Piliç Eti ve Sakatat Örneklerinden İzole Edilen Bacillus cereus Suşlarında Antimikrobiyel Direncin Araştırılması

Öz

Bacillus cereus, doğada yaygın olarak bulunan, emetik ve diyarel formda gıda zehirlenmesine sebep olan patojen bir bakteridir. Bu çalışmada çiğ piliç eti ve sakatat örneklerinde B. cereus varlığı ile elde edilen izolatlarda disk difüzyon yöntemiyle beş farklı antibiyotiğe karşı direnç özelliği belirlenmiştir. Bu amaçla 30 adet piliç baget, 30 adet piliç kanat, 30 adet piliç göğüs ve 30 adet piliç sakatat olmak üzere, toplam 120 adet örnek B. cereus varlığı ve antimikrobiyel dirençliliği yönünden incelendi. Çalışma sonucunda 6’sı piliç baget, 8’i piliç kanat, 10’u piliç göğüs ve 12’si piliç sakatat örneğinde olmak üzere, toplam 120 örneğin 36 (%30)’sının B. cereus ile kontamine olduğu tespit edildi ve sonuçlar Real Time-PCR yöntemi ile doğrulandı. Doğrulanan 36 izolatta disk difüzyon testi ile 5 farklı antibiyotik (penisilin, eritromisin, tetrasiklin, kloramfenikol ve gentamisin) baz alınarak antibiyotik dirençliliği araştırıldı. İzolatların tamamının (%100) penisilin ve eritromisine karşı dirençli olduğu tespit edildi. İzolatlarda tetrasiklin, kloramfenikol ve gentamisine karşı dirençlilik sırasıyla %63,9, %41,7 ve %5,6 düzeyinde bulundu. Çoklu antibiyotik direnci izolatların %80,6’sında gözlendi. Kanatlı eti ve ürünlerinin B. cereus ile kontaminasyon düzeyini azaltmak için, kanatlı hayvanların yetiştirildiği kümes ortamından başlamak üzere, kesime kadar olan zincirde ve kesim sonrası aşamalarda gerekli hijyenik önlemler HACCP planı çerçevesinde yapılmalıdır. Ayrıca, antibiyotik dirençliliğinin önüne geçmek amacıyla kanatlılardaki antibiyotik kullanımı esaslarına dikkat edilmelidir.

Anahtar Kelimeler

• Bacillus cereus
• RT-PCR
• Piliç eti
• Sakatat
• Antimikrobiyel direnç

Investigation of Antimicrobial Resistance in Bacillus cereus Strains Isolated from Broiler Meat and Offal Samples

Abstract

Bacillus cereus is a pathogenic bacterium commonly found in nature that causes emetic and diarrheal food poisoning. In this study, it was aimed to determine the presence of B. cereus in raw chicken meat and offal samples and the resistance properties of the isolates against five different antibiotics using disc diffusion method. In this study, a total of 120 food samples including 30 broiler drumstick, 30 broiler wing, 30 broiler breast, and 30 broiler offal samples were analyzed for the presence of B. cereus and its antimicrobial resistance. As a result of the study, 36 (30%) of a total of 120 food samples including 6 chicken drumsticks, 8 chicken wings, 10 chicken breasts, and 12 chicken offal samples, were found to be contaminated with B. cereus. The results were also confirmed by using the Real-Time PCR method. Antibiotic resistance in the 36 B. cereus isolates was investigated using the disc diffusion method against five different antibiotics: penicillin, erythromycin, tetracycline, chloramphenicol, and gentamicin. All isolates (100%) were found to be resistant to penicillin and erythromycin. Resistance to tetracycline, chloramphenicol, and gentamicin was determined as 63.9%, 41.7%, and 5.6%, respectively. Multidrug resistance was observed in 80.6% of the isolates. To reduce the contamination level with B. cereus in poultry meat and products, appropriate hygienic measures should be implemented within the HACCP framework starting from the poultry farm through to slaughter, and in post-slaughter stages. Furthermore, prudent use of antibiotics in poultry should be ensured to prevent the development of antimicrobial resistance.

