Analyze Diet
Reproduction in domestic animals = Zuchthygiene2009; 44(3); 518-522; doi: 10.1111/j.1439-0531.2008.01267.x

Does the microbial flora in the ejaculate affect the freezeability of stallion sperm?

Abstract: In an attempt to evaluate the possible relationship between the microbial flora in the stallion ejaculate and its ability to freeze,three ejaculates from five stallions were frozen using a standard protocol. Before freezing, an aliquot was removed for bacteriological analysis. Bacterial growth was observed in all the ejaculates studied. The isolated microorganisms were:Staphylococcus spp. and Micrococcus spp. (in all the stallions), beta-haemolytic Streptococcus (in stallions 3 and 4), Corynebacterium spp. (in stallions 1, 3-5), Rhodococcus spp. (in stallion number 2), Pseudomonas spp. (in stallion number 1) and Klebsiella spp. (in stallions 1, 3 and 5). The presence and richness of Klebsiella and beta-haemolytic Streptococcus in the ejaculate were related to two sperm variables post-thaw,namely the proportion of dead spermatozoa (ethidium+ cells; r = 0.55, p < 0.05) and the amplitude of lateral displacement of the sperm head (ALH, microm; r = -0.56, p < 0.05), respectively.The degree of growth of Corynebacterium spp. in the ejaculate was positively correlated with the percentage of spermatozoa showing high caspase activity post-thaw(r = 0.62, p < 0.05). The presence and number of colonies of beta-haemolytic Streptococcus were negatively correlated (r = -0.55, p < 0.05) with low sperm caspase activity. It is concluded that the microbial flora of the equine ejaculate maybe responsible for some of the sublethal damage experimented by the spermatozoa during cryopreservation.
Publication Date: 2009-08-06 PubMed ID: 19655428DOI: 10.1111/j.1439-0531.2008.01267.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.
  • 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 research investigated the link between the microbial flora in stallion semen and the semen’s freezing capacity. It concluded that the microbial flora could cause some sublethal damage to the sperm during cryopreservation.

Research Methodology

  • The researchers collected semen samples from five different stallions. Three ejaculates were retrieved from each stallion.
  • The samples were frozen following a standard protocol. Before the freezing process, a sample (a small portion) of the ejaculate was taken for bacterial analysis.
  • Bacterial growth was observed in all the samples studied.

Findings

  • Various microorganisms were isolated from the stallion ejaculates. These include Staphylococcus spp., Micrococcus spp., beta-haemolytic Streptococcus, Corynebacterium spp., Rhodococcus spp., Pseudomonas spp., and Klebsiella spp.
  • Certain correlations between the presence or richness of specific microbes and post-thaw sperm variables were found. For instance, the existence of Klebsiella and beta-haemolytic Streptococcus in the ejaculate was linked with the proportion of dead sperm cells and the range of lateral displacement of the sperm head (known as the ALH), respectively.
  • A positive correlation was found between Corynebacterium spp. growth and the percentage of spermatozoa that show high caspase (a type of protein that aids in cell death) activity after thawing.
  • An inverse correlation was observed between the colonies of beta-haemolytic Streptococcus and low sperm caspase activity.

Conclusion

  • The researchers concluded that the microbial flora in the ejaculate could be a contributing factor for the sublethal damage sustained by spermatozoa during cryopreservation.
  • Therefore, the microbiological factor should be taken into consideration during semen cryopreservation to ensure higher fertility rates.

Cite This Article

APA
Ortega-Ferrusola C, González-Fernández L, Muriel A, Macías-García B, Rodríguez-Martínez H, Tapia JA, Alonso JM, Peña FJ. (2009). Does the microbial flora in the ejaculate affect the freezeability of stallion sperm? Reprod Domest Anim, 44(3), 518-522. https://doi.org/10.1111/j.1439-0531.2008.01267.x

Publication

ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 44
Issue: 3
Pages: 518-522

Researcher Affiliations

Ortega-Ferrusola, C
  • Veterinary Teaching Hospital, Laboratory of Spermatology, University of Extremadura, Cáceres, Spain.
González-Fernández, L
    Muriel, A
      Macías-García, B
        Rodríguez-Martínez, H
          Tapia, J A
            Alonso, J M
              Peña, F J

                MeSH Terms

                • Animals
                • Bacteria / growth & development
                • Bacteria / isolation & purification
                • Caspases / analysis
                • Corynebacterium / isolation & purification
                • Cryopreservation / veterinary
                • Flow Cytometry
                • Horses / microbiology
                • Klebsiella / isolation & purification
                • Male
                • Micrococcus / isolation & purification
                • Pseudomonas / isolation & purification
                • Rhodococcus / isolation & purification
                • Semen / microbiology
                • Semen Preservation / methods
                • Semen Preservation / veterinary
                • Sperm Motility
                • Spermatozoa / physiology
                • Staphylococcus / isolation & purification
                • Streptococcus / isolation & purification

