Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE (Open Access)

Effect of zinc supplementation on the quality of cooled, stored equine sperm

Patricio D. Palacios https://orcid.org/0000-0003-0194-8373 A , Isabel Ortiz https://orcid.org/0000-0002-2479-498X B , Jesús Dorado https://orcid.org/0000-0002-4310-7663 B , Manuel Hidalgo https://orcid.org/0000-0001-7830-7422 B , Juan Ramón García Díaz https://orcid.org/0000-0002-2966-7824 B C and Andrés Gambini https://orcid.org/0000-0002-3652-2068 A B D *
+ Author Affiliations
- Author Affiliations

A School of Agriculture and Food Sustainability, The University of Queensland, Gatton, Qld 4343, Australia.

B Veterinary Reproduction Group, Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Cordoba, Cordoba 14014, Spain.

C Facultad de Ciencias Agropecuarias, Universidad Central “Marta Abreu” de Las Villas, Santa Clara, Villa Clara 54830, Cuba.

D School of Veterinary Sciences, The University of Queensland, Gatton, Qld 4343, Australia.

* Correspondence to: a.gambini@uq.edu.au

Handling Editor: Chris Rogers

Animal Production Science 64, AN24005 https://doi.org/10.1071/AN24005
Submitted: 16 January 2024  Accepted: 23 May 2024  Published: 13 June 2024

© 2024 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND)

Abstract

Context

Collecting, cooling, and cryopreserving semen is essential for accessing genetically superior stallions. However, preserving stallion sperm presents unique challenges compared with other species.

Aims

This study aimed to investigate the effect of zinc (Zn) supplementation on the quality of equine sperm during cold storage.

Methods

Various factors contributing to sperm quality were assessed at 24 and 48 h after cooling, and after subjecting the sperm samples to a heat-resistance test. In Experiment 1, four experimental groups were examined, each with different concentrations of Zn sulfate, as follows: 0, 1, 2, and 3 mM. Subsequently, Experiment 2 involved testing a wider range of concentrations (0, 0.1, 0.2, 0.4, 0.8, 1.6, and 3.2 mM), including evaluation of samples after incubation for 240 min at 37°C (heat-resistance test).

Key results

The addition of different concentrations of Zn to the extender INRA96 did not yield substantial improvements in sperm-quality parameters for cooling stallion semen after 24 or 48 h. Moreover, no protective benefits were observed when samples underwent a heat-resistance test. Concentrations of Zn surpassing 3 mM had an adverse effect on sperm-quality parameters.

Conclusions and implications

These findings have contributed to the understanding of Zn supplementation as a strategy for improving semen preservation in stallions.

Keywords: equine, fertility, heat-resistance test, horses, sperm quality, stallion, zinc supplementation, ZnSO4 .

References

Allouche-Fitoussi D, Breitbart H (2020) The role of zinc in male fertility. International Journal of Molecular Sciences 21(20), 7796.
| Crossref | Google Scholar | PubMed |

Andrade AFC, Knox RV, Torres MA, Pavaneli APP (2022) What is the relevance of seminal plasma from a functional and preservation perspective? Animal Reproduction Science 246, 106946.
| Crossref | Google Scholar |

Atrian S, Capdevila M (2013) Metallothionein–protein interactions. Biomolecular Concepts 4(2), 143-160.
| Crossref | Google Scholar | PubMed |

Colenbrander B, Gadella BM, Stout TAE (2003) The predictive value of semen analysis in the evaluation of stallion fertility. Reproduction in Domestic Animals 38(4), 305-311.
| Crossref | Google Scholar | PubMed |

Contreras MJ, Treulen F, Arias ME, Silva M, Fuentes F, Cabrera P, Felmer R (2020) Cryopreservation of stallion semen: effect of adding antioxidants to the freezing medium on sperm physiology. Reproduction in Domestic Animals 55(2), 229-239.
| Crossref | Google Scholar | PubMed |

Dorado J, Acha D, Ortiz I, Gálvez MJ, Carrasco JJ, Gómez-Arrones V, Calero-Carretero R, Hidalgo M (2014) Effect of extender and amino acid supplementation on sperm quality of cooled-preserved Andalusian donkey (Equus asinus) spermatozoa. Animal Reproduction Science 146(1–2), 79-88.
| Crossref | Google Scholar | PubMed |

