Morphology and chromatin integrity of stallion spermatozoa prepared by density gradient and single layer centrifugation through silica colloids.

Morrell, JM
·
Johannisson, A
·
Dalin, AM
·
Rodriguez-Martinez, H

Abstract: The objective was to investigate whether it is possible to improve the quality of stallion semen, with respect to sperm morphology and chromatin integrity, both of which have been linked to fertility, using either density gradient centrifugation (DGC) or a new method, hereby named single layer centrifugation (SLC). The two methods of colloidal centrifugation were evaluated using 38 ejaculates from 10 stallions. Sperm morphology, subjective motility and sperm chromatin integrity were compared in uncentrifuged samples and in centrifuged sperm preparations. The proportion of morphologically normal spermatozoa varied between stallions (p < 0.001) and was increased by both methods of colloidal centrifugation (median value before centrifugation 67.5%; after SLC 78%; after DGC 77%; p < 0.001). The incidence of certain abnormalities was reduced, e.g. proximal cytoplasmic droplets were reduced from 12.9% to 8.8% (p < 0.001), and mid-piece defects from 5.3% to 1.4% (p < 0.05). Similarly, sperm motility and chromatin integrity were significantly improved (p < 0.001), with no difference between the two centrifugation methods. Centrifugation through colloids can enrich the proportions of stallion spermatozoa with normal morphology and normal chromatin structure in sperm preparations. The new method, SLC, was as effective as DGC in selecting motile stallion spermatozoa with normal morphology and intact chromatin. SLC, being simpler to use than DGC, would be appropriate for routine use by stud personnel to improve stallion sperm quality in insemination doses.

Get Full Text

Publication Date: 2008-10-16 PubMed ID: 18992087DOI: 10.1111/j.1439-0531.2008.01265.xGoogle Scholar: Lookup

The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.

Facebook X LinkedIn Email

Summary
Cite This
Publication
Affiliations
MeSH
Citations

Comparative Study
Journal Article
Research Support
Non-U.S. Gov't

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.

The research investigates methods to improve qualitative aspects of stallion semen—specifically sperm morphology and chromatin integrity—by using two methods: density gradient centrifugation (DGC) and a new method, single layer centrifugation (SLC). The study demonstrated that both methods increased the proportion of morphologically normal spermatozoa and improved sperm motility and chromatin integrity. Furthermore, the study found no significant difference between the effectiveness of the two centrifugation methods.

Research Method and Materials

The scientists used 38 ejaculates from 10 stallions for the research work.
They analysed the morphology, subjective motility, and chromatin integrity of uncentrifuged samples and compared them to the respective measurements of centrifuged sperm preparations.

Key Findings of the Study

There was a variation in the proportion of morphologically normal spermatozoa across different stallions. However, the median value was found to increase from 67.5% before centrifugation to 77-78% after centrifugation.
The incidence of certain abnormalities in spermatozoa also reduced post-centrifugation. The proximal cytoplasmic droplets reduced from 12.9% to 8.8% and mid-piece defects reduced from 5.3% to 1.4%.
Sperm motility and chromatin integrity were significantly improved post-centrifugation without any noticeable differences between the two centrifugation techniques. Therefore, centrifugation proved effective in enriching the proportion of stallion spermatozoa with normal morphology and normal chromatin structure.
The study concluded that the new method, SLC, is as effective as DGC in filtering motile stallion spermatozoa with normal morphology and intact chromatin . Given that SLC is easier to apply than DGC, it is suitable for routine use to improve the quality of stallion sperm in insemination doses.

Cite This Article

APA

Morrell JM, Johannisson A, Dalin AM, Rodriguez-Martinez H. (2008). Morphology and chromatin integrity of stallion spermatozoa prepared by density gradient and single layer centrifugation through silica colloids. Reprod Domest Anim, 44(3), 512-517. https://doi.org/10.1111/j.1439-0531.2008.01265.x

Publication

Reproduction in domestic animals = Zuchthygiene

ISSN: 1439-0531

NlmUniqueID: 9015668

Country: Germany

Language: English

Volume: 44

Issue: 3

Pages: 512-517

Researcher Affiliations

Morrell, J M

Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden. jane.morrell@kv.slu.se

Johannisson, A

Dalin, A-M

Rodriguez-Martinez, H

MeSH Terms

Animals
Cell Separation / methods
Cell Separation / veterinary
Centrifugation / veterinary
Centrifugation, Density Gradient / veterinary
Chromatin / ultrastructure
Colloids
Horses
Male
Silicon Dioxide
Sperm Count
Sperm Motility
Spermatozoa / abnormalities
Spermatozoa / ultrastructure

Citations

This article has been cited 16 times.

