During Phase I research, BCS, Inc. proposes to develop a molecular, PCR-based test for characterizing human and animal-specific E. faecalis. In addition to characterizing E. faecalis, we will also focus Phase I efforts on characterizing the distribution of additional Enterococcus spp. in the State of Florida, with subsequent efforts directed on genetic analysis of isolated species, specifically E. faecium, E. casseliflavus, E. flavescens and E. durans. APPROACH: The molecular, PCR-based test for characterizing human and animal-specific E. faecalis will be developed based on characterization and identification of genetic differences within isolates collected from different animal hosts. NON-TECHNICAL SUMMARY: Maintenance of the microbiological quality and safety of water systems used for drinking, recreating, and in the harvesting of seafood is imperative as contamination of these systems can exact high risks to human health as well as result in significant economic losses. Waters contaminated with human feces are generally regarded as a greater risk to human health as they are more likely to contain human-specific pathogens. Understanding the origin of fecal pollution is paramount in assessing associated health risks as well as implementing actions necessary to remedy the problem while it still exists. The usefulness of using microbial indicators as tools for risk assessment can be significantly enhanced by the development of methods and analysis techniques that can define specific sources of these organisms. If successful, Phase I research will result in a rapid, PCR-based test that will be useful in identifying specific animal sources of Enterococcus spp. Such a rapid test for use in tracking sources of Enterococcus is currently unavailable and would allow for water quality managers and regulatory agencies to remedy problems associated with fecally-polluted environmental waters used for drinking, swimming, recreating, and in the harvesting of seafood. PCR-based methods are extremely desirable due to the ease in which they are performed as well as the fact that limited laboratory equipment, reagents, and technical expertise are necessary.
Keywords: microbial source tracking; enterococci; molecular markers; pcr; indicators; fecal pollution