Keywords

• Bacillus cereus
• RT-PCR
• Broiler meat
• Offal
• Antimicrobial resistance

Kaynakça

1. Abd El Tawab, A., El-Hofy, F., El-Ekhnawy, K. I. ve El-Shora, H. E. (2019). Detection of some virulence and resistance genes of S. aureus and B. cereus isolated from some meat products. Nature and Science, 17(2), 85–91.
2. Abdelaziz, M. N. S., Zayda, M. G., Maung, A. T., El-Telbany, M., Mohammadi, T. N., Lwin, S. Z. C., Linn, K. Z., Wang, C., Yuan, L., Masuda, Y., Honjoh, K. I. ve Miyamoto, T. (2024). Genetic characterization, antibiotic resistance, and virulence genes profiling of Bacillus cereus strains from various foods in Japan. Antibiotics, 13(8), 774. https://doi.org/10.3390/antibiotics13080774
3. Aklilu, E., Tukimin, E. B., Abu Daud, N. B. ve Kyaw, T. (2016). Enterotoxigenic Bacillus cereus from cooked chicken meat: A potential public health hazard. Malaysian Journal of Microbiology, 12(1), 112–115.
4. Algammal, A. M., Eid, H. M., Alghamdi, S., Ghabban, H., Alatawy, R., Almanzalawi, E. A., Alqahtani, T. M., Elfouly, S. G., Mohammed, G. M., Hetta, H. F. ve El-Tarabili, R. M. (2024). Meat and meat products as potential sources of emerging MDR Bacillus cereus: groEL gene sequencing, toxigenic and antimicrobial resistance. BMC Microbiology, 24(1), 50. https://doi.org/10.1186/s12866-024-03150-9
5. Anonim. (2006). Yem katkıları ve premikslerin üretimi, ithalatı, ihracatı, satışı ve kullanımı hakkındaki tebliğ’de değişiklik yapılmasına dair tebliğ (Tebliğ No: 2006/1). Resmî Gazete (21.01.2006–26056).
6. Anonim. (2026). EFSA (European Food Safety Authority): Antimicrobial resistance. https://www.efsa.europa.eu/en/topics/topic/antimicrobial-resistance
7. Aust, M. O., Godlinski, F., Travis, G. R., Hao, X., McAllister, T. A., Leinweber, P. ve Thiele-Bruhn, S. (2008). Distribution of sulfamethazine, chlortetracycline and tylosin in manure and soil of Canadian feedlots after subtherapeutic use in cattle. Environmental Pollution, 156(3), 1243–1251. https://doi.org/10.1016/j.envpol.2008.01.011
8. Bashir, M., Malik, M. A., Javaid, M., Badroo, G. A., Bhat, M. A. ve Singh, M. (2017). Prevalence and characterization of Bacillus cereus in meat and meat products in and around Jammu Region of Jammu and Kashmir, India. International Journal of Current Microbiology and Applied Sciences, 6(12), 1094–1106. https://doi.org/10.20546/ijcmas.2017.612.123