Gutiu00e9rrez-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
Sharafi M, Borghei-Rad SM, Hezavehei M, Shahverdi A, Benson JD. Cryopreservation of Semen in Domestic Animals: A Review of Current Challenges, Applications, and Prospective Strategies.. Animals (Basel) 2022 Nov 24;12(23).
doi: 10.3390/ani12233271pubmed: 36496792google scholar: lookup
Turri F, Capra E, Lazzari B, Cremonesi P, Stella A, Pizzi F. A Combined Flow Cytometric Semen Analysis and miRNA Profiling as a Tool to Discriminate Between High- and Low-Fertility Bulls.. Front Vet Sci 2021;8:703101.
doi: 10.3389/fvets.2021.703101pubmed: 34355036google scholar: lookup
Hermansson U, Johannisson A, Axnu00e9r E. Cryopreservation of dog semen in a Tris extender with two different 1% soybean preparations compared with a Tris egg yolk extender.. Vet Med Sci 2021 May;7(3):812-819.
doi: 10.1002/vms3.445pubmed: 33570263google scholar: lookup
Nongbua T, Utta A, Am-In N, Suwimonteerabutr J, Johannisson A, Morrell JM. Effects of season and single layer centrifugation on bull sperm quality in Thailand.. Asian-Australas J Anim Sci 2020 Sep;33(9):1411-1420.
doi: 10.5713/ajas.19.0624pubmed: 32054188google scholar: lookup
Trasorras VL, Carretero MI, Neild DM, Chaves MG, Giuliano SM, Miragaya MH. Production, Preservation, and Transfer of South American Camelid Embryos.. Front Vet Sci 2017;4:190.
doi: 10.3389/fvets.2017.00190pubmed: 29181380google scholar: lookup
Capra E, Turri F, Lazzari B, Cremonesi P, Gliozzi TM, Fojadelli I, Stella A, Pizzi F. Small RNA sequencing of cryopreserved semen from single bull revealed altered miRNAs and piRNAs expression between High- and Low-motile sperm populations.. BMC Genomics 2017 Jan 4;18(1):14.
doi: 10.1186/s12864-016-3394-7pubmed: 28052756google scholar: lookup
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.
doi: 10.1186/s40104-016-0088-6pubmed: 27158492google scholar: lookup
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.
doi: 10.1155/2014/325451pubmed: 26464926google scholar: lookup
Morrell JM, van Wienen M, Wallgren M. Single Layer Centrifugation Can Be Scaled-Up Further to Process up to 150u2009mL Semen.. ISRN Vet Sci 2011;2011:183412.
doi: 10.5402/2011/183412pubmed: 23738111google scholar: lookup
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.
doi: 10.5402/2011/128984pubmed: 23738093google scholar: lookup
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.
doi: 10.5402/2011/548385pubmed: 23738091google scholar: lookup
Gutiu00e9rrez-Cepeda L, Fernu00e1ndez A, Crespo F, Ramu00edrez Mu00c1, Gosu00e1lvez 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.
doi: 10.1186/1751-0147-54-72pubmed: 23217215google scholar: lookup
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 6u00b0C prior to artificial insemination.. Acta Vet Scand 2012 Jul 12;54(1):40.
doi: 10.1186/1751-0147-54-40pubmed: 22788670google scholar: lookup
Morrell JM, Rodriguez-Martinez H. Practical applications of sperm selection techniques as a tool for improving reproductive efficiency.. Vet Med Int 2010 Aug 4;2011.
doi: 10.4061/2011/894767pubmed: 20814435google scholar: lookup
Morrell JM, Johannisson A, Juntilla L, Rytty K, Bu00e4ckgren 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.
doi: 10.4061/2010/659862pubmed: 20445788google scholar: lookup