Isolation, Molecular Identification and Antibiotic Sensitivity of Enterococcus faecalis from Cheese Samples Sold in Giresun Public Markets

Year 2023, Volume: 13 Issue: 1, 264 - 274, 15.03.2023

Tuğba Cebeci

https://doi.org/10.31466/kfbd.1247882

Abstract

Enterococci are opportunistic pathogens responsible for food contamination that can pose an epidemic threat in the hospital setting due to their current virulence and multidrug resistance. This study was carried out to investigate the contamination E. faecalis in the samples of tulum cheese sold in the public markets and antibiotic resistance to 17 different antibiotics of the obtained isolates. A total of 50 tulum cheese samples were used as a material obtained from public markets 8 coastal districts of Giresun province. According to the results of the analysis; 15 out of 50 tulum cheese samples (30%) were found contaminated with E. faecalis. Two isolates against (13.3%) fosfomycin, 2 isolates against (13.3%) levofloxacin, 2 isolates against (13.3%) erythromycin, 4 isolates against (26.6%) teicoplanin, 4 isolates against (26.6%) rifampin and 6 isolates against (40%) tetracycline resistance was confirmed by PCR in 15 E. faecalis isolates. Vancomycin resistance was found to be 93.3% in E. faecalis isolates. None of the strains showed resistance to high level streptomycin and high level gentamicin. As a result of the detection of this pathogen in cheese samples, there is a potential risk factor that seriously affects public health. From farm to table, it is recommended to take necessary hygiene measures during processing, consumption, and sale of the products.

Keywords

Tulum cheese, Enterococcus faecalis, MALDI-TOF MS, PCR, antibiotic sensitivity

Giresun Pazarlarında Satışa Sunulan Peynir Örneklerinden Enterococcus faecalis izolasyonu, Moleküler Tanımlanması ve Antibiyotik Duyarlılıklarının Araştırılması

Abstract

Enterokoklar, gıda kontaminasyonundan sorumlu, mevcut virülansları ve çoklu ilaca dirençleri nedeniyle hastane ortamında salgın bir tehdit oluşturabilen fırsatçı patojenlerdir. Bu çalışma, pazarlarda satılan tulum peyniri örneklerindeki Enterococcus faecalis ( E. faecalis) kontaminasyonunu ve elde edilen izolatların 17 farklı antibiyotiğe karşı dirençliliklerini araştırmak amacıyla gerçekleştirilmiştir. Giresun İli, 8 sahil ilçesinde kurulan pazarlardan temin edilen toplam 50 adet tulum peynir örneği materyal olarak kullanılmıştır. Yapılan analiz sonuçlarına göre; 50 adet tulum peyniri örneğinin 15 (%30) tanesinin E. faecalis ile kontamine olduğu belirlenmiştir. PCR ile doğrulanan 15 E. faecalis izolatının, 6(%40)'sının tetrasiklin'e, 4(%26.6)'ünün rifampin'e, 4(%26.6)'ünün teikoplanin'ine, 3(%20)'ünün siprofloksasin'e, 3(%20)'ünün kloramfenikol'e, 2(%13.3)'sinin eritromisin'e, 2(%13.3)'sinin levofloksasin'e ve 2(%13.3)'sinin fosfomisin'e karşı dirençli olduğu belirlenmiştir. E. faecalis izolatlarında vankomisin direnci %93.3 olarak tespit edilmiştir. İzole edilen suşlarda yüksek düzey streptomisin direnci ve yüksek düzey gentamisin direnci saptanmamıştır. Elde edilen veriler sonucunda, peynir örneklerinde bu patojenin saptanması, halk sağlığını ciddi anlamda etkileyen potansiyel risk faktörü olarak karşımıza çıkmaktadır. Çiftlikten sofraya kadar gelen süreçte; ürünlerin işlenmesi, tüketilmesi ve satışı sırasında gerekli hijyen tedbirlerinin alınması önerilmektedir.

