Detection of Pathogens in Drinking Water (SEER 2)EPA Grant Number: R827683E03
Title: Detection of Pathogens in Drinking Water (SEER 2)
Investigators: Hovde Bohach, Carolyn J. , Bohach, Gregory A. , Crawford, Ronald L. , Hartzell, Patricia , Paszczynski, Andrzej J.
Institution: University of Idaho
EPA Project Officer: Hunt, Sherri
Project Period: August 15, 1999 through August 14, 2002
Project Amount: $238,548
RFA: EPSCoR (Experimental Program to Stimulate Competitive Research) (1998) RFA Text | Recipients Lists
Research Category: EPSCoR (The Experimental Program to Stimulate Competitive Research)
The detection of pathogens in drinking water depends largely on traditional approaches, but cellular chemical constituents, i.e., lipids and sugars in cell walls and membranes, are useful as specific markers for microbial species or strains. We propose that it is possible to use chromatographic procedures, including anion-exchange liquid chromatography (AELC), HPLC/MS, GC/MS, and ES/MS/MS, to detect molecules of clinical relevance in water at ng/L or pg/L concentrations. Our primary objectives will be to: (1) develop detection methods based on GC/MS, ES/MS/MS, and AELC for sugar and lipid molecules specific as identifiers for a variety of important pathogenic bacteria that threaten drinking water in the United States; (2) confirm the specificity and sensitivity of each detection method developed; and (3) demonstrate the efficacy of the detection methods with actual drinking water samples under controlled (laboratory) conditions, and in the field.
We will use gas-chromatography/mass-spectrometry (GC/MS), electrospray ionization tandem mass spectrometry (ES/MS/MS), and anion-exchange liquid chromatography (AELC) to analyze genus- and species-specific hydroxy fatty acids and sugars for microbial detection and identification in drinking water. Using GC/MS and selected ion-monitoring mode in ES/MS/MS to identify daughter ion spectra should allow the detection of specific microorganisms in complex mixtures. We will test detection limits and specificity of these MS and ion chromatography methods for detection of sugar moieties and lipids of various pathogens, and work out standard protocols intended to make these procedures the methods of choice for fast analysis of pathogens in drinking water.
We will develop methods that will allow public health agencies to routinely monitor microbial contamination of potable water. We expect to develop detection levels as low as 1-10 cells ml-1 for specific pathogenic bacterial strains.
Improvements in Risk Assessment or Risk Management: By improving our ability to estimate the level of contamination of drinking water supplies by pathogenic microorganisms, this research will significantly improve the risk assessment or risk management capabilities of US communities and public health districts.