SBIR Phase II: Microbial Source Tracking Using Mitochondrial DNA for Identification of Contaminant Sources

This Small Business Innovation Research (SBIR) Phase II project will result in field validated microbial source tracking (MST) assays that provide cost-effective identification of sources of fecal pollution. Despite efforts mandated under the BEACH and Clean Water Acts, beach closures have exceeded 20,000 days in each of the last four years primarily due to fecal pollution. The problem continues because traditional methods cannot identify the sources of fecal contamination (sewage, livestock, domestic animals, wildlife). MST assays employing quantitative polymerase chain reaction (qPCR) were developed to quantify source-specific genetic markers encoded on the mitochondrial DNA (mtDNA) of the source animal (human, cattle, dog, etc.). The Phase I results demonstrated that mtDNA-based assays combined with bacterial source tracking methods will provide conclusive identification of fecal contamination sources allowing implementation of corrective measures to improve water quality and protect human health. Phase II studies will include a modification of the DNA extraction procedure to permit quantification of live fecal bacteria to aid in risk assessment and extended field validation studies at two beaches and two coastal watersheds impaired by unknown sources of fecal pollution. The broader impacts of this research are that the MST assays developed and validated during the Phase II project will empower stakeholders with the type of actionable data required to identify fecal contamination sources, implement appropriate corrective actions, and safeguard the nation?s waters. Fecal contamination of water resources currently exacts a severe toll in terms of increased risks to human health and impacts on coastal economies.