Beyaz Peynirlerde Staphylococcus aureus Varlığı: Enterotoksin ve Antimikrobiyal Direnç Genlerinin Belirlenmesi

Abstract

Bu çalışma hem ambalajlı hem de ambalajsız olmak üzere toplam 100 beyaz peynir örneğinde Staphylococcus aureus kontaminasyonu, enterotoksin varlığı ve antibiyotik direncini değerlendirmeyi amaçlamıştır. Polimeraz Zincir Reaksiyonu (PCR) analizi, 50 ambalajsız örneğin 16'sında (%32) ve 50 ambalajlı örneğin 8'inde (%16) S. aureus varlığını göstermiştir. 24 S. aureus pozitif örnekten 46 izolat elde edilmiştir. Ayrıca, enterotoksin üretimiyle ilişkili sed geni, biri ambalajlı biri ambalajsız beyaz peynir örneklerinden elde edilen iki izolatta tespit edilmiştir. İzolatlar eritromisin (%6.52), gentamisin (%10.87), kloramfenikol (%6.52), trimetoprim-sülfametoksazol (%2.37), rifampisin (%15.21), tetrasiklin (%28.26), sefoksitin (%43.47) ve penisilin G'ye (%34.78) direnç göstermiştir. 46 S. aureus izolatında oksasilin ve vankomisin direnci, minimal inhibisyon konsantrasyonu (MİK) yöntemi kullanılarak incelenmiştir. MİK sonuçlarına göre, izolatların hiçbirinin oksasiline dirençli olmadığı tespit edilmiştir. Ancak, izolatların %47.82'si vankomisine direnç göstermiştir. Genel olarak, izolatlar test edilen 10 antibiyotiğin 9'una direnç göstermiştir. Direnç profillerine ek olarak, direnç genleri de araştırılmıştır. 46 S. aureus izolatının 3'ünün (%13.6) mecA genini, 6'sının (%27.2) mecC genini ve 1'inin (%2.17) hem mecA hem de mecC genlerini taşıdığı tespit edilmiştir. Sonuç olarak, beyaz peynirde S. aureus tespit edilmesi, hijyen koşullarının yetersiz olduğunu göstermektedir. Yüksek antibiyotik direnci, direnç genlerinin varlığı ve bazı izolatlarda enterotoksin genlerinin bulunması, dirençli mikroorganizmaların insanlara bulaşma riskini işaret etmekte olup, bu durum, hastalık tedavisini zorlaştırabilir ve gıda kaynaklı hastalıklara neden olabilir. Bu da şimdi ve gelecekte halk sağlığı açısından endişe verici bir durum oluşturmaktadır.

Keywords

• Antibiyotik dirençliliği
• Enterotoksin
• Halk sağlığı
• Staphylococcus aureus

Project Number

TYL-2021-9501

Presence of Staphylococcus aureus in White Cheese: Determination of Enterotoxin and Antimicrobial Resistance Genes

Abstract

This study aimed to evaluate Staphylococcus aureus contamination, enterotoxin presence, and antibiotic resistance in 100 white cheese samples, both packaged and unpackaged. Polymerase Chain Reaction (PCR) analysis showed S. aureus in 16 (32%) of the 50 unpackaged samples and 8 (16%) of the 50 packaged samples. 46 S. aureus isolates were obtained from 24 S. aureus positive samples. Moreover, the sed gene, associated with enterotoxin production, was detected in two 46 isolates, one from packaged and one from unpackaged white cheese samples. The isolates exhibited resistance to erythromycin (6.52%), gentamicin (10.87%), chloramphenicol (6.52%), trimethoprim-sulfamethoxazole (2.37%), rifampicin (15.21%), tetracycline (28.26%), cefoxitin (43.47%), and penicillin G (34.78%). Oxacillin and vancomycin resistance among the 46 S. aureus isolates was examined using the minimal inhibition concentration (MIC) method. According to the MIC results, none of the isolates were resistant to oxacillin. However, 47.82% of the isolates were resistant to vancomycin. Overall, the isolates exhibited resistance to 9 out of the 10 antibiotics tested. In addition to the resistance profiles, resistance genes were also investigated. It was found that 3 out of the 46 S. aureus isolates (13.6%) carried the mecA gene, 6 (27.2%) carried the mecC gene, and 1 (2.17%) carried both mecA and mecC genes. In conclusion, detecting S. aureus in white cheese indicates poor hygiene. The high antibiotic resistance, presence of resistance genes, and enterotoxin genes in some isolates suggest poses a risk of transmitting resistant microorganisms to humans, potentially complicating disease treatment and causing foodborne illness, posing a public health concern now and in the future.

