Tuğçe GÜRKAN, Meryem Burcu KÜLAHCI, Sumru ÇITAK

Gıda örneklerinden izole edilen Enterococcus türlerinin çeşitli virülans özellikleri, biyofilm oluşumu ve antibiyotik dirençliliklerinin belirlenmesi

Çalışmamızda Eylül -Aralık 2020 tarihlerinde 20 çiğ süt, 20 peynir, 20 çiğ tavuk eti, ve 20 çiğ et olmak üzere 80 adet gıda ürününden izole edilen Enterococcus izolatlarının tanımlanması için temel biyokimyasal testler ve identifikasyon işlemi MALDI-TOF Kütle Spektrofotometresi ile yapılarak 60 izolattan 42 (%70) Enterococcus faecalis ve 18 (%30) Enterococcus faecium türü izole edilmiştir. E.faecalis ve E.faecium türlerinin Kirby-Bauer disk difüzyon metoduyla antibiyotik direnç profilleri, lipaz, jelatinaz, DNaz aktiviteleri, hemolitik aktivitesi, S tabakası varlığı ve biyofilm aktiviteleri incelenmiştir. E.faecalis türlerinin en fazla duyarlı olduğu antibiyotikler %100 kloramfenikol, streptomisin, vankomisin ve teikoplanin, %97,6 gentamisin ve norfloksasin, %95,2 ampisilin, %92,8 siprofloksasin olarak saptanmıştır. En fazla dirençlilik gösterdiği antibiyotik %90,4 tetrasiklindir. E.faecium türlerinin en fazla duyarlı olduğu antibiyotikler %100 ampisilin, kloramfenikol, streptomisin, gentamisin, vankomisin ve teikoplanin, % 77,7 siprofloksasin, %72,2 norfloksasindir. En fazla dirençlilik gösterdiği antibiyotik %61,1 tetrasiklindir. Lipaz ve jelatinaz aktiviteleri tümünde negatif, DNaz aktiviteleri ise tümünde pozitif sonuç vermiştir. S tabakası varlığı E.faecalis’te %83,3 ve E.faecium’da %16,6 bulunmuştur. Biyofilm aktiviteleri Kongo Red Agar’da 60 adet Enterococcus spp. türü bakterinin 2‘si (%3.3) kuvvetli, 22‘si (%36.6) orta kuvvetli, 36’sı (%60) zayıf biyofilm pozitif üretmiştir. Mikroplak yönteminde E.faecalis’te 32’si (%53,3) zayıf, 8’i (%13,3) orta kuvvetli, 2’si (%3,3) kuvvetli biyofilm üretmiştir. E.faecium’da 17’si (%28,3) zayıf, 1’i (%1,6) orta kuvvetli biyofilm üretmiştir. Kuvvetli biyofilm üreten Enterococcus türleri saptanmamıştır.

Anahtar Kelimeler:

Gıda, Enterococcus, Virülans özellikler, Virülans genler, Biyofilm oluşumu, Antibiyotik dirençliliği, Disk difüzyon.

Determination of various virulence properties and biofilm formation antibiotic resistance of enterococcus species isolated from food samples

In our study, basic biochemical tests and identification process for the identification of Enterococcus isolates isolated from 80 food products, including 20 raw milk, 20 cheese, 20 raw chicken meat, and 20 raw meat, between September and December 2020, were performed with MALDI-TOF Mass Spectrophotometer and 42 of 60 isolates were identified. (70%) Enterococcus faecalis and 18 (30%) Enterococcus faecium species were isolated. Antibiotic resistance profiles, lipase, gelatinase, DNase activities, hemolytic activity, presence of S layer and biofilm activities of E.faecalis and E.faecium species were investigated by Kirby-Bauer disc diffusion method. The antibiotics to which E.faecalis species were most susceptible were found to be 100% chloramphenicol, streptomycin, vancomycin and teicoplanin, 97.6% gentamicin and norfloxacin, 95.2% ampicillin, 92.8% ciprofloxacin. The antibiotic with the highest resistance is 90.4% tetracycline. The antibiotics to which E.faecium species are most susceptible are 100% ampicillin, chloramphenicol, streptomycin, gentamicin, vancomycin and teicoplanin, 77.7% ciprofloxacin, 72.2% norfloxacin. The antibiotic with the highest resistance is 61.1% tetracycline. Lipase and gelatinase activities were negative in all, and DNase activities were positive in all. Presence of S layer was found 83.3% in E.faecalis and 16.6% in E.faecium. Biofilm activities of 60 Enterococcus spp. on Congo Red Agar. Of the bacteria, 2 (3.3%) were strong, 22 (36.6%) were medium strong, and 36 (60%) were weak biofilm positive. In the microplate method, 32 (53.3%) weak, 8 (13.3%) medium-strong, 2 (3.3%) strong biofilms were produced in E.faecalis. In E.faecium, 17 (28.3%) produced weak biofilms and 1 (1.6%) produced medium-strength biofilm. Strong biofilm-producing Enterococcus species were not detected.

Keywords:

Food, Enterococcus species Enterococcus, Enterococci, Virulence determinants, Virulence genes, Biofilm formation, Antibiotic resistance, Disc diffusion.,

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