Boğa Spermasında Escherichia coli Kontaminasyonu Sonrası Bazı Spermatolojik Parametrelerin Değerlendirilmesi

Abstract

Bu çalışma, genişlemiş spektrumlu beta laktamaz (ESBL) üreten ve üretmeyen Escherichia coli konsantrasyonlarının boğa sperma motilite parametreleri ve spermatozoa canlılığı üzerindeki etkilerini değerlendirmeyi amaçlamıştır. Aynı Simental boğasından toplam 50 adet donmuş sperma payeti kullanıldı. Tüm sperma payetleri 37°C'de su banyosunda çözdürülerek, her biri 10 sperma örneğinden oluşan 5 gruba ayrıldı. Grup 1 (G1) (n=10), kontamine edilmeyen kontrol grubunu oluşturdu. Grup 2 (G2) (n=10), 100.000 cfu/ml ve Grup 3 (G3) (n=10), 1.000.000 cfu/ml konsantrasyonunda E. coli ATCC 25922 ile, Grup 4 (G4) (n=10), 100.000 cfu/ml konsantrasyonda ve Grup 5 (G5) (n=10), 1.000.000 cfu/ml konsantrasyonda ESBL üreten E. coli BAA-196 ile kontamine edilen grupları oluşturuldu. Çalışmada bu gruplara ait sperma örneklerinde progresif motilite, motilite ve ölü-canlı spermatozoa yüzdelerinin zaman içinde değişimi analiz edildi. Spermatozoa motilitesi ve canlılığında önemli azalmalar gözlemlendi ve en belirgin etkiler, yüksek ESBL üreten E. coli konsantrasyonlarıyla kontamine olan gruplarda (G4 ve G5) görüldü (p<0.05). Bakteriyel kontaminasyonun sperma parametreleri üzerindeki etkisinin doza bağlı olduğu ve daha yüksek konsantrasyonların sperma kalitesinde daha hızlı ve ciddi bozulmaya neden olduğu belirlendi. Bu bulgular, özellikle dirençli suşlarla bakteriyel kontaminasyonun sperma kalitesini azaltmadaki rolünü vurgulamakta ve suni tohumlama uygulamalarında mikrobiyal kontaminasyonun önemine dikkat çekmektedir. Reprodüktif teknolojilerde kontaminasyonu kontrol etmede ve antibiyotik direnciyle mücadele etmeyi sağlamada alternatif yöntemleri keşfetmek için daha fazla araştırmaya ihtiyaç vardır.

Keywords

• Boğa
• Kontaminasyon
• Escherichia coli
• Sperm parametreleri

Evaluation of Some Spermatological Parameters Following Escherichia coli Contamination in Bull Semen

Abstract

This study aimed to evaluate the effects of extended-spectrum beta-lactamase (ESBL) producing and non-producing Escherichia coli concentrations on bull semen motility parameters and spermatozoa viability. A total of 50 frozen semen straws from the same Simmental bull were used. All semen straws were thawed in a water bath at 37°C and divided into 5 groups of 10 semen samples. Group 1 (G1) (n=10) was the uncontaminated control group. Group 2 (G2) (n=10) was contaminated with 100,000 cfu/ml and Group 3 (G3) (n=10) was contaminated with E. coli ATCC 25922 at a concentration of 1,000,000 cfu/ml. Group 4 (G4) (n=10) was contaminated with 100,000 cfu/ml and Group 5 (G5) (n=10) was contaminated with ESBL producing E. coli BAA-196 at a concentration of 1,000,000 cfu/ml. In the study, progressive motility, motility and percentages of dead-live spermatozoa in semen samples of these groups were analyzed over time. Significant decreases in spermatozoa motility and viability were observed and the most significant effects were seen in groups (G4 and G5) contaminated with high concentrations of ESBL-producing E. coli (p<0.05). It was determined that the effect of bacterial contamination on spermatological parameters was dose-dependent, with higher concentrations causing more rapid and severe deterioration in semen quality. These findings highlight the role of bacterial contamination, especially with resistant strains, in reducing semen quality and draw attention to the importance of microbial contamination in artificial insemination practices. Further research is needed to explore alternative methods to control contamination in reproductive technologies and combat antibiotic resistance.

