Veterinary research communications2024; doi: 10.1007/s11259-024-10321-3

Bacterial diversity in semen from stallions in three European countries evaluated by 16S sequencing.

Abstract: The microbiome plays a significant role in shaping the health and functioning of the systems it inhabits. The seminal microbiome of stallions has implications for the health of the reproductive tract, sperm quality during preservation and antibiotic use in semen extenders. Diverse bacteria are present on the external genital tract and a mix of commensal microorganisms populates various parts of the reproductive tract, influencing the seminal bacterial content. Other sources of bacteria include the environment, semen collection equipment, and personnel. The bacterial load can adversely affect sperm quality and fertility, particularly in artificial insemination, where semen is extended and stored before use. Antibiotics are frequently used to inhibit bacterial growth, but their effectiveness varies depending on the bacterial strains present. The aim of this study was to assess the bacterial diversity in semen from 37 healthy stallions across three European nations (Germany, Portugal, and Sweden) using 16S sequencing. Semen samples were collected from individual stallions at three AI centers; DNA extraction, sequencing, and bioinformatic analysis were performed. Differences in bacterial diversity among the stallions were seen; although bacterial phyla were shared across the regions, differences were observed at the genus level. Climate, husbandry practices, and individual variability likely contribute to these differences. These findings underscore the importance of tailoring antibiotic strategies for semen preservation based on regional bacterial profiles. The study presents a comprehensive approach to understanding the intricacies of the stallion seminal microbiome and its potential implications for reproductive technologies and animal health.
Publication Date: 2024-02-02 PubMed ID: 38305959PubMed Central: 7015180DOI: 10.1007/s11259-024-10321-3Google 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

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 the diversity of bacteria in semen from male horses in three European countries, aiming to illuminate how different bacterial strains may influence the health and quality of the reproductive system. Through comprehensive DNA extraction, sequencing, and analysis methods, the study presents a vital understanding of the intricate interactions within the reproductive tract’s microbiome.

Understanding the Microbiome in Stallion Semen

  • This study primarily focuses on the microbiome of stallions’ seminal fluid, the role it plays in reproductive health, the quality of sperm, and the implications for the application of antibiotics. The microbiome is an ecosystem of microorganisms that live on and in our bodies. It significantly contributes to health and physiological functions.
  • The research highlights the presence of diverse bacteria on the external genital tract and a variety of commensal microorganisms within the different parts of the stallion’s reproductive tract. These bacteria can originate from different sources, like the environment, semen collection equipment, or even the personnel that handle the specimens.
  • An essential component mentioned is the bacterial load’s potential negative impact, particularly on sperm quality and fertility, especially in the context of artificial insemination where the semen has to be extended and preserved before it can be used.

Diversity Assessed through 16S Sequencing

  • With the aim of understanding the bacterial diversity within the semen of stallions, researchers collected samples from 37 healthy stallions from Germany, Portugal, and Sweden. These samples were then subjected to DNA extraction, followed by 16S sequencing and bioinformatics analysis.
  • 16S sequencing is a common method for studying bacterial biodiversity, allowing researchers to identify and catalog all the bacterial species present in each semen sample.
  • This unique approach allowed researchers to see differences in the bacterial diversity among the sampled stallions. They found that while certain bacterial phyla were common across samples, variations were observed at the genus level.

Implications for Animal Reproductive Health

  • The research was able to link these variations at the genus level to external environmental factors such as climate, different animal rearing practices, and individual health variations within the animals.
  • Consequently, the study recommends tailoring the use of antibiotics during semen preservation to individual animals and regions, given the variability in bacterial profiles which impacts the effectiveness of antibiotics. By understanding the composition of the microbiome in relation to different factors, the study contributes significantly to the advancement of reproductive technologies and animal health management strategies.

Cite This Article

APA
Malaluang P, Niazi A, Guo Y, Nagel C, Guimaraes T, Rocha A, Aurich C, Morrell JM. (2024). Bacterial diversity in semen from stallions in three European countries evaluated by 16S sequencing. Vet Res Commun. https://doi.org/10.1007/s11259-024-10321-3

