Hayvansal kaynaklı gıdalarda enterokokların varlığı, tür düzeyinde identifikasyonları ve antibiyotik dirençliliklerinin belirlenmesi
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Date
2014
Authors
Muş, Tülay Elal
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Publisher
Uludağ Üniversitesi
Abstract
Bu tez çalışması Nisan – Aralık 2011 tarihleri arasında Güney Marmara Bölgesi'nde satışa sunulan hayvansal gıdalarda enterokokların varlığını belirlemek, tür düzeyinde identifikasyonunu yapmak ve antibiyotik direnç profillerini ortaya koymak amacıyla yapıldı. Bu amaçla 150 adet et ve et ürünü (parça et, kıyma, köfte, sucuk, salam, sosis, pastırma), 150 adet süt ve süt ürünü (peynir, yoğurt, kefir, tereyağ), 75 adet tavuk eti (göğüs, but, kanat) ve işlenmiş kanatlı eti ürünü (piliç burger, piliç köfte, nugget, şinitzel) olmak üzere toplam 375 adet gıda örneği materyal olarak kullanıldı. Klasik mikrobiyolojik tekniklerle, analiz edilen örneklerin % 54.13'ünde (203/375) Enterococcus spp. kontaminasyonu gözlendi. API 20 STREP bakteri tanımlama sistemi kullanılarak, izolatların 92'si E. faecium, 88'i E. faecalis, 20'si E. durans, 2'si E. avium ve 1'i E. gallinarum olarak tür düzeyinde identifiye edildi. İdentifikasyonları tamamlanan tüm izolatların ampisilin (10 µg), penisilin (10 ünite), linezolid (30 µg), kinupristin/dalfopristin (15 µg), vankomisin (30 µg), teikoplanin (30 µg), siprofloksasin (5 µg), tetrasiklin (30 µg), yüksek düzey aminoglikozit direnci için gentamisin (120 µg) ve streptomisin (300 µg) antibiyotiklerine duyarlılık profilleri disk difüzyon tekniği ile test edildi. Vankomisin ve teikoplanin antibiyotiklerine dirençli olduğu görülen suşların, E-test tekniği ile minimum inhibitör konsantrasyon (MIK) değerleri tespit edildi. Antibiyotik duyarlılık testi sonucu tüm suşların % 18,7'sinin (38 suş) bir antibiyotiğe, % 5.9'unun (12 suş) iki antibiyotiğe, % 2.9'unun (6 suş) 3 antibiyotiğe ve % 0.5'inin (1 suş) 6 antibiyotiğe dirençli olduğu belirlendi. E. faecium suşları arasında dirençli suşların oranı tetrasiklin için % 18.5 (17/92) ve penisilin için % 3.3 (3/92) iken; bu oran vankomisin, teikoplanin, ampisilin, kinupristin/dalfopristin ve siprofloksasin antibiyotiklerinin her biri için % 1.1 (1/92) olarak saptandı. E. faecium suşlarının % 6.5'inde (6/92) birden fazla antibiyotiğe dirençlilik ortaya kondu. Linezolid, yüksek düzey streptomisin ve gentamisin antibiyotiklerine karşı tüm E. faecium suşlarının duyarlı olduğu gözlendi. E. faecalis suşlarının % 28.4'ünün (25/88) tetrasiklin, % 25'inin (22/88) kinupristin/dalfopristin, % 3.4'ünün (3/88) siprofloksasin, % 2.3'ünün (2/88) linezolid ve yüksek düzey streptomisine dirençli olduğu tespit edildi. Yine vankomisin, teikoplanin, ampisilin, penisilin ve yüksek düzey gentamisin antibiyotiklerinin her birine direnç gösteren suşların oranı % 1.1 (1/88) olarak belirlendi. E. faecalis suşlarının % 14.8'i (13/88) test edilen antibiyotiklerden en az ikisine dirençliydi. E. durans suşlarından yalnızca 2'sinin (% 10) tetrasiklin dirençliliğine sahip olduğu, kalan E. durans suşları ile E. avium ve E. gallinarum'un tüm suşlarının yukarıda adı geçen antibiyotiklere duyarlılık gösterdiği ortaya kondu. Disk diffizyon yöntemiyle vankomisin ve teikoplanine dirençli olduğu belirlenen E. faecium ve E. faecalis suşlarına uygulanan E-testin sonuçları, MIK değerlerinin 4 µg/ml'nin altında olduğunu, dolayısıyla bu suşların vankomisin ve teikoplanine duyarlı olduğunu gösterdi. Sonuç olarak hayvansal gıdalarda enterokokların varlığı ve dağılımı belirlenmiş ve izole edilen suşlarda farklı antibiyotiklere karşı direnç saptanmıştır. Mevcut tez çalışması, dirençli suşların bu genlerini patojen olan ve olmayan diğer bakterilere aktarabilme özellikleri göz önüne alındığında, antibiyotik dirençli bakterilerin doğada yayılmasında enterokokların önemini ortaya koydu.
