000 2002 2004 2006 2008 2010 Embryo Transfers and % IVP embryos In vivo 109,802 (56.2% frozen) In vitro 5,661 (39.7% frozen) Embryo transfers and % IVP embryos 180000 25 160000 140000 120000 100000 80000 60000 40000 20000 20 15 10 5 numb transf % IVF 0 0
1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 Development of bovine in vitro embryo production 10-13 countries 10000 OPU sessions 9000 8000 7000 6000 5000 4-5 countries 4000 3000 2000 1000 0
Use of OPU in Europe ET teams Italy AVANTEA The Netherlands CRV France: UNCEIA France MIDATEST France: CREAVIA Germany MASTERRIND Minimum age of start OPU in pregnant animals Breeds 6-7 month Yes HF, Brown Swiss, Simmenthal, Chianina etc 12 month Yes HF and Belgium Blue 15 month Yes (between 35 and 60days) 15 month Yes (66% of donors) >10 month Yes, a selection After first heat 8-9 month Yes Montbeliard, HF, Charolaise, local breeds HF, Brown Swiss, Blonde d Aquitaine HF HF All other breeds Source Knijn H, AETE Workshop 2012: Oocyte collection; what is new?
IVP technologies used in 4 countries 2011 Activity The Netherlan ds Germany Italy France Description of OPU activity OPU, twice a week, without superovulation, CRV station (donors) + farm OPU, once a week, without superovulation, Masterrind station (donors + recipients) >> farm OPU, once or twice a week, without superovulation, Avantea station + farm OPU, once a week, with superovulation, 4 stations (donors, CREAVIA, GD, MIDATEST, UNCEIA) Number of OPU sessions (donor cows) Oocytes Number Transferable embryos 3530 28678 3814 (1,1) 1012 4564 3217 (3,2) 168 2434 423 (2,5) 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 1.06 a Cyst. 589 4557 2939 (64.5) b 1569 (34.4) b 1017 1.73 b 63.2% Merton et al., IETS 2006
We have to preserve embryo viability! Author Gonzales et al., 2008 Lacaze et al., 2009 Biopsied embryos 56 day 7 in vivo derived embryos (E) 671 day 7 in vivo derived G1 embryos (F) Biopsy procedure Freezing Pregnancy rate Home-made microblade fixed to a micromanipulator + holding pipette Home-made microblade fixed to a micromanipulator + holding pipette G1 (24), G2 (7) and G3 (23) embryos frozen in 1.5 M EG 2, direct transfer On D 35 and D 90 : G1 : 62.5 54.2 % G2 : 57.1 57.1 % G3 : 52.2 47.8 % Fresh (284) 59 % 1.5 M EG + 40 % FCS 1 (53) or 0.1 M sucrose (340) 50 % In vivo derived embryos : high Pregnancy rates achieved under experimental and farm conditions
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 BULLS AI STATION SLAUGHTERHOUSE FARMS BEST BULLS! BULL CALVES FEMALES POPULATION OF FEMALE CALVES GENOMIC SELECTION GENOMIC SELECTION CALVES AI STATION, GERMANY NUCLEUS 50 60 TOP HEIFERS (BULL DAMS) START WITH 10 MONTH GENOMIC SELECTION TRANSFER (FRESH) 30 40 % RECIPIENTS TRANSFER 2 FLUSHINGS OPU/IVP (UP TO 4TH MONTH PREGN.) (5 EMBRYOS/FLUSH) (3 4 EMBRYOS/OPU SESSION) CONTAINER BEST HEIFERS! 20 60 EMBRYOS PER DONOR FREEZING 60 70% PURCHASED EMBRYOS KNUT ROSCHLAU, MASTERRIND
Bovine In vivo Derived Embryo Activity in 2011 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 %
Table: Bovine In vitro Produced Embryos 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) N. America 8.034 8.034 3.419 11.453 3.06 % 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 Per Cent Change 450.549 315.715 23.970 339.685 0.65 % 8.94 % 24.91 % 10.06 %
Impact of Embryo genotyping on: AI business: early breeding evaluation sexing Preservation of genetic diversity? Handling 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