Analysis of the Persistence of Enteric Markers in Sewage Polluted Water on a Solid Matrix and in Liquid Suspension

Addressing the persistence of bacterial indicators using qPCR and their respective DNA targets under various conditions is a critical part of risk assessment for water quality monitoring. The goal of this study was to examine the persistence of fecal indicator bacteria (FIB) via Escherichia coli uidA , enterococci 23S rDNA and Bacteroides thetataiotaomicron 1,6 alpha mannanase from cells attached to a solid matrix and in suspension. Raw sewage (10% vol/vol) was seeded into autoclaved river water with half of the sample volume in suspension and the other half was filtered onto membranes and stored at 4°, 27° and 37°C for up to 28 days. At various time points, DNA from cells was extracted, markers were quantified, and were fit to linear and non-linear models (first order exponential, biphasic (double) exponential, two-staged, log-logistic, and Gompertz 3-parameter). First order and biphasic exponential models fit 73% of the experimental data. Persistence increased significantly when the cells were stored in an attached state (p < 0.001). Increasing temperature had an inverse effect on persistence for the cells in suspension. Bacterial cells could be stored on a solid matrix at 4°, 27° and 37 °C for up to 27, 18, and 3 days, respectively, with <90% decay. The least stable indicator at 4°, 27° and 37 °C was B. thetataiotaomicron in suspension with T90 = 9.6, 1.8, and 1.1 days, respectively. The most persistent indicator was enterococci, with T90 > 28 days in an attached state at all temperatures.