AUTHOR=Ahmed Warish , Gyawali Pradip , Hamilton Kerry A. , Joshi Sayalee , Aster David , Donner Erica , Simpson Stuart L. , Symonds Erin M. 

TITLE=Antibiotic Resistance and Sewage-Associated Marker Genes in Untreated Sewage and a River Characterized During Baseflow and Stormflow

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.632850

DOI=10.3389/fmicb.2021.632850

ISSN=1664-302X

ABSTRACT=<p>Since sewage is a hotspot for antibiotic resistance genes (ARGs), the identification of ARGs in environmental waters impacted by sewage, and their correlation to fecal indicators, is necessary to implement management strategies. In this study, sewage treatment plant (STP) influent samples were collected and analyzed using quantitative polymerase chain reaction (qPCR) to investigate the abundance and correlations between sewage-associated markers (i.e., <italic>Bacteroides</italic> HF183, <italic>Lachnospiraceae</italic> Lachno3, crAssphage) and ARGs indicating resistance to nine antibiotics (belonging to aminoglycosides, beta-lactams, sulfonamides, macrolides, and tetracyclines). All ARGs, except <italic>bla</italic><sub>VIM</sub>, and sewage-associated marker genes were always detected in untreated sewage, and <italic>ermF</italic> and <italic>sul1</italic> were detected in the greatest abundances. <italic>intl1</italic> was also highly abundant in untreated sewage samples. Significant correlations were identified between sewage-associated marker genes, ARGs and the <italic>intl1</italic> in untreated sewage (τ = 0.488, <italic>p</italic> = 0.0125). Of the three sewage-associated marker genes, the BIO-ENV procedure identified that HF183 alone best maximized correlations to ARGs and <italic>intl1</italic> (τ = 0.590). Additionally, grab samples were collected from peri-urban and urban sites along the Brisbane River system during base and stormflow conditions, and analyzed for <italic>Escherichia coli</italic>, ARGs, the <italic>intl1</italic>, and sewage-associated marker genes using quantitative polymerase chain reaction (qPCR). Significant correlations were identified between <italic>E. coli</italic>, ARGs, and <italic>intl1</italic> (τ = 0.0893, <italic>p</italic> = 0.0032), as well as with sewage-associated marker genes in water samples from the Brisbane River system (τ = 0.3229, <italic>p</italic> = 0.0001). Of the sewage-associated marker genes and <italic>E. coli</italic>, the BIO-ENV procedure identified that crAssphage alone maximized correlations with ARGs and <italic>intl1</italic> in river samples (τ = 0.4148). Significant differences in <italic>E. coli</italic>, ARGs, <italic>intl1</italic>, and sewage-associated marker genes, and by flow condition (i.e., base vs. storm), and site types (peri-urban vs. urban) combined were identified (<italic>R</italic> = 0.3668, <italic>p</italic> = 0.0001), where percent dissimilarities between the multi-factorial groups ranged between 20.8 and 11.2%. Results from this study suggest increased levels of certain ARGs and sewage-associated marker genes in stormflow river water samples compared to base flow conditions. <italic>E. coli</italic>, HF183 and crAssphage may serve as potential indicators of sewage-derived ARGs under stormflow conditions, and this merits further investigation. Data presented in this study will be valuable to water quality managers to understand the links between sewage pollution and ARGs in urban environments.</p>