Grantee Research Project Results
Final Report: Detection of Pathogens in Drinking Water (SEER 2)
EPA Grant Number: R827683E03Title: Detection of Pathogens in Drinking Water (SEER 2)
Investigators: Hovde Bohach, Carolyn J. , Bohach, Gregory A. , Paszczynski, Andrzej J. , Crawford, Ronald L. , Hartzell, Patricia
Institution: University of Idaho
EPA Project Officer: Chung, Serena
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)
Objective:
The objective of this research project was to examine molecular biology-based approaches for determining the probable sources of Escherichia coli O157:H7 contamination in environments such as drinking water. The pathogenic bacterium E. coli O157:H7 is found in fecal matter produced by mammalian sources, and is a serious contaminant of food and drinking water. Traditional methods for tracking the sources of such contamination are lacking.
Summary/Accomplishments (Outputs/Outcomes):
Project investigators developed a polymerase chain reaction (PCR)-based technique
to detect E. coli 0157:H7 cells in environmental samples using previously
reported PCR primers for the specific detection of genes involved in biosynthesis
of 0157 polysaccharide and H7 flagella antigens. They also developed a multiplex
PCR technique to simultaneously detect animal cells (sheep, cow, human, or horse)
and E. coli 0157:H7. PCR primers to detect animal cells were based upon
the known DNA sequences of J region and C region of the regulatory D-loop of
mitochondrial DNA. E. coli 0157:H7 (ATCC 43894) was used as the pathogen
standard for PCR procedure development. E. coli DH5 and non-O157:H7
E. coli strains isolated from cattle feces were used as negative controls.
Experimental samples were taken from water troughs of the various animals being
studied as possible contamination sources. The E. coli 0157:H7 titer
detected by the PCR assay had as few as six cells. Two hundred E. coli
0157:H7 cells were easily detected after 38 amplification cycles. The D-loop
primer sets were able to detect and differentiate the presence of human from
horse or cattle, but not sheep DNA samples.
Conclusions:
These results provide a significant step toward developing methods for tracing sources of drinking water contamination by this bacterium.
Supplemental Keywords:
drinking water, pathogenic bacteria, Escherichia coli O157:H7, E. coli, environmental science, Idaho, EPA Region 10., RFA, Health, Scientific Discipline, INTERNATIONAL COOPERATION, Geographic Area, Water, POLLUTANTS/TOXICS, Health Risk Assessment, Risk Assessments, State, Environmental Chemistry, Microorganisms, Drinking Water, Ecology and Ecosystems, community water system, E. Coli, health effects, Idaho (ID), human exposure, fecal contamination, pathogens, monitoring, drinking water contaminants, drinking water treatment, human health riskRelevant Websites:
http://www.ag.uidaho.edu/mmbb/ Exit
Progress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.