Edirne İlindeki Sığırlarda Escherichia coli O157:H7’nin Moleküler Prevalansı

Öz

Escherichia coli O157:H7, rezervuarı olduğu sığırların bağırsak florasında bulunan ve dışkılarının yayılmasıyla gıda ve suyu kontamine eden zoonotik bir bakteridir. Direkt temas veya indirekt aracılarla bulaşabilen etken insanlarda ciddi hastalıklara sebep olabilmektedir. Bu nedenle, sığırların etken taşıyıcılıklarının araştırılması olası bulaşların önlenmesi açısından önem arz etmektedir. Bu çalışmada, Edirne İlinde yetiştirilen sığırlarda E. coli O157:H7’nin varlığının Real-Time PCR ile araştırılması amaçlanmıştır. Çalışmanın materyalini, Edirne il ve ilçelerindeki köylerde bulunan yarı ekstansif (n=59) ve entansif (n=20) işletmelerden alınan toplam 400 taze dışkı örneği oluşturdu. Bu örnekler, klinik duruma göre asemptomatik (n=260) ve semptomatik (n=140) hayvanlardan; yaş grubuna göre ise buzağı (n=120), düve (n=80) ve erişkin sığırdan (n=200) alınmıştır. Dışkı örneklerinin E. coli O157:H7 yönünden analizi Real-Time PCR yöntemi ile gerçekleştirildi. Biri entansif işletmeye ait halsizlik, iştahsızlık ve hafif kanlı sulu ishalli bir buzağı ve biri yarı ekstansif bir işletmeye ait asemptomatik ergin bir sığır olmak üzere toplam iki (%0,5) örnekte E. coli O157:H7 pozitifliği tespit edildi. Sığırlarda yetiştirme türü, klinik durum ve yaş gibi faktörlerle E. coli O157:H7 pozitifliği arasında istatistiksel olarak anlamlı bir ilişki bulunmamasına rağmen (p>0,05), farklı karakteristikleriyle ile değişen oranlarda rezervuarlık yaptıkları E. coli O157:H7’nin epidemiyolojisini etkileyebilecekleri düşündürmektedir. Ancak, pozitif numunelerin sınırlı sayısı, anlamlı farklılıkları tespit etme gücünü azaltmış olabilir, bu nedenle bu bulgular dikkatle yorumlanmalıdır.

Anahtar Kelimeler

• Escherichia coli O157:H7
• Sığır dışkı örnekleri
• Rezervuar
• Moleküler tanı

Molecular Prevalence of Escherichia coli O157:H7 in Cattle in Edirne Province, Türkiye

Öz

Escherichia coli O157:H7 is a zoonotic bacterium that resides in the intestinal flora of cattle, which serve as reservoirs, and contaminates food and water through the spread of their feces. The causative agent can be transmitted by direct contact or indirect intermediaries and can cause serious diseases in humans. Therefore, it is important to investigate the carrier status of cattle to prevent possible transmission. This study aimed to investigate the presence of E. coli O157:H7 in cattle raised in the Edirne Province (Türkiye) by Real-Time PCR. The study material consisted of a total of 400 fresh fecal samples collected from semi-extensive (n=59) and intensive (n=20) farms located in villages of Edirne province and its districts. These samples were obtained from asymptomatic (n=260) and symptomatic (n=140) cattle based on clinical status, and from calves (n=120), heifers (n=80), and adult cattle (n=200) based on age group. The samples were analyzed for E. coli O157:H7 by Real-Time PCR. E. coli O157:H7 positivity was detected in a total of two (0.5%) samples, one of which was a calf with malaise, anorexia, and mild bloody diarrhea from an intensive farm and one asymptomatic adult cattle from a semi-extensive farm. Although no statistically significant relationship was found between E. coli O157:H7 positivity and factors such as breeding type, clinical condition, and age in cattle (p>0.05), their potential role as reservoirs with different characteristics and varying reservoir rates suggests they may influence the epidemiology of the pathogen. However, the limited number of positive samples may have reduced the power to detect significant differences, so these findings should be interpreted with caution.

