AKSARAY İLİNDE SATIŞA SUNULAN TULUM PEYNİRLERİNDEN İZOLE EDİLEN ENTEROKOKLARIN ANTİBİYOTİK DİRENÇLİLİKLERİ İLE HEMOLİTİK AKTİVİTELERİNİN BELİRLENMESİ

Bu çalışmada, Aksaray ilinde satışa sunulan tulum peyniri örneklerinden izole edilen enterokokların hemolitik aktivitesi ile antibiyotik duyarlılığının değerlendirilmesi amaçlanmıştır. İzolatlar, 16S rRNA gen sekanslaması sonucunda E. faecium (n = 30), E. faecalis (n = 25) ve E. durans (n = 7) olarak tanımlanmıştır. Üç izolat (E. faecalis ATC12, E. faecium ATC49 ve E. faecium ATC54) ß-hemolitik aktivite gösterirken, diğerlerinin hemolitik aktivite göstermediği saptanmıştır. İzolatların toplamda nalidiksik aside (%100), oksasiline (%92) ve steptomisine (%72.6) karşı orta veya yüksek derecede direnç gösterdiği saptanmıştır. E. faecalis suşlarının klinik olarak önem arz eden çeşitli antibiyotiklere E. faecium ve E. durans suşlarından daha dirençli fenotipleri olduğu görülmüştür. Çoklu antibiyotik direncinin %41.93 olduğu saptanmıştır. Sonuçlar değerlendirildiğinde Aksaray'da üretilen Tulum peynirinin gıda yolu ile antibiyotik direncinin yayılmasında tehlikeli bir araç olduğu kanısına varılmıştır.

Anahtar Kelimeler:

Tulum peyniri, Enterococcus, hemolitik aktivite, antibiyotik dirençlik

ANTIBIOTIC RESISTANCE AND HEMOLYTIC ACTIVITY IN ENTEROCOCCI ISOLATED FROM TULUM CHEESE SOLD IN AKSARAY PROVINCE

This study aimed to evaluate the hemolytic activity and antibiotic susceptibility of enterococci isolated from Tulum cheese samples sold in Aksaray province. The isolates were identified as E. faecium (n = 30), E. faecalis (n = 25), and E. durans (n = 7) as a result of 16S rRNA gene sequencing. Three strains (E. faecalis ATC12, E. faecium ATC49, and E. faecium ATC54) exhibited ß-hemolytic activity, whereas others were non-hemolytic. Enterococci were found to have an intermediary or high resistance to nalidixic acid (%100), oxacillin (92%), and streptomycin (72.6%), respectively. E. faecalis strains had more resistant phenotypes to various clinically significant antibiotics than E. faecium and E. durans. Multi-drug resistance was found in 41.93% of the isolates. According to the results, Tulum cheese produced in Aksaray could be a potential vehicle for the transmission of antibiotic resistance via the food chain.

Keywords:

