Pre-freezing Treatment with Butylated Hydroxytoluene and Cholesterol-loaded Methyl-β-cyclodextrin Improves Quality of Cryopreserved Boar Semen

Cryo Letters. 2018 Sep/Oct;39(5):336-344.

Abstract

Background: Pre-freezing treatment of boar sperm with additives improves the quality of post-thaw sperms.

Objectives: The study aimed to determine the efficacy of butylated hydroxy-toluene (BHT) and cholesterol-loaded methyl-β-cyclodextrin (CLC) for the improvement of the frozen-thawed boar sperm quality.

Methods: Split samples of 30 ejaculates from six boars were cryopreserved in lactose-egg yolk-glycerol extender containing BHT (0.2 mM), CLC (5 mg/ 200-240 x 106 sperm) or BHT (0.2 mM) plus CLC (5 mg per 200-240 x 106 sperm). Semen samples were evaluated for motility, membrane integrity, acrosomal status, mitochondrial membrane potential (MMP), DNA integrity and lipid peroxidation after equilibration and after freezing.

Results: The addition of BHT and CLC into the extender significantly improved (P<0.05) plasma membrane integrity and decreased (P<0.05) lipid peroxidation after freezing. Post-thaw motility and live intact acrosome were significantly (P<0.05) higher in the extenders with BHT or BHT plus CLC. The post-thaw MMP of viable spermatozoa and DNA integrity were not affected. BHT plus CLC showed a significant (P<0.05) improvement on motility as compared to BHT and CLC alone.

Conclusion: Treatment of boar spermatozoa with BHT and CLC improved post-thaw sperm quality.

MeSH terms

  • Acrosome
  • Animals
  • Butylated Hydroxytoluene / chemistry*
  • Cholesterol
  • Cryopreservation*
  • Cryoprotective Agents / chemistry*
  • Freezing
  • Male
  • Semen
  • Semen Preservation*
  • Sperm Motility
  • Spermatozoa / physiology*
  • Swine
  • beta-Cyclodextrins / chemistry*

Substances

  • Cryoprotective Agents
  • beta-Cyclodextrins
  • methyl-beta-cyclodextrin
  • Butylated Hydroxytoluene
  • Cholesterol