Inflammatory Biomarkers and Sperm Dysfunction in ‎Bacterial Semen Infections

  • Authors

    • Satyanarayan Samantaray School of Paramedics and Allied Health Sciences; Centurion University of Technology and ‎Management; Odisha
    • Soumya Jal School of Paramedics and Allied Health Sciences; Centurion University of Technology and ‎Management; Odisha
    • Gopal Krishna Purohit Heredity Biosciences, Plot No:818, Mayfair Lagoon Road, Jayadev Vihar, Bhubaneswar, ‎Odisha
    • Soumya Dash Fertility specialist, Ankura Medical and research Center, Near Sisubhavan Bus Stop, Bapuji ‎ Nagar, Bhubaneswar, Odisha 751009
    https://doi.org/10.14419/qwr5b132

    Received date: July 21, 2025

    Accepted date: September 2, 2025

    Published date: November 4, 2025

  • Male Infertility; Semen Microbiota; Inflammatory Biomarkers; 16S rRNA; PCR; Cytokine ‎Gene Expression
  • Abstract

    Male infertility is a global concern because bacterial infections of the reproductive tract play a ‎significant role. These infections trigger inflammatory responses that impair sperm quality. ‎This study explored the relationship between bacterial semen infection, inflammatory ‎biomarkers, and sperm dysfunction using PCR and quantitative real-time PCR (qPCR). ‎Semen samples were collected from 60 male participants attending the infertility clinics. ‎Semen analysis was performed according to the WHO 2021 guidelines to assess ‎concentration, motility, morphology, and viability. Bacterial DNA was isolated from the ‎samples and screened by PCR targeting 16S rRNA genes to detect common seminal ‎pathogens. The expression levels of inflammatory cytokine genes (IL-6, TNF-α, and IL-1β) ‎were quantified using SYBR Green-based qPCR. Gene expression was normalized to that of ‎GAPDH, which was used as a housekeeping gene. Correlations between bacterial presence, ‎cytokine expression, and sperm parameters were analyzed. PCR revealed bacterial DNA in ‎‎40% of the semen samples, with Escherichia coli and Staphylococcus aureus being ‎predominant. Samples positive for bacterial DNA showed significantly higher expression of ‎IL-6, TNF-α, and IL-1β (P < 0.01), indicating an inflammatory response. These elevated ‎cytokine levels correlated with decreased sperm motility, viability, and morphological ‎integrity (r > -0.6, p < 0.05). The study showed that bacterial infections in semen are ‎associated with the upregulation of inflammatory cytokines, contributing to sperm ‎dysfunction. PCR and qPCR effectively detect the presence of bacteria and quantify ‎inflammation at the molecular level. These findings underscore the need for molecular ‎diagnostics for male infertility evaluation and highlight inflammatory biomarkers as potential ‎therapeutic targets‎.