Enterococcus, hemolytic activity, virulence factor, antibiotic resistance,

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Arslaner, A., Turkmen, O. (2020). Erzincan Tulum Cheese. Turkish JAF Sci Tech, 8(4): 932-940, doi: 10.24925/turjaf.v8i4.932-940.3170.
Aydin, F., Ardic, M. (2019). Farklı illerden toplanan sıkma peynirlerinin mikrobiyolojik ve kimyasal özellikleri. GIDA 44(5): 826-836, doi: 10.15237/gida.GD19088.
Aydin, F., Ozer, G., Alkan, M., Cakir, I. (2020). The utility of iPBS retrotransposons markers to analyze genetic variation in yeast. Int J Food Microbiol, 325: 108647, doi: 10.1016/j.ijfoodmicro.2020.108647.
Baker, G.C., Smith, J.J., Cowan, D.A. (2003). Review and re-analysis of domain-specific 16S primers. J Microbiol Methods, 55(3): 541-555, doi: 10.1016/j.mimet.2003.08.009.
Bulajić, S., Tambur, Z. Opačić, D., Miljkovic-Semlimovic, B., Doder, R., Cenic-Milosevic, D. (2015) Characterization of antibiotic resistance phenotypes and resistance genes in Enterococcus spp. isolated from cheeses. Arch Biol Sci, 67(1):139–146, doi:10.2298/ABS140426016B.
Bulut, C., Gunes, H., Okuklu, B., Harsa, S., Kilic, S., Coban, H.S., Yenidunya, A.F. (2005). Homofermentative lactic acid bacteria of a traditional cheese, Comlek peyniri from Cappadocia region, J Dairy Res, 72, 19-24, doi: 10.1017/S0022029904000536.
Cariolato, D., Andrighetto, C., Lombardi, A. (2008). Occurrence of virulence factors and antibiotic resistances in Enterococcus faecalis and Enterococcus faecium collected from dairy and human samples in North Italy. Food Control, 19(9): 886-892, doi: 10.1016/j.foodcont.2007.08.019.
Citak, S., Yucel, N., Orhan, S. (2004). Antibiotic resistance and incidence of Enterococcus species in Turkish white cheese. Int J Dairy Technol, 57(1): 27-31, doi: 10.1111/j.1471-0307.2004.00122.x.
CLSI (2011) Performance Standards for Antimicrobial Susceptibility Testing 21th Informational Supplement. Document M100-S21. Clinical Laboratuary Standarts Institue.
CLSI (2016). Performance Standards for Antimicrobial Susceptibility Testing. 26th Informational Supplement, Document M100-S26. Clinical Laboratuary Standarts Institue.
Dagdemir, E., Ozdemir, S. (2008). Technological characterization of the natural lactic acid bacteria of artisanal Turkish White Pickled cheese. Int J Dairy Technol, 61(2) 133-140, doi: 10.1111/j.1471-0307.2008.00394.x.
Domingos-Lopes, M.F.P, Stanton, C., Ross, P.R., Dapkevicius, M.L.E., Silva, C.C.G. (2017). Genetic diversity, safety and technological characterization of lactic acid bacteria isolated from artisanal Pico cheese. Food Microbiol, 63: 178-190, doi: 10.1016/j.fm.2016.11.014.
EFSA (2013). Scientific Opinion on the maintenance of the list of QPS biological agents intentionally added to food and feed (2013 update). EFSA Journal, 11(11), 3449.
Franz, C.M., Holzapfel, W.H., Stiles, M.E. (1999). Enterococci at the crossroads of food safety?. Int J Food Microbiol, 47(1-2): 1-24, doi: 10.1016/S0168-1605(99)00007-0.
Furlaneto-Maia, L., Rocha, K.R., Henrique, F.C., Giazzi, A., Furlaneto, M.C. (2014). Antimicrobial resistance in Enterococcus sp isolated from soft cheese in Southern Brazil. Adv Microbiol, 4(3): 175-181, doi: 10.4236/aim.2014.43023
Garrido, A.M., Galvez, A., Pulido, R.P. (2014). Antimicrobial resistance in enterococci. J Infect Dis Ther, 2(4):150, doi: 10.4172/2332-0877.1000150.
Giraffa, G. (2003). Functionality of enterococci in dairy products. Int J Food Microbiol, 88(2-3): 215-222, doi: 10.1016/S0168-1605(03)00183-1.
Gomes, B.C., Esteves, C.T., Palazzo, I.C.V., Darini, A.L.C. Felis, G.E., Sechi, L.A., Franco, B.D.G.M., De Martinis, E.C.P. (2008) Prevalence and Characterization of Enterococcus spp. isolated from Brazilian foods. Food Microbiol, 25: 668–675, doi: 10.1016/j.fm.2008.03.008.
Ispirli, H., Demirbas, F., Dertli, E. (2017). Characterization of functional properties of Enterococcus spp. isolated from Turkish white cheese. LWT-Food Sci Technol, 75: 358-365, doi: 10.1016/j.lwt.2016.09.010.
Jahan, M., Zhanel, G.G, Sparling, R., Holley, R.A. (2015). Horizontal transfer of antibiotic resistance from Enterococcus faecium of fermented meat origin to clinical isolates of E. faecium and Enterococcus faecalis. Int J Food Microbiol, 199: 78-85, doi: 10.1016/j.ijfoodmicro.2015.01.013.
Jamet, E., Akary, E., Poisson, M.A., Chamba, J.F., Bertrand, X., Serror, P. (2012). Prevalence and characterization of antibiotic resistant Enterococcus faecalis in French cheeses. Food Microbiol, 31(2), 191-198, doi: 10.1016/j.fm.2012.03.009.
Kankaya, D.A., Tuncer, B.Ö., Tuncer, Y. (2017). Gıda kaynaklı enterokokların potansiyel risk faktörleri. GIDA 42(1): 8-19, doi: 10.15237/gida.GD16048.
Kirmaci, H.A., Ozer, B.H., Akcelik, M., Akcelik. N. (2016). Identification and characterisation of lactic acid bacteria isolated from traditional Urfa cheese. Int J Dairy Technol, 69 (2): 301-307, doi: 10.1111/1471-0307.12260.