Keywords

Tulum peyniri, Enterococcus faecalis, MALDI-TOF MS, antibiyotik duyarlılığı, gıda hijyeni, halk sağlığı

References

• Abouelnaga, M., Lamas, A., Quintela-Baluja, M., Osman, M., Miranda, J. M., Cepeda, A., and Franco, C. M. (2016). Evaluation of the extent of spreading of virulence factors and antibiotic resistance in Enterococci isolated from fermented and unfermented foods. Annals of Microbiology, 66(2), 577-585.
• Aragão, B. B., Trajano, S. C., de Oliveira, R. P., da Conceição Carvalho, M., de Carvalho, R. G., Juliano, M. A., Junior, J. W. P., and Mota, R. A. (2022). Occurrence of emerging multiresistant pathogens in the production chain of artisanal goat coalho cheese in Brazil. Comparative Immunology, Microbiology and Infectious Diseases, 84, 101785.
• Ayhan, Ş., Kahve, H. İ., Aydın, F., and Ardıç, M. (2020). Antibiotic resistance and hemolytic activity in enterococci isolated from tulum cheese sold in Aksaray province. Gıda, 45(4), 689-698.
• Chajęcka-Wierzchowska, W., Zadernowska, A., and García-Solache, M. (2020). Ready-to-eat dairy products as a source of multidrug-resistant Enterococcus strains: Phenotypic and genotypic characteristics. Journal of dairy science, 103(5), 4068-4077.
• Chotinantakul, K., Chansiw, N., and Okada, S. (2018). Antimicrobial resistance of Enterococcus spp. isolated from Thai fermented pork in Chiang Rai Province, Thailand. Journal of Global Antimicrobial Resistance, 12, 143-148.
• Clinical and Laboratory Standard Institute (2022) Performance Standards for Antimicrobial Susceptibility Testing. 32th Informational Supplement CLSI supplement M100-A32. Wayne, PA: Clinical and Laboratory Standards Institute.
• Colak, H., Hampikyan, H., Bingol, E. B., and Ulusoy, B. (2007). Prevalence of L. monocytogenes and Salmonella spp. in tulum cheese. Food Control, 18(5), 576-579.
• Del Rio, B., Sánchez-Llana, E., Martínez, N., Fernández, M., Ladero, V., and Alvarez, M. A. (2021). Isolation and characterization of Enterococcus faecalis-infecting bacteriophages from different cheese types. Frontiers in Microbiology, 3409.
• Economou, V., Sakkas, H., Delis, G., and Gousia, P. (2016). Antibiotic resistance in Enterococcus spp. Friend or foe?. Foodborne pathogens and antibiotic resistance, 365-395.
• Elmalı, M., and Can, H. Y. (2018). The prevalence, vancomycin resistance and virulence gene profiles of Enterococcus species recovered from different foods of animal origin. Veterinarski arhiv, 88(1), 111-124.
• EUCAST (2022): The European Committee on Antimicrobial Susceptibility Testing. EUCAST supplement 2022: Breakpoint tables for interpretation of MICs and zone diameters, version 12.0, Växjö, Sweden.
• Giraffa, G. (2003). Functionality of enterococci in dairy products. International journal of food microbiology, 88(2-3), 215-222.
• Han, D., Unno, T., Jang, J., Lim, K., Lee, S. N., Ko, G., Sadowsky, M. J., and Hur, H. G. (2011). The occurrence of virulence traits among high-level aminoglycosides resistant Enterococcus isolates obtained from feces of humans, animals, and birds in South Korea. International journal of food microbiology, 144(3), 387-392.
• Hussein, W. E., Abdelhamid, A. G., Rocha-Mendoza, D., García-Cano, I., and Yousef, A. E. (2020). Assessment of safety and probiotic traits of Enterococcus durans OSY-EGY, isolated from Egyptian artisanal cheese, using comparative genomics and phenotypic analyses. Frontiers in microbiology, 11, 608314.
• Jahansepas, A., Sharifi, Y., Aghazadeh, M., and Ahangarzadeh Rezaee, M. (2020). Comparative analysis of Enterococcus faecalis and Enterococcus faecium strains isolated from clinical samples and traditional cheese types in the Northwest of Iran: Antimicrobial susceptibility and virulence traits. Archives of microbiology, 202, 765-772.
• Jahansepas, A., Aghazadeh, M., Rezaee, M. A., Heidarzadeh, S., Mardaneh, J., Mohammadzadeh, A., and Pouresmaeil, O. (2022). Prevalence, antibiotic resistance and virulence of Enterococcus spp. isolated from traditional cheese types. Ethiopian Journal of Health Sciences, 32(4), 799-808.
