Soğutulmuş ve Dondurulmuş Köftelerde Clostridioides Difficile Kontaminasyonunun Araştırılması
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Dosyalar
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Clostridioides difficile anaerobik, spor oluşturan bir nozokomiyal enfeksiyon etmenidir. Son
yıllarda yapılan çalışmalar C. difficile’in gıda kaynaklı bir enfeksiyon etmeni olabileceği yönünde şüpheleri
gündeme getirmektedir. C. difficile çeşitli hayvansal gıdalarda bulunabilir. Bu çalışmanın amacı, ısıl işlem
uygulanmamış soğutulmuş ve dondurulmuş köftelerde C. difficile varlığını araştırmaktadır. Bu amaçla,
Konya il merkezinde satışa sunulan 36 adet soğutulmuş ve 36 adet dondurulmuş köftede C. difficile varlığı
incelendi. Çalışmada köfte örnekleri farklı kasap ve marketlerden farklı zaman diliminde alınarak incelendi.
C. difficile’i tespit etmek amacıyla kültürel izolasyon yöntemi ile multiplex Polymerase Chain
Reaction (mPCR) tekniği kullanıldı. Numuneler 50 ml Phosphate Buffered Saline ile homojenize edildi.
C.difficile’in kültürel izolasyonunda selektif zenginleştirme için LCC Selective Supplement ilaveli
Clostridium difficile Mannitol Taurocholate Broth kullanıldı. 1 ml homojenat 9 ml Clostridium difficile
Mannitol Taurocholate Broth’a eklenerek 37°C’de 7 gün anaerobik şekilde inkübe edildi. İnkübasyon
sonunda % 96’lik alkol ile 1:1 oranında karıştırılarak 45 dk inkübe edildi ve 4400xg’de santrifüj edildikten
sonra dipteki peletten alınarak kültivasyon için %7 koyun kanı ilaveli Clostiridium difficile Agar’a ekim
yapıldı. Ekim yapılan Clostiridium difficile Agar’lar 37°C’de 2 gün süreyle anaerobik inkübasyona bırakıldı.
Şüpheli kolonilere Gram boyama yapıldı. Ardından koloniler Blood Agar’da 37 °C’de 2 gün süreyle
anaerobik inkübe edilerek subkültive edildi. Subkültive edilen koloniler 0,2 ml’lik Brucella Broth’a ve 0,2
ml lik PBS’e aktarılarak -20°C’de kullanılacağı süreye kadar muhafaza edildi. mPCR ile triose phosphat
isomerase (tpi), tcdA, tcdB gen bölgeleri ile cdtA ve cdtB gen bölgelerinin varlığının saptanması amaçlandı.
İncelenen tüm örneklerde kültürel yöntemlerle tespit edilen dört şüpheli izolat (3 soğutulmuş köfte,
1 dondurulmuş köfte) mPCR ile negatif bulundu. Bu çalışma, köftelerin üretim aşamalarında uygulanan
hijyen koşullarının ve üretimde kullanılan etlerin mikrobiyolojik kalitesinin C. difficile kontaminasyonu
riskini önemli ölçüde azaltabileceğini göstermektedir. Bu bulgular, gıda güvenliği uygulamalarının
etkinliğini değerlendirmede bir temel oluşturmaktadır.
