Microbiota of semen from stallions in Sweden identified by MALDI-TOF.
Abstract: Stallion semen is known to contain environmental bacteria and normal commensals, and in some cases may contain opportunistic pathogens. These bacteria may negatively influence sperm quality during storage before artificial insemination. The bacteria isolated depend on the culture conditions and method of identification; therefore, the aim of this study was to identify as many of the bacteria present in stallion semen as possible by culturing aliquots of semen under a variety of conditions. Eleven semen samples were available: five extended semen samples from one stud together with a sample of the extender, and six raw semen samples from another stud. Aliquots of semen samples were cultured on different agars and under specialized conditions; individual bacterial colonies were identified using Matrix-assisted laser desorption ionization time of flight mass spectrometry. Approximately 55% of the bacteria could be identified, with 20 bacterial taxa being isolated from semen samples from the five stallions on the first stud and 11 taxa from the semen samples from six stallions on the second stud. spp. were present in all samples, and spp. were present in all of the extended semen samples although they were also isolated from the extender. The number of bacteria in colony forming units per mL varied considerably among samples. Only one microbe known to be associated with equine infertility, spp., was isolated from three samples, albeit in low numbers. In conclusion, bacterial culture followed by MALDI-TOF does not identify all bacteria present in stallion semen samples. In-depth knowledge of which microbes are likely to be present is useful in determining their effects on sperm quality and, where appropriate, developing protocols for effectively controlling microbial growth.
© 2020 The Author(s).
Publication Date: 2020-10-01 PubMed ID: 33145452PubMed Central: PMC7593612DOI: 10.1016/j.vas.2020.100143Google Scholar: Lookup
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- Journal Article
Summary
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The research article is a study of the various bacteria present in stallion semen in Sweden, conducted to understand their effects on sperm quality using a method known as Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF).
Objective of the Study
- This research was aimed at identifying as many bacteria as possible present in stallion semen by culturing semen under different conditions. The goal is to determine any potential negative influence these bacteria might have on sperm quality, crucial in the context of storage before artificial inseparation.
Methodology
- Eleven semen samples were used in the study. Five were extended semen samples from one stud along with a extender sample, and six were raw semen samples from another stud.
- The researchers went ahead to culture aliquots of these semen samples on different agars and under specialized conditions.
- Identification of individual bacterial colonies was achieved using Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF), a cutting edge technology that allows for precise identification of bacteria.
Results and Findings
- About 55% of the bacteria present in the samples were identifiable with the methods employed in this study.
- 20 bacterial species were isolated from semen samples of the five stallions from the first stud and 11 species from semen samples of six stallions from the second stud.
- The number of bacteria in colony-forming units per mL varied significantly among samples.
Conclusions
- The research concluded that the bacterial culture followed by MALDI-TOF technique does not identify every bacteria present in stallion semen samples.
- The study emphasized that in-depth knowledge of the potential microbes present is crucial in understanding their effects on sperm quality and effectively designing protocols for controlling microbial growth where necessary.
Cite This Article
APA
Al-Kass Z, Eriksson E, Bagge E, Wallgren M, Morrell JM.
(2020).
Microbiota of semen from stallions in Sweden identified by MALDI-TOF.
Vet Anim Sci, 10, 100143.
https://doi.org/10.1016/j.vas.2020.100143 Publication
Researcher Affiliations
- Department of Clinical Sciences, Swedish University of Agricultural Sciences, Box 7054, SE-75007 Uppsala, Sweden.
- Department of Surgery and Theriogenology, College of Veterinary Medicine, university of Mosul, Mosul, Iraq.
- Department of Bacteriology, National Veterinary Institute SE-751 89, Uppsala, Sweden.
- Department of Bacteriology, National Veterinary Institute SE-751 89, Uppsala, Sweden.
- Department of Clinical Sciences, Swedish University of Agricultural Sciences, Box 7054, SE-75007 Uppsala, Sweden.
- Department of Clinical Sciences, Swedish University of Agricultural Sciences, Box 7054, SE-75007 Uppsala, Sweden.
Conflict of Interest Statement
The authors do not have any conflict of interest to declare.
References
This article includes 38 references
- https://en.wikipedia.org/wiki/Fusobacterium.
- https://microbewiki.kenyon.edu/index.php/Psychrobacter; accessed 25 September (2020).
- https://en.wikipedia.org/wiki/Neisseria, accessed 25 September (2020).
- 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.
