Genetic Diversity of E. coli O157:H7 Isolated from Some Leafy Greens, Irrigated by Aleppo River, Using Random Amplified Polymorphic DNA (RAPD) Marker

Abstract

In recent years, fresh fruits and vegetables have been linked to numerous foodborne illness outbreaks in different regions of the world. In Syria, there is not a lot of research that study E. coli and its serotypes by PCR technology. In this study, we have fulfilled a total count of bacteria, the census total coliform group, and Escherichia coli, as well as the serotype E. coli O157:H7 on some leafy greens (Spinacia oleracea, Beta vulgaris) irrigated by Aleppo River. The molecular characterization was done for ten strains of E. coli isolated from collected samples. The samples were inoculated on blood agar and suspicious colonies, then transferred to EMB and MacConkey agar using a primer (COL 1) in RAPD technic. Molecular characterization also performed ten strains of serotype E. coli O157:H7 isolated in medium (Sorbitol-MacCONKEY Agar), then by primers (OPA-03, OPA-13 OPC-12, OPE-20) in RAPD technic. The results showed significant differences between collected samples. The total count of bacteria in the first site (Handarat) for spinach and chard were the lowest, but in the fourth site (Jezraya) it was the highest among all samples. The results showed also the existence of E. coli in all sites except the first one in riverbed Handarat, and E. coli O157:H7 was found only in Jezraya village. Extracted DNA from samples was amplified by RAPD. after electrophoresis in the gel agarose, eleven different bands were detected from isolated strains of E. coli. These results refer to the great genetic diversity of Escherichia coli. For serotype E. coli O157:H7 thirty-four different bands were detected. RAPD analysis had the high discriminatory capacity for typing E. coli isolates. Because of its simplicity and rapidity, RAPD analysis appears to be a highly valuable tool for studying E. coli molecular epidemiology.