The Assessment of Escherichia coli as an Indicator of Microbial Quality of Irrigation Waters used for Produce

Abstract

Escherichia coli is a bacterial species that lives in the gut of all warm-blooded animals, fish, birds as well as reptiles and is commonly used as an indicator of fecal contamination in water. This project assessed currently used culture based media for the detection of E. coli in irrigation waters used in Arizona and California, and will present recommendations towards the most reliable media for the evaluation of irrigation waters used for produce. Currently, no microbial indicator standards exist for irrigation waters used for produce production in the United States. The produce industry suggests that the recreational water standard guideline (126 E. coli/100 ml) established by the United States Environmental Protection Agency (USEPA) be used. There is concern that the false positive rate of E. coli detection may be high in these waters giving false indications of the level of risk from enteric pathogens. This project evaluated three commercially available media for E. coli detection to test irrigation waters from three agricultural areas (Yuma and Maricopa, AZ and Imperial Valley, CA) and then assessed false positive rates by utilizing Polymerase Chain Reaction (PCR) and DNA sequencing of the bacterial isolates. The media that were chosen for evaluation were (1) MI Agar, (2) IDEXX Colilert Quanti-Tray® and (3) m-ColiBlue24® broth, all evaluation media accepted by the USEPA and widely used in the monitoring of irrigation water quality by the produce industry. Four hundred and fifty 1-L irrigation water grab samples were collected between March 2012 and November 2012. The samples were analyzed for both cultural counts and water quality parameters including temperature, salinity and pH. Isolates positive and negative for E. coli were then selected and assessed utilizing PCR and DNA sequencing. The false positive rate of each method was found to be high, with MI Agar, m-ColiBlue24® broth and the IDEXX Colilert Quanti-Tray® at an accuracy of 67%, 72%, and 51% respectively. A false positive result is reported when presumptive E. coli sub cultured from the media is found to be non-E. coli through molecular analysis. Overall the IDEXX Colilert Quanti-Tray® performed at a greater rate of accuracy than the other two media evaluated, however, high false positive rates may lead to inaccurate assessment of water quality.