Colloidal centrifugation with Androcoll-E prolongs stallion sperm motility, viability and chromatin integrity.
Abstract: The objective was to investigate the changes in stallion sperm quality (sperm motility, viability, membrane integrity and chromatin integrity) occurring during cool storage, and to study the effect of sperm selection by single layer colloidal centrifugation on these parameters of sperm quality. Spermatozoa from 3 stallions (10 ejaculates, 3-4 per stallion) were selected by centrifugation through a single layer of colloid (SLC). The resulting sperm preparations and the control samples (extended but unselected semen samples) were stored at 5 degrees C for 48h. Assessments of sperm quality, such as sperm motility, viability (SYBR-14/PI staining), membrane stability (Annexin-V/PI staining) and chromatin integrity, were performed on aliquots of the selected sperm preparations and unselected samples on the day of collection (3h) and after 24 and 48h of storage. In the SLC-selected sperm samples, sperm motility, sperm viability, proportions of spermatozoa with normal morphology and with intact chromatin were significantly better than in unselected samples (motility: 77+/-4% vs. 64+/-8% at 3h; P<0.001; viability: 79.5+/-9% vs. 64.7+/-9%, P<0.001; normal morphology 89+/-6% vs. 69+/-9%; chromatin integrity DFI 11.3+/-5% vs. 22.1+/-10%). Membrane stability, however, was not different in the SLC-selected and unselected samples (74.6+/-8% vs. 69.3+/-8%). The deterioration seen in sperm quality in the unselected samples was prevented by SLC, so that sperm viability, membrane stability and chromatin integrity were unchanged in the selected samples by 48h compared to 3h (P<0.001), whereas the unselected samples were significantly worse by 48h (P<0.001). Furthermore, it should be possible to send an aliquot of a normal insemination dose (i.e. unselected spermatozoa) overnight to a reference laboratory for analysis of both plasma membrane and chromatin integrity. In conclusion, centrifugation of stallion spermatozoa through a single layer of colloid is a useful technique for selecting the best spermatozoa from an ejaculate and, moreover, sperm quality is maintained during storage.
Publication Date: 2009-01-24 PubMed ID: 19217226DOI: 10.1016/j.anireprosci.2009.01.008Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article investigates how the use of colloidal centrifugation with Androcoll-E can improve and extend the life of stallion sperm. It specifically looks at factors relating to sperm quality and shows that this method yields significantly improved results compared to control samples.
Objective and Methodology
- The primary goal of this study was to examine the impacts various quality parameters of stallion sperm had during cool storage. These traits included sperm motility, viability, membrane integrity and chromatin integrity.
- The researchers also aimed to evaluate the effects of sperm selection via single layer colloidal centrifugation on these sperm quality parameters.
- Sperm from three stallions were gathered through centrifugation through a single layer of colloid (SLC). The samples prepared from this method were then stored at 5 degrees Celsius for 48 hours. Comparisons were made against a control group, which consisted of semen samples that were extended but not selected.
Assessment and Major Findings
- Quality assessments, including tests for sperm motility, viability, membrane stability and chromatin integrity were carried out on sections of both, the selected and unselected samples. These assessments were completed at the time of collection, and after 24 and 48 hours.
- The results showed the samples selected via the SLC method had superior sperm motility, viability, normal morphology proportions, and chromatin integrity compared to the unselected samples.
- However, the membrane stability between the SLC-selected sperm samples and the control samples remained statistically indistinguishable.
- Interestingly, the deterioration of sperm quality seen in the unselected samples was averted by the SLC, which managed to uphold sperm viability, membrane stability, and chromatin integrity in the selected samples. Conversely, the unselected samples’ quality significantly declined by 48 hours.
Conclusion
- Based on the findings, the researchers concluded that centrifugation of stallion sperm through a single layer of colloid is a useful technique for choosing the best sperm from an ejaculate.
- The study summarizes the colloidal centrifugation technique’s ability not only to select superior quality stallion sperm but also to maintain that quality during storage. This technique could be of great use in animal breeding programs and equivalent contexts.
Cite This Article
APA
Johannisson A, Morrell JM, Thorén J, Jönsson M, Dalin AM, Rodriguez-Martinez H.
(2009).
Colloidal centrifugation with Androcoll-E prolongs stallion sperm motility, viability and chromatin integrity.
