Abstract

BackgroundMicroorganisms in activated sludge (AS) play key roles in the wastewater treatment processes. However, their ecological behaviors and differences from microorganisms in other environments have mainly been studied using the 16S rRNA gene that may not truly represent in situ functions.ResultsHere, we present 2045 archaeal and bacterial metagenome-assembled genomes (MAGs) recovered from 1.35 Tb of metagenomic data generated from 114 AS samples of 23 full-scale wastewater treatment plants (WWTPs). We found that the AS MAGs have obvious plant-specific features and that few proteins are shared by different WWTPs, especially for WWTPs located in geographically distant areas. Further, we developed a novel machine learning approach that can distinguish between AS MAGs and MAGs from other environments based on the clusters of orthologous groups of proteins with an accuracy of 96%. With the aid of machine learning, we also identified some functional features (e.g., functions related to aerobic metabolism, nutrient sensing/acquisition, and biofilm formation) that are likely vital for AS bacteria to adapt themselves in wastewater treatment bioreactors.ConclusionsOur work reveals that, although the bacterial species in different municipal WWTPs could be different, they may have similar deterministic functional features that allow them to adapt to the AS systems. Also, we provide valuable genome resources and a novel approach for future investigation and better understanding of the microbiome of AS and other ecosystems.8riZfkhYByJu5oh1qMcWxqVideo Abtract.

Highlights

  • Microorganisms in activated sludge (AS) play key roles in the wastewater treatment processes

  • 2045 metagenome-assembled genomes (MAGs) were obtained from AS of different wastewater treatment plants (WWTPs) Approximately 1.35 Tb of metagenomic sequencing data generated from 114 AS samples of 23 municipal WWTPs located in eight countries were used to construct MAGs (Additional file 1: Figure S1, Table S1, Table S2)

  • Phylogeny and functional features cannot well separate MAGs from AS and MAGs from other environments In addition to comparing MAGs among different WWTPs, we explored whether the 2024 bacterial AS MAGs obtained in this study could be distinguished from the 7164 MAGs of other non-engineered environments [20]

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Summary

Introduction

Microorganisms in activated sludge (AS) play key roles in the wastewater treatment processes. Their ecological behaviors and differences from microorganisms in other environments have mainly been studied using the 16S rRNA gene that may not truly represent in situ functions. Metagenomics aimed at recovering population genomes and annotating genetic potentials have been applied to AS and uncovered individual microorganisms or functions that are challenging to study using other methods [12,13,14], demonstrating that this approach is promising for revealing greater diversity at the functional level than the analysis of 16S rRNA gene sequences. Comparing the populations in AS and various non-AS ecosystems could provide insights into how the AS microbial community is assembled and whether the AS populations possess unique functional features that are vital to the adaption to the conditions of wastewater treatment bioreactors

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