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Ankara Üniversitesi Fen Bilimleri Enstitüsü Yüksek ANKARA ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ YÜKSEK LİSANS TEZİ ET VE ET ÜRÜNLERİNİN Listeria spp. VARLIĞI BAKIMINDAN ARAŞTIRILMASI Raşit KESKİN GIDA MÜHENDİSLİĞİ ANABİLİM DALI ANKARA 2020 Her hakkı saklıdır ÖZET Yüksek Lisans Tezi ET VE ET ÜRÜNLERİNİN Listeria spp. VARLIĞI BAKIMINDAN ARAŞTIRILMASI Raşit KESKİN Ankara Üniversitesi Fen Bilimleri Enstitüsü Gıda Mühendisliği Anabilim Dalı Danışman: Doç. Dr. Pınar ŞANLIBABA Bu çalışmada Ankara’daki farklı satış yerlerinden temin edilen birbirinden farklı çiğ ve işlem görmüş et ürünlerinin Listeria spp. varlığı bakımından taranması amaçlanmıştır. 116 gıda örneğinin taraması, ISO 11290-1 protokolü doğrultusunda yapılmıştır. Gram (+), katalaz (+), oksidaz (-) ve hareketlilik (+) sonuç veren 40 izolatın, tür düzeyinde biyokimyasal olarak tanımlamasında API® Listeria test kiti kullanılmıştır. Bu kapsamda, 40 izolattan 15’i L. monocytogenes, 12’si L. innocua, 8’i L. welshimeri ve 4’ü ise L. grayi olarak tanımlanmıştır. 1 izolatın tanımlaması yapılamamıştır. Listeria spp. ve L. monocytogenes varlığı bakımından kıyma en riskli gıda olarak saptanmıştır. Çiğ etin %67,50’si ve işlem görmüş etin ise %32,50’si Listeria spp. ile kontamine olmuştur. Kırmızı et ve et ürünlerinin, beyaz et ve et ürünlerine oranla daha riskli olduğu saptanmıştır. Ankara’da satışa sunulan et ve et ürünleri, L. monocytogenes bakımından riskli bulunmuştur. 2020, 73 sayfa Anahtar Kelimeler: Listeria, et ve et ürünleri, izolasyon, biyokimyasal tanımlama iv ABSTRACT Master Thesis AN INVESTIGATION ON THE PRESENCE OF Listeria spp. IN MEAT AND MEAT PRODUCTS Raşit KESKİN Ankara University Graduate School of Natural and Applied Sciences Department of Food Engineering Supervisor: Assoc. Prof. Dr. Pınar ŞANLIBABA In this study, it was aimed to screen different raw and processed meat products obtained from different markets in Ankara for Listeria spp. A total of 116 food samples were analyzed in accordance with ISO 11290-1 protocol. API® Listeria test kit was used for biochemical characterization at species level of 40 isolates showed gram (+), catalase (+), oxidase (-) and mobility (+). In this study, 15 isolates were identified as L. monocytogenes, 12 as L. innocua, 8 as L. welshimeri and 4 as L. grayi. One isolate could not be identified. Minced meat was the the most risky food for Listeria spp. and L. monocytogenes. Further, 67.50% of raw meat and 32.50% processed meat were contaminated with Listeria spp. Red meat and meat products were found to be more risky than chicken meat products. Meat and meat products retailed in Ankara were found to be at risk for L. monocytogenes. 2020, 73 pages Key Words: Listeria, meat and meat products, isolation, biochemical characterization v TEŞEKKÜR Yüksek Lisans eğitimimin gerek ders döneminde gerekse tez döneminde her türlü destek ve yardımlarını benden esirgemeyen hem akademik hem de diğer konulardaki yönlendirmeleriyle yüksek lisans hayatımı tamamlamamı sağlayan danışman hocam Sayın Doç. Dr. Pınar ŞANLIBABA’ya (Ankara Üniversitesi Mühendislik Fakültesi Gıda Mühendisliği Bölümü), Hem lisans eğitimim hem de yüksek lisans eğitimim süresince her türlü desteğini yanımda hissettiğim, analizlerim sırasında da her türlü bilgi ve tecrübelerini benimle paylaşan ve çalışmalarım için gerekli olan referans suşlarımın temininde de yardımcı olan Sayın Prof. Dr. A. Kadir HALKMAN’a (Ankara Üniversitesi Mühendislik Fakültesi Gıda Mühendisliği Bölümü), Geçen iki yıllık süre zarfında aynı laboratuvarı paylaştığım, her türlü yardım ve desteklerini benimle paylaşan çalışma arkadaşları Esra ŞENTÜRK ve Gürcü Aybige ÇAKMAK’a, Yüksek lisans çalışmalarına başlamama vesile olan Dr. C. Haluk ÖZALP’e, eğitimim boyunca yanımda olan ve desteklerini esirgemeyen değerli anneme, babama ve eşime, Sonsuz teşekkürlerimi sunuyorum. Bu yüksek lisans çalışması ‘Listeria Türleri ile Fajlarının Farklı Materyallerden İzolasyonu ve Genotipik Tanımlanması ve Listeria monocytogenes’in Gıda Sistemlerinde Faj Uygulaması ile Biyokontrolü’ başlıklı ve 15B0443010 nolu Bağımsız Projesi kapsamında Ankara Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından desteklenmiştir. Raşit KESKİN Ankara, 2020 vi İÇİNDEKİLER TEZ ONAY SAYFASI ETİK .................................................................................................................................... i ÖZET .................................................................................................................................. ii ABSTRACT ...................................................................................................................... iii TEŞEKKÜR ..................................................................................................................... iv SİMGELER VE KISALTMALAR DİZİNİ ................................................................. vii ŞEKİLLER DİZİNİ ....................................................................................................... viii ÇİZELGELER DİZİNİ ................................................................................................... ix 1. GİRİŞ ............................................................................................................................. 