Anahtar Kelimeler

• Escherichia coli O157:H7
• Bovine fecal samples
• Reservoir
• Molecular diagnostics

Kaynakça

1. Ağralı, M. Ş., & Doğan, M. (2021). Investigation of bacterial gastroenteritis agents in patients who admitted to pediatric clinic with diarrhea. Klimik Derg, 34(3), 219-224.
2. Aksoy, A., Yıldırım, M., Kaçmaz, B., Apan, T. Z., & Göçmen, J. S. (2007). Verotoxin production in strains of Escherichia coli isolated from cattle and sheep, and their resistance to antibiotics. Turk J Vet Anim Sci, 31, 225-231.
3. Ameer, M. A., Wasey, A., & Salen, P. (2025). Escherichia coli (E. coli 0157:H7) [Updated 2023 Aug 8]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan. Available from: https://www.ncbi.nlm.nih.gov/books/NBK507845/
4. Ashenaf, A., Dereje, E., & Haben, F. (2020). Isolation and antimicrobial susceptibility profile of Escherichia coli O157:H7 from raw milk of dairy cattle in Holeta District, Central Ethiopia. Hindawi Int J Microbiol 1, 6626488.
5. Aslantaş, Ö., Erdogan, S., Cantekin, Z., Gulactı, I., & Evrendilek, G. A. (2006). Isolation and characterization of verocytotoxin-producing Escherichia coli O157 from Turkish cattle. Int J Food Microbiol, 106, 338-342.
6. Babacan, O. (2024). Investigating the presence and antibiotic susceptibilities of Escherichia coli O157 and Listeria monocytogenes in ruminant feces and feed in Balıkesir province. Ankara Univ Vet Fak Derg, 71, 31-39.
7. Bach, S. J., McAllister, T. A., Veira, D. M., Gannon, V. P. J., & Holley, R.A. (2002). Transmission and control of Escherichia coli O157:H7: A review. Can J Anim Sci, 82, 475-490.
8. Bai, Z., Xu, X., Wang, C., Wang, T., Sun, C., Liu, S., & Li, D. (2022). A comprehensive review of detection methods for Escherichia coli O157:H7. TrAC, 152, 116646.

