Analysis of the persistence of enteric markers in sewage polluted water on a solid matrix and in liquid suspension. (1st June 2015) - Brooks, Yolanda
Abstract: 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. Graphical abstract: Highlights: Persistence of 3 enteric markers measured in water samples stored at 4, 27 and 37 °C. Evaluated persistence in liquid suspension and attached to a solid matrix for ≤28 days. Most to least persistent indicators: enterococci, E. coli, and B. thetataiotaomicron. Indicator persistence increased on a solid matrix, and at low temperature. Indicators stored at 4 °C and attached to solid matrix had estimated T90 ≥ 27 days.