  • References

    1. Agarwal, A., Mulgund, A., Hamada, A., & Chyatte, M. R. (2015). A unique view on male infertility around the globe. Reproductive Biology and Endocrinology, 13, 37. https://doi.org/10.1186/s12958-015-0032-1.
    2. Acharya, P. P., Jal, S., Purohit, G. K., & Pati, P. K. (2025). Molecular identification of uropathogenic bacteria via 16S rRNA gene amplification and sequencing. Journal of Neonatal Surgery, 14(32s), 467–475.
    3. Aitken, R. J., & Baker, M. A. (2004). Oxidative stress and male reproductive biology. Reproduction, Fertility and Development, 16(5), 581–588. https://doi.org/10.1071/RD03089.
    4. Akinloye, O., Azenabor, A., & Ekun, A. O. (2015). Impact of inflammation on male reproductive tract. Journal of Reproduction & Infertility, 16(3), 123–129.
    5. Azenabor, A., Ekun, A. O., & Akinloye, O. (2015). Impact of inflammation on male reproductive tract. Journal of Reproduction & Infertility, 16(3), 123–129. PMID: 26913230; PMCID: PMC4508350.
    6. Babakhanzadeh, E., Ghasemifar, S., Nazari, M., & Khodadadian, A. (2020). Some of the factors involved in male infertility: A prospective review. International Journal of General Medicine, 13(8), 29–41. https://doi.org/10.2147/IJGM.S241099.
    7. Comhaire, F. H., Mahmoud, A., Depuydt, C. E., & Zalata, A. A. (1999). Mechanisms of the spermotoxic effect of leukocytes in semen. Fertility and Sterility, 72(3), 816–820.
    8. Cooper, T. G., Noonan, E., von Eckardstein, S., et al. (2010). World Health Organization reference values for human semen characteristics. Human Reproduction Update, 16(3), 231–245. https://doi.org/10.1093/humupd/dmp048.
    9. Dutta, S., & Sengupta, P. (2025). Bacterial infections and male fertility. In Bentham Science (pp. 122–138). https://doi.org/10.2174/9789815305302125010009.
    10. Fraczek, M., & Kurpisz, M. (2007). Inflammatory mediators exert toxic effects of reactive oxygen species on human spermatozoa. Journal of An-drology, 28(2), 325–333. https://doi.org/10.2164/jandrol.106.001149.
    11. Hedger, M. P. (2011). Immunophysiology and pathology of inflammation in the testis and epididymis. Journal of Andrology, 32(6), 625–640. https://doi.org/10.2164/jandrol.111.012989.
    12. Hirano, T. (2021). IL-6 in inflammation, autoimmunity and cancer. International Immunology, 33(3), 127–148. https://doi.org/10.1093/intimm/dxaa078.
    13. Huleihel, M., & Lunenfeld, E. (2004). Regulation of spermatogenesis by cytokines. International Journal of Andrology, 27(4), 198–199.
    14. Kaltsas, A., Zachariou, A., Markou, E., Dimitriadis, F., Sofikitis, N., & Pournaras, S. (2023). Microbial dysbiosis and male infertility: Understand-ing the impact and exploring therapeutic interventions. Journal of Personalized Medicine, 13(10), 1491. https://doi.org/10.3390/jpm13101491.
    15. Khan, M. S., Ullah, A., & Mehmood, A. (2024). Male infertility: Its diagnosis and management. BMC Journal of Medical Sciences, 5(1), 1–2. https://doi.org/10.70905/bmcj.05.01.0283.
    16. Khodamoradi, K., Kuchakulla, M., Narasimman, M., Khosravizadeh, Z., Ali, A., Brackett, N., Ibrahim, E., & Ramasamy, R. (2020). Laboratory and clinical management of leukocytospermia and hematospermia: A review. Therapeutic Advances in Reproductive Health, 14(Suppl 4), 263349412092251. https://doi.org/10.1177/2633494120922511.
    17. La Vignera, S., Vicari, E., Condorelli, R., D’Agata, R., & Calogero, A. E. (2011). Male accessory gland infection and sperm parameters. Interna-tional Journal of Andrology, 34(5 Pt 2), e330–e347. https://doi.org/10.1111/j.1365-2605.2011.01200.x.
    18. Loveland, K. L., Klein, B., Pueschl, D., Indumathy, S., Bergmann, M., Loveland, B. E., Hedger, M. P., & Schuppe, H. C. (2017). Cytokines in male fertility and reproductive pathologies: Immunoregulation and beyond. Frontiers in Endocrinology, 8, 307. https://doi.org/10.3389/fendo.2017.00307.