Keywords

• Bull
• Contamination
• Escherichia coli
• Sperm parameters

References

1. Cojkic, A., Niazi, A., Guo, Y., Hallap, T., Padrik, P., & Morrell, J. M. (2021). Identification of bull semen microbiome by 16S sequencing and possible relationships with fertility. Microorganisms, 9(12), 2431.
2. Cottell, E., McMorrow, J., Lennon, B., Fawsy, M., Cafferkey, M., & Harrison, R. F. (1996). Microbial contamination in an in vitro fertilization-embryo transfer system. Fertility and Sterility, 66(5), 776-780.
3. Diemer, T., Huwe, P., Ludwig, M., Schroeder‐Printzen, I., Michelmann, H. W., Schiefer, H. G., & Weidner, W. (2003). Influence of autogenous leucocytes and Escherichia coli on sperm motility parameters in vitro. Andrologia, 35(2), 100-105.
4. Ďuračka, M., Belić, L., Tokárová, K., Žiarovská, J., Kačániová, M., Lukáč, N., & Tvrdá, E. (2021). Bacterial communities in bovine ejaculates and their impact on the semen quality. Systems Biology in Reproductive Medicine, 67(6), 438-449.
5. Folliero, V., Santonastaso, M., Dell’Annunziata, F., De Franciscis, P., Boccia, G., Colacurci, N., De Filippis, A., Galdiero, M., Franci, G. (2022). Impact of Escherichia coli outer membrane vesicles on sperm function. Pathogens, 11(7), 782.
6. Gaczarzewicz, D., Udala, J., Piasecka, M., Blaszczyk, B., & Stankiewicz, T. (2016). Bacterial contamination of boar semen and its relationship to sperm quality preserved in commercial extender containing gentamicin sulfate. Polish Journal of Veterinary Sciences, 19(3), 451-459.
7. Garba, S. I., Abubakar, M. M., Macha, Y. P., Maı, M. H., Garba, I. (2023). Present of bacterial contamination on semen quality of selected Nigerian breeds of rams. International Journal of Science and Applied Research (IJSAR), 6(1), 1-10
8. Khalili, M. A., Pourshafiei, M. R., Saifi, M., & Khalili, M. B. (2000). Bacterial infection of the reproductive tract of infertile men in Iran. Middle East Fertility Society Journal, 5, 126–131.

9. Kuster, C. E., & Althouse, G. C. (2016). The impact of bacteriospermia on boar sperm storage and reproductive performance. Theriogenology, 85(1), 21-26.
10. Marcus, S., Bernstein, M., Ziv, G., Glickman, A., Gipps, M. (1994). Norfloxacin nicotinate in the treatment of Pseudomonas aeruginosa infection in the genital tract of a bull. Veterinary Research Communications, 18(5):331–336.
11. Mayer, F., Weidner, W., Diemer, T., Huwe, P., Michelmann, H. W., & Schiefer, H. G. (2000). Escherichia coli-induced alterations of human spermatozoa. An electron microscopy analysis. International Journal of Andrology, 23(3), 178-186.
12. Mitra, J., Chowdhury, S., Panda, S., Chakraborty, M., Singha, A. (2016). Microbiological evaluation of bovine frozen semen samples in West Bengal, India. Exploratory Animal and Medical Research, 6(2),185–191.
13. Monga, M., & Roberts J. A. (1994). Spermagglutination by bacteria: receptor‐specific interactions. Journal of Andrology, 15(2), 151-156.
14. Moretti, E., Capitani, S., Figura, N., Pammolli, A., Federico, M. G., Giannerini, V., & Collodel, G. (2009). The presence of bacteria species in semen and sperm quality. Journal of Assisted Reproduction and Genetics, 26, 47-56.
15. Oghbaei, H., Rezaei, Y. R., Nikanfar, S., Zarezadeh, R., Sadegi, M., Latifi, Z., & Bleisinger, N. (2020). Effects of bacteria on male fertility: Spermatogenesis and sperm function. Life Sciences, 256, 117891.
16. Peirano, G., & Pitout J. D. (2010). Molecular epidemiology of Escherichia coli producing CTX-M beta-lactamases: the worldwide emergence of clone ST131 O25:H4. International Journal of Antimicrobial Agents, 35, 316-321.
17. R Core Team. (2023). R: A language and environment for statistical computing. R Foundation for Statistical Computing. https://www.R-project.org/
18. Rana, N., Vaid, R. K., Phulia, S. K., & Singh, P. (2012). Assessment of bacterial diversity in fresh bubaline semen. Indian Journal of Animal Sciences, 82(6), 596.
19. Schmid, A., Hörmansdorfer, S., Messelhäusser, U., Käsbohrer, A., Sauter-Louis, C., & Mansfeld, R. (2013). Prevalence of extended-spectrum β-lactamase-producing Escherichia coli on Bavarian dairy and beef cattle farms. Applied and Environmental Microbiology, 79(9), 3027-3032.
20. Sorkytė, Š., Šiugždinienė, R., Virgailis, M., Vaičiulienė, G., Wysokińska, A., Wójcik, E., ... & Sutkevičienė, N. (2024). The interaction between canine semen bacteria and semen quality parameters. Animals, 14(15), 2151.
21. Stringfellow, D. A., & Givens, M. D. (2000). Infectious agents in bovine embryo production: hazards and solutions. Theriogenology, 53(1), 85-94.
22. Thibier M, Guerin B. (2000). Hygienic aspects of storage and use of semen for artificial insemination. Animal Reprodoction Science, 62(1–3):233–251.
23. Wolff, H., Panhans, A., Stolz, W., & Meurer, M. (1993). Adherence of Escherichia coli to sperm: a mannose mediated phenomenon leading to agglutination of sperm and E. coli. Fertility and Sterility, 60(1), 154-158.
24. Yániz, J. L., Marco-Aguado, M. A., Mateos, J. A., & Santolaria, P. (2010). Bacterial contamination of ram semen, antibiotic sensitivities, and effects on sperm quality during storage at 15 C. Animal Reproduction Science, 122(1-2), 142-149.