With increased water scarcity, it is critical that safe water supplies be maintained, including wastewater for environmental discharge or reuse. Currently microbial (bacterial and viral) water quality is measured using standard microbiological and biotechnological culture methods to establish safe water quality. This proposed EAGER is transformative as the study aims to explore the use of a newly developed detailed and sensitive metagenomic sequence analysis to understand and identify microbial communities for accurate determination of biological water quality in water, wastewater, biofilms, and water reuse systems.
With the proposed metagenomic analysis system, microorganisms, including bacteria, viruses, fungi, and parasites, will be accurately identified to species and strains. Additionally, antibiotic resistance and pathogenicity properties are also identified to evaluate diversity and abundance of the genes and microorganisms in-situ. By assessing the microbial communities and comparing their composition, a better understanding of the microbial communities and their ecology is developed to provide improved methods for assessing microbial water quality for indirect potable reuse and direct potable reuse. Culture-dependent methods and indicator microorganism measurements have proven to be slow and biased because they do not represent all species, often not even the dominant species present during advanced water and wastewater treatment. Molecular techniques have provided insights into microbial community structure; however, these methods are seriously limited because they target single microorganisms that are often the less dominant species. Therefore, with metagenomic analysis using next generation sequencing, accurate resolution of the microbial communities and their diversity is achieved, resulting in significant benefits for environmental and public health applications. Metagenomic whole DNA sequencing involves shearing all DNA in a sample that is then sequenced by next generation sequencing. Next generation sequencing is non-discriminatory, identifying all organisms, even the underrepresented, while identifying functional and resistant genes, pathogenicity, and providing taxonomic structure based on phylogenetic trees, all in a single analysis and within approximately one day. Therefore, next generation sequencing provides a more complete identification, with greater resolution of the microbial community and its ecology. The results from this study have the potential of defining microbial communities in water systems at each step of the process and will transform the ability to understand and design better indicators, validate surrogates, and also define sustainable microbial communities beneficial for drinking water and water reuse. The long term societal impact will be paramount as the proposed project will yield a more defined microbial community structure for water quality and safe water supplies and will demonstrate the utility of this important, emerging approach for the water and water reuse industry. During the course of this study, a doctoral student will gain fundamental and applied knowledge in the area of bioinformatics and water reuse that encompasses advanced treatment technologies that are employed in water reuse. Student interns will have the opportunity to learn field sampling techniques. This study will establish a good foundation to achieve notable broader impacts for the environment.
