Metagenomic Insights into AMR Gene Prevalence in Municipal Wastewater: A One Health Approach
Michael Anthony Hajkowski
Department of Biology
Faculty Supervisor: Archana Anand
With the evolution and emergence of novel bacterial and viral pathogens, the identification and classification of the threats they may pose is the first step toward understanding associated impacts on environmental and public health. Wastewater Treatment Plants (WWTP) play a major role in disseminating antimicrobial resistance (AMR), further circulating their infectivity range. To identify targeted microorganisms and functional genes, traditional wastewater-based epidemiology approaches rely on membrane filtration concentration and PCR-based molecular methods. Nanotrap-based concentration methods paired with next-generation sequencing (NGS) for metagenomic analysis can provide higher species resolution with the ability to detect AMR presence and abundance. While the identification of AMR-associated genes is important, the integrative “One Health” approach recommended by the World Health Organization (WHO) elucidates the risk severity of AMR-associated genes with the interconnections of human, animal, and plant health and their shared environments.
To address these gaps, I propose to 1) develop and validate an antimicrobial resistance (AMR) surveillance workflow by conducting a 16S rRNA analysis to assess overall wastewater microbial diversity and performing shotgun sequencing on a mock community to evaluate method effectiveness, 2) compare microbial taxa and AMR gene prevalence in influent samples collected during winter and summer months and apply an integrated risk assessment framework to evaluate seasonal variations in AMR dissemination, and 3) compare microbial taxa and AMR gene prevalence between influent and effluent samples from the same time period to assess anthropogenic pollution contributions and track AMR dissemination patterns using the integrated risk assessment. This study will elucidate the presence of pathogenic shedding and AMR dissemination in wastewater influent while illuminating wastewater treatment efficiency by comparing wastewater taxa and AMR frequencies to effluent. I expect to develop a WBE workflow to monitor clinically relevant AMR-associated genes in municipal wastewater.