                Citations

                This article has been cited 17 times.
                1. Ullah A, Chen W, Shi L, Wang M, Geng M, Na J, Akhtar MF, Khan MZ, Wang C. Challenges and Enhancing Strategies of Equine Semen Preservation: Nutritional and Genetic Perspectives. Vet Sci 2025 Aug 25;12(9).
                  doi: 10.3390/vetsci12090807pubmed: 41012733google scholar: lookup
                2. Zabala SM, Serres C, Montero N, Crespo F, Lorenzo PL, Pérez-Aguilera V, Oliet A, Hijón V, Moreno S, González-Zorn B, Gutiérrez-Cepeda L. Innovative Approaches to Avoid Antibiotic Use in Equine Semen Cryopreservation: Advancing Sustainable Reproductive Technologies. Animals (Basel) 2025 May 9;15(10).
                  doi: 10.3390/ani15101368pubmed: 40427246google scholar: lookup
                3. Al-Kass Z, Morrell JM. Freezing Stallion Semen-What Do We Need to Focus on for the Future?. Vet Sci 2024 Feb 2;11(2).
                  doi: 10.3390/vetsci11020065pubmed: 38393083google scholar: lookup
                4. Malaluang P, Niazi A, Guo Y, Nagel C, Guimaraes T, Rocha A, Aurich C, Morrell JM. Bacterial diversity in semen from stallions in three European countries evaluated by 16S sequencing. Vet Res Commun 2024 Jun;48(3):1409-1421.
                  doi: 10.1007/s11259-024-10321-3pubmed: 38305959google scholar: lookup
                5. Zabala SM, Serres C, Montero N, Crespo F, Lorenzo PL, Pérez-Aguilera V, Galán C, Domínguez-Gimbernat M, Oliet A, Moreno S, González-Zorn B, Gutiérrez-Cepeda L. Strategies to Reduce the Use of Antibiotics in Fresh and Chilled Equine Semen. Animals (Basel) 2024 Jan 5;14(2).
                  doi: 10.3390/ani14020179pubmed: 38254348google scholar: lookup
                6. Quiñones-Pérez C, Martínez A, Ortiz I, Crespo F, Vega-Pla JL. The Semen Microbiome and Semen Parameters in Healthy Stallions. Animals (Basel) 2022 Feb 22;12(5).
                  doi: 10.3390/ani12050534pubmed: 35268102google scholar: lookup
                7. Malaluang P, Wilén E, Lindahl J, Hansson I, Morrell JM. Antimicrobial Resistance in Equine Reproduction. Animals (Basel) 2021 Oct 22;11(11).
                  doi: 10.3390/ani11113035pubmed: 34827768google scholar: lookup
                8. Quiñones-Pérez C, Hidalgo M, Ortiz I, Crespo F, Vega-Pla JL. Characterization of the seminal bacterial microbiome of healthy, fertile stallions using next-generation sequencing. Anim Reprod 2021;18(2):e20200052.
                  doi: 10.1590/1984-3143-AR2020-0052pubmed: 34394753google scholar: lookup
                9. Al-Kass Z, Eriksson E, Bagge E, Wallgren M, Morrell JM. Microbiota of semen from stallions in Sweden identified by MALDI-TOF. Vet Anim Sci 2020 Dec;10:100143.
                  doi: 10.1016/j.vas.2020.100143pubmed: 33145452google scholar: lookup
                10. Quiñones-Pérez C, Martínez A, Crespo F, Vega-Pla JL. Comparative Semen Microbiota Composition of a Stallion in a Taylorella equigenitalis Carrier and Non-Carrier State. Animals (Basel) 2020 May 17;10(5).
                  doi: 10.3390/ani10050868pubmed: 32429567google scholar: lookup
                11. 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
                12. Al-Kass Z, Eriksson E, Bagge E, Wallgren M, Morrell JM. Bacteria detected in the genital tract, semen or pre-ejaculatory fluid of Swedish stallions from 2007 to 2017. Acta Vet Scand 2019 May 30;61(1):25.
                  doi: 10.1186/s13028-019-0459-zpubmed: 31146786google scholar: lookup
                13. Banday MN, Lone FA, Rasool F, Rather HA, Rather MA. Does natural honey act as an alternative to antibiotics in the semen extender for cryopreservation of crossbred ram semen?. Iran J Vet Res 2017 Fall;18(4):258-263.
                  pubmed: 29387098
                14. Meena GS, Raina VS, Gupta AK, Mohanty TK, Bhakat M, Abdullah M, Bishist R. Effect of preputial washing on bacterial load and preservability of semen in Murrah buffalo bulls. Vet World 2015 Jun;8(6):798-803.
                15. Gibb Z, Aitken RJ. The Impact of Sperm Metabolism during In Vitro Storage: The Stallion as a Model. Biomed Res Int 2016;2016:9380609.
                  doi: 10.1155/2016/9380609pubmed: 26881234google scholar: lookup
                16. 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
                17. 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.
                  doi: 10.1371/journal.pone.0030688pubmed: 22292020google scholar: lookup