Skip to main content
SpringerLink
Log in

Mitigating microbiological risks of potential pathogens carrying antibiotic resistance genes and virulence factors in receiving rivers: Benefits of wastewater treatment plant upgrade

  • Research Article
  • Published:
Frontiers of Environmental Science & Engineering Aims and scope Submit manuscript

Abstract

Wastewater treatment plants (WWTPs) with additional tertiary ultrafiltration membranes and ozonation treatment can improve water quality in receiving rivers. However, the impacts of WWTP upgrade (WWTP-UP) on pathogens carrying antibiotic resistance genes (ARGs) and virulence factors (VFs) in rivers remain poorly understood. In this study, ARGs, VFs, and their pathogenic hosts were investigated in three rivers impacted by large-scale WWTP-UP. A five-year sampling campaign covered the periods before and after WWTP-UP. Results showed that the abundance of total metagenome-assembled genomes (MAGs) containing both ARGs and VFs in receiving rivers did not decrease substantially after WWTP-UP, but the abundance of MAGs belonging to pathogenic genera that contain both ARGs and VFs (abbreviated as PAVs) declined markedly. Genome-resolved metagenomics further revealed that WWTP-UP not only reduced most types of VFs and ARGs in PAVs, but also effectively eliminated efflux pump and nutritional VFs carried by PAVs in receiving rivers. WWTP-UP narrowed the pathogenic host ranges of ARGs and VFs and mitigated the cooccurrence of ARGs and VFs in receiving rivers. These findings underline the importance of WWTP-UP for the alleviation of pathogens containing both ARGs and VFs in receiving rivers.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Alneberg J, Bjarnason B S, de Bruijn I, Schirmer M, Quick J, Ijaz U Z, Lahti L, Loman N J, Andersson A F, Quince C (2014). Binning metagenomic contigs by coverage and composition. Nature Methods, 11(11): 1144–1146

    Article  CAS  Google Scholar 

  • Balcázar J L, Subirats J, Borrego C M (2015). The role of biofilms as environmental reservoirs of antibiotic resistance. Frontiers in Microbiology, 6: 1216

    Article  Google Scholar 

  • Biswal B K, Mazza A, Masson L, Gehr R, Frigon D (2014). Impact of wastewater treatment processes on antimicrobial resistance genes and their co-occurrence with virulence genes in Escherichia coli. Water Research, 50: 245–253

    Article  CAS  Google Scholar 

  • Cheng X, Xu J, Smith G, Nirmalakhandan N, Zhang Y (2021). Metagenomic profiling of antibiotic resistance and virulence removal: Activated sludge vs. Algal wastewater treatment system. Journal of Environmental Management, 295: 113129

    Article  CAS  Google Scholar 

  • Chu B T T, Petrovich M L, Chaudhary A, Wright D, Murphy B, Wells G, Poretsky R (2018). Metagenomics reveals the impact of wastewater treatment plants on the dispersal of microorganisms and genes in aquatic sediments. Applied and Environmental Microbiology, 84(5): e02168–17

    Article  Google Scholar 

  • Das D, Bordoloi A, Achary M P, Caldwell D J, Suri R P S (2022). Degradation and inactivation of chromosomal and plasmid encoded resistance genes/ARBs and the impact of different matrices on UV and UV/H2O2 based advanced oxidation process. Science of the Total Environment, 833: 155205

    Article  CAS  Google Scholar 

  • Eaves D J, Ricci V, Piddock L J V (2004). Expression of acrB, acrF, acrD, marA, and soxS in salmonella enterica serovar typhimurium: Role in multiple antibiotic resistance. Antimicrobial Agents and Chemotherapy, 48(4): 1145–1150

    Article  CAS  Google Scholar 

  • Eggen R I L, Hollender J, Joss A, Schärer M, Stamm C (2014). Reducing the discharge of micropollutants in the aquatic environment: the benefits of upgrading wastewater treatment plants. Environmental Science & Technology, 48(14): 7683–7689

