Effects of glycerol, equilibration time and antioxidants on post-thaw functional integrity of bovine spermatozoa directly obtained from epididymis

Andrologia. 2017 Apr;49(3). doi: 10.1111/and.12623. Epub 2016 Jun 2.

Abstract

This work aimed to evaluate the effect of stabilisation times, glycerol concentration, and the catalase and superoxide dismutase supplementation of diluent on parameters of frozen-thawed spermatozoa from epididymis of Nelore bulls: Experiment 1: spermatozoa diluted in Tris-egg yolk with glycerol (3%, 5% or 7%) and stabilisation times (0, 2 or 4 hr at 5°C); Experiment 2: Tris-egg yolk only, Tris-egg yolk with catalase (CAT, 50 or 100 U ml-1 ) or superoxide dismutase (SOD, 50 or 100 U ml-1 ). Frozen-thawed spermatozoa were evaluated for kinetic parameters, plasma membrane and acrosome integrity, mitochondrial activity and IVF capacity. ALH and BCF were affected (p < .05) by glycerol at 3% after 4-hr equilibration time and 7% after 2-hr equilibration time. Glycerol 3% had lower (p < .05) iPM and iAc after 4 hr. Glycerol 5% had greater (p < .05) hPMM after 4 hr and iAc after 2 hr than at 0 hr. SOD 100 U ml-1 had lower (p < .05) linearity and wobble compared to control group. No was observed differences to fertilisation rate (p < .05) among groups. In conclusion, glycerol 5% in Tris-egg yolk extender for 4 hr is suitable for the preservation of sperm kinetics and membrane integrity. CAT (50 and 100 U ml-1 ) or SOD (50-100 U ml-1 ) had no beneficial effects on sperm kinetics, plasma and acrosomal membrane integrity, mitochondrial activity or the capacity for IVF of frozen-thawed spermatozoa from epididymis of Nelore bulls.

Keywords: bull; catalase; cryopreservation; spermatozoa; superoxide dismutase.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Antioxidants / pharmacology*
  • Catalase / pharmacology
  • Cattle
  • Cryopreservation / veterinary
  • Cryoprotective Agents / pharmacology*
  • Epididymis / cytology*
  • Freezing / adverse effects
  • Glycerol / pharmacology*
  • Insemination, Artificial / veterinary*
  • Male
  • Semen Preservation / veterinary*
  • Specimen Handling / veterinary*
  • Sperm Motility / drug effects
  • Spermatozoa / drug effects*
  • Superoxide Dismutase / pharmacology
  • Time Factors

Substances

  • Antioxidants
  • Cryoprotective Agents
  • Catalase
  • Superoxide Dismutase
  • Glycerol