Adherence Pheno-genotypic of Escherichia coli O157:H7 Isolated from Beef, Feces of Cattle, Chicken and Human
https://doi.org/10.22146/ijbiotech.15990 I Wayan Suardana(1*), Wayan Tunas Artama(2), Widya Asmara(3), Budi Setiadi Dayono(4)
(1) Department of Veterinary Publich Health, Faculty of Veterinary Medicine, Udayana University, Denpasar, Indonesia Study Program of Biotechnology, The Graduate School of Universitas Gadjah Mada, Yogyakarta, Indonesia
(2) Department of Biochemistry, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
(3) Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
(4) Department of Genetic, Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia
(*) Corresponding Author
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
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
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PDF DOI: https://doi.org/10.22146/ijbiotech.15990
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