Detection of The Presence of Leptospirosis in Horses by ELISA Method

Öz

Leptospirosis is one of the common zoonotic diseases worldwide. It significantly affects humans and animals, causing socio-economic losses. Equine leptospirosis often manifests itself with recurrent genital tract infections, reproductive disorders, abortion, embryonic death, and birth of weak foals. Studies on Leptospirosis, especially in horses, are quite limited. For this reason, our study was planned to investigate the transmission cycle in terms of farm animals and public health, and also to determine the presence of Leptospirosis in horses. For our research, 187 horse blood serum samples were collected from different settlements in Turkey. In the collected samples, Leptospira spp. ELISA test was used to investigate the presence of antibodies. A total of 8 samples (4.27%) were detected as seropositive in horse serum samples. The agent, a significant pathogen for both human and animal health, was identified as seropositive. In addition, the high rate of transmission of leptospirosis through contact with infected horses and the potential for spread through different animals should be taken into consideration. Leptospirosis was determined as seropositivity by ELISA in the horses sampled. Therefore, it is important to diagnose and control the infection rapidly.

Anahtar Kelimeler

• ELISA
• Horse
• Leptosipirosis
• Serologic Test
• Zoonoses

Atlarda Leptospirosis Varlığının ELISA Yöntemi ile Tespiti

Öz

Leptospirosis dünya çapında yaygın zoonotik hastalıklardan biridir. İnsanları ve hayvanları önemli ölçüde etkileyerek sosyo-ekonomik kayıplara neden olur. At leptospirozu sıklıkla tekrarlayan genital sistem enfeksiyonları, üreme bozuklukları, abort, embriyonik ölüm ve zayıf tayların doğumu ile kendini gösterir. Özellikle atlarda Leptospirozis üzerine yapılan çalışmalar oldukça kısıtlıdır. Bu sebeple, çiftlik hayvanları ve halk sağlığı açısından bulaşma döngüsünü araştırmak, aynı zamanda atlarda Leptospirozis’in varlığını belirlemek amacıyla çalışmamız planlanmıştır. Araştırmamız için Türkiye’nin farklı yerleşim yerlerinden 187 adet at kan serumu örneği toplandı. Toplanan örneklerde, Leptospira spp. antikorlarının varlığını araştırmak için ELISA testi kullanıldı. At serum örneklerinde toplam 8 örnek (%4,27) seropozitif olarak tespit edildi. İnsan ve hayvan sağlığı için önemli bir patojen olan etken seropozitif olarak teşhis edilmiştir. Ayrıca, leptospirozis hastalığının enfekte atlarla temas yoluyla bulaşma oranının yüksek olması ve farklı hayvanlar ile yayılma potansiyeli göz önüne alınmalıdır. Örnek alınan atlarda Leptospirosis ELISA ile seropoziti olarak belirlenmiştir. Bu nedenle enfeksiyonun hızla teşhis edilerek kontrol altına alınması önem taşımaktadır.