Keywords

• Antibiotic resistance
• Enterotoxin
• Public health
• Staphylococcus aureus

Supporting Institution

the Research Fund of Van Yüzüncü Yıl University

Project Number

TYL-2021-9501

Thanks

This study was supported by the Research Fund of Van Yüzüncü Yıl University with the project number of TYL-2021-9501. We want to thank the institution.

References

1. Alghizzi M, Shami A (2021). The prevalence of Staphylococcus aureus and methicillin resistant Staphylococcus aureus in milk and dairy products in Riyadh, Saudi Arabia. Saudi J Biological Sci, 28 (12), 7098-7104.
2. Asao T, Kumeda Y, Kawai T et al. (2003). An extensive outbreak of staphylococcal food poisoning due to low-fat milk in Japan: estimation of enterotoxin A in the incriminated milk and powdered skim milk. Epidemiol Infect, 130, 33-40.
3. Balaban N, Rasooly A (2000). Staphylococcal enterotoxins. Int J Food Microbiol, 61, 1-10.
4. Bendahou A, Lebbadi M, Ennanei L, Essadqui FZ, Abid M (2008). Characterization of Staphylococcus species isolated from raw milk and milk products (lben and jben) in North Morocco. J Infect Develop Count, 2 (03), 218-225.
5. Bostan K, Çetin Ö, Büyükünal S, Ergün Ö (2006). The Presence of Staphylococcus aureus and Staphylococcal Enterotoxins in Ready-to-Cook Meatballs and White Pickled Cheese. İstanbul Üniv Vet Fak Derg, 32 (3), 31-39.
6. Bulajic S, Colovic S, Misic D et al. (2017). Enterotoxin production and antimicrobial susceptibility in Staphylococci isolated from traditional raw milk cheeses in Serbia. J Env Sci Healt, 52 (12), 864-870.
7. Cai H, Kou X, Ji H et al. (2021). Prevalence and characteristics of Staphylococcus aureus isolated from Kazak cheese in Xinjiang, China. Food Control, 123, 107759.
8. CLSI (2020). Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing. 30th ed. CLSI supplement M100. Clinical and Laboratory Standards Institute, USA.