Clostridioides difficile is an anaerobic, spore-forming nosocomial infectious factor. Recent studies have raised concerns that C. difficile may be a foodborne infectious factor. C. difficile can be found in various animal foods. The aim of this study was to determine the presence of C. difficile in not heat treated refrigerated and frozen meatballs. For this purpose, 36 refrigerated meatballs and 36 frozen meatballs sold in Konya city center were examined for the presence of C. difficile. The meatball samples were obtained from different butcher shops and grocery stores at two different time periods. The cultural isolation method and multiplex Polymerase Chain Reaction (mPCR) technique were used to detect C. difficile. Samples were homogenized with 50 ml Phosphate Buffered Saline. Clostridium difficile Mannitol Taurocholate Broth with LCC Selective Supplement was used for selective enrichment in the cultural isolation of C. difficile. 1 ml homogenate was added to 9 ml Clostridium difficile Mannitol Taurocholate Broth and incubated anaerobically at 37°C for 7 days. At the end of incubation, it was mixed 1:1 with 96% alcohol and incubated for 45 min. After centrifugation at 4400xg, the bottom pellet was taken from the pellet and inoculated onto Clostiridium difficile Agar with 7% sheep blood for cultivation. The inoculated Clostiridium difficile Agar’s were left for anaerobic incubation at 37°C for 2 days. Gram staining was performed on suspicious colonies. The colonies were then subcultivated by anaerobic incubation in Blood Agar at 37°C for 2 days. Subcultivated colonies were transferred to 0.2 ml Brucella Broth and 0.2 ml PBS and stored at -20°C until further use. It was aimed to determine the presence of triose phosphate isomerase (tpi), tcdA, tcdB gene regions and cdtA and cdtB gene regions by mPCR. Four suspicious isolates (3 chilled meatballs, 1 frozen meatball) detected by cultural methods in all samples examined with mPCR were found negative. This study shows that the hygiene conditions applied during the production stages of meatball production and the meat used in production can significantly reduce the risk of C. difficile contamination. These findings provide a basis for evaluating the effectiveness of food safety practices.
Clostridioides difficile is an anaerobic, spore-forming nosocomial infectious factor. Recent studies have raised concerns that C. difficile may be a foodborne infectious factor. C. difficile can be found in various animal foods. The aim of this study was to determine the presence of C. difficile in not heat treated refrigerated and frozen meatballs. For this purpose, 36 refrigerated meatballs and 36 frozen meatballs sold in Konya city center were examined for the presence of C. difficile. The meatball samples were obtained from different butcher shops and grocery stores at two different time periods. The cultural isolation method and multiplex Polymerase Chain Reaction (mPCR) technique were used to detect C. difficile. Samples were homogenized with 50 ml Phosphate Buffered Saline. Clostridium difficile Mannitol Taurocholate Broth with LCC Selective Supplement was used for selective enrichment in the cultural isolation of C. difficile. 1 ml homogenate was added to 9 ml Clostridium difficile Mannitol Taurocholate Broth and incubated anaerobically at 37°C for 7 days. At the end of incubation, it was mixed 1:1 with 96% alcohol and incubated for 45 min. After centrifugation at 4400xg, the bottom pellet was taken from the pellet and inoculated onto Clostiridium difficile Agar with 7% sheep blood for cultivation. The inoculated Clostiridium difficile Agar’s were left for anaerobic incubation at 37°C for 2 days. Gram staining was performed on suspicious colonies. The colonies were then subcultivated by anaerobic incubation in Blood Agar at 37°C for 2 days. Subcultivated colonies were transferred to 0.2 ml Brucella Broth and 0.2 ml PBS and stored at -20°C until further use. It was aimed to determine the presence of triose phosphate isomerase (tpi), tcdA, tcdB gene regions and cdtA and cdtB gene regions by mPCR. Four suspicious isolates (3 chilled meatballs, 1 frozen meatball) detected by cultural methods in all samples examined with mPCR were found negative. This study shows that the hygiene conditions applied during the production stages of meatball production and the meat used in production can significantly reduce the risk of C. difficile contamination. These findings provide a basis for evaluating the effectiveness of food safety practices.
Açıklama
Anahtar Kelimeler
Meatballs, Cold storage method, Clostridioides difficile, Köfte, Soğuk muhafaza yöntemi, mPCR
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Gürdemir, E., (2023). Soğutulmuş ve Dondurulmuş Köftelerde Clostridioides Difficile Kontaminasyonunun Araştırılması. (Doktora Tezi). Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü, Konya.