    19. Oghbaei, H., Rastgar Rezaei, Y., Nikanfar, S., Zarezadeh, R., Sadegi, M., Latifi, Z., Nouri, M., Fattahi, A., Ahmadi, Y., & Bleisinger, N. (2020). Effects of bacteria on male fertility: Spermatogenesis and sperm function. Life Sciences, 256, 117891. https://doi.org/10.1016/j.lfs.2020.117891.
    20. Ochsendorf, F. R. (1999). Infections in the male genital tract and reactive oxygen species. Human Reproduction Update, 5(5), 399–420. https://doi.org/10.1093/humupd/5.5.399.
    21. Potiris, A., Moustakli, E., Trismpioti, E., Drakaki, E., Mavrogianni, D., Matsas, A., Zikopoulos, A., Sfakianakis, A., Tsakiridis, I., Dagklis, T., et al. (2025). From inflammation to infertility: How oxidative stress and infections disrupt male reproductive health. Metabolites, 15(4), 267. https://doi.org/10.3390/metabo15040267.
    22. Prabha, V., Sandhu, R., Kaur, S., et al. (2010). Characterization of E. coli isolated from semen samples of infertile males. Indian Journal of Medical Research, 131, 83–87.
    23. Raghuwanshi, P. T., & Kanwal, M. R. (2025). Causes and associated factors with male infertility among couples visiting infertility/andrologist clin-ics. Journal of Drug Delivery and Therapeutics, 15(3), 21–29. https://doi.org/10.22270/jddt.v15i3.7008.
    24. Ricchi, M., Bertasio, C., Boniotti, M. B., Vicari, N., Russo, S., Tilola, M., Bellotti, M. A., & Bertasi, B. (2017). Comparison among the quantifica-tion of bacterial pathogens by qPCR, dPCR, and cultural methods. Frontiers in Microbiology, 8, 1174. https://doi.org/10.3389/fmicb.2017.01174 .
    25. Ricci, G., Perticarari, S., Fragonas, E., et al. (2002). Cytokines and male fertility: IFN-γ and IL-6 in human semen. European Journal of Obstetrics & Gynecology and Reproductive Biology, 100(2), 165–170.
    26. Saing, T., Valdivia, A., Hussain, P., Ly, J., Gonzalez, L., Guilford, F. T., Pearce, D., & Venketaraman, V. (2016). Data on pro-inflammatory cyto-kines IL-1β, IL-17, and IL-6 in the peripheral blood of HIV-infected individuals. Data in Brief, 8(11), 1044–1047. https://doi.org/10.1016/j.dib.2016.07.023.
    27. Said, T. M., & Agarwal, A. (2006). Role of sperm oxidative stress in male infertility: A review. Journal of Andrology, 27(6), 737–748.
    28. Samantaray, S., Jal, S., & Purohit, G. K. (2024). Microbial contaminants in semen and their effects on sperm motility, morphology, and fertility out-comes. Journal of Neonatal Surgery, 53(3). https://doi.org/10.48047/4st0qc93.
    29. Shen, C.-C., Peng, H., & Zhu, X. (2011). Male infertility due to Ureaplasma urealyticum of semen analysis. International Medicine and Health Guidance News, 17(2), 151–154.
    30. Syafrizayanti, S., Nurhayati, N., & Safni, S. (2022). Detection and absolute quantification of porcine DNA in sausages using quantitative polymer-ase chain reaction (qPCR) method. Jurnal Kimia Unand, 11(2), 5–13. https://doi.org/10.25077/jku.11.2.5-13.2022.
    31. Tremellen, K. (2008). Oxidative stress and male infertility—a clinical perspective. Human Reproduction Update, 14(3), 243–258. https://doi.org/10.1093/humupd/dmn004.
    32. Wang, S., Zhang, K., Yao, Y., Li, J., & Deng, S. (2021). Bacterial infections affect male fertility: A focus on the oxidative stress-autophagy axis. Frontiers in Cell and Developmental Biology, 9, 727812. https://doi.org/10.3389/fcell.2021.727812.
    33. Weng, S. L., Chiu, C. M., Lin, F. M., et al. (2014). Bacterial communities in semen from men of infertile couples: Metagenomic sequencing reveals relationships with semen quality. PLoS ONE, 9(10), e110152. https://doi.org/10.1371/journal.pone.0110152.
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  • How to Cite

    Samantaray, S. ., Jal , S. ., Purohit, G. K. . ., & Dash , S. . (2025). Inflammatory Biomarkers and Sperm Dysfunction in ‎Bacterial Semen Infections. International Journal of Basic and Applied Sciences, 14(7), 123-129. https://doi.org/10.14419/qwr5b132