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Home / Equine Research Bank / Reprod Domest Anim / Variation among stallions in sperm quality after single laye...
Reproduction in domestic animals = Zuchthygiene2021; 56(6); 848-856; doi: 10.1111/rda.13924

Variation among stallions in sperm quality after single layer centrifugation.

Abstract: Although single layer centrifugation (SLC) selects robust spermatozoa from stallion semen, the effect of individual variation has not been studied in detail. The objective of this study was to determine the variation among stallions in the effects of SLC on sperm quality during cooled storage for up to 48 hr. Semen samples from seven stallions (18 ejaculates) were split, with one portion being used for SLC and the other serving as a control (CON). Sperm quality (kinematics, reactive oxygen species (ROS) production, membrane integrity (MI) and chromatin integrity) were analysed at 0, 24 and 48 hr using computer-assisted sperm analysis and flow cytometry. Sperm quality was better in SLC than in CON at all timepoints, especially chromatin integrity and MI (p < .0001 for both), and some categories of ROS production (e.g. proportion of live hydrogen peroxide negative spermatozoa, p < .0001), but the degree of improvement varied among stallions and type of ROS (p < .05-p < .0001). Total and progressive motility were also better in SLC samples than in CON at 24 and 48 hr (p < .0001), although the effect on sperm kinematics varied. The interaction of treatment, time and stallion was not significant. In conclusion, sperm quality was better in SLC samples than in CON, although there was considerable individual variation among stallions. The improvement in sperm quality, particularly in chromatin integrity, was clearly beneficial, and therefore the use of this technique would be warranted for all stallion semen samples.
© 2021 The Authors. Reproduction in Domestic Animals published by Wiley-VCH GmbH.
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Publication Date: 2021-03-19 PubMed ID: 33706415DOI: 10.1111/rda.13924Google 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.

This study explores the variability in sperm quality among stallions after undergoing a specific procedure called single-layer centrifugation (SLC). Results revealed that SLC led to improved sperm quality across samples, with notable benefits in chromatin integrity, though the degree of improvement varied between individual stallions.

Objective of the Study

  • This research aimed to investigate the effects, and the level of individual variation, of single-layer centrifugation (SLC) on stallion sperm quality during cooled storage for up to 48 hours.

Methodology

  • Semen samples were obtained from seven stallions, totalling 18 ejaculates.
  • Each sample was split into two, with one part being processed using SLC, and the other serving as a control sample.
  • The researchers examined sperm quality, specifically kinematics, reactive oxygen species (ROS) production, membrane integrity (MI), and chromatin integrity, across different time intervals (0, 24, and 48 hours).
  • Data collection and analysis were done using computer-assisted sperm analysis and flow cytometry.

Results

  • The study found that sperm quality was generally better in samples treated with SLC compared to control samples at all time points.
  • Significant improvements were seen in chromatin integrity and membrane integrity.
  • SLC also effectively reduced some types of ROS production. This is crucial as ROS can be harmful to sperm cells, causing both structural damage and potential DNA damage.
  • SLC-treated samples displayed improved total and progressive motility compared to control samples.
  • However, the degree of improvement in these measures varied among stallions, indicating individual variation in response to SLC.
  • The interaction between the treatment, time, and individual stallion did not show significance, affirming the dominant role of the treatment itself in the results.

Conclusions

  • The study concluded that SLC significantly improves sperm quality, especially chromatin integrity, across stallion samples, even though the level of improvement can vary between individuals.
  • Given the above results, the researchers suggested that SLC should be utilized for all stallion semen samples to enhance their quality.

Cite This Article

APA
Al-Kass Z, Brown A, Johannisson A, Ntallaris T, Morrell JM. (2021). Variation among stallions in sperm quality after single layer centrifugation. Reprod Domest Anim, 56(6), 848-856. https://doi.org/10.1111/rda.13924

Publication

Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 56
Issue: 6
Pages: 848-856

Researcher Affiliations

Al-Kass, Ziyad
  • Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
  • Department of Surgery and Theriogenology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq.
Brown, Alexander
  • Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
Johannisson, Anders
  • Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
Ntallaris, Theodoros
  • Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
Morrell, Jane M
  • Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.