    Article  CAS  Google Scholar 

  • Gao T W, Xiao K, Zhang J, Xue W C, Wei C H, Zhang X P, Liang S, Wang X M, Huang X (2022). Techno-economic characteristics of wastewater treatment plants retrofitted from the conventional activated sludge process to the membrane bioreactor process. Frontiers of Environmental Science & Engineering, 16(4): 49

    Article  Google Scholar 

  • Gu D, Dong N, Zheng Z, Lin D, Huang M, Wang L, Chan E W C, Shu L, Yu J, Zhang R, Chen S (2018). A fatal outbreak of ST11 carbapenem-resistant hypervirulent Klebsiella pneumoniae in a Chinese hospital: a molecular epidemiological study. Lancet. Infectious Diseases, 18(1): 37–46

    Article  Google Scholar 

  • Harnisz M, Kiedrzyńska E, Kiedrzyński M, Korzeniewska E, Czatzkowska M, Koniuszewska I, Jóźwik A, Szklarek S, Niestępski S, Zalewski Mm (2020). The impact of WWTP size and sampling season on the prevalence of antibiotic resistance genes in wastewater and the river system. Science of the Total Environment, 741: 140466

    Article  CAS  Google Scholar 

  • Ho Sui S J, Fedynak A, Hsiao W W L, Langille M G I, Brinkman F S L (2009). The association of virulence factors with genomic islands. PLoS One, 4(12): e8094

    Article  Google Scholar 

  • Hu Y, Yang X, Qin J, Lu N, Cheng G, Wu N, Pan Y, Li J, Zhu L, Wang X, Meng Z, Zhao F, Liu D, Ma J, Qin N, Xiang C, Xiao Y, Li L, Yang H, Wang J, Yang R, Gao G F, Wang J, Zhu B (2013). Metagenome-wide analysis of antibiotic resistance genes in a large cohort of human gut microbiota. Nature Communications, 4(1): 2151

    Article  Google Scholar 

  • Hu Y R, Zhang T Y, Jiang L, Luo Y, Yao S J, Zhang D, Lin K, Cui C (2019). Occurrence and reduction of antibiotic resistance genes in conventional and advanced drinking water treatment processes. Science of the Total Environment, 669: 777–784

    Article  CAS  Google Scholar 

  • Hyatt D, Chen G L, LoCascio P F, Land M L, Larimer F W, Hauser L J (2010). Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinformatics, 11(1): 119

    Article  Google Scholar 

  • Iakovides I C, Michael-Kordatou I, Moreira N F F, Ribeiro A R, Fernandes T, Pereira M F R, Nunes O C, Manaia C M, Silva A M T, Fatta-Kassinos D (2019). Continuous ozonation of urban wastewater: Removal of antibiotics, antibiotic-resistant Escherichia coli and antibiotic resistance genes and phytotoxicity. Water Research, 159: 333–347

    Article  CAS  Google Scholar 

  • Jäger T, Hembach N, Elpers C, Wieland A, Alexander J, Hiller C, Krauter G, Schwartz T (2018). Reduction of antibiotic resistant bacteria during conventional and advanced wastewater treatment, and the disseminated loads released to the environment. Frontiers in Microbiology, 9: 2599

    Article  Google Scholar 

  • Jain C, Rodríguez-R L, Phillippy M, Konstantinidis A T K, Aluru S (2018). High throughput ANI analysis of 90K prokaryotic genomes reveals clear species boundaries. Nature Communications, 9(1): 5114

    Article  Google Scholar 

  • Jiao Y N, Chen H, Gao R X, Zhu Y G, Rensing C (2017). Organic compounds stimulate horizontal transfer of antibiotic resistance genes in mixed wastewater treatment systems. Chemosphere, 184: 53–61

    Article  CAS  Google Scholar 

  • Josenhans C, Suerbaum S (2002). The role of motility as a virulence factor in bacteria. International Journal of Medical Microbiology, 291(8): 605–614

