Quantification of Survival of Escherichia coli O157:H7 on Plants Affected by Contaminated Irrigation Water

Enterohemorrhagic E. coli O157: H7 (EHEC) is a major foodborne pathogen capable of causing diarrhea and vomiting, with further complications such as hemolytic-uremic syndrome (HUS). The aim of this study was to use the real-time PCR method to quantify the survival of Escherichia coli O157:H7/pGFP in phyllosphere (leaf surface), rhizosphere (volume of soil tightly held by plant roots), and non-rhizosphere soils (sand and clay) irrigated with contaminated water and compare the results obtained between real-time PCR method and conventional plate counts. The real-time PCR probe was designed to hybridize with the (eae) gene of E. coli O157:H7. The probe was incorporated into real-time PCR containing DNA extracted from the phyllosphere, rhizosphere, and non-rhizosphere soils irrigated with water artificially contaminated with E. coli O157:H7. The detection limit for E. coli O157:H7 quantification by real-time PCR was 2.3 × 10 in the rhizosphere and phyllosphere samples. E. coli O157:H7 survived longer in rhizosphere soil than the non-rhizosphere soil. The concentration of E. coli O157:H7/pGFP in rhizosphere soils was ≥ 10 CFU/g in both soils at day 12 based on both plate count and real time PCR, with the clay soil significantly (P = 0.05) higher than the sandy soil. This data showed that E. coli O157H:7 can persist in the environment for more than 50 d, and this may pose some risk for both animal and human infection and provides a very significant pathway for pathogen recontamination in the environment.