9. Bekar, E. (2019). Etlerde Escherichia coli O157:H7 varlığının araştırılması. Kafkas Üniversitesi, Sağlık Bilimleri Enstitüsü, Yüksek Lisans Tezi, Kars, Türkiye. (In Turkish).
10. Birdal, E., & Ak, S. (2018). Prevalence of E. coli O157:H7 in dairy cattle and the farm environment. J Anatol Environ Animal Sci, 3(2), 85-90.
11. Blanco, J. E., Blanco, M., Alonso, M. P., Mora, A., Dahbi, G., Coira, M. A., & Blanco, J. (2004). Serotypes, Virulence genes, and intimin types of shiga toxin (verotoxin)- producing Escherichia coli isolates from human patients: prevalence in lugo, spain, from 1992 through 1999. J Clin Microbiol, 42, 311-319.
12. Bosilevac, J. M., Dwivedi, H. P., Chablain, P., Ullery, M., Bailey, J. S., & Dutta, V. (2019). Comparative performance evaluation of Real-Time PCR and dual-labeled fluorescence resonance energy transfer probe-based melt peak analysis for the detection of Escherichia coli O157:H7 in beef products. J Food Prot, 82(3), 507-512.
13. Caprioli, A., Morabito, S., Brugère, H., & Oswald, E. (2005). Enterohaemorrhagic Escherichia coli: Emerging issues on virulence and modes of transmission. Vet Res, 36(3), 289-311.
14. Chapman, P. A. (2000). Sources of Escherichia coli O157 and experiences over the past 15 years in Sheffield, UK. J Appl Microbiol, 88, 51-60.
15. Chapman, P. A., Siddons, A., Wright, D. J., Norman, P., Fox, J., & Crick, E. (1993). Cattle as a possible source of verocytotoxin-producing Escherichia coli O157:H7 infections in man. Epidemiol Infect, 111, 439-447.
16. Coura, F. M., Freitas, M. D., Ribeiro, J, de Leme, R. A., de Souza, C., Alfieri, A. A., Facury Filho, E. J., de Carvalho, A. Ú., Silva, M. X., Lage, A. P., & Heinemann, M. B. (2015). Longitudinal study of Salmonella spp., diarrheagenic Escherichia coli, Rotavirus, and Coronavirus isolated from healthy and diarrheic calves in a Brazilian dairy herd. Trop Anim Health Prod, 47(1), 3-11.
17. Çabalar, M., Boynukara, B., Gülhan, T., & Ekin, H. (2001). Prevalence of Rotavirus, Escherichia coli K99 and O157:H7 in healthy dairy cattle herds in Van, Turkey. Turk J Vet Anim Sci, 25, 191-196.
18. Çiçek, E. (2008). Ege Bölgesindeki Sığırların Süt ve Dışkı Örneklerinden Escherichia coli O157:H7 İzolasyonu ve Verotoksinlerinin Belirlenmesi. Sağlık Bilimleri Enstitüsü, Mikrobiyoloji Anabilim Dalı. Yüksek Lisans tezi, Adnan Menderes Üniversitesi, Aydın, Türkiye. (In Turkish).
19. European Centre for Disease Prevention and Control (ECDC). (2024). STEC infection. In: ECDC. Annual epidemiological report for 2022. Stockholm: ECDC.
20. Gambushe, S. M., Zishiri, O. T., & El Zowalaty, M. E. (2022). Review of Escherichia coli O157:H7 prevalence, pathogenicity, heavy metal and antimicrobial resistance, African Perspective. Infect Drug Resist, 15, 4645-4673.
21. Guy, R. A., Tremblay, D., Beausoleil, L., Harel, J., & Champagne, M. J. (2014). Quantification of E. coli O157 and STEC in feces of farm animals using direct multiplex real time PCR (qPCR) and a modified most probable number assay comprised of immunomagnetic bead separation and qPCR detection. J Microbiol Methods, 99, 44-53.
22. Hunduma, D., Amenu, K., Desta, H., Grace, D., Agga, G. E., & Kerro Dego, O. (2024). Prevalence and antimicrobial resistance of Escherichia coli O157:H7 and Salmonella, and the prevalence of Staphylococcus aureus in dairy cattle and camels under pastoral production system. Antibiotics, 13(1), 26.
23. ISO. (2012). ISO/TS-13136:2012: Microbiology of Food and Animal Feed. Real-Time Polymerase Chain Reaction (PCR)- Based Method for the Detection of Food-Borne Pathogens. Horizontal Method for the Detection of Shiga Toxin Producing Escherichia coli (STEC) and the Determination of O157, O111, O26, O103 and O145 Serogroups. Geneva, Switzerland: International Organization for Standardization.