Publication

ISSN: 1573-7446
NlmUniqueID: 8100520
Country: Switzerland
Language: English

Researcher Affiliations

Malaluang, Pongpreecha
  • Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, 75007, Sweden.
  • Faculty of Veterinary Sciences, Mahasarakham University, Maha Sarakham, 40000, Thailand.
Niazi, Adnan
  • SLU-Global Bioinformatics Centre, Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences (SLU), Uppsala, SE-750 07, Sweden.
  • Science for Life Laboratory, National Bioinformatics Infrastructure Sweden (NBIS), Uppsala University, Uppsala, SE-752 36, Sweden.
Guo, Yongzhi
  • Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, 75007, Sweden.
Nagel, Christina
  • Graf Lehndorff Institute for Equine Science, University of Veterinary Medicine, Vienna, Austria.
Guimaraes, Tiago
  • School of Medicine and Biomedical Sciences (ICBAS), University of Porto (UP), Porto, Portugal.
  • Center for the Study of Animal Sciences (CECA), ICETA, University of Porto, Campus Agru00e1rio de Vairu00e3o, Vairu00e3o, Portugal.
Rocha, Antonio
  • School of Medicine and Biomedical Sciences (ICBAS), University of Porto (UP), Porto, Portugal.
  • Center for the Study of Animal Sciences (CECA), ICETA, University of Porto, Campus Agru00e1rio de Vairu00e3o, Vairu00e3o, Portugal.
Aurich, Christine
  • Artificial Insemination and Embryo Transfer, Department for Small Animals and Horses, University of Veterinary Medicine, Vienna, Austria.
Morrell, Jane M
  • Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, 75007, Sweden. jane.morrell@slu.se.