- Al-Kass Z, Spergser J, Aurich C, Kuhl J, Schmidt K, Morrell JM. Effect of presence or absence of antibiotics and use of modified single layer centrifugation on bacteria in pony stallion semen.. Reprod Domest Anim 2019 Feb;54(2):342-349.
- Althouse G.C., Skaife J., Loomis P.. Prevalence and types of contaminant bacteria in extended, chilled equine semen.. Anim. Reprod. Sci. 2010;121S:224–225.
- Aurich C, Spergser J. Influence of bacteria and gentamicin on cooled-stored stallion spermatozoa.. Theriogenology 2007 Mar 15;67(5):912-8.
- Ben-Amor K, Heilig H, Smidt H, Vaughan EE, Abee T, de Vos WM. Genetic diversity of viable, injured, and dead fecal bacteria assessed by fluorescence-activated cell sorting and 16S rRNA gene analysis.. Appl Environ Microbiol 2005 Aug;71(8):4679-89.
- Biswas S, Rolain JM. Use of MALDI-TOF mass spectrometry for identification of bacteria that are difficult to culture.. J Microbiol Methods 2013 Jan;92(1):14-24.
- Bizzini A, Greub G. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry, a revolution in clinical microbial identification.. Clin Microbiol Infect 2010 Nov;16(11):1614-9.
- Blanchard TL, Kenney RM, Timoney PJ. Venereal disease.. Vet Clin North Am Equine Pract 1992 Apr;8(1):191-203.
- Braga RM, Dourado MN, Araújo WL. Microbial interactions: ecology in a molecular perspective.. Braz J Microbiol 2016 Dec;47 Suppl 1(Suppl 1):86-98.
- Carter G.R.. Isolation and Identification of bacteria.. In: Carter G.R., Cole J.R. Jr, editors. Diagnostic procedure in veterinary bacteriology and mycology. Academic Press, 5th Edition, Harcourt Brace Jovanovich, Publishers; San Diego: 1990. pp. 19–39.
- Christoffersen M, Troedsson M. Inflammation and fertility in the mare.. Reprod Domest Anim 2017 Aug;52 Suppl 3:14-20.
- Corona A, Cherchi R. Microbial quality of equine frozen semen.. Anim Reprod Sci 2009 Oct;115(1-4):103-9.
- Corona A., Cossu I., Bertulu A., Cherchi R.. Characterisation of bacteria in fresh semen of stallions during the breeding season.. Anim. Reprod. Sci. 2006;94:85–88.
- Croxatto A, Prod'hom G, Greub G. Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology.. FEMS Microbiol Rev 2012 Mar;36(2):380-407.
- Gunasekera TS, Attfield PV, Veal DA. A flow cytometry method for rapid detection and enumeration of total bacteria in milk.. Appl Environ Microbiol 2000 Mar;66(3):1228-32.
- Harrigan W.. Laboratory methods in food microbiology.. 3rd ed. Academic Press, San Diego; California, USA: 1998. Cultivation of microorganisms; pp. 43–70.
- Heitmann J, Kirchhoff H, Petzoldt K, Sonnenschein B. Isolation of acholeplasmas and mycoplasmas from aborted horse fetuses.. Vet Rec 1979 Apr 14;104(15):350.
- Kirchhoff H, Heitmann J, Bisping W. [Mycoplasmas isolated from the genital tract of mares (author's transl)].. Zentralbl Bakteriol A 1980 Mar;246(2):228-35.
- Kuczynski J, Lauber CL, Walters WA, Parfrey LW, Clemente JC, Gevers D, Knight R. Experimental and analytical tools for studying the human microbiome.. Nat Rev Genet 2011 Dec 16;13(1):47-58.
- Morrell JM, Wallgren M. Alternatives to antibiotics in semen extenders: a review.. Pathogens 2014 Dec 15;3(4):934-46.
- Morrell JM, Klein C, Lundeheim N, Erol E, Troedsson MH. Removal of bacteria from stallion semen by colloid centrifugation.. Anim Reprod Sci 2014 Feb;145(1-2):47-53.
- Neto R.C., Silva Y., Resende H.L., Guasti P.N., Monteiro G.A., Papa P.M.. Control methods and evaluation of bacterial growth on fresh and cooled stallion semen.. J. Eq. Vet. Sci. 2015;35:277–282.
- Niehaus L, Boland I, Liu M, Chen K, Fu D, Henckel C, Chaung K, Miranda SE, Dyckman S, Crum M, Dedrick S, Shou W, Momeni B. Microbial coexistence through chemical-mediated interactions.. Nat Commun 2019 May 3;10(1):2052.
