Skip to main content
SpringerLink
Log in

Microbiological Analysis of Two Deep Constructed Wetlands with Special Emphasis on the Removal of Pathogens and Antibiotic-Resistant Bacteria

  • Published:
Water, Air, & Soil Pollution Aims and scope Submit manuscript

Abstract

With the global concern on the role of wastewater treatment technologies in manifesting the emergence and dissemination of antibiotic-resistant bacteria (ARB), it has now become imperative to analyze the emerging technologies for handling them. This study assesses the efficiency of two deep constructed wetlands (CWs) receiving partially treated sewage from a residential complex and a hospital designed for the removal of organics and pathogens. These systems were further analyzed for the presence of major ARB to identify the role of CWs in mitigating antibiotic resistance among microbial communities. The bacterial community responsible for metabolic conversions was analyzed by metagenomic sequencing. Finally, the efficiencies of deep CWs were analyzed for the removal of specific bacteria resistant to three antibiotics—piperacillin, colistin (polymixin E), and cefoperazone. The overall removal of extended spectrum beta lactamase producers and carbapenemase producers was also studied. Our results indicate that CWs offer decent BOD and COD removal efficiencies of 74.04–78.71% and 53.85–64.37% respectively. However, a zero-order reaction between loading rate and removal rate was obtained after loading rate of 170 g/m2.day indicating the organic loading capacity of the system. Metagenomic analysis revealed the presence of bacteria with diverse metabolic potentials for substrate conversion. Removal of fecal coliforms was high in the CWs, but the most interesting observation was the attenuation in ARB, which was found to be comparable to, or even better, than the reported values from conventional moving bed bioreactor. This observation may be attributed to the high retention times offered by CWs compared to that of conventional systems making them an attractive future alternative for treating domestic as well as hospital sewage for emerging pollutants.

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

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Aga, D., Davies, J., Gandra, S., et al. (2018). (2018) Initiatives for addressing antimicrobial resistance in the environment: Current situation and challenges. Wellcome Trust.

  • Anderson, J. C., Carlson, J. C., Low, J. E., et al. (2013). Performance of a constructed wetland in Grand Marais, Manitoba, Canada: Removal of nutrients, pharmaceuticals, and antibiotic resistance genes from municipal wastewater. Chem Cent J, 7, 54. https://doi.org/10.1186/1752-153X-7-54.

    Article  CAS  Google Scholar 

  • APHA. (2005). Standard methods for the examination of water and wastewater (21st ed.). American Public Health Association.

  • Appling, D., Habteselassie, M. Y., Radcliffe, D., & Bradshaw, J. K. (2013). Preliminary study on the effect of wastewater storage in septic tank on E. coli concentration in summer. Water, 5, 1141–1151. https://doi.org/10.3390/w5031141.

    Article  CAS  Google Scholar 

  • Arenskötter, M., Bröker, D., & Steinbüchel, A. (2004). Biology of the metabolically diverse genus Gordonia. Appl Environ Microbiol, 70, 3195–3204. https://doi.org/10.1128/AEM.70.6.3195-3204.2004.

    Article  CAS  Google Scholar 

  • Arias, C. A., Cabello, A., Brix, H., & Johansen, N. H. (2003). Removal of indicator bacteria from municipal wastewater in an experimental two-stage vertical flow constructed wetland system. Water Sci Technol, 48, 35–41. https://doi.org/10.2166/wst.2003.0274.

    Article  CAS  Google Scholar 

  • Auvinen, H, Du Laing, G, Meers, E, Rousseau, DPL (2016) Constructed wetlands treating municipal and agricultural wastewater – An overview for Flanders, Belgium. In: Natural and constructed wetlands. Springer International Publishing, pp 179–207

  • Barancheshme, F, Munir, M (2018) Strategies to combat antibiotic resistance in the wastewater treatment plants. Front. Microbiol. 8

  • Berendonk, T. U., Manaia, C. M., Merlin, C., et al. (2015). Tackling antibiotic resistance: The environmental framework. Nat Rev Microbiol., 13, 310–317. https://doi.org/10.1038/nrmicro3439.

    Article  CAS  Google Scholar 

  • Börjesson, S., Matussek, A., Melin, S., et al. (2010). Methicillin-resistant Staphylococcus aureus (MRSA) in municipal wastewater: An uncharted threat? J Appl Microbiol, 108, 1244–1251. https://doi.org/10.1111/j.1365-2672.2009.04515.x.

    Article  Google Scholar 

  • Brown, D. G., & Jaffé, P. R. (2001). Effects of nonionic surfactants on bacterial transport through porous media. Environ Sci Technol, 35, 3877–3883. https://doi.org/10.1021/es010577w.

    Article  CAS  Google Scholar 

  • Centers for Disease Control and Prevention. (2013). Antibiotic resistancethreats in the United States, 2013. CDC.

  • Chandy, S., Thomas, K., Mathai, E., et al. (2013). Patterns of antibiotic use in the community and challenges of antibiotic surveillance in a lower-middle-income country setting: A repeated cross-sectional study in Vellore, South India. J Antimicrob Chemother, 68, 229–236.

    Article  CAS  Google Scholar 

  • Chen, J., Wei, X. D., Liu, Y. S., et al. (2016). Removal of antibiotics and antibiotic resistance genes from domestic sewage by constructed wetlands: Optimization of wetland substrates and hydraulic loading. Sci Total Environ, 565, 240–248. https://doi.org/10.1016/j.scitotenv.2016.04.176.