    Article  CAS  Google Scholar 

  • Ju F, Li B, Ma L P, Wang Y B, Huang D P, Zhang T (2016). Antibiotic resistance genes and human bacterial pathogens: co-occurrence, removal, and enrichment in municipal sewage sludge digesters. Water Research, 91: 1–10

    Article  CAS  Google Scholar 

  • Kang D W D, Li F, Kirton E, Thomas A, Egan R, An H, Wang Z (2019). MetaBAT 2: an adaptive binning algorithm for robust and efficient genome reconstruction from metagenome assemblies. PeerJ, 7: e7359

    Article  Google Scholar 

  • Keum J W, Bermudez H (2009). Enhanced resistance of DNA nanostructures to enzymatic digestion. Chemical Communications (Cambridge), 7(45): 7036–7038

    Article  Google Scholar 

  • Le T H, Ng C, Tran N, Chen H, Gin K Y (2018). Removal of antibiotic residues, antibiotic resistant bacteria and antibiotic resistance genes in municipal wastewater by membrane bioreactor systems. Water Research, 145: 498–508

    Article  CAS  Google Scholar 

  • Leclerc H, Moreau A (2002). Microbiological safety of natural mineral water. FEMS Microbiology Reviews, 26(2): 207–222

    Article  CAS  Google Scholar 

  • Lemos L N, Pereira R V, Quaggio R B, Martins L F, Moura L M S, da Silva A R, Antunes L P, da Silva A M, Setubal J C (2017). Genome-centric analysis of a thermophilic and cellulolytic bacterial consortium derived from composting. Frontiers in Microbiology, 8: 644

    Article  Google Scholar 

  • Li D, Liu C M, Luo R, Sadakane K, Lam T W (2015). MEGAHIT: an ultra-fast single-node solution for large and complex metagenomics assembly via succinct de Bruijn graph. Bioinformatics (Oxford, England), 31(10): 1674–1676

    CAS  Google Scholar 

  • Liang J, Mao G, Yin X, Ma L, Liu L, Bai Y, Zhang T, Qu J (2020). Identification and quantification of bacterial genomes carrying antibiotic resistance genes and virulence factor genes for aquatic microbiological risk assessment. Water Research, 168: 115160

    Article  CAS  Google Scholar 

  • Lin Z J, Zhou Z C, Zhu L, Meng L X, Shuai X Y, Sun Y J, Chen H (2021). Behavior of antibiotic resistance genes in a wastewater treatment plant with different upgrading processes. Science of the Total Environment, 771: 144814

    Article  CAS  Google Scholar 

  • Liu B, Zheng D D, Jin Q, Chen L H, Yang J (2019). VFDB 2019: a comparative pathogenomic platform with an interactive web interface. Nucleic Acids Research, 47(D1): D687–692

    Article  CAS  Google Scholar 

  • Lüddeke F, Heß S, Gallert C, Winter J, Güde H, Löffler H (2015). Removal of total and antibiotic resistant bacteria in advanced wastewater treatment by ozonation in combination with different filtering techniques. Water Research, 69: 243–251

    Article  Google Scholar 

  • Majeed H J, Riquelme M V, Davis B C, Gupta S, Angeles L, Aga D S, Garner E, Pruden A, Vikesland P J (2021). Evaluation of metagenomic-enabled antibiotic resistance surveillance at a conventional wastewater treatment plant. Frontiers in Microbiology, 12: 657954

    Article  Google Scholar 

  • Mao D Q, Luo Y, Mathieu J, Wang Q, Feng L, Mu Q H, Feng C Y, Alvarez P J J (2014). Persistence of extracellular DNA in river sediment facilitates antibiotic resistance gene propagation. Environmental Science & Technology, 48(1): 71–78

    Article  CAS  Google Scholar 

  • Nguyen A Q, Vu H P, Nguyen L N, Wang Q L, Djordjevic S P, Donner E, Yin H B, Nghiem L D (2021). Monitoring antibiotic resistance genes in wastewater treatment: Current strategies and future challenges. Science of the Total Environment, 783: 146964

