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Improved Quality of Cryopreserved Cheetah (Acinonyx jubatus) Spermatozoa after Centrifugation Through Accudenz

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Abstract

  • Sperm cryopreservation, in combination with assisted reproductive techniques, is a valuable tool for the genetic management of endangered felids. However, the acrosome of the cheetah spermatozoon is especially sensitive to cryopreservation, with {approx}40% of spermatozoa experiencing acrosomal damage immediately after thawing and then another {approx}15% loss during the next 4 h in vitro. Additionally, thawing causes a reduction in sperm motility by {approx}20% with another decrease of {approx}12% during subsequent incubation in vitro. We hypothesized that slow removal of glycerol from cryopreserved cheetah spermatozoa using an Accudenz gradient would improve acrosomal integrity, sperm motility longevity and structural morphology. Accudenz was compared to traditional cheetah sperm processing methods for glycerol removal that involved washing, multi-step resuspension and swim-up processing. Electroejaculates (n = 21; n = 8 males) were washed in Ham's F10 medium (HF10) and sperm pellets resuspended in TEST Yolk-Buffer (TYB) with 0% glycerol. Samples were cryopreserved in straws in 4% final glycerol, thawed and assessed for percent intact acrosomes (% IA), percent motility (% M) and forward progressive status (FPS; scale 0-5). Sperm motility index (SMI) was calculated as [% M (FPS x 20)] / 2. In Study 1, glycerol removal by centrifugation through an Accudenz gradient (4%, 10%) was compared to traditional sperm washing (control) and multi-step resuspension protocols. At each time after centrifugation (hourly for 4 h), % IA was improved (P 10% improvement in overall sperm motility and, more importantly, allows retaining {approx}40% or more of sperm with intact acrosomes.

Publication Date

  • 2008

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