Mapping heterotrophic functions to microbial population structures in wastewater resource recovery processes for optimization and prediction of antimicrobial resistance dissemination.

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Microbial communities in sewer systems and in the Wastewater Resource Recovery Facilities (WRRFs) are connected through immigration driven by the influent wastewater. How sewer immigration shapes the microbial communities in WRRFs is still debated, but it is likely modulated by the specific bioprocess (e.g., aerobic vs. anaerobic) and the metabolic functions of the populations of interest. These considerations are important for the development of new bioprocesses that promote resource recovery, which increasingly utilize anaerobic technologies. They are also significant for the prediction of the dissemination of antimicrobial resistance genes (ARGs) through WRRFs. Municipal wastewater is recognized as a hotspot of ARGs, and the current antimicrobial resistance crisis demands to revise the operation of wastewater facilities that are not designed to reduce the dissemination of ARGs. These issues are at central to my research program with the long-term objective of developing novel wastewater resource recovery bioprocesses based on an understanding of their microbial ecology and to provide the theoretical framework for the rational design and optimization of the urban wastewater systems as a whole. In the short-term, this proposal aims to look in more details at the immigration interface between sewer and WRRF bioprocesses. Several tools will be developed and adapted to reach the following short-term goals including: (1) the development of new synthetic wastewater to manipulate the bioprocesses microbial community independently of immigration, (2) the use new flow cytometry-based techniques to reveal the metabolic functions heterotrophs across the immigration interface, (3) the use of a multiplex ARG amplicon sequencing tools to trace fate of specific allelic sequence variants across the interface. The results will provide information to optimize bioprocesses in function of the microbial communities immigrating with the influent wastewater, and to predict the fate of emerging pollutants like ARGs. Impact This work will train 2 PhDs, 2 Master's and 5 undergraduate students. Their training in both wastewater engineering and microbial community analyses/metagenomics will equip them well to enter an increasingly broad interdisciplinary workforce with opportunities in process development and public health. From a technical perspective, this work will provide the platform to evaluate a number of questions with respect to the immigration interface between sewers and WRRFs. The WRRF industry is not only charged with the development of more efficient bioprocesses, but also has to content with an increasing awareness of WRRFs as hotspots for the dissemination and recombination of ARGs. Microbial communities in sewers and WRRFs are critical for both these aspects. In this context, the framework of this proposal will inform the mechanism at play during immigration, and allow a wider prediction of microbial community responses to changing WRRF procedures.