In this Ph D thesis, the presence of Enterococcus spp. in animal originated foods that were collected from Southern Marmara Region between April and December 2011 and their antibiotic resistance profiles were undertaken. For this purpose, totally 375 food samples consisting of 150 meat and meat products (veal cubes, minced meat, meatball, soujouk, salami, sausage, pastrami), 150 milk and dairy products (cheese, yoghurt, kefir, butter), 75 chicken meats (breast, thigh, wing) and poultry products (burger, meatball, nugget, schnitzel) were used as material. The contamination of Enterococcus spp. was observed in 54.13 % (203/375) of the samples analysed by classical microbiological techniques. By using API 20 STREP bacteria identification system, 92 isolates were identified as E. faecium, 88 as E. faecalis, 20 as E. durans, 2 as E. avium and 1 as E. gallinarum. Antibiotic susceptibility profile of isolates were tested by disc diffusion technique for ampicillin (10 µg), penicilin (10 unit), linezolid (30 µg), quinupristin/dalfopristin (15 µg), vancomycin (30 µg), teicoplanin (30 µg), ciprofloxacin (5 µg), tetracycline (30 µg), high level aminoglycosid resistance gentamicin (120 µg) and streptomycin (300 µg) antibiotics. Minimum inhibitory consantrations (MIC) for vancomycin and teicoplanin resistant strains were determined by E-test method. It has been pointed out that 18.7 % (38 strains) of strains were resistant to one antibiotic, 5.9 % (12 strains) to two antibiotics, 2.9 % (6 strains) to three antibiotics and 0.5 % (1 strain) to six antibiotics. The percentage of resistance among E. faecium strains was 18.5 % (17/92) for tetracycline and 3.3 % (3/92) for penicilin; meanwhile, this value was 1.1 % (1/92) for each of vancomycin, teicoplanin, quinupristin/dalfopristin, and ciprofloxacin antibiotics. 6.5 % (6/92) of E. faecium strains showed the resistance to more than one antibiotic. All of E. faecium strains were susceptible to linezolid, high level streptomycin and gentamicin. Variable level of resistance was noted among the E. faecalis strains. Their resistance to tetracycline was 28.4 % (25/88), to quinupristin/dalfopristin was 25 % (22/88), to ciprofloxacin was 3.4 % (3/88), to linezolid and high level streptomycin was 2.3 % (2/88). Moreover, the resistance to each of vancomycin, teicoplanin, ampicillin, penicilin, and high level gentamicin antibiotics was 1.1 % (1/88). 14.8 % (13/88) of E. faecalis strains were resistant to at least two of antibiotics tested. It was observed that two (10 %) of E. durans strain had resistance to tetracycline. Other E. durans, E. avium and E. gallinarum strains were susceptible to any antibiotics mentioned above. E. faecium (one strain) and E. faecalis (one strain) strains exhibiting resistance to vancomycin and teicoplanin by disc diffusion method assays were checked by E- test and they were confirmed to be susceptible for these antibiotics because their MIC levels were lower than 4 µg/ml. As a result, the presence and the incidence of enterococci in animal originated foods and, their resistance to different antibiotics was determined. Data obtained by the current thesis may be an indication of the possible role and importance of enterococci in the spread of antibiotic resistant bacteria in nature, notably by taking into account their ability to transfer these genetic characters to other pathogen and non-pathogen bacteria.
