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Related Experiment Videos

Salvaging urospermic ejaculates from brown bear (Ursus arctos).

S Gomes-Alves1, M Alvarez1, M Nicolas1

  • 1ITRA-ULE, INDEGSAL, University of León, 24071 León, Spain; Animal Reproduction and Obstetrics, University of León, 24071 León, Spain.

Animal Reproduction Science
|October 4, 2014
PubMed
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Pre-freezing washing did not improve sperm quality in urine-contaminated ejaculates. Density gradient centrifugation, however, enhanced sperm motility and viability, improving cryopreservation outcomes.

Area of Science:

  • Reproductive biology
  • Cryopreservation science
  • Animal reproduction

Background:

  • Urine contamination in ejaculates causes osmotic stress, negatively impacting sperm quality.
  • Effective sperm preparation techniques are crucial for successful artificial insemination and cryopreservation.

Purpose of the Study:

  • To evaluate pre-freezing washing and density gradient centrifugation for reversing osmotic stress in urine-contaminated bear ejaculates.
  • To determine the impact of these methods on sperm motility, viability, and acrosomal status before and after cryopreservation.

Main Methods:

  • Experiment 1: Sperm washing with varying osmolality extenders (200-700 mOsm/kg) and centrifugation.
  • Experiment 2: Density gradient centrifugation (PureSperm) and centrifugation control.
Keywords:
Brown bearOsmotic stress reversionPureSperm(®)SpermatozoaUrospermic ejaculates

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  • Sperm analysis included motility, viability (SYBR-14/PI), and acrosomal status (PNA-FITC/PI) pre- and post-freezing.

    • Main Results:

    • Pre-freezing washing, except at 700 mOsm/kg, showed no significant improvement in sperm viability compared to controls.
    • Washing with 700 mOsm/kg solutions had detrimental effects on sperm viability.
    • Density gradient centrifugation significantly improved pre-freezing sperm motility and viability.
    • Sperm processed via density gradient centrifugation exhibited better cryopreservation capacity.

    Conclusions:

    • Density gradient centrifugation is a superior method for preparing urine-contaminated ejaculates for cryopreservation.
    • Pre-freezing washing is not effective in mitigating osmotic stress in these samples.
    • Optimized sperm preparation is vital for preserving reproductive potential in challenging samples.