FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Equine veterinary journal2009; 41(1); 53-58; doi: 10.2746/042516408x322139

Comparison of density gradient and single layer centrifugation of stallion spermatozoa: yield, motility and survival.

Abstract: A new, simpler, technique of colloidal centrifugation has recently been developed, designated single layer centrifugation (SLC). This technique requires evaluation by comparison with a density gradient for its ability to select the best quality spermatozoa and its practicality of use on studfarms. Objective: To compare the effect of 2 methods of colloidal centrifugation, density gradient centrifugation and single layer centrifugation, on stallion sperm motility, yield and survival, using freshly collected extended stallion semen. Methods: Aliquots of extended stallion semen from 10 stallions (38 ejaculates) were processed by the 2 methods of colloidal centrifugation. For both uncentrifuged and centrifuged samples, sperm yield was calculated and subjective sperm motility assessed over several days to provide an estimate of sperm survival. Some stored semen samples, held at 4 degrees C overnight, were also available for testing. Results: For fresh, extended semen, a similar recovery yield of motile spermatozoa was seen for the 2 methods of preparation for single layers and density gradients, respectively. Sperm motility and survival rate were significantly improved by colloidal centrifugation compared to unprocessed ejaculate, without any significant difference between methods (SLC vs. gradient). However, the yield was reduced by 18-20% when cold-stored semen was used for centrifugation compared to fresh semen; and more variation between ejaculates was observed than for fresh ejaculates. Again, sperm motility and sperm survival were improved in the centrifuged sperm preparations compared to stored, unprocessed ejaculates. Conclusions: The 2 colloid centrifugation techniques produce equivalent sperm preparations in terms of sperm quality. However, the SLC method would be more practical and convenient for use in the field.
Get Full Text
Publication Date: 2009-03-24 PubMed ID: 19301582DOI: 10.2746/042516408x322139Google 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.
LinkedInCopy
  • 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 article evaluates a new technique for increasing the quality of stallion sperm known as single layer centrifugation (SLC), comparing it to the traditional method of density gradient centrifugation. The authors found both methods produce similar results in terms of sperm quality, but suggest that the new SLC method may be more practical for use.

Objective and Methods

  • The main goal of this study was to compare two methods of colloidal centrifugation, namely density gradient centrifugation and single layer centrifugation (SLC), to see their effects on stallion sperm motility, output, and longevity.
  • The scientists used semen samples from ten stallions, processing them using both methods.
  • They calculated the sperm yield and assessed motility subjectively over several days to estimate how long the sperm would survive.
  • They also tested some semen samples stored at 4 degrees Celsius overnight.

Results

  • The researchers found that for fresh, extended semen, both methods resulted in similar recovery yields of motile spermatozoa.
  • The sperm’s movement and survival rates significantly improved with BOTH methods, compared to unprocessed ejaculate, without any significant difference between the two centrifugation methods.
  • However, they noticed a decrease of 18-20% in sperm yield when using cold-stored semen for centrifugation compared to fresh semen. They also observed more variation between ejaculates when using cold-stored semen.
  • Despite the reduced yield with cold-stored semen, they found that sperm motility and sperm survival still improved in the centrifuged preparations compared to stored, unprocessed ejaculates.

Conclusions

  • The study concluded that both colloid centrifugation techniques produce the equivalent sperm preparations in terms of sperm quality.
  • However, the SLC method would be more convenient and practical for use in real-world applications, such as studfarms, due to its simplicity.

Cite This Article

APA
Morrell JM, Dalin AM, Rodriguez-Martinez H. (2009). Comparison of density gradient and single layer centrifugation of stallion spermatozoa: yield, motility and survival. Equine Vet J, 41(1), 53-58. https://doi.org/10.2746/042516408x322139

Publication

Equine veterinary journal
ISSN: 0425-1644
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 41
Issue: 1
Pages: 53-58

Researcher Affiliations

Morrell, J M
  • Swedish University of Agricultural Sciences-SLU, Department of Clinical Sciences, Division of Reproduction, Box 7054, SE-75007 Uppsala, Sweden.
Dalin, A M
    Rodriguez-Martinez, H

      MeSH Terms

      • Animals
      • Centrifugation, Density Gradient / methods
      • Centrifugation, Density Gradient / veterinary
      • Ejaculation
      • Horses / physiology
      • Male
      • Population Density
      • Semen Preservation / methods
      • Semen Preservation / veterinary
      • Specimen Handling / veterinary
      • Sperm Count / veterinary
      • Sperm Motility / physiology
      • Spermatozoa / cytology
      • Spermatozoa / physiology
      • Time Factors
      • Tissue and Organ Harvesting / methods
      • Tissue and Organ Harvesting / veterinary

      Citations

      This article has been cited 13 times.
      1. Pinto SZ, Aneck-Hahn N. Effect of in vitro exposure of first-line antiretrovirals on healthy human spermatozoa on kinematics and motility. Int Urol Nephrol 2025 Jun;57(6):1715-1735.
        doi: 10.1007/s11255-024-04340-xpubmed: 39753908google scholar: lookup
      2. Bisiau C, Moffett P, Graham J, McCue P. Comparison of Nanoparticles and Single-Layer Centrifugation for Separation of Dead from Live Stallion Spermatozoa. Vet Sci 2024 Jul 10;11(7).
        doi: 10.3390/vetsci11070307pubmed: 39057991google scholar: lookup
      3. Alves NC, Diniz SA, Viegas RN, Arigoni AL, Freitas MM, Lana ÂQ, Lagares MA. Action of swim-up and caffeine on equine frozen sperm. Anim Reprod 2022;19(4):e20220056.
        doi: 10.1590/1984-3143-AR2022-0056pubmed: 36504918google scholar: lookup
      4. Papas M, Catalán J, Recuero S, Morrell JM, Yeste M, Miró J. Single Layer Centrifugation Improves the Quality of Fresh Donkey Semen and Modifies the Sperm Ability to Interact with Polymorphonuclear Neutrophils. Animals (Basel) 2020 Nov 16;10(11).
        doi: 10.3390/ani10112128pubmed: 33207812google scholar: lookup
      5. Gómez-Quispe OE, Gutiérrez-Reynoso GA, Gallegos-Cardenas A, Fumuso FG, Asparrin M, Asparrin-Del Carpio M, Jara CW, Ponce D, Miguel M, Youngs CR, Vivanco HW. Motility and recovery of alpaca (Vicugna pacos) spermatozoa after centrifugation in a density gradient solution. Iran J Vet Res 2019 Spring;20(2):96-104.
        pubmed: 31531031
      6. 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
      7. Jatesada J, Elisabeth P, Anne-Marie D. Seminal plasma did not influence the presence of transforming growth factor-β1, interleukine-10 and interleukin-6 in porcine follicles shortly after insemination. Acta Vet Scand 2013 Sep 10;55(1):66.
        doi: 10.1186/1751-0147-55-66pubmed: 24020676google scholar: lookup
      8. 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
      9. 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
      10. 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
      11. 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
      12. 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
      13. 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
      Subscribe to our newsletter
      Join 99,824 horse owners like you who receive our email updates on horse health and nutrition.