MeSH Terms

  • Animals
  • Cell Membrane
  • Centrifugation / methods
  • Centrifugation / veterinary
  • DNA Fragmentation
  • Horses
  • Male
  • Reactive Oxygen Species / analysis
  • Semen Analysis / veterinary
  • Semen Preservation / veterinary
  • Sperm Motility
  • Spermatozoa / physiology

Grant Funding

  • NA / Sveriges Lantbruksuniversitet
  • NA / Ministry of Higher Education and Scientific Research, Iraq.
  • vacation scholarship / Society for Reproduction and Fertility

References

This article includes 25 references
  1. Akbarinejad V, Fathi R, Shahverdi A, Esmaeili V, Rezagholizadeh A, Ghaleno LR. The relationship of mitochondrial membrane potential, reactive oxygen species, adenosine triphosphate content, sperm plasma membrane integrity, and kinematic properties in warmblood stallions. Journal of Equine Veterinary Science 94, 103267.
    doi: 10.1016/j.jevs.2020.103267google scholar: lookup
  2. Al-Essawe EM, Johannisson A, Wulf M, Aurich C, Morrell JM. Improved cryosurvival of stallion spermatozoa after colloid centrifugation is independent of the addition of seminal plasma. Cryobiology 81, 145-152.
    doi: 10.1016/j.cryobiol.2018.01.009google scholar: lookup
  3. Al-Kass Z. Characterization, quantification and removal of potential pathogens from stallion semen (doctoral thesis). Sveriges lantbruksuniv., Acta Universitatis Agriculturae Sueciae 1652-6880; 2019:17.
  4. Al-Kass Z, Spergser J, Aurich C, Kuhl J, Schmidt K, Johannisson A, Morrell JM. Sperm quality during storage is not affected by the presence of antibiotics in EquiPlus semen extender but is improved by single layer centrifugation. Antibiotics 7, 1.
    doi: 10.3390/antibiotics7010001google scholar: lookup
  5. Aurich J, Aurich C. Developments in European horse breeding and consequences for veterinarians in equine reproduction. Reproduction in Domestic Animals 41, 275-279.
    doi: 10.1111/j.1439-0531.2006.00719.xgoogle scholar: lookup
  6. Colenbrander B, Gadella BM, Stout TAE. The predictive value of semen analysis in the evaluation of stallion fertility. Reproduction in Domestic Animals 38(4), 305-311.
    doi: 10.1046/j.1439-0531.2003.00451.xgoogle scholar: lookup
  7. Costa AL, Martins-Bessa A, de Andrade AR, Guimaraes T, Rebordao MR, Gamboa S, Bravo PP, Correia MJ, Colaco J, Gaivao I, Rocha A. Single layer centrifugation with Androcoll-E™ improved progressive motility and percentage of live spermatozoa with intact acrosome of chilled stallion semen but did not have an effect on DNA integrity. Open Journal of Animal Sciences 2, 159-165.
    doi: 10.4236/ojas.2012.23022google scholar: lookup
  8. Crespo F, Gutierrez-Cepeda L, Gosalvez J, Serres C, Johanston SD. Colloidal centrifugation of stallion semen results in a reduced rate of sperm DNA fragmentation. Reproduction in Domestic Animals 48, e23-25.
    doi: 10.1111/j.1439-0531.2012.02140.xgoogle scholar: lookup
  9. Guimaraes T, Lopes G, Pinto M, Silva E, Miranda C, Correia MJ, Damasio L, Thompson G, Rocha A. Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen- thawed semen without affecting seminal kinetics. Theriogenology 83, 186-191.
    doi: 10.1016/j.theriogenology.2014.09.003google scholar: lookup
  10. Halliwell B, Gutteridge JMC. Free radicals in biology and medicine, 3rd ed. Oxford University Press .
  11. Hossain MS, Johannisson A, Wallgren M, Nagy S, Siqueira AP, Rodriguez-Martinez H. Flowcytometry for the assessment of animal sperm integrity and functionally: State of the art. Asian Journal of Andrology 13, 406-419.
    doi: 10.1038/aja.2011.15google scholar: lookup
  12. Johannisson A, Lundgren A, Humblot P, Morrell JM. Naturally and stimulated levels of reactive oxygen species in cooled stallion semen destined for artificial insemination. Animal 8, 1706-1714.
    doi: 10.1017/s1751731114001499google scholar: lookup
  13. Johannisson A, Morrell JM, Thoren J, Jonsson M, Dalin AM, Rodriguez-Matinez H. Colloidal centrifugation with Androcoll-E™ prolongs stallion sperm motility, viability and chromatin integrity. Animal Reproduction Science 116, 119-128.
