A procedure for Poitou jackass sperm cryopreservation

摘要

The suitability of 80 mM L-glutamine and 10% quail egg yolk as cryoprotectants of spermatozoa from Poitou donkeys and the effect of post-thaw glycerol removal on sperm motility and fertilizing ability were evaluated. Glutamine at 80 mM and 10% (v/v)quail egg yolk in a basal medium containing 4% (v/v) glycerol (T2-94 medium) improved the post-thaw total and progressive motility and velocity assessed with the automated analyser ATS-M. The T2-94 medium also preserved the sperm nuclear, acrosome, andplasma membrane integrity as assessed with the acridine orange method, fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA) lectin procedure, and hypo-osmotic swelling test, respectively. Semen frozen-thawed in T2-94 medium was used to inseminate 13 Poitou females from the beginning of oestrus to ovulation during 4 cycles at a rate of 1 insemination per day. The pregnancy rate was 62% (8/13) and 3 foals were born but only when the glycerol had been removed from spermatozoa before AI. It is concluded that the cryopreservation of Poitou semen for sperm banking may succeed by using the T2-94 medium and removing the glycerol after thawing.