References

This article includes 41 references
  1. Al-Kass Z, Guo Y, Vinnere Pettersson O, Niazi A, Morrell JM (2020) Metagenomic analysis of bacteria in stallion semen. Anim Reprod Sci 221:106568. https://doi.org/10.1016/J.ANIREPROSCI.2020.106568
  2. Al-Kass Z, Spergser J, Aurich C, Kuhl J, Schmidt K, Morrell JM (2019) Effect of presence or absence of antibiotics and use of modified single layer centrifugation on bacteria in pony stallion semen. Reprod Domest Anim 54:342u2013349.
  3. Althouse GC (2008) Sanitary procedures for the production of Extended Semen. Reprod Domest Anim 43:374u2013378. https://doi.org/10.1111/j.1439-0531.2008.01187.x
  4. Andrews S (2010) FastQC:A Quality Control tool for High Throughput Sequence Data. In: Babraham Bioinformatics. https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ . Accessed 30 Jun 2023
  5. Aurich C, Spergser J (2007) Influence of bacteria and gentamicin on cooled-stored stallion spermatozoa. Theriogenology 67:912u2013918. https://doi.org/10.1016/j.theriogenology.2006.11.004
  6. Bermingham EN, Young W, Butowski CF, Moori CD, Maclean PH, Rosendale D, Cave NJ, Thomas DG (2018) The fecal microbiota in the domestic cat (Felis catus) is influenced by interactions between age and diet; a five year longitudinal study. Front Microbiol 9:365211. https://doi.org/10.3389/FMICB.2018.01231/BIBTEX
  7. Bolyen E, Rideout JR, Dillon MR, Bokulich NA, Abnet CC, Al-Ghalith GA, Alexander H, Alm EJ, Arumugam M, Asnicar F, Bai Y, Bisanz JF, Bittinger K, Brejnrod A, Brislawn CJ, Brown CT, Callahan BJ, Caraballo-Rodru00edguez AM, Chase J, Cope EK, Da Silva R, Diener C, Dorrestein PC, Douglas GM, Caporaso JG (2019) Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nat Biotechnol 37(8 37):852u2013857. https://doi.org/10.1038/s41587-019-0209-9
    doi: 10.1038/s41587-019-0209-9pubmed: 31341288pmc: 7015180google scholar: lookup
  8. Bu00f6ttger EC (1989) Rapid determination of bacterial ribosomal RNA sequences by direct sequencing of enzymatically amplified DNA. FEMS Microbiol Lett 65. https://doi.org/10.1016/0378-1097(89)90386-8
  9. Bowen JM, Tobin N, Simpson RB, Ley WB, Ansari MM (1982) Effects of washing on the bacterial flora of the stallionu2019s penis. J Reprod Fertil Suppl 32:41u201345 PMID: 6820065
    pubmed: 6820065
  10. Bradley AJ, Leach KA, Green MJ, Gibbons J, Ohnstad IC, black DH, Payne, B, Prout VE, Breen JE (2018) The impact of dairy cowsu2019bedding material and its microbial content on the quality and safety of milk u2013A cross sectional study of UK farms. Int J Food Microbiol 269:36u201345. https://doi.org/10.1016/J.IJFOODMICRO.2017.12.022
  11. Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP (2016) DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods 2016 13:7. https://doi.org/10.1038/nmeth.3869
    doi: 10.1038/nmeth.3869google scholar: lookup
  12. Cojkic A, Hansson I, Johannisson A, Axnu00e9r E, Morrell JM (2023) Single layer centrifugation as a method for bacterial reduction in bull semen for assisted reproduction. Vet Res Commun 2023:1u201310. https://doi.org/10.1007/S11259-023-10178-Y
  13. Corona A, Cherchi R (2009) Microbial quality of equine frozen semen. Anim Reprod Sci 115:103u2013109. https://doi.org/10.1016/j.anireprosci.2008.11.016
  14. Goldberg AMG, Argenti LE, Faccin JE, Linck L, Santi M, Bernardi ML, Cardoso MRL, Wentz I, Bortolozzo FP (2013) Risk factors for bacterial contamination during boar semen collection. Res Vet Sci 95:362u2013367. https://doi.org/10.1016/j.rvsc.2013.06.022
    doi: 10.1016/j.rvsc.2013.06.022pubmed: 23891384google scholar: lookup
  15. Guimaru00e3es T, Lopes G, Pinto M, Silva E, Miranda C, Correia MJ, Damu00e1sio L, Thompson G, Rocha A (2015) Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics. Theriogenology 83:186u2013191. https://doi.org/10.1016/J.THERIOGENOLOGY.2014.09.003
  16. Hogan JS, Smith KL, Hoblet KH, Todhunter DA, Schoenberger PS, Hueston WD, Pritchar DE, Bowman GL, Heider LE, Brockett BL, Conrad HR (1989) Bacterial counts in bedding materials used on nine commercial dairies. J Dairy Sci 72:250u2013258. https://doi.org/10.3168/JDS.S0022-0302(89)79103-7
  17. Jang J, Hur H-G, Sadowsky MJ, Byappanahalli MN, Yan T, Ishii S (2017) Environmental Escherichia coli: ecology and public health implicationsu2014a review. J Appl Microbiolo 123:570u2013581
    doi: 10.1111/jam.13468google scholar: lookup
  18. Kenney RM, Bergman RV, Cooper WL, Morse FW (1975) Minimal contamination techniques for breeding mares: techniques and preliminary findings. Proc Am Association Equine Practitioners 21:327u2013336
  19. Kuczynski J, Lauber CL, Walters WA, Parfrey LW, Clemente JC, Gevers D, Knight R (2012) Experimental and analytical tools for studying the human microbiome. Nat Rev Genet 13.
  20. Martin M (2011) Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J 17:10u201312. https://doi.org/10.14806/EJ.17.1.200 .
  21. McMurdie PJ, Holmes S (2013) Phyloseq: an R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data. PLoS ONE 8:e61217. https://doi.org/10.1371/JOURNAL.PONE.0061217
    doi: 10.1371/JOURNAL.PONE.0061217pubmed: 23630581pmc: 3632530google scholar: lookup