In this Ph D thesis, the presence of Enterococcus spp. in animal originated foods that were collected from Southern Marmara Region between April and December 2011 and their antibiotic resistance profiles were undertaken. For this purpose, totally 375 food samples consisting of 150 meat and meat products (veal cubes, minced meat, meatball, soujouk, salami, sausage, pastrami), 150 milk and dairy products (cheese, yoghurt, kefir, butter), 75 chicken meats (breast, thigh, wing) and poultry products (burger, meatball, nugget, schnitzel) were used as material. The contamination of Enterococcus spp. was observed in 54.13 % (203/375) of the samples analysed by classical microbiological techniques. By using API 20 STREP bacteria identification system, 92 isolates were identified as E. faecium, 88 as E. faecalis, 20 as E. durans, 2 as E. avium and 1 as E. gallinarum. Antibiotic susceptibility profile of isolates were tested by disc diffusion technique for ampicillin (10 µg), penicilin (10 unit), linezolid (30 µg), quinupristin/dalfopristin (15 µg), vancomycin (30 µg), teicoplanin (30 µg), ciprofloxacin (5 µg), tetracycline (30 µg), high level aminoglycosid resistance gentamicin (120 µg) and streptomycin (300 µg) antibiotics. Minimum inhibitory consantrations (MIC) for vancomycin and teicoplanin resistant strains were determined by E-test method. It has been pointed out that 18.7 % (38 strains) of strains were resistant to one antibiotic, 5.9 % (12 strains) to two antibiotics, 2.9 % (6 strains) to three antibiotics and 0.5 % (1 strain) to six antibiotics. The percentage of resistance among E. faecium strains was 18.5 % (17/92) for tetracycline and 3.3 % (3/92) for penicilin; meanwhile, this value was 1.1 % (1/92) for each of vancomycin, teicoplanin, quinupristin/dalfopristin, and ciprofloxacin antibiotics. 6.5 % (6/92) of E. faecium strains showed the resistance to more than one antibiotic. All of E. faecium strains were susceptible to linezolid, high level streptomycin and gentamicin. Variable level of resistance was noted among the E. faecalis strains. Their resistance to tetracycline was 28.4 % (25/88), to quinupristin/dalfopristin was 25 % (22/88), to ciprofloxacin was 3.4 % (3/88), to linezolid and high level streptomycin was 2.3 % (2/88). Moreover, the resistance to each of vancomycin, teicoplanin, ampicillin, penicilin, and high level gentamicin antibiotics was 1.1 % (1/88). 14.8 % (13/88) of E. faecalis strains were resistant to at least two of antibiotics tested. It was observed that two (10 %) of E. durans strain had resistance to tetracycline. Other E. durans, E. avium and E. gallinarum strains were susceptible to any antibiotics mentioned above. E. faecium (one strain) and E. faecalis (one strain) strains exhibiting resistance to vancomycin and teicoplanin by disc diffusion method assays were checked by E- test and they were confirmed to be susceptible for these antibiotics because their MIC levels were lower than 4 µg/ml. As a result, the presence and the incidence of enterococci in animal originated foods and, their resistance to different antibiotics was determined. Data obtained by the current thesis may be an indication of the possible role and importance of enterococci in the spread of antibiotic resistant bacteria in nature, notably by taking into account their ability to transfer these genetic characters to other pathogen and non-pathogen bacteria.
Description
Keywords
Enterokok, Hayvansalgıda, Antibiyotikdirenci, Enterococci, Animal originated food, Antibiotic resistance
Citation
Muş, T. E. (2014). Hayvansal kaynaklı gıdalarda enterokokların varlığı, tür düzeyinde identifikasyonları ve antibiyotik dirençliliklerinin belirlenmesi. Yayınlanmamış doktora tezi. Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.