9. Çiftçi İH, Altındiş M, Çetinkaya Z, Aşık G, Aktepe OC (2009). Klinik örneklerden izole edilen stafilokoklarda mecA varlığının araştırılması. Kocatepe Tıp Dergisi, 10 (1), 17-20.
10. El-Jakee JK, Aref NE, Gomaa A et al. (2013). Emerging of coagulase negative staphylococci as a cause of mastitis in dairy animals: An environmental hazard. Int J Vet Sci Med, 1-5.
11. Ertas N, Gonulalan Z, Yıldırım Y, Kum E (2010). Detection of Staphylococcus aureus enterotoxins in sheep cheese and dairy desserts by multiplex PCR technique. Int J Food Microbiol, 142 (1-2), 74-77.
12. EUCAST (2021). European Committee on Antimicrobial Susceptibility Testing (EUCAST) Breakpoint tables for interpretation of MICs and zone diameters Version 11.0. Acces Data: 01 January 2021. Erişim adresi: https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v_ 11.0_Breakpoint_Tables.pdf
13. EFSA and ECDC (2019). European Food Safety Authority and European Centre for Disease Prevention and Control. The European Union one health 2018 zoonoses report. EFSA Journal, 17 (12), 5926.
14. Giacinti G, Carfora V, Caprioli A et al. (2017). Prevalence and characterization of methicillin-resistant Staphylococcus aureus carrying mecA or mecC and methicillin-susceptible Staphylococcus aureus in dairy sheep farms in central Italy. J Dairy Sci, 100 (10), 7857-7863.
15. Godek Z, Mustafa N, Semerci AB, Tunç K (2021). Determination of Microbial Quality of White Cheese That Sale in Sakarya Bazaars. ALKÜ Fen Bilimleri Dergisi, 3 (1), 44-49.
16. Gücükoğlu A, Onur Kevenk T, Uyanik T, Çadirci Ö, Terzi G (2012). Detection of enterotoxigenic Staphylococcus aureus in raw milk and dairy products by multiplex PCR. J Food Sci, 77 (11), M620-M623.
17. Güngören A, Demircioğlu A, Saytekin AM (2022). Beyaz peynir örneklerinden Staphylococcus aureus suşlarının izolasyonu, makrolid-linkozamid-streptogramin B (MLSB) direnç fenotipleriyle, metisilin ve vankomisin duyarlılıklarının belirlenmesi. Harran Üniv Vet Fak Derg, 11 (1), 1-1.
18. Hennekinne JA, De Buyser ML, Dragacci S (2012). Staphylococcus aureus and its food poisoning toxins: characterization and outbreak investigation. FEMS Microbiol Rev, 36 (4), 815-836.
19. Jarraud S, Cozon G, Vandenesch F, Bes M, Etienne J (1999). Involvement of enterotoxins G and I in staphylococcal toxic shock syndrome and staphylococcal scarlet fever. J Clin Microbiol, 37 (8), 2446-2449.
20. Jay JM, Loessner MJ, Golden DA (2005). Modern Food Microbiology. 7th Edition. Springer, USA.
21. Jevons MP, Coe AW, Parker MT (1963). Methicillin resistance in staphylococci. Lancet, 904-907.
22. Kayılı E (2020). Investigation of coagulase positive Staphylococcus aureus in white cheese and determination of their antibiotic resistance. Mater Thesis. Ankara University Graduate School of Natural and Applied Sciences Department of Food Engineering, Ankara.
23. Kızanlık PK, Goksoy EO (2024). The prevalence, enterotoxigenic properties and antimicrobial susceptibility of Staphylococcus aureus isolated from various foods of animal origin. Vet Arhiv, 94, 43-54.
24. Kocak P (2014). The microbiological investigation of white, tulum, kasar and lor cheese produced at dairy processing premises located in Aydin. Master Thesis. Adnan Menderes University Health Sciences Institute, Aydın.
25. Lee HJ (2006). Occurence of methicillin-resistant Staphylococcus aureus strains from cattle and chicken, and analyses of their mecA, mecR1 and mecI genes. Vet Microbiol, 114, 155-159.
26. Lee JH (2003). Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Appl Environ Microbiol, 69 (11), 6489-6494.
27. Lindsay JA, Holden MT (2004). Staphylococcus aureus: superbug, super genome? Trend Microbiol, 12 (8), 378-385.
28. Mehrotra M, Wang G, Johnson WM (2000). Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1, and methicillin resistance. J Clin Microbiol, 38 (3), 1032-1035.
29. Monecke S, Gavier-Widen D, Mattsson R et al. (2012). Detection of mecC-positive Staphylococcus aureus (CC130-MRSA-XI) in diseased European hedgehogs (Erinaceus europaeus) in Sweden. PloS one, 8 (6), e66166.
30. Monday SR, Bohach GA (1999). Use of multiplex PCR to detect classical and newly described pyrogenic toxin genes in staphylococcal isolates. J Clin Microbiol, 37 (10), 3411-3414.
31. Normanno G, La Salandra G, Dambrosio A et al. (2007). Occurrence, characterization and antimicrobial resistance of enterotoxigenic Staphylococcus aureus isolated from meat and dairy products. Int J Food Microbiol, 115 (3), 290-296.
32. Obaidat MM, Bani Salman AE, Roess AA (2018). High prevalence and antimicrobial resistance of mecA Staphylococcus aureus in dairy cattle, sheep, and goat bulk tank milk in Jordan. Trop Anim Health Pro, 50 (2), 405-412.
