Adherence Pheno-genotypic of Escherichia coli O157:H7 Isolated from Beef, Feces of Cattle, Chicken and Human

Abstract

Generally, adherence of micro-organisms to host cells is the frst step of the colonization to host surfaces. Escherichia coli O157:H7 can colonize to the intestine and induce attaching-effacing (AE) lessions. The capacity of inducing AE lesions is encoded by a pahtogenicity island, the locus of enterocyte effacement (LEE) that contains genes involved in generation of attaching and effaching (A/E) lesions. Among which that, the eae gene is encoding intimin, an outer membrane protein that is responsible to intimate attachment to the intestinal epithelial cells. A total of 20 local isolates obtained from human clinically, beef, cattle, chicken, and human non-clinically were tested to adherence pheno-genotypic of E. coli O157:H7. The eae gene was identifed using polymerase chain reaction with a specifc primer i.e AE19 forward and AE20 reverse. To confrm phenotypic of gene, further study was performed by culturing the bacteria in vero cell, followed by Giemsa staining and Acridine Orange Fluorescent staining 3 h and 6 h after incubation, respectivelly. Result of study showed that there were 19 out of 20 (95%) isolates identifed positive eae gene. Giemsa staining appeared that the bacteria with positive eae gene performed a cluster arroud of cell (localized adherence). On the other hand, the negative eae gene appeared as a diffuse adherence (DA). The study indicated that almost all of E. coli O157:H7 local isolates which was positive eae gene had potency to colonize to the intestine and induce attching-effaching lessions, and also cause cytopahatic effects in intestinal epithelial cell

Keywords

• E.coli O157:H7
• eae gene
• intimin