24. Kuyucuoğlu, Y., Şeker, E., Sareyyüpoğlu, B., & Konak, S. (2011). Virulence genes of Shiga toxin-producing Escherichia coli O157:H7 strains isolated from calves and cattle. Ankara Üniver Vet Fak Derg, 58(4), 255-260.
25. Mohseni, M., Djawadi, B., & Khazaei, N. (2023). Escherichia coli O157:H7 and its effect on human health. IntechOpen. doi: 10.5772/intechopen.101825
26. Moxley, R. A., & Smith, D. R. (2010). Attaching-effacing Escherichia coli infections in cattle. Vet Clin North Am Food Anim Pract, 26(1), 29-56.
27. Nitecki, S., Teape, N., Carney, B., Slater, J., & Brück, W. (2015). A duplex qPCR for the simultaneous detection of Escherichia coli O157:H7 and Listeria monocytogenes using LNA probes. Letters Appl Microbiol, 61(1), 20-27.
28. Noll, L. W., Chall, R., Shridhar, P. B., Liu, X., Bai, J., Delannoy, S., Fach, P., & Nagaraja, T. G. (2018). Validation and application of a Real-Time PCR assay based on the CRISPR array for serotype-specific detection and quantification of Enterohemorrhagic Escherichia coli O157:H7 in cattle feces. J Food Prot, 81(7), 1157-1164.
29. Omisakin, F., MacRae, M., Ogden, I. D., & Strachan, N. J. (2003). Concentration and prevalence of Escherichia coli O157 in cattle feces at slaughter. Appl Environ Microbiol, 69(5), 2444-2447.
30. Pearce, M. C., Fenlon, D., Low, J. C., Smith, A. W., Knight, H. I., Evans, J., Foster, G., Synge, B. A., & Gunn, G. J. (2004). Distribution of Escherichia coli O157 in bovine fecal pats and its impact on estimates of the prevalence of fecal shedding. Appl Environ Microbiol, 70(10), 5737-5743.
31. Sargeant, J. M., Gillespie, J. R., Oberst, R. D., Phebus, R. K., Hyatt, D. R., Bohra, L. K., & Galland, J. C. (2000). Results of a longitudinal study of the prevalence of Escherichia coli O157:H7 on cow-calf farms. Am J Vet Res, 61(11), 1375-1379.
32. Seker, E., & Yardimci, H. (2008). First isolation of Escherichia coli O157:H7 from faecal and milk specimens from anatolian water buffaloes (Bubalus bubalus) in Turkey. J S Afr Vet Assoc, 79(4), 167-170.
33. Somda, N. S., Adesoji, T. O., Tetteh-Quarcoo, P. B., & Donkor, E. S. (2025). A systematic review and meta-analysis on the presence of Escherichia coli O157:H7 in Africa from a one health perspective. Microorganisms, 13(4), 902.
34. Suardana, I. W., Widiasih, D. A., Nugroho, W. S., Wibowo, M. H., & Suyasa, I. N. (2017). Frequency and risk-factors analysis of Escherichia coli O157:H7 in Bali-cattle. Acta Trop, 172, 223-228.
35. Taskin, B. (2017). Novel approaches for monitoring viable pathogenic microorganisms in environmental samples. YYÜ Tar Bı̇l Derg, 27(2), 285-291.
36. U.S. Food and Drug Administration (FDA). (2025). Bacteriological Analytical Manual (BAM), Chapter 4 and 4A, updated 07.2020; Available from (https://www.fda.gov/food/laboratory-methods-food/bacteriological-analytical-manual-bam). Access Date: 01.04.2025.
37. Wirth, J. S., Leeper, M. M., Smith, P. A., Vasser, M., Katz, L. S., Vidyaprakash, E., Carleton, H. A., Chen, J. C. (2025). Genomic characterization of Escherichia coli O157:H7 associated with multiple sources, United States. Emerg Infect Dis, 31(13), 109-116.
38. Yılmaz, A., Gün, H., & Yılmaz, H. (2002). Frequency of Escherichia coli O157:H7 in Turkish cattle. J Food Protect, 65, 1637-1640.
39. Zaheer, R., Dugat-Bony, E., Holman, D. B., Cousteix, E., Xu, Y., Munns, K., Selinger, L. J., Barbieri, R., Alexander, T., McAllister, T. A., & Selinger, L. B. (2017). Changes in bacterial community composition of Escherichia coli O157:H7 super-shedder cattle occur in the lower intestine. PLoS One, 12(1), e0170050.
40. Zhao, T., Doyle, M. P., Shere, J., & Garber, L. (1995). Prevalence of enterohemorrhagic Escherichia coli O157:H7 in a survey of dairy herds. Appl Environ Microbiol, 61, 1290-1293.