Anahtar Kelimeler

• Serolojik Test
• At
• ELISA
• Leptospirozis
• Zoonozlar

Kaynakça

1. Abbexa. (2024). Horse Leptospira spp. Antibody ELISA Kit. https://www.abbexa.com/horse-leptospira-antibody-lep-ab-elisa-kit
2. Andersen-Ranberg, E. U., Pipper, C., & Jensen, P. M. (2016). Global patterns of Leptospira prevalence in vertebrate reservoir hosts. Journal of wildlife diseases, 52(3), 468-477.
3. Ayçiçek A. (2004). 7 Yaşındaki bir kız çocuğunda Echinococcus multilocularis, Selcuk medical journal, 20,2.
4. Babür, C., Ozdemir, V., Kiliç, S., Erol, E., & Esen, B. (2003). Anti-Leptospira antibodies in slaughterhouse workers in Ankara. Mikrobiyoloji Bulteni, 37(2-3), 143-150.
5. Behera, S. K., Parija, S. C., Sahu, P. S., Khatua, S. P., & Dash, M. (2022). Evaluation of recombinant LigA/B antigen-based IgM Dot ELISA and latex agglutination test for the diagnosis of leptospirosis: A comparative study with the MAT. Diagnostics, 12(6), 1455. https://doi.org/10.3390/diagnostics12061455
6. Bourhy, P., Collet, L., Brisse, S., & Picardeau, M. (2014). Leptospira mayottensis sp. nov., a pathogenic species of the genus Leptospira isolated from humans. International journal of systematic and evolutionary microbiology, 64(12), 4061.
7. Costa, F., Martinez-Silveira, M. S., Hagan, J. E., Hartskeerl, R. A., Reis, M. G. D., & Ko, A. I. (2012). Surveillance for leptospirosis in the Americas, 1996-2005: a review of data from ministries of health. Revista Panamericana de Salud Publica, 32(3), 169-177.
8. Day N. (2021). Leptospirosis: Epidemiology, Microbiology, Clinical Manifestations, and Diagnosis. UpToDate® Home Page, Topic 5527, Version 27.0. Available https://www.uptodate.com/contents/leptospirosis-epidemiology-microbiology-clinical-manifestations-and-diagnosis

