Abstract

ABSTRACTWastewater treatment plants (WWTPs) release treated effluent containing mobile genetic elements (MGEs), antibiotic resistance genes (ARGs), and microorganisms into the environment, yet little is known about their influence on nearby microbial communities and the retention of these factors in receiving water bodies. Our research aimed to characterize the genes and organisms from two different WWTPs that discharge into Lake Michigan, as well as from surrounding lake sediments to determine the dispersal and fate of these factors with respect to distance from the effluent outfall. Shotgun metagenomics coupled to distance-decay analyses showed a higher abundance of genes identical to those in WWTP effluent genes in sediments closer to outfall sites than in sediments farther away, indicating their possible WWTP origin. We also found genes attributed to organisms, such as those belonging to Helicobacteraceae, Legionellaceae, Moraxellaceae, and Neisseriaceae, in effluent from both WWTPs and decreasing in abundance in lake sediments with increased distance from WWTPs. Moreover, our results showed that the WWTPs likely influence the ARG composition in lake sediments close to the effluent discharge. Many of these ARGs were located on MGEs in both the effluent and sediment samples, indicating a relatively broad propensity for horizontal gene transfer (HGT). Our approach allowed us to specifically link genes to organisms and their genetic context, providing insight into WWTP impacts on natural microbial communities. Overall, our results suggest a substantial influence of wastewater effluent on gene content and microbial community structure in the sediments of receiving water bodies.IMPORTANCE Wastewater treatment plants (WWTPs) release their effluent into aquatic environments. Although treated, effluent retains many genes and microorganisms that have the potential to influence the receiving water in ways that are poorly understood. Here, we tracked the genetic footprint, including genes specific to antibiotic resistance and mobile genetic elements and their associated organisms, from WWTPs to lake sediments. Our work is novel in that we used metagenomic data sets to comprehensively evaluate total gene content and the genetic and taxonomic context of specific genes in environmental samples putatively impacted by WWTP inputs. Based on two different WWTPs with different treatment processes, our findings point to an influence of WWTPs on the presence, abundance, and composition of these factors in the environment.

Highlights

  • IMPORTANCE Wastewater treatment plants (WWTPs) release their effluent into aquatic environments

  • Before looking for evidence of specific genes released from wastewater treatment plant (WWTP) to Lake Michigan, we assessed the impact of WWTP effluent on the overall genetic profile of sediment samples in the lake

  • To examine whether these genes were present in the environment with limited or no WWTP influence, we included three reference samples that were collected in the middle of Lake Michigan

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Summary

Introduction

IMPORTANCE Wastewater treatment plants (WWTPs) release their effluent into aquatic environments. Evidence for the impact of wastewater treatment effluent on the receiving aquatic environment has been investigated using culture-dependent methods that isolate ARB and PCR or quantitative PCR (qPCR) methods to assess the presence and abundance of specific ARB, ARGs, or MGEs [21, 22]. Recent metagenomic approaches have provided evidence for the impact of WWTP on effluent-receiving water bodies, none have simultaneously tracked microorganisms, ARGs, and other genes from a WWTP into the environment by linking organisms and genes. We present a comprehensive analysis of the abundances of ARGs, MGEs, and microbial communities, including putative pathogenic microorganisms and their virulence factors, in the effluent of two full-scale WWTPs that use two different treatment processes (trickling filter and activated sludge). Our characterization of ARGcarrying taxa and ARGs associated with plasmids provides evidence for a mechanism of dispersal and proliferation of WWTP genetic elements in the environment

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