Kumar, S., Stecher, G., Li, M., Knyaz, C., Tamura, K. (2018). MEGA X: molecular evolutionary genetics analyses across computing platforms. Mol Biol Evol, 35: 1547–1549, doi: 10.1093/molbev/msy096.
Maia, L.F., Giazzi, A., Brandalize, C., Katsuda, M.S., Rocha, K.R., Terra, M.R., Furlaneto, M. C. (2017). Isolation and characterization of potential probiotic enterococci strains from soft cheese flora. Afr J Microbiol Res, 11(12): 482-487, doi: 10.5897/AJMR2017.8429.
Martino, G.P., Quintana, I.M., Espariz, M., Blancato, V.S., Magni, C. (2016). Aroma compounds generation in citrate metabolism of Enterococcus faecium: Genetic characterization of type I citrate gene cluster. Int J Food Microbiol, 218, 27-37, doi: 10.1016/j.ijfoodmicro.2015.11.004.
Miller, W.R., Munita, J.M., Arias, C.A. (2014). Mechanisms of antibiotic resistance in enterococci. Expert Rev Anti-Infe, 12(10): 1221-1236, doi: 10.1586/14787210.2014.956092.
Miller, W.R., Murray, B.E., Rice, L.B., Arias, C. A. (2016). Vancomycin-resistant enterococci: therapeutic challenges in the 21st century. Infect Dis Clin, 30(2), 415-439, doi: 10.1016/j.idc.2016.02.006.
Oner, Z., Sagdic, O., Simsek, B. (2004). Lactic acid bacteria profiles and tyramine and tryptamine contents of Turkish tulum cheeses. Eur Food Res Technol, 219(5): 455-459, doi: 10.1007/s00217-004-0962-x.
Ozdemir, R., Tuncer, Y. (2020). Detection of antibiotic resistance profiles and aminoglycoside-modifying enzyme (AME) genes in high-level aminoglycoside-resistant (HLAR) enterococci isolated from raw milk and traditional cheeses in Turkey. Mol Biol Rep, 1-10, doi: 10.1007/s11033-020-05262-4.
Patterson, A.J., Colangeli, R., Spigaglia, P., Scott, K.P. (2007). Distribution of specific tetracycline and erythromycin resistance genes in environmental samples assessed by macroarray detection. Environ Microbiol, 9(3): 703-715, doi: 10.1111/j.1462-2920.2006.01190.x
Pérez-Pulido, R., Abriouel, H., Omar, N. B., Lucas, R., Martinez-Canamero, M., & Galvez, A. (2006). Safety and potential risks of enterococci isolated from traditional fermented capers. Food Chem Toxicol, 44(12): 2070-2077, doi: 10.1016/j.fct.2006.07.008.
Pesavento, G., Calonico, C., Ducci, B., Magnanini, A., Nostro, A.L. (2014). Prevalence and antibiotic resistance of Enterococcus spp. isolated from retail cheese, ready-to-eat salads, ham, and raw meat. Food Microbiol, 41: 1-7, doi: 10.1016/j.fm.2014.01.008.
Psoni, L., Kotzamanides, C., Andrighetto, C., Lombardi, A., Tzanetakis, N., Litopoulou-Tzanetaki, E. (2006). Genotypic and phenotypic heterogeneity in Enterococcus isolates from Batzos, a raw goat milk cheese. Int J Food Microbiol, 109(1-2): 109-120, doi: 10.1016/j.ijfoodmicro.2006.01.027.
Russo, N., Caggia, C., Pino, A., Coque, T.M., Arioli, S., Randazzo, C.L. (2018). Enterococcus spp. in ragusano PDO and pecorino siciliano cheese types: a snapshot of their antibiotic resistance distribution. Food Chem Toxicol, 120: 277-286, doi: 10.1016/j.fct.2018.07.023.
Sanlibaba, P., Senturk, E. (2018). Prevalence, characterization and antibiotic resistance of enterococci from traditional cheeses in Turkey. Int J Food Prop, 21(1): 1955-1963, doi: 10.1080/10942912.2018.1489413.
Semedo, T., Santos, M.A., Martins, P., Lopes, M.F.S., Marques, J.J.F., Tenreiro, R., Crespo, M.T.B. (2003). Comparative study using type strains and clinical and food isolates to examine hemolytic activity and occurrence of the cyl operon in enterococci. J Clin Microbiol, 41(6): 2569-2576, doi: 10.1128/JCM.41.6.2569-2576.2003.
Tekinsen, K.K., Akar, D. (2017). Erzincan tulum peyniri. Atatürk University J Vet Sci, 12(2): 218-226, doi: 10.17094/ataunivbd.347980.
Thompson, J.D., Higgins, D.G., Gibson, T.J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res, 22: 4673–4680, doi: 10.1093/nar/22.22.4673.
Togay, S.O., Temiz, A. (2011). Gıda kaynaklı enterokokların gıda ve insan sağlığı yönünden önemi. GIDA 36(5): 303-310.
Trivedi, K., Cupakova, S., Karpiskova, R. (2011). Virulence factors and antibiotic resistance in enterococci isolated from food-stuffs. Vet Med, 56(7): 352-357.
Wegener, H.C. (2003). Antibiotics in animal feed and their role in resistance development. Curr Opin Microbiol, 6(5): 439-445, doi: 10.1016/j.mib.2003.09.009.
Yerlikaya, O., Akbulut, N. (2020). In vitro characterisation of probiotic properties of Enterococcus faecium and Enterococcus durans strains isolated from raw milk and traditional dairy products. Int J Dairy Technol, 73 (1): 98-107, doi: 10.1111/1471-0307.12645.
Yogurtcu, N.N., Tuncer, Y. (2013). Antibiotic susceptibility patterns of Enterococcus strains isolated from Turkish Tulum cheese. Int J Dairy Technol, 66(2): 236-242, doi: 10.1111/1471-0307.12014.
Yuksel, F.N., Akcelik, N., Akcelik, M. (2015). Incidence of antibiotic resistance and virulence determinants in Enterococcus faecium and Enterococcus faecalis strains, isolated from traditional cheeses in Turkey. Mol Genet Microbiol Virol 30(4,): 206-215, doi: 10.3103/S089141681504014X.