9. Divers, T. J., Chang, Y. F., Irby, N. L., Smith, J. L., & Carter, C. N. (2019). Leptospirosis: Kuzey Amerika atlarında önemli bir bulaşıcı hastalık. At Veteriner. J., 51: 287–292. doi: 10.1111/evj.13069.
10. Ellis, W. A. (2014). Animal leptospirosis. Leptospira and leptospirosis, 99-137.
11. Ellis, W. A. (2023). Animal leptospirosis: a review of current knowledge. Veterinary Microbiology, 279, 109602. https://doi.org/10.1016/j.vetmic.2022.109602
12. Goris, M. G. A., Leeflang, M. M. G., Boer, K. R., Goeijenbier, M., Van Gorp, E. C. M., Wagenaar, J. F. P., & Hartskeerl, R. A. (2012). Establishment of Valid Laboratory Case Definition for Human Leptospirosis. J. Bacteriol Parasitol. 3, 132.
13. Haake, D. A., & Levett, P. N. (2015). Leptospirosis in humans. Curr. Top. Microbiol. Immunol. , 387, 65–97. Indicus cattle in Cameroon. PLoS One. 2010;5:e8623. Interrogans Serovar hardjo is not a major cause of bovine abortion in Victoria. Aust Vet J. 1989;66:330–3.
14. Ibrahim, N. A., Alrashdi, B. M., Elnaker, Y. F., Elmahallawy, E. K., Alblihed, M. A., Daib, M. S., ... & Elbaz, E. (2022). Serological investigation and epidemiological analysis of bovine leptospirosis in Egypt. Tropical Medicine and Infectious Disease, 7(9), 208.
15. Kaya M. (2017). The comparison of brucella gel agglutination test with other serological tests for the diagnosis of brucellosis. Selcuk Medical Journal, 34(1), 1-5.
16. Kepenek E. (2021). Üçüncü Basamak Bir Hastanede İzlenen Brusellozis Olgularının Değerlendirilmesi. Selçuk Tıp Dergisi, 37(2), 130-136.
17. Lau, C. L., Smythe, L. D., Craig, S. B., & Weinstein, P. (2010). Climate change, flooding, urbanisation and leptospirosis: fuelling the fire? Transactions of the Royal Society of Tropical Medicine and Hygiene, 104(10), 631-638.
18. Levett P. N. (2001). Leptospirosis. Clin Microbiol Rev. 14:296–326.
19. Lowe R. C. (2010). Equine üveiti: Birleşik Krallık perspektifi. At Veteriner. J. Ek. doi: 10.1111/j.2042-3306.2010.tb05634.x.
20. Mulla, S., Chakraborty, T., Patel, M., Pandya, H. P., Dadhaniya, V., & Vaghela, G. (2006). Diagnosis of leptospirosis and comparison of ELISA and MAT techniques. Indian journal of pathology & microbiology, 49(3), 468-470.
21. Niloofa, R., Fernando, N., de Silva, N. L., Karunanayake, L., Wickramasinghe, H., Dikmadugoda, N., ... & Agampodi, S. B. (2015). Diagnosis of leptospirosis: comparison between microscopic agglutination test, IgM-ELISA and Leptocheck-WB. PLOS ONE, 10(6), e0129236. https://doi.org/10.1371/journal.pone.0129236
22. Pissawong, T., Maneewatchararangsri, S., Ritthisunthorn, N., Soonthornworasiri, N., Reamtong, O., Adisakwattana, P., Kalambaheti, T., Chaisri, U., & Doungchawee, G. (2020). Immunodominance of LipL3293–272 peptides revealed by leptospirosis sera and therapeutic monoclonal antibodies. J. Microbiol. Immunol. Infect, 53, 11–22.
23. Rosa, M. I., Reis, M. F. D., Simon, C., Dondossola, E., Alexandre, M. C., Colonetti, T., & Meller, F. O. (2017). IgM ELISA for leptospirosis diagnosis: a systematic review and meta-analysis. Ciencia & saude coletiva, 22, 4001-4012.
24. Saito, M., Villanueva, S. Y., Chakraborty, A., Miyahara, S., Segawa, T., Asoh, T., & Yoshida, S. I. (2013). Comparative analysis of Leptospira strains isolated from environmental soil and water in the Philippines and Japan. Applied and environmental microbiology, 79(2), 601-609.
25. Šimpraga, M., Kusak, J., Kurilj, A., Matanović, K., & Đuras, M. (2024). Leptospira kirschneri serogroup Pomona serovar Mozdok as a cause of abortion in mares. Veterinary Sciences, 11(1), 37. https://pubmed.ncbi.nlm.nih.gov/38930421
26. Spiess, B. M. (2010). Atların tekrarlayan üveiti: Avrupa bakış açısı. At Veteriner. J. Ek. doi: 10.1111/j.2042-3306.2010.tb05635.x.
27. Tan, X. T., Amran, F., Cheong, K. C., & Ahmad, N. (2014). In-house ELISA screening using a locallyisolated Leptospira in Malaysia: Determination of its cut-off points. BMC Infect. Dis. 14, 563.
28. Temel H. (2020). Enfeksiyöz Mononükleoz Tanısı Alan Çocuk Hastaların Klinik Ve Laboratuvar Verilerinin Değerlendirilmesi. Selçuk Tıp Dergisi, 36(3), 184-191.
29. World Health Organization and International Leptospirosis Society. Annex 10, Serological Techniques (MAT and ELISA). In Human Leptospirosis: Guidance for Diagnosis, Surveillance and Control; World Health Organisation, Ed.; WHO Library Cataloguing-in-Publication Data: Geneva, Switzerland, 2003; pp. 63–75. ISBN 92 4 154589 5. https://www.who.int/publications/i/item/human-leptospirosis-guidance-for-diagnosis-surveillance-and-control (accessed on 08 January 2023).
30. Wasiński, B., Paschalis-Trela, K., Trela, J., Czopowicz, M., Kita, J., Żychska, M., & Witkowski, L. (2021). Serological Survey of Leptospira Infection in Arabian Horses in Poland. Pathogens, 10(6), 688.
31. Yamaguchi, T., Higa, N., Okura, N., Matsumoto, A., Hermawan, I., Yamashiro, T., & Toma, C. (2018). Characterizing interactions of Leptospira interrogans with proximal renal tubule epithelial cells. BMC microbiology, 18, 1-11.
32. Ye, C., Yan, W., McDonough, P. L., McDonough, S. P., Mohamed, H., Divers, T. J., & Yang, Z. (2014). Serodiagnosis of equine leptospirosis by enzyme-linked immunosorbent assay using four recombinant protein markers. Clinical and Vaccine Immunology, 21(4), 478-483.