    Article  CAS  Google Scholar 

  • Pan Y, Zeng J, Li L, Yang J, Tang Z, Xiong W, Li Y, Chen S, Zeng Z (2020). Coexistence of antibiotic resistance genes and virulence factors deciphered by large-scale complete genome analysis. mSystems, 5(3): e00821–19

    Article  CAS  Google Scholar 

  • Parks D H, Chuvochina M, Waite D W, Rinke C, Skarshewski A, Chaumeil P A, Hugenholtz P (2018). A standardized bacterial taxonomy based on genome phylogeny substantially revises the tree of life. Nature Biotechnology, 36(10): 996–1004

    Article  CAS  Google Scholar 

  • Penttinen R, Kinnula H, Lipponen A, Bamford J K, Sundberg L R (2016). High nutrient concentration can induce virulence factor expression and cause higher virulence in an environmentally transmitted pathogen. Microbial Ecology, 72(4): 955–964

    Article  CAS  Google Scholar 

  • Piotrowska M, Popowska M (2014). The prevalence of antibiotic resistance genes among aeromonas species in aquatic environments. Annals of Microbiology, 64(3): 921–934

    Article  CAS  Google Scholar 

  • Riquelme Breazeal M V, Novak J T, Vikesland P J, Pruden A (2013). Effect of wastewater colloids on membrane removal of antibiotic resistance genes. Water Research, 47(1): 130–140

    Article  CAS  Google Scholar 

  • Stamm C, Rasanen K, Burdon F J, Altermatt F, Jokela J, Joss A, Ackermann M, Eggen R I L (2016). Unravelling the impacts of micropollutants in aquatic ecosystems: interdisciplinary studies at the interface of large-scale ecology. Advances in Ecological Research, 55: 183–223

    Article  Google Scholar 

  • Stange C, Sidhu J P S, Toze S, Tiehm A (2019). Comparative removal of antibiotic resistance genes during chlorination, ozonation, and UV treatment. International Journal of Hygiene and Environmental Health, 222(3): 541–548

    Article  CAS  Google Scholar 

  • Su D, Ben W, Strobel B W, Qiang Z (2021). Impacts of wastewater treatment plant upgrades on the distribution and risks of pharmaceuticals in receiving rivers. Journal of Hazardous Materials, 406: 124331

    Article  CAS  Google Scholar 

  • Tang J Y, Bu Y Q, Zhang X X, Huang K L, He X W, Ye L, Shan Z J, Ren H Q (2016). Metagenomic analysis of bacterial community composition and antibiotic resistance genes in a wastewater treatment plant and its receiving surface water. Ecotoxicology and Environmental Safety, 132: 260–269

    Article  CAS  Google Scholar 

  • Tavares R D S, Tacao M, Figueiredo A S, Duarte A S, Esposito F, Lincopan N, Manaia C M, Henriques I (2020). Genotypic and phenotypic traits of blaCTX-M-carrying Escherichia coli strains from an UV-C-treated wastewater effluent. Water Research, 184: 116079

    Article  CAS  Google Scholar 

  • Varela A R, Manaia C M (2013). Human health implications of clinically relevant bacteria in wastewater habitats. Environmental Science and Pollution Research International, 20(6): 3550–3569

    Article  CAS  Google Scholar 

  • Wang H, Xu J, Tang W, Li H, Xia S, Zhao J, Zhang W X, Yang Y (2019). Removal efficacy of opportunistic pathogens and bacterial community dynamics in two drinking water treatment trains. Small, 15:1804436

    Google Scholar 

  • Wang Q, Liang J, Zhao C, Bai Y, Liu R, Liu H, Qu J (2020). Wastewater treatment plant upgrade induces the receiving river retaining bioavailable nitrogen sources. Environmental Pollution, 263:114478

    Article  CAS  Google Scholar 

  • Wu Y W, Simmons B A, Singer S W (2016). MaxBin 2.0: an automated binning algorithm to recover genomes from multiple metagenomic datasets. Bioinformatics (Oxford, England), 32(4): 605–607

