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Home / Equine Research Bank / Reprod Domest Anim / Pre- and Post-Thaw Addition of L-Carnitine and Pyruvate: Eff...
Reproduction in domestic animals = Zuchthygiene2024; 59(9); e14720; doi: 10.1111/rda.14720

Pre- and Post-Thaw Addition of L-Carnitine and Pyruvate: Effect on Stallion Sperm Parameters.

Abstract: The addition of antioxidants to cryopreservation media reportedly improves sperm post-thaw quality and reproductive performance after artificial insemination. Therefore, the objectives of this study were to evaluate if the addition of L-carnitine and pyruvate to freezing media, or their addition to samples after thawing, improves the post-thaw quality of equine spermatozoa. Thus, in Experiment 1, stallion semen samples were cryopreserved in: (1) EDTA-glucose-based extender with 20% egg yolk and 5% dimethylformamide (EDTA control); (2) skim milk-based extender with 20% egg yolk and 5% dimethylformamide (milk control); (3) Extender 1 supplemented with 50 mM L-carnitine and 10 mM pyruvate (EDTA-carnitine-pyruvate); and (4) Extender 2 supplemented with 50 mM L-carnitine and 10 mM pyruvate (milk-carnitine-pyruvate). In Experiment 2, 50 mM L-carnitine and 10 mM pyruvate were added post-thaw to samples cryopreserved with extenders 1 and 2 (EDTA control and milk control). Sperm kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability were evaluated after thawing. No significant differences (p > 0.05) were observed for most of the kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability of spermatozoa, between the samples frozen in the presence or absence of L-carnitine and pyruvate, nor between the samples after the post-thaw addition of these components. A higher (p < 0.05) mean velocity and higher (p < 0.05) amplitude of lateral head displacement were observed in the samples frozen in the milk-based extender with the addition of L-carnitine and pyruvate after thawing. The addition of 50 mM L-carnitine and 10 mM pyruvate, either to the freezing extenders or after thawing, was not deleterious for sperm; however, it did not improve equine sperm motility, viability, acrosome and DNA integrity, nor decrease membrane lipid peroxidation after thawing.
© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.
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Publication Date: 2024-09-15 PubMed ID: 39267414DOI: 10.1111/rda.14720Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Overview

  • This study investigated whether adding antioxidants L-carnitine and pyruvate before freezing or after thawing improves the quality of stallion sperm following cryopreservation.
  • The research found that these additions did not significantly enhance most sperm quality parameters post-thaw, though some minor improvements in specific motility parameters were noted with post-thaw addition in one extender type.

Objective and Background

  • Adding antioxidants to sperm freezing media has been proposed to protect sperm during cryopreservation, potentially enhancing survival, motility, and fertility after artificial insemination.
  • L-carnitine and pyruvate are antioxidants thought to support cell metabolism and reduce oxidative damage.
  • The study aimed to evaluate the effects of these compounds added either before freezing (in the extender) or immediately after thawing on stallion sperm quality.

Experimental Design

  • Two experiments were conducted:
    • Experiment 1: Semen samples were frozen using four different protocols:
      • EDTA-glucose-based extender (control)
      • Skim milk-based extender (control)
      • EDTA-based extender supplemented with 50 mM L-carnitine and 10 mM pyruvate
      • Milk-based extender supplemented with 50 mM L-carnitine and 10 mM pyruvate
    • Experiment 2: L-carnitine (50 mM) and pyruvate (10 mM) were added post-thaw to samples originally frozen in the two control extenders (EDTA and milk).

Parameters Measured

  • Post-thaw sperm quality assessments included:
    • Kinematic parameters (e.g., velocity, amplitude of lateral head displacement)
    • DNA fragmentation (indicator of genetic integrity)
    • Membrane lipid peroxidation (indicator of oxidative damage to cell membranes)
    • Acrosome status (important for fertilization capacity)
    • Viability (percentage of live sperm)

Results

  • Most kinematic parameters and all evaluated sperm quality markers (DNA fragmentation, lipid peroxidation, acrosome status, viability) showed no significant differences when comparing samples frozen with or without antioxidants, or after post-thaw antioxidant addition (p > 0.05).
  • One exception was that samples frozen in milk-based extender and supplemented with L-carnitine and pyruvate, post-thaw, exhibited significantly higher mean velocity and lateral head displacement amplitude (p < 0.05), indicating some improvement in sperm movement characteristics.
  • The antioxidants neither improved nor harmed sperm parameters overall.

Conclusions

  • The addition of L-carnitine (50 mM) and pyruvate (10 mM), whether included in the freezing extender or added immediately after thawing, was safe but did not broadly enhance stallion sperm quality post-thaw.
  • The slight improvement in motility parameters observed only with post-thaw addition in the milk-based extender suggests a limited context-dependent benefit.
  • Therefore, routine supplementation with these antioxidants in stallion semen cryopreservation protocols may not provide meaningful improvements in sperm viability or integrity.

Cite This Article

APA
Caldevilla M, Ferrante A, Neild DM. (2024). Pre- and Post-Thaw Addition of L-Carnitine and Pyruvate: Effect on Stallion Sperm Parameters. Reprod Domest Anim, 59(9), e14720. https://doi.org/10.1111/rda.14720

Publication

Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 59
Issue: 9
Pages: e14720

Researcher Affiliations

Caldevilla, Mariana
  • Facultad de Ciencias Veterinarias, INITRA, Cátedra de Teriogenología, Universidad de Buenos Aires, Buenos Aires, Argentina.
Ferrante, Alejandro
  • Facultad de Ciencias Veterinarias, INITRA, Cátedra de Teriogenología, Universidad de Buenos Aires, Buenos Aires, Argentina.
Neild, Débora M
  • Facultad de Ciencias Veterinarias, INITRA, Cátedra de Teriogenología, Universidad de Buenos Aires, Buenos Aires, Argentina.

MeSH Terms

  • Animals
  • Male
  • Horses
  • Cryopreservation / veterinary
  • Cryopreservation / methods
  • Carnitine / pharmacology
  • Carnitine / administration & dosage
  • Semen Preservation / veterinary
  • Semen Preservation / methods
  • Spermatozoa / drug effects
  • Spermatozoa / physiology
  • Pyruvic Acid / pharmacology
  • Cryoprotective Agents / pharmacology
  • Lipid Peroxidation / drug effects
  • DNA Fragmentation / drug effects
  • Semen Analysis / veterinary
  • Acrosome / drug effects
  • Sperm Motility / drug effects
  • Antioxidants / pharmacology

Grant Funding

  • 1535 / Agencia Nacional de Promoción Científica y Tecnológica, Préstamo BID-PICT-2014
  • 20020150100101BA / UBACYT project

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