Addition of seminal plasma to thawed stallion spermatozoa did not repair cryoinjuries.
Abstract: Freezing and thawing processes induce structural and functional damage to sperm plasma membranes and internal organelles. Adding seminal plasma (SP) has been found to minimize or repair the cryoinjuries in some species. The objective of this study was to investigate whether adding SP from stallions of known freezability after thawing could repair cryoinjuries. Semen was collected from warmblood stallions (n = 8, three ejaculates/stallion) and processed by Single Layer Centrifugation (SLC) to remove SP prior to freezing. Pooled SP (5%) from bad freezer (BF) or good freezer (GF) stallions was added after thawing. Post-thaw sperm quality was assessed by flow cytometry in terms of chromatin integrity (%DFI), membrane integrity, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and MitoSOX. Sperm kinematics were also assessed by computer-assisted sperm analysis. The %DFI was lower in SLC control (C) than in BF or GF (P < 0.0001, P < 0.0003 respectively). The proportion of viable spermatozoa with intact cell membranes was higher in C than in SP treated groups (C vs. BF, P = 0.02; C vs GF, P = 0.05). There were fewer spermatozoa with low MMP and more with high MMP for C than GF (P = 0.006). The spermatozoa treated with SP from good freezers produced more ROS than when treated with SP from bad freezers (P = 0.007). Motility parameters were not affected by adding SP. In conclusion, adding SP after thawing does not have a beneficial effect on sperm quality, suggesting an inability to repair stallion sperm cryoinjuries, regardless of whether the SP originated from stallions semen, which has good or bad quality after thawing.
Copyright © 2018 Elsevier B.V. All rights reserved.
Publication Date: 2018-06-25 PubMed ID: 30041960DOI: 10.1016/j.anireprosci.2018.06.018Google Scholar: Lookup
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- Journal Article
Summary
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The research investigated whether adding seminal plasma (SP) from stallions to thawed sperm could repair damage caused by freezing and thawing processes. They found that adding SP, whether from stallions with high or low-quality semen after thawing, did not provide any beneficial effects, suggesting that it couldn’t repair the cryoinjuries to the sperm.
Research Context
- The freezing and thawing process of sperm for preservation purposes often results in structural and functional damages to the sperm plasma and internal organelles. This is referred to as ‘cryoinjury’.
- In some species, adding seminal plasma (SP) has been shown to reduce or repair these cryoinjuries.
- Previous studies have shown varying results on the effects of SP addition to thawed stallion spermatozoa. This study sought to provide clarity by testing the effect of adding SP from stallions known for good and bad freezability.
Methodology
- Semen was collected from eight warmblood stallions, each providing three ejaculates.
- The semen was processed through Single Layer Centrifugation (SLC) to separate seminal plasma from the spermatozoa.
- Post-thawing, the spermatozoa were treated with pooled SP (5%) from either stallions known for good or bad freezing characteristics.
- The quality of the post-threw sperm was assessed using flow cytometry to examine various parameters such as chromatin integrity, membrane integrity, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and MitoSOX.
- Sperm movement was also analyzed using computer-assisted sperm analysis.
Findings
- The study found that the proportion of spermatozoa with intact cell membranes and a high MMP was higher in the control group than in the SP treated groups.
- Spermatozoa treated with SP from stallions with good freezing qualities produced more ROS than those treated with SP from stallions with poor freezing qualities.
- Adding SP to the spermatozoa post-thawing did not improve motility parameters.
Conclusion
- This research concluded that the addition of SP after thawing did not have a positive impact on sperm quality.
- It appears to demonstrate that SP is unable to repair the cryoinjuries that occur as a result of the freezing and thawing process in stallion spermatozoa.
- This finding prevailed regardless of whether the SP came from stallions with good or bad freezing qualities.
Cite This Article
APA
Al-Essawe EM, Johannisson A, Wulf M, Aurich C, Morrell JM.
(2018).
Addition of seminal plasma to thawed stallion spermatozoa did not repair cryoinjuries.
Anim Reprod Sci, 196, 48-58.
https://doi.org/10.1016/j.anireprosci.2018.06.018 Publication
Researcher Affiliations
- Swedish University of Agricultural Sciences, Clinical Sciences, Uppsala, Sweden; Al-Nahrain University - High Institute of Infertility Diagnosis and Assisted Reproductive Technologies, Clinical Reproductive Physiology, Baghdad, Iraq. Electronic address: essraa.m.al.essawe@slu.se.
- Swedish University of Agricultural Sciences, Clinical Sciences, Uppsala, Sweden. Electronic address: anders.johannisson@slu.se.
- Graf Lehndorff Institute for Equine Sciences, Vetmeduni Vienna, Austria and Neustadt (Dosse), Germany. Electronic address: manuela.Wulf@Neustaedter-Gestuete.de.
- Artificial Insemination and Embryo Transfer, Department for Small Animals and Horses, Vetmeduni Vienna, Vienna, Austria. Electronic address: christine.aurich@vetmeduni.ac.at.
- Swedish University of Agricultural Sciences, Clinical Sciences, Uppsala, Sweden. Electronic address: jane.morrell@slu.se.
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryopreservation / veterinary
- Horses / physiology
- Male
- Semen
- Semen Analysis
- Semen Preservation / adverse effects
- Semen Preservation / veterinary
- Sperm Motility
- Spermatozoa / physiology
Citations
This article has been cited 2 times.- Ortiz I, Dorado J, Morrell JM, Diaz-Jimenez MA, Pereira B, Consuegra C, Hidalgo M. Comparison of sperm selection techniques in donkeys: motile subpopulations from a practical point of view.. Anim Reprod 2019 Oct 23;16(2):282-289.
- Loureiro KC, Lima-Verde IB, Johannisson A, Ntallaris T, Jager A, Štěpánek P, da Costa Mendonça M, Severino P, Morrell JM. Effects of cashew gum and nanoparticles on cooled stallion semen.. Acta Vet Scand 2020 Jun 18;62(1):31.
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