    CAS  Google Scholar 

  • Yang L, Wen Q X, Chen Z Q, Duan R, Yang P (2019). Impacts of advanced treatment processes on elimination of antibiotic resistance genes in a municipal wastewater treatment plant. Frontiers of Environmental Science & Engineering, 13(3): 32

    Article  CAS  Google Scholar 

  • Yin X, Jiang X T, Chai B, Li L, Yang Y, Cole J R, Tiedje J M, Zhang T (2018). ARGs-OAP v2.0 with an expanded SARG database and Hidden Markov Models for enhancement characterization and quantification of antibiotic resistance genes in environmental metagenomes. Bioinformatics (Oxford, England), 34(13): 2263–2270

    Article  CAS  Google Scholar 

  • Yuan Q B, Huang Y M, Wu W B, Zuo P, Hu N, Zhou Y Z, Alvarez P J J (2019). Redistribution of intracellular and extracellular free & adsorbed antibiotic resistance genes through a wastewater treatment plant by an enhanced extracellular DNA extraction method with magnetic beads. Environment International, 131: 104986

    Article  CAS  Google Scholar 

  • Zhang B, Xia Y, Wen X H, Wang X H, Yang Y F, Zhou J Z, Zhang Y (2016). The composition and spatial patterns of bacterial virulence factors and antibiotic resistance genes in 19 wastewater treatment plants. PLoS One, 11(12): e0167422

    Article  Google Scholar 

  • Zhang J, Yu D, Dian L, Hai Y, Xin Y, Wei Y (2022). Metagenomics insights into the profiles of antibiotic resistome in combined sewage overflows from reads to metagenome assembly genomes. Journal of Hazardous Materials, 429: 128277

    Article  CAS  Google Scholar 

  • Zhao J, Li B, Lv P, Hou J, Qiu Y, Huang X (2022). Distribution of antibiotic resistance genes and their association with bacteria and viruses in decentralized sewage treatment facilities. Frontiers of Environmental Science & Engineering, 16(3): 35

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (42101128, 51578537, and 51778603) and Chinese Academy of Sciences (QYZDY-SSW-DQC004). The authors thank the Beijing Genomics Institute Central China for providing high-throughput sequencing services.

Author information

Authors and Affiliations

  1. Key Laboratory of Drinking Water Science and Technology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China

    Guannan Mao, Donglin Wang, Yaohui Bai & Jiuhui Qu

  2. State Key Laboratory of Tibetan Plateau Earth System, Environment and Resources (TPESER), Institute of Tibetan Plateau Research, Chinese Academy of Sciences, Beijing, 100101, China

    Guannan Mao

  3. University of Chinese Academy of Sciences, Beijing, 100049, China

    Donglin Wang

Authors
  1. Guannan Mao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Donglin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yaohui Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jiuhui Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yaohui Bai.

Additional information

Highlights

• Abundance of MAGs carrying ARG-VF pairs unchanged in rivers after WWTP upgrade.

• Upgrade of WWTPs significantly reduced diversity of pathogenic genera in rivers.

• Upgrade of WWTPs reduced most VF (ARG) types carried by potential pathogens in rivers.

• Upgrade of WWTPs narrowed the pathogenic host ranges of ARGs and VFs in rivers.

Supporting Information

11783_2023_1682_MOESM1_ESM.pdf

Mitigating microbiological risks of potential pathogens carrying antibiotic resistance genes and virulence factors in receiving rivers: Benefits of wastewater treatment plant upgrade

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mao, G., Wang, D., Bai, Y. et al. Mitigating microbiological risks of potential pathogens carrying antibiotic resistance genes and virulence factors in receiving rivers: Benefits of wastewater treatment plant upgrade. Front. Environ. Sci. Eng. 17, 82 (2023). https://doi.org/10.1007/s11783-023-1682-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11783-023-1682-4

Keywords

Search

Navigation