The Water Microbiome Through a Pilot Scale Advanced Treatment Facility for Direct Potable Reuse

Advanced treatment facilities for potable water reuse of wastewater are designed to achieve high removal levels of specific pathogens, as well as many other constituents. However, changes to the microbial community throughout treatment, storage, and distribution of this water have not been well characterized. We applied high-throughput amplicon sequencing, read-based, assembly-based, and genome-resolved metagenomics, and flow cytometry to investigate the microbial communities present in a pilot-scale advanced water treatment facility. Advanced treatment of secondary-treated wastewater consisted of ozonation, chloramination, microfiltration, reverse osmosis (RO), advanced oxidation (UV/H₂O₂), granular activated carbon (GAC) filtration, and chlorination. Treated water was fed into bench-scale simulated distribution systems (SDS). Cell counts and microbial diversity in bulk water decreased until GAC filtration, and the bacterial communities were significantly different following each treatment step. Bacteria grew within GAC media and contributed to a consistent microbial community in the filtrate, which included members of the Rhizobiales and Mycobacteriaceae. After chlorination, some of the GAC filtrate community was maintained within the SDS, and community shifts were associated with stagnation. Putative antibiotic resistance genes and potential opportunistic pathogens were identified before RO and after advanced oxidation, although few if any members of the wastewater microbial community passed through these treatment steps. These findings can contribute to improved design of advanced treatment trains and management of microbial communities in post-treatment steps.