Leukocyte profiles in bull semen and their relationship with sperm quality parameters
DOI:
https://doi.org/10.33448/rsd-v14i2.48156Keywords:
Bovine; Post-thawing; Leukocytospermia; Neutrophils; Sperm-defect.Abstract
The role of leukocytes in semen remains controversial, with both beneficial effects, such as the removal of microorganisms and abnormal sperm, and harmful effects, including reactive oxygen species (ROS) production and reduced fertility potential. Research on this topic in farm animals is limited. This study investigated correlations between differential leukocyte counts in bull semen and sperm parameters, both fresh and post-thaw. A total of 44 ejaculates from 36 adult bulls were collected using the artificial vagina method. Semen cytology was performed with Diff-Quick staining, and leukocytes were quantified under an optical microscope. Sperm parameters analyzed included ejaculate volume, motility (fresh and post-thaw), concentration, and morphology (major, minor, and total defects). Lymphocytes predominated (82%), followed by macrophages (13%), neutrophils (4%), and eosinophils (1%). Neutrophils showed weak negative correlations with major defects (r = -0.36, P = 0.027) and total defects (r = -0.328, P = 0.047), as well as a moderate negative correlation with post-thaw motility (r = -0.515, P = 0.0012) in Spearman’s correlation analysis. In contrast, lymphocytes, macrophages, and eosinophils showed no significant correlations with sperm parameters. These findings suggest that among leukocyte populations, only neutrophils negatively impact sperm morphology in fresh semen and motility in thawed semen.
References
Aitken, R. J., Buckingham, D. W., Brindle, J., Gomez, E., Baker, H. G., & Irvine, D. S. (1995). Andrology: Analysis of sperm movement in relation to the oxidative stress created by leukocytes in washed sperm preparations and seminal plasma. Human Reproduction, 10(8), 2061-2071.
Aitken, R. J., Drevet, J. R., Moazamian, A., & Gharagozloo, P. (2022). Male infertility and oxidative stress: a focus on the underlying mechanisms. Antioxidants, 11(2), 306.
De Bellabarba, G. A., Tortolero, I., Villarroel, V., Molina, C. Z., Bellabarba, C., & Velazquez, E. (2000). Nonsperm cells in human semen and their relationship with semen parameters. Archives of andrology, 45(3), 131-136.
Aziz, N., Agarwal, A., Lewis-Jones, I., Sharma, R. K., & Thomas Jr, A. J. (2004). Novel associations between specific sperm morphological defects and leukocytospermia. Fertility and Sterility, 82(3), 621-627.
Arjadi, F., Indriani, V., Siswandari, W., Priyanto, E., & Pangestu, M. (2022). Semen Leucocytes Affect Sperm Quality of Infertility Patient. Medical Laboratory Technology Journal, 8(2), 159-167.
Baumber, J. V. O. A. et al. Generation of reactive oxygen species by equine neutrophils and their effect on motility of equine spermatozoa. Theriogenology, v. 57, n. 3, p. 1025-1033, 2002.
Blom, E. (1973). The ultrastructure of some characteristic sperm defects and a proposal for a new classification of the bull spermiogram.
Castellini, C., D'andrea, S., Martorella, A., Minaldi, E., Necozione, S., Francavilla, F., ... & Barbonetti, A. (2020). Relationship between leukocytospermia, reproductive potential after assisted reproductive technology, and sperm parameters: a systematic review and meta‐analysis of case–control studies. Andrology, 8(1), 125-135.
Chenoweth, P. J., & McPherson, F. J. (2016). Bull breeding soundness, semen evaluation and cattle productivity. Animal reproduction science, 169, 32-36.
Doria-Filho, U. Introdução à bioestatística: para simples mortais. (2.ed.). Editora Negócio, 1999. p.152.
Enciso, M., Cisale, H., Johnston, S. D., Sarasa, J., Fernández, J. L., & Gosálvez, J. (2011). Major morphological sperm abnormalities in the bull are related to sperm DNA damage. Theriogenology, 76(1), 23-32.
Ferrer, M. S., Hurley, D. J., Norton, N., & Ellerbrock, R. E. (2023). Analytical validation of five diagnostic tests for the detection of polymorphonuclear cells in stallion semen. Journal of Equine Veterinary Science, 122, 104185.
Ferrer, M. S., Palomares, R., Heins, B., Xavier, P., Fyke, H., Hurley, D. J., & Gordon, J. (2024). Analytical and clinical validation of diagnostic tests for the detection of leucospermia in beef bulls. Theriogenology, 230, 46-53.
Gürler, H., Malama, E., Heppelmann, M., Calisici, O., Leiding, C., Kastelic, J. P., & Bollwein, H. (2016). Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology, 86(2), 562-571.HENKEL, R. et al. Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients. Fertil Steril, v.83, n.3, p.635-42. Mar, 2005.
Kiessling, A. A., Lamparelli, N., Yin, H. Z., Seibel, M. M., & Eyre, R. C. (1995). Semen leukocytes: friends or foes?. Fertility and Sterility, 64(1), 196-198.
Kortebani, G., Gonzales, G. F., Barrera, C., & Mazzolli, A. B. (1992). Leucocyte populations in semen and male accessory gland function: relationship with antisperm antibodies and seminal quality. Andrologia, 24(4), 197-204.
Kovalski, N. N., de Lamirande, E., & Gagnon, C. (1992). Reactive oxygen species generated by human neutrophils inhibit sperm motility: protective effect of seminal plasma and scavengers. Fertility and sterility, 58(4), 809-816.
