Determination of the Presence and Antimicrobial Susceptibility of Clostridioides difficile in Chicken Doner Kebabs Sold in Istanbul

Abstract

Clostridioides difficile is a gram (+), anaerobic, spore-forming, rod-shaped bacterium that causes pseudomembranous colitis, toxic megacolon, intestinal perforation and diarrhea, especially due to long-term usage of antibiotics. The hospital-acquired agent has been detected in soil, water, seafood, slaughtered animals and poultry, and these foods are thought to be potential new reservoirs for C. difficile. Especially in recent years, the detection of C. difficile strains isolated from humans in butchery and poultry animals has raised concerns that this agent may pose a serious risk to public health. Accordingly, 128 chicken doner kebab samples offered for sale in Istanbul were analyzed for the presence of C. difficile and 12 of them were identified as C. difficile suspects. Only 2 (1.56%) out of the 12 suspected colonies were confirmed as C. difficile, and both isolates were resistant to vancomycin and cefotaxime. In conclusion, C. difficile strains isolated from chicken doner samples showed that the agent can be found in poultry carcasses as well as in animal-originated foods obtained from these carcasses and can also be resistant to cooking temperatures.

Keywords

• Antimicrobial susceptibility
• chicken doner kebab
• Clostridioides difficile
• poultry
• public health

İstanbul’da Satışa Sunulan Tavuk Dönerlerinde Clostridioides difficile Varlığının ve Antimikrobiyal Duyarlılıklarının Belirlenmesi

Öz

Clostridioides difficile, gram (+), anaerob, sporlu, çomak şeklinde bakteri olup özellikle uzun süreli antibiyotik kullanımı sonucunda psödomembranoz kolit, toksik megakolon, intestinal perforasyon ve diareye sebep olmaktadır. Hastane kaynaklı olan etken toprakta, suda, su ürünlerinde, kasaplık hayvanlarda ve kanatlılarda tespit edilmiş olup, bu durum gıdaların C. difficile için potansiyel yeni rezervuarlar olabileceğini düşündürmektedir. Özellikle son yıllarda insanlardan izole edilen C. difficile suşlarının kasaplık ve kanatlı hayvanlarda da saptanması bu etkenin halk sağlığı yönünden ciddi bir risk oluşturabileceği kaygısını doğurmuştur. Bu doğrultuda, İstanbul’da satışa sunulan 128 tavuk döner örneği C. difficile varlığı yönünden analiz edilmiş ve bunlardan 12’si C. difficile şüpheli olarak tespit edilmiştir. Şüpheli 12 örnekten sadece 2’si (%1.56) C. difficile olarak doğrulanırken, her iki izolatın da vankomisin ve sefotaksime dirençli olduğu saptanmıştır. Sonuç olarak, tavuk döner örneklerinden izole edilen C. difficile suşları, etkenin kanatlı hayvan karkaslarının yanı sıra bunlardan elde edilen hayvansal kökenli gıdalarda da bulunabildiğini ve pişirme sıcaklıklarına da dirençli olabildiğini göstermiştir.

Anahtar Kelimeler

• Antimikrobiyal duyarlılık
• Clostridioides difficile
• Halk sağlığı
• Kanatlı eti
• Tavuk döner

Kaynakça

1. Pelaez T, Alcalá L, Blanco JL. et al. (2013). Characterization of Swine Isolates of Clostridium difficile in Spain: A Potential Source of Epidemic Multidrug Resistant Strains? Anaerobe. 22: 45-49.
2. Troiano T, Harmanus C, Sanders IMJG. et al. (2015). Toxigenic Clostridium difficile PCR Ribotypes in Edible Marine Bivalve Molluscs in Italy. Int J Food Microbiol. 208: 30-34.
3. De Boer E, Zwartkruis-Nahuis A, Heuvelink AE, Hurmanus C, Kuijper EJ. (2011). Prevalance of Clostridium difficile in Retailed Meat in the Netherlands. Int J Food Microbiol. 144: 561-564.
4. Rodriguez C, Taminiau B, Van Broeck J, Avesani V, Delmée M, Daube G. (2012). Clostridium difficile in Young Farm Animals and Slaughter Animals in Belgium. Anaerobe. 18: 621-625.
5. Thitaram SN, Frank JF, Siragusa GR. et al. (2016). Antimicrobial Susceptibility of Clostridium difficile Isolated from Food Animals on Farms. Int J Food Microbiol. 227: 1-5.
6. Simango C, Mwakurudza S. (2008). Clostridium difficile in Broiler Chickens Sold at Market Places in Zimbabwe and their Antimicrobial Susceptibility. Int J Food Microbiol. 124: 268-270.
7. Weese JS, Reid-Smith RJ, Avery BP, Rousseau J. (2010). Detection and Characterization of Clostridium difficile in Retail Chicken. Lett Appl Microbiol. 50: 362-365.
8. Romano V, Albanese F, Dumontet S, Krovacek K, Petrini O, Pasquale V. (2012). Prevalence and Genotypic Characterization of Clostridium difficile from Ruminants in Switzerland. Zoonoses Public Hlth. 59: 545-548.

