Kanatlı Orijinli Escherichia coli suşlarının virülens özelliklerinin fenotipik ve moleküler metotlarla belirlenmesi

Öz

Bu çalışmada, kolibasillozisli kanatlı hayvanlardan izole edilen 200 adet Escherichia (E.) coli suşunda, özellikle ekstra-intestinal infeksiyonlar için önemli olan virülens faktörlerinden serum direnci, aerobaktin demir elde etme sistemleri, çeşitli adezinler (Tip 1 fimbrialar, P fimbrialar, S fimbrialar ve afimbrial adezinler), hemoliz ve sitotoksik nekrotizan faktör 1 gibi virülens özellikleri fenotipik yöntemlerle ve moleküler yöntemlerle araştırıldı. Çalışmada, suşlar koyun kanlı agar kullanılarak yapılan testle hemoliz üretimi ve moleküler olarak hemoliz özelliğini kodlayan gen yönünden negatif bulundu. Suşların tamamının demir kısıtlayıcı besiyerinde ürediği ve bu suşların %88’inin aerobaktin özelliğini kodlayan İucD genine sahip oldukları saptandı. Vero hücre kültürleri kullanılarak yapılan toksin analizlerinde suşların %40’ında ısıya duyarlı toksin belirlenirken suşların hiç birinde fenotipik ve moleküler olarak sitotoksik nekrotizan faktör 1 belirlenmedi. İnsan O grubu ve değişik hayvanlara ait eritrositler kullanılarak yapılan hemaglutinasyon testi ile suşların %90’ında hemaglutinasyon aktivitesi saptandı ve bunların %26’sı mannoz resistans hemaglutinasyon özelliğinde olduğu belirlendi. Tip 1 fimbriaları kodlayan FimH geni suşların %88’inde, P fimbriaları kodlayan PapEF geni %26’sında bulundu ve bu izolatların %14’ü aynı zamanda PapC genini de bulundurduğu saptandı. Ancak, suşlarda S fimbrialar ve afimbrial adezinleri kodlayan Sfa ve Afa genleri belirlenemedi. Değişik hayvanlara ait serumlarla yapılan testte suşların tamamında serum direnci fenotipik olarak saptandı ve serum direncinden sorumlu TraT geni suşların %90’ında PCR ile belirlendi. Bu çalışmada elde edilen sonuçlara göre, kanatlı orijinli E. coli suşlarında en önemli virülens özelliklerinin aerobaktin demir elde etme sistemleri, fimbrial adezinler ve serum direnci olduğu belirlenmiştir

Anahtar Kelimeler

• Escherichia coli, Kanatlı hayvanlar, Virülens özellikleri

Detection of Virulence Properties Escherichia coli Originated From Poultry by Phenotypic and Molecular Methods

Öz

In this study, virulence factors important for extra-intestinal infections such as serum resistance, aerobactin iron uptake systems, some adhesins (type 1 fimbria, P fimbria, S fimbria, afimbrail adhesins), haemolysis and cytotoxic necrotizing factor 1 in 200 Escherichia coli isolates from poultry with colibacillosis, were investigated by both phenotypic methods and molecular methods. Strains were detected to be negative in haemolysin synthesis tests performed with sheep blood agar, and they were negative in molecular tests investigating the haemolysin gene. All strains grew on iron deficient medium and 88% of these were found to harbor iucD gene encoding aerobactin. Heat-labile toxin was detected in 40% of the isolates in the toxin analyses performed in Vero cells, however, cytotoxic necrotizing factor 1 could not be determined in the strains with phenotypic and molecular tests. Haemagglutination activity was determined in 90% of strains in haemagglutination tests performed with human O group erythrocytes and erythrocytes derived from different animals. Twenty six percent of these strains were found to have MRHA activity. FimH gene encoding type 1 fimbria, papEF gene encoding P fimbria, were detected respectively in 88% and 26% of the strains, and 14% of all isolates were also found to harbor the papC gene also encoding the P fimbria. However, sfa and afa genes encoding S fimbria and afimbrial adhesins, respectively, could not be detected in the study. Serum resistance was phenotypically determined in all strains in the tests performed with sera derived from different animals and traT gene encoding serum resistance were detected in 90% of strains by PCR. In the study, serum resistance, aerobactin iron uptake systems and the existence of fimbrial adhesins were determined as the most important virulence factors of avian pathogenic E. coli isolates

Anahtar Kelimeler

• Escherichia coli, Poultry, Virulence Properties

Kaynakça

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