1 2. KURAMSAL TEMELLER VE KAYNAK ÖZETLERİ ......................................... 3 2.1 Listeria spp.’nin Genel Özellikleri ............................................................................ 3 2.2 Listeria spp.’nin Sınıflandırılması ............................................................................. 4 2.3 Listeria monocytogenes ............................................................................................... 7 2.4 Listeriosis .................................................................................................................... 8 2.5 Listeria spp.’nin Tanımlanması .............................................................................. 10 2.5.1 Kültürel yöntemlerle tanımlama ......................................................................... 11 2.6 Et ve Et Ürünlerinde Listeria spp. Varlığı ............................................................. 11 3. MATERYAL VE METOT ........................................................................................ 21 3.1 Materyal ................................................................................................................... 21 3.1.1 Gıda numuneleri ................................................................................................... 21 3.1.2 Referans bakteriler .............................................................................................. 21 3.1.3 Çalışmada kullanılan besiyerleri ve kimyasallar ............................................... 21 3.1.4 Stok kültürlerin hazırlanması .............................................................................. 23 3.2 Yöntem ...................................................................................................................... 23 3.2.1 Et ve et ürünlerinin analize hazırlanması ve muhtemel Listeria spp.’nin izolasyonu .............................................................................................................. 23 3.2.2 Morfolojik ve biyokimyasal analizler .................................................................. 26 3.2.2.1 Gram boyama ..................................................................................................... 26 3.2.2.2 Katalaz testi ........................................................................................................ 26 3.2.2.3 Oksidaz testi ........................................................................................................ 26 vii 3.2.2.4 Hareketlilik testi ................................................................................................. 26 3.2.2.5 Listeria izolatlarının stok kültürlerinin hazırlanması ..................................... 27 3.2.2.6 API®Listeria testi ............................................................................................... 27 4. BULGULAR ve TARTIŞMA ................................................................................... 29 4.1 Et ve Et Ürünlerinden Listeria spp.’nin İzolasyonu .............................................. 29 4.2 Listeria İzolatlarının Tanımlanması ....................................................................... 36 4.2.1 Morfolojik ve kültürel testler .............................................................................. 36 4.2.2 Suşların biyokimyasal tanımlanması .................................................................. 40 5. SONUÇ ........................................................................................................................ 50 KAYNAKLAR ............................................................................................................... 52 ÖZGEÇMİŞ .................................................................................................................... 62 viii SİMGELER DİZİNİ pH Asitlik Değeri β Beta dk Dakika h Saat s Saniye g Gram kob Koloni Oluşturan Birim µm Mikrometre mL Mililitre L Litre mm Milimetre vb. Ve benzeri % Yüzde vd. Ve diğerleri oC Santigrat Derece Kısaltmalar AB Avrupa Birliği (European Union) API Application Programming Interface ATCC Amerikan Tipi Kültür Koleksiyonu (American Type Culture Collection) BLEB Buffered Listeria Enrichment Broth CDC Hastalık Koruma ve Kontrol Merkezi (Centers For Disease Control And Prevention) FDA Gıda ve İlaç İdaresi (US Food And Drug Administration) ISO Uluslararası Standardizasyon
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