Dorostkar K, Shoushtari SMA, Khaki A (2014) Effects of in vitro zinc sulphate additive to the semen extender on water buffalo (bubalusbubalis) spermatozoa before and after freezing. International Journal of Fertility & Sterility 8(3), 325.
| Google Scholar | PubMed |

Fallah A, Mohammad-Hasani A, Colagar AH (2018) Zinc is an essential element for male fertility: a review of zn roles in men’s health, germination, sperm quality, and fertilization. Journal of Reproduction & Infertility 19(2), 69.
| Google Scholar | PubMed |

Fayez E, El Sayed MAI, Rawash ZM, Salama A (2023) Influence of the addition of zinc oxide nanoparticles to cryopreservation medium for dog epididymal spermatozoa. Topics in Companion Animal Medicine 52, 100736.
| Crossref | Google Scholar | PubMed |

Ghallab ARM, Shahat AM, Fadl AM, Ayoub MM, Moawad AR (2017) Impact of supplementation of semen extender with antioxidants on the quality of chilled or cryopreserved Arabian stallion spermatozoa. Cryobiology 79, 14-20.
| Crossref | Google Scholar | PubMed |

Gibb Z, Aitken RJ (2016) The impact of sperm metabolism during in vitro storage: the stallion as a model. BioMed Research International 2016, 9380609.
| Crossref | Google Scholar |

Gualtieri R, Barbato V, Fiorentino I, Braun S, Rizos D, Longobardi S, Talevi R (2014) Treatment with zinc, d-aspartate, and coenzyme Q10 protects bull sperm against damage and improves their ability to support embryo development. Theriogenology 82(4), 592-598.
| Crossref | Google Scholar | PubMed |

Hansen PJ (2014) Current and future assisted reproductive technologies for mammalian farm animals. In ‘Current and future reproductive technologies and world food production’. (Eds GC Lamb, N DiLorenzo) pp. 1–22. (Springer)

Henkel R, Bittner J, Weber R, Hüther F, Miska W (1999) Relevance of zinc in human sperm flagella and its relation to motility. Fertility and Sterility 71(6), 1138-1143.
| Crossref | Google Scholar | PubMed |

Höfner L, Luther A-M, Waberski D (2020) The role of seminal plasma in the liquid storage of spermatozoa. Animal Reproduction Science 220, 106290.
| Crossref | Google Scholar | PubMed |

Isaac AV, Kumari S, Nair R, Urs DR, Salian SR, Kalthur G, Adiga SK, Manikkath J, Mutalik S, Sachdev D, Pasricha R (2017) Supplementing zinc oxide nanoparticles to cryopreservation medium minimizes the freeze-thaw-induced damage to spermatozoa. Biochemical and Biophysical Research Communications 494(3–4), 656-662.
| Crossref | Google Scholar | PubMed |

Kandiel MMM, El Khawagah ARM (2018) Evaluation of semen characteristics, oxidative stress, and biochemical indices in Arabian horses of different ages during the hot summer season. Iranian Journal of Veterinary Research 19(4), 270-275.
| Google Scholar | PubMed |

Khodaei-Motlagh M, Masoudi R, Karimi-Sabet MJ, Hatefi A (2022) Supplementation of sperm cooling medium with zinc and zinc oxide nanoparticles preserves rooster sperm quality and fertility potential. Theriogenology 183, 36-40.
| Crossref | Google Scholar | PubMed |

Kotdawala AP, Kumar S, Salian SR, Thankachan P, Govindraj K, Kumar P, Kalthur G, Adiga SK (2012) Addition of zinc to human ejaculate prior to cryopreservation prevents freeze-thaw-induced DNA damage and preserves sperm function. Journal of Assisted Reproduction and Genetics 29, 1447-1453.
| Crossref | Google Scholar | PubMed |

Lee SR (2018) Critical role of zinc as either an antioxidant or a prooxidant in cellular systems. Oxidative Medicine and Cellular Longevity 2018, 9156285.
| Crossref | Google Scholar |

Leemans B, Gadella BM, Stout TAE, De Schauwer C, Nelis H, Hoogewijs M, Van Soom A (2016) Why doesn’t conventional IVF work in the horse? The equine oviduct as a microenvironment for capacitation/fertilization. Reproduction 152(6), R233-R245.
| Crossref | Google Scholar | PubMed |

