Microbial Load Reduction and Sperm Quality Improvement Through Modified Gradient-Based Protocols
Keywords:
Bacteriospermia, bacterial clearance, assisted reproductive technology, sperm motility, sperm DNA fragmentationAbstract
Infertility affects 15% of couples worldwide, with male infertility accounting for half of cases. Bacteriospermia, involving bacteria in semen, can compromise sperm quality and fertilization. In assisted reproductive technologies (ART), effective sperm preparation methods are crucial to improve sperm quality and reduce bacterial contamination. This research evaluated sperm preparation methods for bacterial clearance. Ninety semen samples were categorized into four groups: density-gradient centrifugation method (DGCM1), swim-up technique (SUT2), combination of density-gradient and swim-up technique (DGCM1SUT3), and unprocessed raw semen (UPRS4) as control. Bacterial cultures were grown on MacConkey Agar and Blood Agar, with Colony-Forming Unit (CFU) counting, biochemical identification, and antimicrobial testing. Initially, 22.5% (25/90) of samples were bacteria-free. Post-preparation, bacterial clearance rates were 19.8% for DGCM1, 10.8% for SUT2, and 27.9% for DGCM1SUT3, showing the combined method's effectiveness. Among 37 bacterial strains identified, Staphylococcus sp. and Streptococcus sp. were most common. In DGCM1, 24.3% of staphylococci and 71.2% of streptococci remained, while SUT2 showed 14.3% and 25.1% persistence. DGCM1SUT3 eliminated Staphylococcus sp. and reduced Streptococcal presence to 1.3%, proving most effective. Sperm DNA fragmentation analysis showed lower fragmentation index for DGCM1SUT3, indicating better sperm integrity. These results highlight Bacteriospermia's impact on fertility and need for optimized sperm processing in ART. The combined method demonstrated highest bacterial clearance and DNA integrity preservation, making it preferred in fertility clinics. Future studies should examine bacterial load, sperm DNA fragmentation, ART success rates, and explore targeted antibiotic treatments for severe Bacteriospermia
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References
Agarwal A, Baskaran S, Parekh N, Cho CL, Henkel R, Vij S, Arafa M. Male infertility. Lancet. 2021;397(10271):319–33. https://doi.org/10.1016/S0140-6736(21)00347-4
Mendz GL, Kaakoush NO, Quinlivan JA. Bacteria and male infertility. J Assist Reprod Genet. 2022;39(2):321–37. https://doi.org/10.1007/s10815-021-02277-1
Sanocka-Maciejewska D, Ciupińska M, Kurpisz M. Bacterial infections and male infertility: Mechanisms and treatments. Reprod Biol. 2020;20(3):391–400. https://doi.org/10.1016/j.repbio.2020.06.001
Weng SL, Chiu CM, Lin FM, Huang HD, Lu JJ, Chang TH, et al. Bacteriospermia and male infertility: Current insights. Andrology. 2020;8(5):1106–17. https://doi.org/10.1111/andr.12838
Esteves SC, Roque M, Bedoschi G, Haahr T, Humaidan P. Sperm selection methods and assisted reproduction outcomes: A systematic review and meta-analysis. Hum Reprod Update. 2022;28(2):135–56. https://doi.org/10.1093/humupd/dmab045
Bibi R, Jahan S, Afsar T, Almajwal A, Hammadeh ME, Amor H, et al. Analyzing the differential impact of semen preparation methods on the outcomes of assisted reproductive techniques. Biomedicines. 2023;11(2):467. https://doi.org/10.3390/biomedicines11020467
Montjean D, Belloc S, Benkhalifa M. Bacterial contamination and sperm quality: Effects and ART implications. Fertil Steril. 2021;115(2):283–95. https://doi.org/10.1016/j.fertnstert.2020.11.010