9. Rahimi E, Afzali ZS, Baghbadorani ZT. (2015). Clostridium difficile in Ready-to-eat Foods in Isfahan and Shahrekord, Iran. Asian Pac J Trop Biomed. 5: 128-131.
10. Hampikyan H, Bingol EB, Muratoglu K., Akkaya E, Cetin O, Colak H. (2018). The Prevalence of Clostridium difficile in Cattle and Sheep Carcasses and the Antibiotic Susceptibility of Isolates. Meat Sci. 139: 120-124.
11. Rodriguez-Palacios A, Reid-Smith RJ, Staempfli HR. et al. (2009). Possible Seasonality of Clostridium difficile in Retail Meat, Canada. Emerg Infect Dis. 15: 802-805.
12. Metcalf DS, Costa MC, Dew WMV, Weese J.S. (2010). Clostridium difficile in Vegetables. Canada Lett Appl Microbiol. 51: 600-602.
13. Rodriguez C, Avesani V, Van Broeck J, Taminiau B, Delmée M, Daube G. (2013). Presence of Clostridium difficile in Pigs and Cattle Intestinal Contents and Carcass Contamination at the Slaughterhouse in Belgium. Int J Food Microbiol. 166: 256-262.
14. Zidaric V, Zemljic M, Janezic S, Kocuvan A, Rupnik M. (2008). High Diversity of Clostridium difficile Genotypes Isolated from a Single Poultry Farm Producing Replacement Laying Hens. Anaerobe. 14: 325-327.
15. Guran HS, Ilhak OI. (2015). Clostridium difficile in Retail Chicken Meat Parts and Liver in the Eastern Region of Turkey. J Verbr Lebensm. 10: 359-364.
16. Abdel-Glil MY, Thomas P, Schmoock G. et al. (2018). Presence of Clostridium difficile in Poultry and Poultry meat in Egypt. Anaerobe. 51: 21-25.
17. Ersöz ŞŞ, Coşansu S. (2018). Prevalence of Clostridium difficile Isolated from Beef and Chicken meat products in Turkey. Korean J Food Sci Anim Resour. 38: 759-767.
18. Bingol EB, Hampikyan H, Muratoglu K, Akkaya E, Cetin O, Colak H. (2020). Characterisation and Antibiotic Susceptibility Profile of Clostridioides (Clostridium) difficile Isolated from Chicken Carcasses. J Vet Res. 64: 407-412.
19. Muratoglu K, Akkaya E, Hampikyan H, Bingol EB, Cetin O, Colak H. (2020). Detection, Characterization and Antibiotic Susceptibility of Clostridioides (Clostridium) difficile in Meat Products. Food Sci Anim Resour. 40(4): 578-587.
20. Lemee L, Dhalliun A, Testelin S. et al. (2004). Multiplex PCR Targeting tpi (triose phosphate isomerase), tcdA (Toxin A), and tcdB (Toxin B) Genes for Toxigenic Culture of Clostridium difficile. J Clin Microbiol. 42(12): 5710-5714.
21. Stubbs S, Rupnik M, Gibert M, Brazier J, Duerden B, Popoff M. (2000). Production of Actin-Specific ADP-Ribosyltransferase (Binary Toxin) by Strains of Clostridium difficile. FEMS Microbiol Lett. 186: 307-312.
22. CLSI. (2024). Performance standards for antimicrobial susceptibility testing. 34th ed. CLSI supplement M100. Clinical and Laboratory Standards Institute.
23. EUCAST. (2023). European Committee for Antimicrobial Susceptibility Testing. EUCAST clinical breakpoint table v. 2.0. 62.
24. Indra A, Lassnig H, Baliko N. et al. (2009). C. difficile: A New Zoonotic Agent? Wien Klin Wochenschr. 121: 91-95.
25. Heise J, Witt P, Maneck C, Wichmann-Schauer H, Maurischat S. (2021). Prevalence and Phylogenetic Relationship of Clostridioides difficile Strains in Fresh Poultry Meat Samples Processed in Different Cutting Plants. Int J Food Microbiol. 339.
26. Tkalec V, Jamnikar-Ciglenecki U, Rupnik M, Vadnjal S, Zelenik K, Biasizzo M. (2020). Clostridioides difficile in National Food Surveillance, Slovenia, 2015 to 2017. Eurosurveill. 25.
27. Songer, JG, Trinh HT, Killgore GE, Thompson AD, McDonald LC, Limbago BM. (2009). Clostridium difficile in Retail Meat Products, USA, 2007. Emerg Infect Dis. 15: 819-821.