Anim Reprod Sci, 116(1-2), 119-128.
https://doi.org/10.1016/j.anireprosci.2009.01.008 Publication
Researcher Affiliations
- Swedish University of Agricultural Sciences - SLU, Dept. of Anatomy, Physiology & Biochemistry, Uppsala, Sweden.
MeSH Terms
- Androgens / pharmacology
- Animals
- Cell Membrane / drug effects
- Cell Membrane / physiology
- Cell Survival / drug effects
- Chromatin / drug effects
- Chromatin / ultrastructure
- Female
- Horses
- Insemination, Artificial / methods
- Insemination, Artificial / veterinary
- Male
- Pregnancy
- Sperm Count
- Sperm Motility / drug effects
- Spermatozoa / cytology
- Spermatozoa / drug effects
- Spermatozoa / physiology
Citations
This article has been cited 16 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.
- Johannisson A, Al-Essawe EM, Al-Saffar AK, Karkehabadi S, Lima-Verde I, Wulf M, Aurich C, Morrell JM. Season does not have a deleterious effect on proportions of stallion seminal plasma proteins. J Reprod Dev 2020 Jun 12;66(3):215-221.
- Gamboa S, Quaresma A, Castro F, Bravo P, Rebordão MR, Oom MDM, Rocha A. In vivo fertilizing ability of stallion spermatozoa processed by single layer centrifugation with Androcoll-E™. Saudi J Biol Sci 2017 Nov;24(7):1489-1496.
- 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 (Basel) 2017 Dec 21;7(1).
- Gloria A, Carluccio A, Wegher L, Robbe D, Befacchia G, Contri A. Single and double layer centrifugation improve the quality of cryopreserved bovine sperm from poor quality ejaculates. J Anim Sci Biotechnol 2016;7:30.
- Morrell JM, Johannisson A. Comparison of the Effect of Heterologous and Homologous Seminal Plasma on Motility and Chromatin Integrity of Stallion Spermatozoa Selected by Single Layer Centrifugation. J Vet Med 2014;2014:325451.
- Aitken RJ, Baker MA, Nixon B. Are sperm capacitation and apoptosis the opposite ends of a continuum driven by oxidative stress?. Asian J Androl 2015 Jul-Aug;17(4):633-9.
- Morrell JM, van Wienen M, Wallgren M. Single Layer Centrifugation Can Be Scaled-Up Further to Process up to 150 mL Semen. ISRN Vet Sci 2011;2011:183412.
- Morrell JM, Johannisson A, Rodriguez-Martinez H. Effect of Osmolarity and Density of Colloid Formulations on the Outcome of SLC-Selection of Stallion Spermatozoa. ISRN Vet Sci 2011;2011:128984.
- van Wienen M, Johannisson A, Wallgren M, Parlevliet J, Morrell JM. Single layer centrifugation with androcoll-p can be scaled-up to process larger volumes of boar semen. ISRN Vet Sci 2011;2011:548385.
- Gutiérrez-Cepeda L, Fernández A, Crespo F, Ramírez MÁ, Gosálvez J, Serres C. The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA. Acta Vet Scand 2012 Dec 5;54(1):72.
- 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 Vet Scand 2012 Jul 12;54(1):40.
- Gallardo Bolaños JM, Miró Morán Á, Balao da Silva CM, Morillo Rodríguez A, Plaza Dávila M, Aparicio IM, Tapia JA, Ortega Ferrusola C, Peña FJ. Autophagy and apoptosis have a role in the survival or death of stallion spermatozoa during conservation in refrigeration. PLoS One 2012;7(1):e30688.
- Morrell JM, Johannisson A, Juntilla L, Rytty K, Bäckgren L, Dalin AM, Rodriguez-Martinez H. Stallion Sperm Viability, as Measured by the Nucleocounter SP-100, Is Affected by Extender and Enhanced by Single Layer Centrifugation. Vet Med Int 2010;2010:659862.
- Johannisson A, Morrell JM, Ntallaris T. A combination of biomarkers for predicting stallion sperm fertility. Vet Res Commun 2024 Aug;48(4):2157-2169.
- Cojkic A, Hansson I, Johannisson A, Axner E, Morrell JM. Single layer centrifugation as a method for bacterial reduction in bull semen for assisted reproduction. Vet Res Commun 2024 Feb;48(1):39-48.
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