The long-term objective of the principal investigator is to significantly reduce bacteria pollution on Lake Erie. The objective of this proposal, which is a critical step for meeting this long-term objective, is to collect preliminary data to demonstrate that precursor 16S rRNA can be used as an effective screening technology to identify viable waterborne pathogens and pathogenc indicators in the environment.

The principal investigator proposes to accomplish the overall objective of this project by pursuing the following specific aims: Specific Aim 1: Assess the value of "molecular fingerprinting" with precursor 16S rRNA to track Escherichia coli in the environment; Specific Aim 2: Assess the value of "molecular fingerprinting" with precursor 16S rRNA to track Enterococci in the environment; and Specific Aim 3: Assess the value of "molecular fingerprinting" with precursor 16S rRNA to track alternative indicators of pathogenic bacteria pollution. The principal investigator expects to use recent advances in molecular biotechnology to track viable bacteria pollution on Lake Erie. The molecular biotechnologies that will be employed in this study have been classified broadly as "library-independent bacteria source tracking methods". These biotechnologies are based upon the molecular-level determination of bacteria identity using precursor 16S rRNA-targeted forensics.

Reducing bacteria pollution from municipal sewage required a significant investment in sewage treatment plants to eliminate bacteria pollution at the end-of-the-pipe. The diffuse nature of non-point source bacteria pollution is such that the cost for treating all potential sources is prohibitive. Instead, science-based information is needed to rank the importance of remaining sources of bacteria pollution on Lake Erie. The rationale for this project is that molecular biotechnologies provide the critical science-based information needed to track viable abundant sources of bacteria pollution including pathogens and pathogen indicators on Lake Erie.