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Theriogenology2009; 72(6); 879-884; doi: 10.1016/j.theriogenology.2009.05.015

Single-layer centrifugation with Androcoll-E can be scaled up to allow large volumes of stallion ejaculate to be processed easily.

Abstract: The objective of the current study was to optimize the volumes of Androcoll-E and sperm sample used in various sizes of centrifuge tube to scale up single-layer centrifugation (SLC) for routine use in the field. Although sperm suspensions of equivalent quality were produced using Androcoll-E in small and large tubes, the sperm yield was much lower in the latter (P<0.001). In contrast, in 200-mL tubes (XL), the yields were approximately 25% higher than those for the small tubes. An increased volume (4.5 mL) of extended ejaculate in small tubes (SLC-Inc) or 15 to 18 mL extended ejaculate on 15 mL of colloid of a reduced density, Androcoll-E-Large (SLC-Large), in 50-mL tubes were both found to give similar yields of motile spermatozoa as that of the SLC-Small method (SLC-Small, 49.7+/-18.6%; SLC-Inc, 53.3+/-17.1%; SLC-Large, 44.9+/-18.3%) and were found to be equivalent in quality (motility: 88.0+/-8.8%, 84.0+/-3.5%, 90.0+/-5.4%; normal morphology: 69.4+/-17.0%, 69.4+/-12.7%, 63.9+/-15.6%; viability: 78+/-16.7%, 83.8+/-12.5%, 80.05+/-14.6%; DNA fragmentation index: 14.7+/-10.9%, 12.8+/-8.1%, 11.6+/-7.6%, respectively). The processing of an "average" stallion Equus caballus ejaculate in approximately twenty-seven 10-mL tubes (SLC-Inc) or eight 50-mL tubes (SLC-Large) is feasible, the latter being considered more practical for on-stud use.
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Publication Date: 2009-07-14 PubMed ID: 19604571DOI: 10.1016/j.theriogenology.2009.05.015Google Scholar: Lookup
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  • 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.

This study focuses on optimizing the centrifugation process for stallion semen using a substance called Androcoll-E, with the aim to make the process more feasible for use in typical field settings. The results showed that varying volumes and methods led to similar sperm yields, but using larger tubes (200-mL) yielded 25% more sperm compared to smaller tubes.

Overview of the Research

  • The study’s main purpose was to enhance the volumes of stallion sperms and Androcoll-E deployed in various centrifuge tube sizes. The ultimate goal was to upscale the single-layer centrifugation (SLC) technique to make it commonplace and more effective in various situations.
  • The researchers used different volumes of Androcoll-E and sperm samples in tubes of various sizes to conduct this research.

Findings of the Research

  • Although the use of this substance in large and small tube sizes resulted in the production of sperm suspensions of equivalent quality, the yield of sperm was significantly lesser in the larger tubes.
  • In contrast, the research discovered that 200-mL tubes yielded about 25% higher sperm volumes compared to their smaller counterparts.
  • Additionally, the use of an increased sample volume of 4.5 mL in the small tubes or 15 to 18 mL sample on 15 mL of colloid of reduced density in 50-mL tubes gave similar yields of motile sperms. These methods are known as SLC-Inc and SLC-Large respectively.
  • Overall sperm yield and quality were found to be similar across the SLC-Small, SLC-Inc, and SLC-Large methods.
  • All three methods showed similar sperm quality, evaluating factors such as normal morphology, motility, viability, and DNA fragmentation index.

Practical Applications and Conclusion

  • The study demonstrated that it is feasible to process an “average” stallion ejaculate using either twenty-seven 10-mL tubes (SLC-Inc method) or eight 50-mL tubes (SLC-Large method).
  • The researchers conclude the latter is more practical for on-stud use thanks to providing similar yields of motile sperm while being less time-consuming and easier to use in the field.

Cite This Article

APA
Morrell JM, Johannisson A, Dalin AM, Rodriguez-Martinez H. (2009). Single-layer centrifugation with Androcoll-E can be scaled up to allow large volumes of stallion ejaculate to be processed easily. Theriogenology, 72(6), 879-884. https://doi.org/10.1016/j.theriogenology.2009.05.015

Publication

Theriogenology
ISSN: 1879-3231
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 72
Issue: 6
Pages: 879-884

Researcher Affiliations

Morrell, J M
  • Department of Clinical Sciences, Division of Reproduction, Swedish University of Agricultural Sciences, Uppsala, Sweden. jane.morrell@kv.slu.se
Johannisson, A
    Dalin, A-M
      Rodriguez-Martinez, H

        MeSH Terms

        • Animals
        • Centrifugation
        • Chromatin / chemistry
        • Colloids
        • Horses
        • Male
        • Silicon Dioxide
        • Sperm Motility
        • Spermatozoa / physiology
        • Tissue and Organ Harvesting

        Citations

        This article has been cited 14 times.
        1. Lima-Verde I, Hurri E, Ntallaris T, Johannisson A, Stålhammar H, Morrell JM. Sperm Quality in Young Bull Semen Can Be Improved by Single Layer Centrifugation. Animals (Basel) 2022 Sep 15;12(18).
          doi: 10.3390/ani12182435pubmed: 36139296google scholar: lookup
        2. 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.
          doi: 10.1186/s13028-020-00530-6pubmed: 32552825google scholar: lookup
        3. 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
        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 (Basel) 2017 Dec 21;7(1).
          doi: 10.3390/antibiotics7010001pubmed: 29267226google scholar: lookup
        5. Morrell JM, Wallgren M. Alternatives to antibiotics in semen extenders: a review. Pathogens 2014 Dec 15;3(4):934-46.
          doi: 10.3390/pathogens3040934pubmed: 25517429google scholar: lookup
        6. 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.
          doi: 10.5402/2011/183412pubmed: 23738111google scholar: lookup
        7. 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
        8. 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.
          doi: 10.1186/1751-0147-54-72pubmed: 23217215google scholar: lookup
        9. 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.
          doi: 10.1186/1751-0147-54-40pubmed: 22788670google scholar: lookup
        10. 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
        11. 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.
          doi: 10.4061/2010/659862pubmed: 20445788google scholar: lookup
        12. Jochems R, Kristiansen CO, Kommisrud E, Morrell JM. Preliminary study of pregnancy rates and litter sizes following artificial insemination of boar spermatozoa prepared by colloid centrifugation and hypothermic storage. Sci Rep 2025 Nov 19;15(1):40863.
          doi: 10.1038/s41598-025-24586-4pubmed: 41258329google scholar: lookup
        13. Mikkola M, Desmet KLJ, Kommisrud E, Riegler MA. Recent advancements to increase success in assisted reproductive technologies in cattle. Anim Reprod 2024;21(3):e20240031.
          doi: 10.1590/1984-3143-AR2024-0031pubmed: 39176005google scholar: lookup
        14. Andrei CR, Posastiuc FP, Constantin NT, Mitrea IL. New insights into semen separation techniques in buffaloes. Front Vet Sci 2023;10:1347482.
          doi: 10.3389/fvets.2023.1347482pubmed: 38269362google scholar: lookup
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