- Ortega-Ferrusola C, González-Fernández L, Muriel A, Macías-García B, Rodríguez-Martínez H, Tapia JA, Alonso JM, Peña FJ. Does the microbial flora in the ejaculate affect the freezeability of stallion sperm?. Reprod Domest Anim 2009 Jun;44(3):518-22.
- Parlevliet J.M., Samper J.C.. Equine breeding management and artificial insemination.. W.B. Saunders Co.; Philadelphia: 2000. Disease transmission through semen; pp. 133–140.
- Pasing SS, Aurich C, von Lewinski M, Wulf M, Krüger M, Aurich JE. Development of the genital microflora in stallions used for artificial insemination throughout the breeding season.. Anim Reprod Sci 2013 Jun;139(1-4):53-61.
- Patel JB. 16S rRNA gene sequencing for bacterial pathogen identification in the clinical laboratory.. Mol Diagn 2001 Dec;6(4):313-21.
- Roussau R., Kersters K., Falsen E., Jantzen E., Segers P., Union J.A., d.e Ley J. O.. Oligella, a new genus including Oligella urethralis comb. nov. (formerly Moraxella urethralis) and Oligella ureolytica sp. nov. (formerly CDC Group IVe: relationship to Taylorella equigenitalis and related taxa.. Int. J. System. Bact. 1987;37:198–210.
- Samper JC, Tibary A. Disease transmission in horses.. Theriogenology 2006 Aug;66(3):551-9.
- Seng P, Rolain JM, Fournier PE, La Scola B, Drancourt M, Raoult D. MALDI-TOF-mass spectrometry applications in clinical microbiology.. Future Microbiol 2010 Nov;5(11):1733-54.
- Speck ML, Ray B, Read RB Jr. Repair and enumeration of injured coliforms by a plating procedure.. Appl Microbiol 1975 Apr;29(4):549-50.
- Varner DD, Scanlan CM, Thompson JA, Brumbaugh GW, Blanchard TL, Carlton CM, Johnson L. Bacteriology of preserved stallion semen and antibiotics in semen extenders.. Theriogenology 1998 Sep;50(4):559-73.
- vetbact website https://www.vetbact.org/?startpage=1, accessed 25 September (2020).
- Wu L., Wang S., Song Y., Wang X., Yan X.. Applications and challenges for single-bacteria analysis by flow cytometry (Review). Biol. Sens. Imag. 2016;59(1):30–39.
- Xu J.. Next generation sequencing: Current technologies and application. 2014; pp. 419–420. Ontario, CanadaISBN: 978-1-908230-95-9.
Citations
This article has been cited 8 times.- Tvrdá E, Petrovičová M, Ďuračka M, Benko F, Slanina T, Galovičová L, Kačániová M. Short-Term Storage of Rooster Ejaculates: Sperm Quality and Bacterial Profile Differences in Selected Commercial Extenders. Antibiotics (Basel) 2023 Aug 3;12(8).
- Luecke SM, Webb EM, Dahlen CR, Reynolds LP, Amat S. Seminal and vagino-uterine microbiome and their individual and interactive effects on cattle fertility. Front Microbiol 2022;13:1029128.
- Morrell JM, Rocha A. A Novel Approach to Minimising Acute Equine Endometritis That May Help to Prevent the Development of the Chronic State. Front Vet Sci 2021;8:799619.
- Cojkic A, Niazi A, Guo Y, Hallap T, Padrik P, Morrell JM. Identification of Bull Semen Microbiome by 16S Sequencing and Possible Relationships with Fertility. Microorganisms 2021 Nov 25;9(12).
- Medo J, Žiarovská J, Ďuračka M, Tvrdá E, Baňas Š, Gábor M, Kyseľ M, Kačániová M. Core Microbiome of Slovak Holstein Friesian Breeding Bulls' Semen. Animals (Basel) 2021 Nov 22;11(11).
- Navarrete-López P, Asselstine V, Maroto M, Lombó M, Cánovas Á, Gutiérrez-Adán A. RNA Sequencing of Sperm from Healthy Cattle and Horses Reveals the Presence of a Large Bacterial Population. Curr Issues Mol Biol 2024 Sep 19;46(9):10430-10443.
- Kilama J, Dahlen CR, Reynolds LP, Amat S. Contribution of the seminal microbiome to paternal programming. Biol Reprod 2024 Aug 15;111(2):242-268.
- 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).
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