Kumar, A., & Sridharan, T. B. (2020). Oxidative stress-induced changes in fertility status of cryopreserved sperm: a diagnosis based on sperm chromatin dispersion assay. Cryoletters, 41(5), 297-302.
Li, X., Ni, M., Xing, S., Yu, Y., Zhou, Y., Yang, S., ... & Han, M. (2020). Reactive oxygen species secreted by leukocytes in semen induce self-expression of interleukin-6 and affect sperm quality. American Journal of Men's Health, 14(5), 1557988320970053.
Menegassi, S. R. O., Barcellos, J. O. J., Lampert, V. D. N., Borges, J. B. S., & Peripolli, V. (2011). Bioeconomic impact of bull breeding soundness examination in cow-calf systems. Revista Brasileira de Zootecnia, 40, 441-447.
Menkveld, R. (2004, April). Leukocytospermia. In International Congress Series (Vol. 1266, pp. 218-224). Elsevier.
Al Makhzoomi, A. K., Lundeheim, N., Håård, M., & Rodríguez-Martínez, H. (2008). Sperm morphology and fertility of progeny-tested AI dairy bulls in Sweden. Theriogenology, 70(4), 682-691.
Mupfiga, C., Fisher, D., Kruger, T., & Henkel, R. (2013). The relationship between seminal leukocytes, oxidative status in the ejaculate, and apoptotic markers in human spermatozoa. Systems Biology in Reproductive Medicine, 59(6), 304-311.
Omu, A. E., Al-Qattan, F., Al-Abdul-Hadi, F. M., Fatinikun, M. T., & Fernandes, S. (1999). Seminal immune response in infertile men with leukocytospermia: effect on antioxidant activity. European Journal of Obstetrics & Gynecology and Reproductive Biology, 86(2), 195-202.
Pereira A. S. et al. (2018). Metodologia da pesquisa científica. [free e-book]. Santa Maria/RS. Ed. UAB/NTE/UFSM.
Plante, M., de Lamirande, E., & Gagnon, C. (1994). Reactive oxygen species released by activated neutrophils, but not by deficient spermatozoa, are sufficient to affect normal sperm motility. Fertility and sterility, 62(2), 387-393.
Saleh, R. A., Agarwal, A., Kandirali, E., Sharma, R. K., Thomas Jr, A. J., Nada, E. A., ... & Alvarez, J. G. (2002). Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa. Fertility and sterility, 78(6), 1215-1224.
Silva, M. K. F., Borges, M. S. A., de Paula, R. R., de Souza, L. F. C., Afonso, F. A., Alves, L. A., ... & Crespilho, A. M. (2022). Comparação de diferentes métodos para avaliação da concentração espermática bovina: existem opções a serem comparadas ao NucleoCounter®? Comparison of different methods to access bull sperm concentration: are there options to be compared to NucleoCounter®?. Brazilian Journal of Development, 8(6), 47934-47944.
Shitsuka, R. et al. (2014). Matemática fundamental para tecnologia. 2ed. Editora Erica.
Söderquist, L., Janson, L., Larsson, K., & Einarsson, S. (1991). Sperm morphology and fertility in AI bulls. Journal of Veterinary Medicine Series A, 38(1‐10), 534-543.
Sprecher, D. J., Coe, P. H., & Walker, R. D. (1999). Relationships among seminal culture, seminal white blood cells, and the percentage of primary sperm abnormalities in bulls evaluated prior to the breeding season. Theriogenology, 51(6), 1197-1206.
Stanislavov, R. (1999). Leukocytes in human seminal fluid. Akusherstvo i Ginekologiia, 38(3), 20-21.
Thomas, J., Fishel, S. B., Hall, J. A., Green, S., Newton, T. A., & Thornton, S. J. (1997). Increased polymorphonuclear granulocytes in seminal plasma in relation to sperm morphology. Human reproduction, 12(11), 2418-2421.
Tomlinson, M. J., White, A., Barratt, C. L. R., Bolton, A. E., & Cooke, I. D. (1992). The removal of morphologically abnormal sperm forms by phagocytes: a positive role for seminal leukocytes?. Human Reproduction, 7(4), 517-522.
Tortolero, I., JM, D. O., Arata-Bellabarba, G., & Regadera, J. (2004). The effect of seminal leukocytes on semen quality in subfertile males with and without varicocele. Archivos Espanoles De Urologia, 57(9), 921-928.
Tremellen, K. (2008). Oxidative stress and male infertility—a clinical perspective. Human reproduction update, 14(3), 243-258.
Vieira, S. (2021). Introdução à bioestatística. Editora GEN/Guanabara Koogan.
Wang, A. W., Zhang, H., Ikemoto, I., Anderson, D. J., & Loughlin, K. R. (1997). Reactive oxygen species generation by seminal cells during cryopreservation. Urology, 49(6), 921-925.
World Health Organisation. (1999). WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. Cambridge university press.
Zart, A. L., Jurgielewicz, V. C. L., & Fernandes, C. E. (2014). Seminal leucocytary profile in beef bulls. Reproduction in Domestic Animals, 49(5), 719-724.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Yasmin El Hayek-Martins; Mariana Karla Francolino da Silva; Paulina Betancur Guerra; Fabiana Ferreira de Souza; André Maciel Crespilho
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
1) Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
2) Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
3) Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.