• Kang, H. J., Yoon, S., Kim, K., and Lee, Y. J. (2021). Characteristics of high-level aminoglycoside-resistant Enterococcus faecalis isolated from bulk tank milk in Korea. Animals, 11(6), 1724.
• Kousta, M., Mataragas, M., Skandamis, P., and Drosinos, E. H. (2010). Prevalence and sources of cheese contamination with pathogens at farm and processing levels. Food control, 21(6), 805-815.
• Kürekci, C., Önen, S. P., Yipel, M., Aslantaş, Ö., and Gündoğdu, A. (2016). Characterisation of phenotypic and genotypic antibiotic resistance profile of enterococci from cheeses in Turkey. Korean Journal for Food Science of Animal Resources, 36(3), 352.
• Levy, S. B., and Marshall, B. (2004). Antibacterial resistance worldwide: causes, challenges and responses. Nature medicine, 10(Suppl 12), S122-S129.
• Moon, H. W., Lee, S. H., Chung, H. S., Lee, M., and Lee, K. (2013). Performance of the Vitek MS matrix-assisted laser desorption ionization time-of-flight mass spectrometry system for identification of Gram-positive cocci routinely isolated in clinical microbiology laboratories. Journal of medical microbiology, 62(9), 1301-1306.
• Mrkonjic Fuka, M., Zgomba Maksimovic, A., Tanuwidjaja, I., Hulak, N., and Schloter, M. (2017). Characterization of enterococcal community isolated from an artisan Istrian raw milk cheese: Biotechnological and safety aspects. Food Technology and Biotechnology, 55(3), 368-380.
• Palmeri, M., Mancuso, I., Gaglio, R., Arcuri, L., Barreca, S., Barbaccia, P., and Scatassa, M. L. (2020). Identification and evaluation of antimicrobial resistance of enterococci isolated from raw ewes’ and cows’ milk collected in western Sicily: A preliminary investigation. Italian Journal of Food Safety, 9(4).
• Ramos, S., Silva, V., Dapkevicius, M. D. L. E., Igrejas, G., and Poeta, P. (2020). Enterococci, from harmless bacteria to a pathogen. Microorganisms, 8(8), 1118.
• Russo, N., Caggia, C., Pino, A., Coque, T. M., Arioli, S., and Randazzo, C. L. (2018). Enterococcus spp. in ragusano PDO and pecorino siciliano cheese types: a snapshot of their antibiotic resistance distribution. Food and Chemical Toxicology, 120, 277-286.
• Rubinstein, E., and Keynan, Y. (2013). Vancomycin-resistant enterococci. Critical care clinics, 29(4), 841-852. Sadek, O. A., and Koriem, A. M. (2022). Multidrug Resistance and Virulence Factors of Enterococci Isolated from Milk and Some Dairy Desserts. Journal of food quality and hazards control.
• Sanlibaba, P., and Senturk, E. (2018). Prevalence, characterization and antibiotic resistance of enterococci from traditional cheeses in Turkey. International Journal of Food Properties, 21(1), 1955-1963.
• Smith, D. L., Harris, A. D., Johnson, J. A., Silbergeld, E. K., and Morris Jr, J. G. (2002). Animal antibiotic use has an early but important impact on the emergence of antibiotic resistance in human commensal bacteria. Proceedings of the National Academy of Sciences, 99(9), 6434-6439.
• Soni, M. M., Anjaria, P. A., Bhavsar, P. P., Nayak, J. B., Chaudhary, J. H., and Brahmbhatt, M. N. (2022). Isolation and Molecular Detection of Enterococcus faecalis from Cow Milk of Anand City with Special Reference to Biofilm Production and Multiple Drug Resistance. Indian Journal of Veterinary Sciences & Biotechnology, 18(4), 6-9.
• Výrostková, J., Regecová, I., Dudriková, E., Marcinčák, S., Vargová, M., Kováčová, M., and Maľová, J. (2021). Antimicrobial resistance of Enterococcus spp. isolated from sheep and goat cheeses. Foods, 10(8), 1844.
• Yuksel, F. N., Akcelik, N., and Akcelik, M. (2015). Incidence of antibiotic resistance and virulence determinants in Enterococcus faecium and Enterococcus faecalis strains, isolated from traditional cheeses in Turkey. Molecular Genetics, Microbiology and Virology, 30, 206-215.