  22. Mignard S, Flandrois JP (2006) 16S rRNA sequencing in routine bacterial identification: a 30-month experiment. J Microbiol Methods 67. https://doi.org/10.1016/j.mimet.2006.05.009
  23. Moretti E, Capitani S, Figura N, Pammolli A, Federico MG, Giannerini V, Collodel G (2009) The presence of bacteria species in semen and sperm quality. J Assist Reprod Genet 26:47u201356
    doi: 10.1007/s10815-008-9283-5pubmed: 19089609google scholar: lookup
  24. Morrell JM, Wallgren M (2014) Alternatives to antibiotics in semen extenders: a review. Pathogens 3:943u2013946
    doi: 10.3390/pathogens3040934google scholar: lookup
  25. Morrell JM, Malaluang P, Cojkic A, Hansson I (2022) Alternatives to antibiotics in semen extenders used in artificial insemination. Ch in The Global Antimicrobial Resistance Epidemic u2013 Innovative Approaches and Cutting-Edge Solutions, ISBN 978-1-80356-042-7. https://doi.org/10.5772/intechopen.98171
  26. Nedwell DB (1999) Effect of low temperature on microbial growth: lowered affinity for substrates limits growth at low temperature. FEMS Microbiol Ecol 30:101u2013111. https://doi.org/10.1111/J.1574-6941.1999.TB00639.X
  27. Ortega-Ferrusola C, Gonzau00edez-Fernandez L, Muriel A, Macias-Garcia B, Rodriguez-Martinez H, Tapia JA, Alonso JM, Pena FJ (2009) Does the microbial flora in the ejaculate affect the freezeability of stallion sperm? Wiley Online Libr 44:518u2013522. https://doi.org/10.1111/j.1439-0531.2008.01267.x
  28. Pasing SS, Aurich C, von Lewinski M, Wulf M, Kru00fcger M, Aurich JE (2013) Development of the genital microflora in stallions used for artificial insemination throughout the breeding season. Anim Reprod Sci 139:53u201361. https://doi.org/10.1016/j.anireprosci.2013.03.009
  29. Pickett BW, Voss JL, Jones RL (1999) Control of bacteria in stallions and their semen. J Equine Vet Sci 19:424u2013469. https://doi.org/10.1016/S0737-0806(99)80254-8
  30. Qiu Y, Zhou Y, Chang Y, Liang X, Zhang H, Lin X, Qing K, Zhou X, Luo Z (2022) The effects of Ventilation, Humidity, and temperature on bacterial growth and bacterial genera distribution. Int J Environ Res Public Health 19:15345. https://doi.org/10.3390/ijerph192215345 PMID: 36430064; PMCID: PMC9691097
    doi: 10.3390/ijerph192215345pubmed: 36430064pmc: 9691097google scholar: lookup
  31. Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glu00f6ckner FO (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:D590u2013D596. https://doi.org/10.1093/NAR/GKS1219
    doi: 10.1093/NAR/GKS1219pubmed: 23193283google scholar: lookup
  32. Quiu00f1ones-Pu00e9rez C, Hidalgo M, Ortiz I, Crespo F, Vega-Pia JL (2021) Characterization of the seminal bacterial microbiome of healthy, fertile stallions using next-generation sequencing. Anim Reprod 18:e20200052. https://doi.org/10.1590/1984-3143-AR2020-0052
    doi: 10.1590/1984-3143-AR2020-0052pubmed: 34394753pmc: 8356074google scholar: lookup
  33. Ramires Neto C, Sancler da Silva YFR, Resende HL, Nascimento Guasti P, Montiero GA, Melo Papa P, Dellu00e1qua JA, Junior P, Filho JNP, Alvarenga MA, Papa FO (2015) Control methods and evaluation of bacterial growth on fresh and cooled stallion semen. J Equine Vet Sci 35:277u2013282. https://doi.org/10.1016/j.jevs.2015.01.014
  34. Rota A, Calicchio E, Nardoni S, Fratini F, Ebani VV, Sgorbini M, Panzani D, Camillo F, Mancianti F (2011) Presence and distribution of fungi and bacteria in the reproductive tract of healthy stallions. Theriogenology 76:464u2013470. https://doi.org/10.1016/j.theriogenology.2011.02.023
  35. u015eahin D, Bau015ftan I, Beste u00c7il, Tekin K, Aku00e7ay E, Dau015fku0131n A, Stelletta C (2020) The number of false mounting affects the quality of semen in bulls. Livest Stud 60:9u201315
  36. Samper JC (2008) Equine breeding management and artificial insemination. Elsevier Health Sciences
  37. Samper JC, Tibary A (2006) Disease transmission in horses. Theriogenology 66:551u2013559. https://doi.org/10.1016/j.theriogenology.2006.04.019
  38. van der Straaten T (2015) Next-Generation Sequencing:Current Technologies and Applications. Edited by Jianping Xu. ChemMedChem 10:. https://doi.org/10.1002/cmdc.201402456
  39. Varela E, Rey J, Plaza E, Munoz de Propios P, Ortiz-Rodriguez JM, Alvarez M, Anel-Lopez L, Anel l, De paz P, Gil MC, Morrell JM, Ortega-Ferrusola C (2018) How does the microbial load affect the quality of equine cool-stored semen? Theriogenology 114:212u2013220. https://doi.org/10.1016/j.theriogenology.2018.03.028
  40. Vartoukian SR, Downes J, Palmer RM, Wade WG (2013) Fretibacterium Fastidiosum gen. nov., sp. nov., isolated from the human oral cavity. Int J Syst Evol Microbiol 63:458u2013463. https://doi.org/10.1099/IJS.0.041038-0
    doi: 10.1099/IJS.0.041038-0pubmed: 22493171google scholar: lookup
  41. Wu L, Wen C, Qin Y, van Nostrand JD, Ning D, Sun B, Xue K, Liu F, Deng Y, Liang Y, Zhou J (2015) Phasing amplicon sequencing on Illumina Miseq for robust environmental microbial community analysis. BMC Microbiol 15:125. https://doi.org/10.1186/s12866-015-0450-4t
    doi: 10.1186/s12866-015-0450-4tpubmed: 26084274pmc: 4472414google scholar: lookup

Citations

This article has been cited 0 times.