Investigation of Escherichia coli O157:H7 By IMS-PCR Tecniques in Poultry Slaughtering Stages

Öz

The present study was conducted to investigate the prevalence of Escherichia coli (E. coli) O157:H7 and to

determined its virulence genes (stx1 and stx2, eaeA and ehlyA) in different samples collected from a poultry cutting satages

in Samsun province, Turkey. For this purpose, 40 viscera, 40 intestinal content, 20 scalding tank water and 100 swap samples

taken from various poultry processing units (plucking machine, cutting board, removing unit of intestinal organs and

personnel hands and broiler carcass) were analysed. All samples were analyzed for the isolation of E. coli O157:H7 through

pre-enrichment in modified tryptone soy broth supplemented with novobiocin (mTSB+N) followed by immunomagnetic

separation (IMS) and selective plating on CHROM agar with Cefixim – Tellurite. Presumptive E. coli O157:H7 isolates were

tested with anti-O157 and H7 antisera. Polymerase Chain Reaction (PCR) was performed on isolates for the detection of

rfbO157, fliCH7,stx1, stx2, eaeA and ehylA genes. In this study, two intestinal content (1%) isolates were identified as E. coli

O157 while were found negative for H7. Both isolates contained stx1, stx2, eaeA and ehylA genes. As a result of this study, it

was concluded that E. coli O157 isolates detected at low levels in the broiler intestinal contents might constitute a significant

hazard for human health due to the presence of toxin genes.

Anahtar Kelimeler

• Broiler,Contamination,Escherichia coli O157:H7,Slaughterhouse

Kanatlı Kesimi Aşamalarında E. Coli O157:H7 Varlığının IMS-PZR Yöntemleri ile İncelenmesi

Öz

Bu çalışma, Samsun ilinde kanatlı kesim aşamalarından toplanan farklı örneklerde Escherichia coli (E. coli) O157: H7

prevalansını araştırmak ve virülans genlerini (stx1 ve stx2, eaeA ve ehlyA) belirlemek amacıyla yapıldı. Bu amaçla, 40 iç organ, 40 bağırsak içeriği, 20 haşlama tank suyu ve kanatlı kesimhanesinin farklı birimlerinden (tüy yolma makinesi, kesme tahtası, iç organ çıkarma ünitesi ve personel eli ve 40 broiler karkas örneği) alınan 100 svap örneği analiz edildi. Tüm örnekler, novobiosin içeren selektif modifiye Tryptic Soy Broth (mTSB+N)’da ön zenginleştirmenin ardından immünomanyetik seperasyon (IMS) ve Cefixim – Tellurite’li CHROM agara ekilmesi yoluyla E. coli O157: H7 açısından analiz edildi. Şüpheli koloniler O157 ve H7 antiserumları ile incelendi. rfbO157, fliCH7, stx1, stx2, eaeA ve ehylA genlerinin varlığı Polimeraz Zincir Reaksiyonu (PZR) ile belirlendi. Çalışmada, 2 (%1) bağırsak içeriği örneğinden E. coli O157 identifiye edilmesine karşın, H7 tespit edilemedi. Her iki izolatın da stx1, stx2, eaeA ve ehylA geni içerdiği tespit edildi. Bu çalışma sonucunda, broiler bağırsak içeriğinde düşük oranda tespit edilen E. coli O157: H7 izolatların hepsinin anılan toksin genlerini içermesinden dolayı insan sağlığı için önemli bir tehlike oluşturabileceği kanısına varıldı.

Anahtar Kelimeler

• Broiler,Escherichia coli O157: H7,Kesimhane,Kontaminasyon

Kaynakça

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