Yamanaka M, Tomita K, Hashimoto S, Matsumoto H, Satoh M, Kato H, et al. Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms. J Reprod Dev. 2016;62(6):599–606. https://doi.org/10.1262/jrd.2016-112
Ali AH, Ajina T, Ali MB, et al. Efficacy of density gradient centrifugation technique (DGC) in enhancing sperm cell DNA quality for assisted reproductive technique. Middle East Fertil Soc J. 2022;27:22. https://doi.org/10.1186/s43043-022-00108-4
González-Marín C, Gosálvez J, Roy R. Types, causes, detection, and repair of sperm DNA fragmentation. Reprod Biol Endocrinol. 2020;18(1):95. https://doi.org/10.1186/s12958-020-00652-6
Aitken RJ. Reactive oxygen species and sperm DNA damage. Reproduction. 2021;161(2):F63–70. https://doi.org/10.1530/REP-20-0380
Barbieri C, Montjean D, Belloc S. DNA fragmentation testing and ART outcomes: A review. Hum Reprod Update. 2022;28(1):1–15. https://doi.org/10.1093/humupd/dmab028
Kilama J, Dahlen CR, Reynolds LP, Amat S. Contribution of the seminal microbiome to paternal programming. Biol Reprod. 2024;111(2):242–68. https://doi.org/10.1093/biolre/ioae068
Moretti E, Capitani S, Figura N, Pammolli A, Federico MG, Giannerini V, et al. The presence of bacteria species in semen and sperm quality. J Assist Reprod Genet. 2009;26(1):47–56. https://doi.org/10.1007/s10815-008-9283-5
Eini F, Kutenaei MA, Zareei F, et al. Effect of bacterial infection on sperm quality and DNA fragmentation in subfertile men with leukocytospermia. BMC Mol Cell Biol. 2021;22:42. https://doi.org/10.1186/s12860-021-00380-8
Fraczek M, Kurpisz M. Mechanisms of the harmful effects of bacterial semen infection on ejaculated human spermatozoa: potential inflammatory markers in semen. Folia Histochem Cytobiol. 2015;53(3):201–17. https://doi.org/10.5603/fhc.a2015.0019
Tvrdá E, Ďuračka M, Benko F, Lukáč N. Bacteriospermia - A formidable player in male subfertility. Open Life Sci. 2022;17(1):1001–29. https://doi.org/10.1515/biol-2022-0097
Paribok J, Shi X, Dai Y, Zhang Y, Sun X, Chen D, et al. The role of microbiota in male infertility: A systematic review. Asian J Androl. 2022;24(6):605–13.
Monteiro C, Marques PI, Cavadas B, Damião I, Almeida V, Barros N, et al. Characterization of microbiota in male infertility: Impact of urogenital tract infections and semen quality. J Reprod Immunol. 2018;128:16–22.
Santi D, De Vincentis S, Magnani E, Spaggiari G. Impairment of sperm DNA methylation in male infertility: A meta-analytic study. Andrology. 2018;6(6):845–52.
Weng SL, Chiu CM, Lin FM, Huang WC, Liang C, Yang T, et al. Bacterial communities in semen from men of infertile couples: Metagenomic sequencing reveals relationships of seminal microbiota to semen quality. PLoS One. 2014;9(10):e110152. https://doi.org/10.1371/journal.pone.0110152
Kiessling AA, Desmarais B, Yin HZ, Loverde J, Eyre RC. Detection and identification of bacterial DNA in semen. Fertil Steril. 2008;90(5):1744–56. https://doi.org/10.1016/j.fertnstert.2007.09.046
Whelan S, Lucey B, Finn K. Uropathogenic Escherichia coli (UPEC)-Associated Urinary Tract Infections: The Molecular Basis for Challenges to Effective Treatment. Microorganisms. 2023;11(9):2169. https://doi.org/10.3390/microorganisms11092169
Sanocka-Maciejewska D, Ciupińska M, Kurpisz M, Sanocka D. Bacterial infection and oxidative stress in male infertility. J Reprod Immunol. 2013;100(1):76–82.
Ricci G, Perticarari S, Boscolo R, Montico M, Guaschino S, Presani G. Semen preparation methods and sperm apoptosis: Swim-up versus gradient-density centrifugation technique. Fertil Steril. 2009;91(2):632-638.
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