Backup not only for computers! 27th Annual Meeting of DAGENE, Germany © SZV OÖ Backup not only for computers! Berger B. Knapp P. Gallnböck M.

Introduction Frozen boar semen – since > 40 years Thalheim > 25 years Special uses – not for routine AI Scientific Export Transport Planned mating Genebank – e.g. backup for cattle AI since 1997 Mangalica, Turopolje in Austria since 2000

Frozen boar semen …but long-time storage is possible! About 30% of boars not freezable Non-return rates not really satisfying Smaller litter size Huge amount of sperm cells Less if deep intra-uterine insemination Handling of semen more complicated Processing and freezing Thawing and re-dilution …but long-time storage is possible!

Why another cryo-project? Biggest Austrian pig AI-centre 8 km from Genebank 270 boars, 5 breeds, PRRS-free status Owned by pig breeding association PRRS outbreak 2012, all boars slaughtered Huge financial loss to centre and producers Re-stocked boars not first quality Loss of breeding progress Safeguard against loss of Top-Genetics needed

The project Cooperation between Pig Breeders Association and Genebank Screen all TOP-Genetics boars for freezability of semen Conserve freezable boars in Genebank Do not disrupt semen collection routine at AI centre!

Material & Methods 1 Screening: 62 ejaculates, 28 TOP-boars (15 Piétrain, 7 Landrace, 4 Large White, 1 Duroc, 1 Schwäbisch-Hällisch) Collection within routine of AI centre, 20ml of sperm-rich fraction of ejaculate Dilution 1:1 with BTS extender (DiluPorcTM) Fresh semen analysis CASA (SpermVision, minitube): Motility (mot), Density Transport to Genebank Acrosome integrity (acr) – smears stained acc. KOVACS & FOOTE (1992)

Material & Methods 2 Conservation: 5 to 9 double AI doses/boar Cryo-conservation at Genebank acc. WESTENDORF et al. (1975) mod. EGERSZEGI et al. ( 2009) Extender Androstar Cryo Plus (minitube) 0,5 ml medium French straws, concentration 1x109 cells/ml Thawing at AI centre 37°C for 20 sec. Re-dilution with BTS-extender, CASA, smears Minimal requirements: 30% intact acr, 30% progressive mot Conservation: 5 to 9 double AI doses/boar One whole ejaculate needed

Results – Screening 30% ejaculates discarded! 3 boars not freezable! 70% blocks Minimum/Maximum Median acr mot 30% ejaculates discarded! 3 boars not freezable! fresh thawed

Results – Genebanking 0 ejaculates discarded! All boars freezable! fresh thawed

Discussion Success rate in line with literature Freezability of semen Still 30% negative ejaculates in screening Still 13 boars at least 1 negative result Freezability of semen Individual trait – genetic determination? Fresh semen quality allows no predictions! Sexual resting time important (< 5 days) 5 x 109 sperm cells per dose! Much semen needed for Cryo (one whole ejaculate) Intra-uterine insemination not used by breeders rf

Conclusions Enhanced security for breeders and breeders association Screening well integrated into routine Backup collection owned by breeders association stored at Genebank (extra tank) for 5 years After 5 years 2 double doses into core collection – owned by state, rest destroyed Export orders pending

Thank you for listening! 11 piglets * 10.09.2014 from semen frozen 17.07.1993!