Metagenomic-enabled rapid development of multiple biomarkers for trace organic contaminant biodegradation through Functional gene Amplicon Sequencing tests (FASt)

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Trace organic contaminants (TOrCs) are widespread in US water systems. Among the most prevalent TOrCs are personal care products, antibiotics, flame-retardants, pesticides, and herbicides. Toxicological studies suggest that chronic exposure to many TOrCs may cause cancer and negatively affect human fetal development. TOrCs are difficult to remove in conventional wastewater treatment plants, resulting in widespread dispersal in receiving waters. Biological treatment, which uses bacteria that degrade TOrCs, is a promising technology that could be implemented within existing treatment plants to remove TOrCs and prevent their release. However, no tools are currently available to monitor these specific bacteria capable of degrading TOrCs. The goal of this project is to address the critical need for next-generation monitoring tools to validate and optimize the performance of biological wastewater treatment systems. To achieve this goal, researchers will develop FASt (Functional gene Amplicon Sequencing tests), a next-generation gene sequencing technology to identify specific “functional genes” found only in TOrC degrading bacteria. Successful completion of this project will benefit society through the development of new knowledge and tools to monitor and optimize the removal efficiency of TOrCs in wastewater treatment systems. Further benefits to society will be achieved through education, research training, and outreach to teachers through a Research Experience for Future Teachers program.Nearly all wastewater, storm water, and surface water systems in the Nation have some levels of trace organic contaminants (TOrCs) like personal care products, antibiotics, pesticides, and herbicides. One promising solution for tackling TOrC pollution is through modification of conventional municipal wastewater treatment facilities to employ biological treatment systems to remove TOrCs. However, progress towards employing this technology has been slow as current approaches often use a “black box” approach to establish and manage TOrC treatment reactor microbiomes. This is due to the bottleneck created by the lack of biomarkers that can directly probe genetic potential and activity for TOrC biodegradation. The goal of this project is to address this bottleneck by developing a suite of biomarkers for aerobic TOrC biodegradation using a new approach based on Functional gene Amplicon Sequencing tests (FASt). A key novel feature of this project is the implementation of a functional gene amplicon generation method known as rhAmp polymerase chain reaction (rhAmp PCR), which can amplify hundreds of potential biomarker genes simultaneously. Specific research objectives to achieve this goal are to 1) identify functional gene biomarkers for TOrC biodegradation using FASt; 2) validate the new biomarkers using metabolomics; and 3) translate project findings into field-deployable assays. Successful completion of this project will transform our ability to rapidly monitor and optimize the performance of TOrC-degrading microorganisms in wastewater treatment systems through the development of new biomarker-based tools.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.