Len J, Beehan D, Eilts B, Ebrahimie E, Lyle S (2020) Stallion sperm integrity after centrifugation to reduce seminal plasma concentration and cool storage for 4 days. Journal of Equine Veterinary Science 85, 102819.
| Crossref | Google Scholar | PubMed |

Maitan P, Bromfield EG, Stout TAE, Gadella BM, Leemans B (2022) A stallion spermatozoon’s journey through the mare’s genital tract: in vivo and in vitro aspects of sperm capacitation. Animal Reproduction Science 246, 106848.
| Crossref | Google Scholar | PubMed |

Ortiz I, Felix M, Resende H, Ramírez-Agámez L, Love CC, Hinrichs K (2021) Flow-cytometric analysis of membrane integrity of stallion sperm in the face of agglutination: the ‘zombie sperm’ dilemma. Journal of Assisted Reproduction and Genetics 38(9), 2465-2480.
| Crossref | Google Scholar | PubMed |

Peña FJ, Ortiz-Rodríguez JM, Gaitskell-Phillips GL, Gil MC, Ortega-Ferrusola C, Martín-Cano FE (2022) An integrated overview on the regulation of sperm metabolism (glycolysis–Krebs cycle–oxidative phosphorylation). Animal Reproduction Science 246, 106805.
| Crossref | Google Scholar | PubMed |

Pesch S, Bergmann M, Bostedt H (2006) Determination of some enzymes and macro- and microelements in stallion seminal plasma and their correlations to semen quality. Theriogenology 66(2), 307-313.
| Crossref | Google Scholar | PubMed |

Schulze M, Mohammadpour F, Schröter F, Jakop U, Hönicke H, Hasenfuss T, Henne H, Schön J, Müller K (2021) Suitability of semen stress tests for predicting fertilizing capacity of boar ejaculates. Theriogenology 176, 73-81.
| Crossref | Google Scholar | PubMed |

Soltani L, Samereh S, Mohammadi T (2022) Effects of different concentrations of zinc oxide nanoparticles on the quality of ram cauda epididymal spermatozoa during storage at 4°C. Reproduction in Domestic Animals 57(8), 864-875.
| Crossref | Google Scholar | PubMed |

Sørensen MB, Stoltenberg M, Danscher G, Ernst E (1999) Chelation of intracellular zinc ions affects human sperm cell motility. Molecular Human Reproduction 5(4), 338-341.
| Crossref | Google Scholar | PubMed |

Squires EL (2005) Integration of future biotechnologies into the equine industry. Animal Reproduction Science 89(1–4), 187-198.
| Crossref | Google Scholar | PubMed |

Sutovsky P, Kerns K, Zigo M, Zuidema D (2019) Boar semen improvement through sperm capacitation management, with emphasis on zinc ion homeostasis. Theriogenology 137, 50-55.
| Crossref | Google Scholar | PubMed |

Veeramachaneni DNR, Moeller CL, Sawyer HR (2006) Sperm morphology in stallions: ultrastructure as a functional and diagnostic tool. Veterinary Clinics of North America: Equine Practice 22(3), 683-692.
| Google Scholar | PubMed |

Yánez-Ortiz I, Catalán J, Rodríguez-Gil JE, Miró J, Yeste M (2022) Advances in sperm cryopreservation in farm animals: cattle, horse, pig and sheep. Animal Reproduction Science 246, 106904.
| Crossref | Google Scholar | PubMed |

Yates DJ, Whitacre MD (1988) Equine artificial insemination. Veterinary Clinics of North America: Equine Practice 4(2), 291-304.
| Google Scholar | PubMed |

Zhao J, Dong X, Hu X, Long Z, Wang L, Liu Q, Sun B, Wang Q, Wu Q, Li L (2016) Zinc levels in seminal plasma and their correlation with male infertility: a systematic review and meta-analysis. Scientific Reports 6(1), 22386.
| Crossref | Google Scholar |

Zigo M, Kerns K, Sen S, Essien C, Oko R, Xu D, Sutovsky P (2022) Zinc is a master-regulator of sperm function associated with binding, motility, and metabolic modulation during porcine sperm capacitation. Communications Biology 5(1), 538.
| Crossref | Google Scholar | PubMed |