28. Varshney JB, Very KJ, Williams JL. et al. (2014). Characterization of Clostridium difficile Isolates from Human Fecal Samples and Retail Meat from Pennsylvania. Foodborne Pathog Dis. 11: 822-829.
29. Barezi AA, Shakerian A, Rahimi E, Esfandiari Z. (2023). Examining the Extent of Contamination, Antibiotic Resistance, and Genetic Diversity of Clostridioides (Clostridium) difficile Strains in Meat and Feces of Some Native Birds of Iran. Biomed Res Int. 3524091.
30. Limbago B, Thompson AD, Greene SA. et al. (2012). Development of a Consensus Method for Culture of Clostridium difficile from Meat and its Use in a Survey of U.S. Retail Meats. Food Microbiol. 32: 448-451.
31. Mooyottu S, Flock G, Kollanoor-Johny A, Upadhyaya I, Jayarao B, Venkitanarayanan K. (2015). Characterization of a Multidrug Resistant C. difficile Meat Isolate. Int J Food Microbiol. 192: 111-116.
32. Von Abercron SMM, Karlsson F, Wigh GT, Wierup M, Krovacek K. (2009). Low Occurrence of Clostridium difficile in Retail Ground Meat in Sweden. J Food Prot. 72: 1732-1734.
33. Limbago B, Thompson AD, Greene SA. et al. (2012). Development of a Consensus Method for Culture of Clostridium difficile from Meat and its Use in a Survey of U.S. Retail Meats. Food Microbiol. 32: 448-451.
34. Rahimi E, Khaksar F. (2015). Detection of Toxigenic Clostridium difficile Strains Isolated from Meat and Meat Products in Iran. Bulg J Vet Med. 18: 277-281.
35. Simango C. (2006). Prevalence of Clostridium difficile in the Environment in a Rural Community in Zimbabwe. Trans R Soc Trop Med Hyg. 100: 1146-1150.
36. Lund BM, Peck MW. (2015). A possible route for foodborne transmission of Clostridium difficile? Foodborne Pathog Dis. 12: 177-182.
37. Blanco JL, Álvarez-Pérez S, García ME. (2013). Is the Prevalence of Clostridium difficile in Animals Underestimated? Vet J. 197: 694-698.
38. Knight DR, Thean S, Putsathit P, Fenwick S, Riley TV. (2013). Cross-Sectional Study Reveals High Prevalence of Clostridium difficile Non-PCR Ribotype 078 Strains in Australian Veal Calves at Slaughter. Appl Environ Microbiol. 79, 2630-2635.
39. Rodriguez-Palacios A, Reid-Smith RJ, Staempfli HR, Weese JS. (2010). Clostridium difficile Survives Minimal Temperature Recommended for Cooking Ground Meats. Anaerobe. 16: 540-542.
40. Kouassi KA, Dadie AT, N’Guessan KF, Dje KM, Loukou YG. (2014). Clostridium perfringens and Clostridium difficile in Cooked Beef Sold in CÔte d’Ivoire and Their Antimicrobial Susceptibility. Anaerobe. 28: 90-94.
41. Lawley TD, Croucher NJ, Yu L. et al. (2009). Proteomic and Genomic Characterization of Highly Infectious Clostridium difficile Spores. J Bacteriol. 191: 5377-5386.
42. Saad J, Lendormi T, Le Marechal C, Pourcher A-M, Druilhe C, Lanoiselle J-L. (2023). Kinetic Study of Thermal Inactivation of Enterococci and Clostridial Spores. MATEC Web of Conferences. 379: 05004.
43. Rodriguez-Palacios A, LeJeune JT. (2011). Moist-heat Resistance, Spore Aging, and Superdormancy in Clostridium difficile. Appl Environ Microbiol. 77: 3085-3091.
44. Deng K, Plaza-Garrido A, Torres JA, Paredes-Sabja D. (2015). Survival of Clostridium difficile Spores at Low Temperatures. Food Microbiol. 46: 218-221.
45. Bakri M. (2018). Prevalence of Clostridium difficile in Raw Cow, Sheep, and Goat Meat in Jazan, Saudi Arabia. Saudi J Biol Sci. 25: 783-785.
46. Harvey RB, Norman KN, Andrews K. et al. (2011). Clostridium difficile in Retail Meat and Processing Plants in Texas. J Vet Diagn Invest. 23: 807-811.