    doi: 10.1016/j.anireprosci.2009.01.008google scholar: lookup
  14. Lindahl J, Dalin AM, Stuhtmann G, Morrell JM. Stallion spermatozoa selected by single layer centrifugation are capable of fertilization after storage for up to 96 h at 6°C prior to artificial insemination. Acta Veterinaria Scandinavica 54, 40-44.
    doi: 10.1186/1751-0147-54-40google scholar: lookup
  15. Loomis PR, Graham JK. Commercial semen freezing: Individual male variation in cryosurvival and the response of stallion sperm to customized freezing protocols. Animal Reproduction Science 105, 119-128.
    doi: 10.1016/j.anireprosci.2007.11.010google scholar: lookup
  16. Morrell JM, Dalin AM, Rodriguez-Martinez H. Comparison of density gradient and single layer centrifugation of stallion spermatozoa: yield, motility and survival. Equine Veterinary Journal 41, 53-58.
    doi: 10.2746/042516408x322139google scholar: lookup
  17. Morrell JM, Johannisson A, Anne-Marie D, Hammar L, Sandebert T, Rodriguez-Martinez H. Sperm morphology and chromatin integrity in Swedish warmblood stallions and their relationship to pregnancy rates. Acta Veterinaria Scandinavica 50, 2.
    doi: 10.1186/1751-0147-50-2google scholar: lookup
  18. Morrell JM, Johannisson A, Dalin AM, Rodriguez-Martinez H. Single-layer centrifugation with Androcoll-E can be scaled up to allow large volumes of stallion ejaculate to be processed easily. Theriogenology 72, 879-884.
    doi: 10.1016/j.theriogenology.2009.05.015google scholar: lookup
  19. Morrell JM, Lagerqvist A, Humblot P, Johannisson A. Effect of single layer centrifugation on reactive oxygen species and sperm mitochondrial membrane potential in cooled stallion semen. Reproduction, Fertility and Development 29, 1039-1045.
    doi: 10.1071/rd15440google scholar: lookup
  20. Morrell JM, Richter J, Martinsson G, Stuhtmann G, Hoogewijs M, Roels K, Dalin AM. Pregnancy rates after artificial insemination with cooled stallion spermatozoa either with or without single layer centrifugation. Theriogenology 82, 1102-1105.
    doi: 10.1016/j.theriogenology.2014.07.028google scholar: lookup
  21. Morrell JM, Rodriguez-Martinez H, Johannisson A. Single layer centrifugation of stallion spermatozoa consistently selects the most robust spermatozoa from the rest of the ejaculate in a large sample size. Equine Veterinary Journal 42(7), 579-585.
    doi: 10.1111/j.2042-3306.2010.00101.xgoogle scholar: lookup
  22. Morrell JM, Stuhtmann G, Meurling S, Lundgren A, Winblad C, Macias Garcia B, Johannisson A. Sperm yield after single layer centrifugation with Androcoll-E is related to the potential fertility of the original ejaculate. Theriogenology 81, 1005-1011.
    doi: 10.1016/j.theriogenology.2014.01.013google scholar: lookup
  23. Pessoa GA, Martini AP, Trentin JM, Mimela T, Rubin MIB. Effect of plasma separation techniques and sperm selection on sperm recovery and viability of cooled pony stallion semen for 48h at 5°C. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 72(6), 2017-2026.
    doi: 10.1590/1678-4162-11986google scholar: lookup
  24. Rodriguez-Martinez H. Semen evaluation techniques and their relationship with fertility. Animal Reproduction Science 10, 148-159.
  25. . SAS/ACCESS 9.4 Interface to ADABAS: Reference. SAS Institute Inc. .

Citations

This article has been cited 3 times.
  1. Gutiérrez-Cepeda L, Crespo F, Blazquez JC, Serres C. Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation. Animals (Basel) 2023 Jan 22;13(3).
    doi: 10.3390/ani13030382pubmed: 36766271google scholar: lookup
  2. Al-Kass Z, Morrell JM, Ntallaris T. Effect of Centrifugation of Stallion Semen Through a Low Density Colloid Prior to Freezing on Sperm Cryosurvival. Animals (Basel) 2025 Jun 25;15(13).
    doi: 10.3390/ani15131881pubmed: 40646780google scholar: lookup
  3. Khan IU, Khairullah AR, Khan AY, Rehman AU, Mustofa I. Strategic approaches to improve equine breeding and stud farm outcomes. Vet World 2025 Feb;18(2):311-328.
    doi: 10.14202/vetworld.2025.311-328pubmed: 40182817google scholar: lookup
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