33. Omoe K, Hu DL, Takahashi-Omoe H, Nakane A, Shinagawa K (2005). Comprehensive analysis of classical and newly described staphylococcal superantigenic toxin genes in Staphylococcus aureus isolates. FEMS microbiol Lett, 246 (2), 191-198.
34. Özpınar N (2011). The Determination of Antibiotic Resistance and Biofilm Creating Properties of Staphylococcus Aureus İsolates From Erzincan Tulum Cheese as Phenotypic and Genotypic. Mater Thesis. Erciyes University Health Sciences Institute, Kayseri.
35. Palavecino EL (2020). Clinical, Epidemiologic, and Laboratory Aspects of Methicillin-Resistant Staphylococcus aureus Infections. Yinduo Ji editor. Methicillin-Resistant Staphylococcus aureus (MRSA) Protocols Cutting-Edge Technologies and Advancements. Third edition. Humana Pres, USA.
36. Petersen A, Stegger M, Heltberg O et al. (2013). Epidemiology of methicillin-resistant Staphylococcus aureus carrying the novel mecC gene in Denmark corroborates a zoonotic reservoir with transmission to humans. Clin Microbiol Infect, 19 (1), E16-E22.
37. Prabakusuma AS, Zhu J, Shi Y et al. (2022). Prevalence and antimicrobial resistance profiling of Staphylococcus aureus isolated from traditional cheese in Yunnan, China. 3 Biotech, 12 (1), 1-15.
38. Riffon R, Sayasith K, Khalil H, Dubreuil P, Drolet M (2001). Development of a rapid and sensitive test for identification of major pathogens in bovine mastitis by PCR. J Clin Microbiol, 39 (7), 2584-2589.
39. Sağun E, Sancak H, Durmaz H (2001). Van'da kahvaltı salonlarında tüketime sunulan süt ürünlerinin mikrobiyolojik ve kimyasal kaliteleri üzerine bir araştırma. YYÜ Vet Fak Derg, 12 (1-2), 108-112.
40. Saha B, Singh AK, Ghosh A, Bal M (2008). Identification and characterization of a vancomycin-resistant Staphylococcus aureus isolated from Kolkata (South Asia). J Med Microbiol, 57 (1), 72-79.
41. Shanehbandi D, Baradaran B, Sadigh-Eteghad S, Zarredar H (2014). Occurrence of methicillin resistant and enterotoxigenic Staphylococcus aureus in traditional cheeses in the northwest of Iran. ISRN Microbiology, (1), 129580.
42. Spanu V, Scarano C, Cossu F et al. (2014). De Santis EP. Antibiotic resistance traits and molecular subtyping of Staphylococcus aureus isolated from raw sheep milk cheese. J Food Sci, 79 (10), 2066- 2071.
43. Spanu V, Virdis S, Scarano C et al. (2010). Antibiotic resistance assessment in Staphylococcus aureus strains isolated from raw sheep’s milk cheese. Vet Res Commun, 34 (1), 87-90.
44. SPSS (2006). IBM SPSS statistics version 13.0 for Windows. IBM, New York.
45. Srinivasan V, Sawant AA, Gillespie BE, Headrick SJ, Ceasaris L (2006). Prevalence of enterotoxin and toxic shock syndrome toxin genes in Staphylococcus aureus isolated from milk of cows with mastitis. Foodborne Pathog Dis, 3 (3), 274-283.
46. Sundsfjord A, Simonsen GS, Haldorsen BC et al. (2004). Genetic methods for detection of antimicrobial resistance. Apmis, 112 (11‐12), 815-837.
47. Taban BM, Hassankhani A, Aytac SA (2021). Investigation of mecA-and mecC-positive Staphylococcus aureus from raw milk and traditional artisanal dairy foods. Int J Food Prop, 24 (1), 954-964.
48. Thomas D, Chou S, Dauwalder O, Lina G (2007). Diversity in Staphylococcus aureus enterotoxins. Chem Immunol Allergy, 93, 24-41.
49. Titouche Y, Hakem A, Houali K et al. (2019). Emergence of methicillin-resistant Staphylococcus aureus (MRSA) ST8 in raw milk and traditional dairy products in the Tizi Ouzou area of Algeria. J Dairy Sci, 102 (8), 6876-6884.
50. TS (2021). Turkish Standard. Microbiology of the food chain - Horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species) Part 1: Method using Baird-Parker agar medium. (TS EN ISO 6888-1), Turkish Standards Institution, Ankara, Türkiye.
51. Turkish Food Codex (TFC) (2011). Microbiological Criteria Regulation. Access Date: 20 March 2021. Access address: https://www.resmigazete.gov.tr/eskiler/2011/12/20111229M3-6.htm.
52. Wongboot W, Chomvarin, C, Engchanil C, Chaimanee P (2013). Multiplex PCR for detection of superantigenic toxin genes in methicillin-sensitive and methicillin-resistant Staphylococcus aureus isolated from patients and carriers of a hospital in northeast Thailand. Southeast Asian J Trop Med Public Health, 44 (4), 660-671.
53. Yücel N, Anıl Y (2011). Identification and antimicrobial susceptibility of Staphylococcus aureus and coagulase negative staphylococci isolated from raw milk and cheese sample. Türk Hijyen ve Deneysel Biyoloji Dergisi, 68 (2), 73-78.
54. Zayda MG, Masuda Y, Hammad AM et al. (2020). Molecular characterisation of methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) Staphylococcus aureus isolated from bovine subclinical mastitis and Egyptian raw milk cheese. Int Dairy J, 104, 104646.