9. Can, H. Y., Elmali, M., Karagoz, A. ve Disli, H. B. (2022). Psychrotrophic properties, toxigenic characteristics, and PFGE profiles of Bacillus cereus isolated from different foods and spices. Ciencia Rural, 52(4), e20210166. https://doi.org/10.1590/0103-8478cr20210166
10. Can, H. Y. ve Sarı, K. B. (2023). Malatya ilinde üretilen peynirlerden izole edilen Bacillus cereus suşlarında enterotoksin kodlayan genler ile antibiyotik direncinin araştırılması. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 6(2), 1500–1512.
11. Carlin, F., Brillard, J., Broussolle, V., Clavel, T., Duport, C., Jobin, M., Guinebretière, M.-H., Auger, S., Sorokine, A. ve Nguyen-Thé, C. (2010). Adaptation of Bacillus cereus, an ubiquitous worldwide-distributed foodborne pathogen, to a changing environment. Food Research International, 43(7), 1885–1894. https://doi.org/10.1016/j.foodres.2010.03.028
12. Caro-Astorga, J., Pérez-García, A., de Vicente, A. ve Romero, D. (2015). A genomic region involved in the formation of adhesin fibers in Bacillus cereus biofilms. Frontiers in Microbiology, 5, 745. https://doi.org/10.3389/fmicb.2014.00745
13. Casewell, M., Friis, C., Marco, E., McMullin, P. ve Phillips, I. (2003). The European ban on growth-promoting antibiotics and emerging consequences for human and animal health. Journal of Antimicrobial Chemotherapy, 52(2), 159–161. https://doi.org/10.1093/jac/dkg313
14. Chon, J. W., Kim, J. H., Lee, S. J., Hyeon, J. Y. ve Seo, K. H. (2012). Toxin profile, antibiotic resistance, and phenotypic and molecular characterization of Bacillus cereus in Sunsik. Food Microbiology, 32(1), 217–222. https://doi.org/10.1016/j.fm.2012.06.003
15. Eglezos, S., Huang, B., Dykes, G. A. ve Fegan, N. (2010). The prevalence and concentration of Bacillus cereus in retail food products in Brisbane, Australia. Foodborne Pathogens and Disease, 7(7), 867–870. https://doi.org/10.1089/fpd.2009.0479
16. Gdoura-Ben Amor, M., Siala, M., Zayani, M., Grosset, N., Smaoui, S., Messadi-Akrout, F., Baron, F., Jan, S., Gautier, M. ve Gdoura, R. (2018). Isolation, identification, prevalence, and genetic diversity of Bacillus cereus group bacteria from different foodstuffs in Tunisia. Frontiers in Microbiology, 9, 447. https://doi.org/10.3389/fmicb.2018.00447
17. Gomes, F., Saavedra, M. J. ve Henriques, M. (2016). Bovine mastitis disease/pathogenicity: Evidence of the potential role of microbial biofilms. Pathogens and Disease, 74(3), ftw006. https://doi.org/10.1093/femspd/ftw006
18. Guo, H., Yu, P., Yu, S., Wang, J., Zhang, J., Zhang, Y., Liao, X., Wu, S., Ye, Q., Yang, X., Lei, T., Zeng, H., Pang, R., Zhang, J., Wu, Q. ve Ding, Y. (2021). Incidence, toxin gene profiling, antimicrobial susceptibility, and genetic diversity of Bacillus cereus isolated from quick-frozen food in China. LWT, 140, 110824. https://doi.org/10.1016/j.lwt.2020.110824
19. Kotiranta, A., Lounatmaa, K. ve Haapasalo, M. (2000). Epidemiology and pathogenesis of Bacillus cereus infections. Microbes and Infection, 2(2), 189–198. https://doi.org/10.1016/S1286-4579(00)00269-0
20. Landoni, M. F. ve Albarellos, G. (2015). The use of antimicrobial agents in broiler chickens. The Veterinary Journal, 205(1), 21–27. https://doi.org/10.1016/j.tvjl.2015.04.016
21. Mancuso, G., Midiri, A., Gerace, E. ve Biondo, C. (2021). Bacterial antibiotic resistance: The most critical pathogens. Pathogens, 10(10), 1310. https://doi.org/10.3390/pathogens10101310
22. Mehdi, Y., Létourneau-Montminy, M. P., Gaucher, M. L., Chorfi, Y., Suresh, G., Rouissi, T., Brar, S. K., Côté, C., Ramirez, A. A. ve Godbout, S. (2018). Use of antibiotics in broiler production: Global impacts and alternatives. Animal Nutrition, 4(2), 170–178. https://doi.org/10.1016/j.aninu.2018.03.002
23. Nakayama, T., Yamaguchi, T., Jinnai, M., Yamamoto, S., Li, H. T., Ngo, P. T., Tran, D. N. M., Nguyen, O. T. H., Hoang, P. H., Do Nguyen, P., Van Dang, C., Kumeda, Y. ve Hase, A. (2021). Untargeted phylogenetic group III of multi-drug-resistant Bacillus cereus isolated using Fraser medium from retail chickens in Ho Chi Minh City. Current Microbiology, 78(8), 3115–3123. https://doi.org/10.1007/s00284-021-02563-0
24. Osman, K. M., Kappell, A. D., Orabi, A., Al-Maary, K. S., Mubarak, A. S., Dawoud, T. M., Hemeg, H. A., Moussa, I. M. I., Hessain, A. M., Yousef, H. M. Y. ve Hristova, K. R. (2018). Poultry and beef meat as potential seedbeds for antimicrobial resistant enterotoxigenic Bacillus species: A materializing epidemiological and potential severe health hazard. Scientific Reports, 8(1), 11600. https://doi.org/10.1038/s41598-018-29932-3
25. Park, Y. B., Kim, J. B., Shin, S. W., Kim, J. C., Cho, S. H., Lee, B. K., Ahn, J., Kim, J. M., ve Oh, D. H. (2009). Prevalence, genetic diversity, and antibiotic susceptibility of Bacillus cereus strains isolated from rice and cereals collected in Korea. Journal of Food Protection, 72(3), 612–617. https://doi.org/10.4315/0362-028X-72.3.612
26. Rather, M. A., Aulakh, R. S., Gill, J. P. S., ve Ghatak, S. (2012). Enterotoxin gene profile and antibiogram of Bacillus cereus strains isolated from raw meats and meat products. Journal of Food Safety, 32(1), 22–28. https://doi.org/10.1111/j.1745-4565.2011.00363.x
27. Smith, D. P., Berrang, M. E., Feldner, P. W., Phillips, R. W. ve Meinersmann, R. J. (2004). Detection of B. cereus on selected retail chicken products. Journal of Food Protection, 67(8), 1770–1773. https://doi.org/10.4315/0362-028X-67.8.1770
28. Tallent, S. M., Kotewicz, K. M., Strain, E. A. ve Bennett, R. W. (2012). Efficient isolation and identification of Bacillus cereus group. Journal of AOAC International, 95(2), 446–451. https://doi.org/10.5740/jaoacint.11-251
29. Tewari, A. ve Abdullah, S. (2015). Bacillus cereus food poisoning: International and Indian perspective. Journal of Food Science and Technology, 52(5), 2500–2511. https://doi.org/10.1007/s13197-014-1344-4
30. Wright, G. D. ve Poinar, H. (2012). Antibiotic resistance is ancient: Implications for drug discovery. Trends in Microbiology, 20(4), 157–159. https://doi.org/10.1016/j.tim.2012.01.002
31. Yu, P., Yu, S., Wang, J., Guo, H., Zhang, Y., Liao, X., Zhang, J., Wu, S., Gu, Q., Xue, L., Zeng, H., Pang, R., Lei, T., Zhang, J., Wu, Q. ve Ding, Y. (2019). Bacillus cereus isolated from vegetables in China: Incidence, genetic diversity, virulence genes, and antimicrobial resistance. Frontiers in Microbiology, 10, 948. https://doi.org/10.3389/fmicb.2019.00948