    Article  CAS  Google Scholar 

  • Clinical and Laboratory Standards Institute (2016). In: Performance standards for antimicrobial susceptibility testing. 26th ed. M100S. Clinical and Laboratory Standards Institute

  • Clinical and Laboratory Standards Institute (2017). In: Performance standards for antimicrobial susceptibility testing. 27th ed. M100. Clinical and Laboratory Standards Institute

  • Coates ARM, Halls G (2012) Antibiotics in phase ii and iii clinical trials. pp 167–183

  • Coenen, S., Muller, A., Adriaenssens, N., et al. (2009). European surveillance of antimicrobial consumption (ESAC): Outpatient parenteral antibiotic treatment in Europe. J Antimicrob Chemother, 64, 200–205. https://doi.org/10.1093/jac/dkp135.

    Article  CAS  Google Scholar 

  • Collado, L., Levican, A., Perez, J., et al. (2011). Arcobacter defluvii sp . nov ., Isolated from sewage samples. Int J Syst Evol Microbiol, 61, 2155–2161. https://doi.org/10.1099/ijs.0.025668-0.

    Article  Google Scholar 

  • Costa, S., Ganzerli, S., Rugiero, I., et al. (2017). Potential of Rhodobacter capsulatus grown in anaerobic-light or aerobic-dark conditions as bioremediation agent for biological wastewater treatments. Water (Switzerland), 9. https://doi.org/10.3390/w9020108.

  • Cydzik-Kwiatkowska, A., & Zielińska, M. (2016). Bacterial communities in full-scale wastewater treatment systems. World J Microbiol Biotechnol, 32, 1–8. https://doi.org/10.1007/s11274-016-2012-9.

    Article  CAS  Google Scholar 

  • Cyprowski, M., Stobnicka-Kupiec, A., Ławniczek-Wałczyk, A., et al. (2018). Anaerobic bacteria in wastewater treatment plant. Int Arch Occup Environ Health, 91, 571–579. https://doi.org/10.1007/s00420-018-1307-6.

    Article  CAS  Google Scholar 

  • Dires, S., Birhanu, T., Ambelu, A., & Sahilu, G. (2018). Antibiotic resistant bacteria removal of subsurface flow constructed wetlands from hospital wastewater. J Environ Chem Eng, 6, 4265–4272. https://doi.org/10.1016/j.jece.2018.06.034.

    Article  CAS  Google Scholar 

  • Frazer-Williams, R. A. D. (2010). A review of the influence of design parameters on the performance of constructed wetlands. J Chem Eng C, 29–42. https://doi.org/10.3329/jce.v25i0.7237.

  • García, M., Soto, F., González, J. M., et al. (2008). A comparison ofbacterial removal efficiencies in constructed wetlands and algae-based systems. Ecol Eng, 32, 238–243.

    Article  Google Scholar 

  • Gómez, P., Lozano, C., Benito, D., et al. (2016). Characterization of staphylococci in urban wastewater treatment plants in Spain, with detection of methicillin resistant Staphylococcus aureus ST398. Environment Pollut, 212, 71–76.

    Article  Google Scholar 

  • Gonzalez-Martinez, A., Rodriguez-Sanchez, A., Lotti, T., et al. (2016). Comparison of bacterial communities of conventional and A-stage activated sludge systems. Sci Rep, 6, 1–11. https://doi.org/10.1038/srep18786.

    Article  CAS  Google Scholar 

  • Guan, W., Yin, M., He, T., & Xie, S. (2015). Influence of substrate type on microbial community structure in vertical-flow constructed wetland treating polluted river water. Environ Sci Pollut Res, 22, 16202–16209. https://doi.org/10.1007/s11356-015-5160-9.

    Article  CAS  Google Scholar 

  • Hayward, J. L., Jackson, A. J., Yost, C. K., et al. (2018). Fate of antibiotic resistance genes in two Arctic tundra wetlands impacted by municipal wastewater. Sci Total Environ, 642, 1415–1428. https://doi.org/10.1016/j.scitotenv.2018.06.083.

    Article  CAS  Google Scholar 

  • He, Y., Nurul, S., Schmitt, H., et al. (2018). Evaluation of attenuation of pharmaceuticals, toxic potency, and antibiotic resistance genes in constructed wetlands treating wastewater effluents. Sci Total Environ, 631–632, 1572–1581. https://doi.org/10.1016/j.scitotenv.2018.03.083.

    Article  CAS  Google Scholar 

  • Kadlec, R. H., & Wallace, S. (2008). Treatment wetlands 508–515. CRC Press.

  • Karathanasis, A. D., Potter, C. L., & Coyne, M. S. (2003). Vegetation effects on fecal bacteria, BOD, and suspended solid removal in constructed wetlands treating domestic wastewater. Ecol Eng, 20, 157–169. https://doi.org/10.1016/S0925-8574(03)00011-9.

    Article  Google Scholar 

  • Kickuth, R (1977) Degradation and incorporation of nutrients from rural wastewaters by plant rhizosphere under limnic conditions. Utilization of manure by land spreading, pp.335-343.

  • King, T., Schmidt, S., & Environment SE-S of TT. (2020). Undefined Antibiotic resistant Klebsiella spp. from a hospital, hospital effluents and wastewater treatment plants in the uMgungundlovu District, KwaZulu-Natal, South. Sci Tot Environ, 712, 135550.

    Article  CAS  Google Scholar 

  • Knowlton, M. F., Cuvellier, C., & Jones, J. R. (2002). Initial performance of a high capacity surface-flow treatment wetland. Wetlands, 22, 522–527. https://doi.org/10.1672/0277-5212(2002)022[0522:IPOAHC]2.0.CO;2.

    Article  Google Scholar 

  • Kotwani, A., & Holloway, K. (2011). Trends in antibiotic use among outpatients in New Delhi, India. BMC Infect Dis, 11, 99. https://doi.org/10.1186/1471-2334-11-99.

    Article  Google Scholar 

  • Kotwani, A., Wattal, C., Joshi, P. C., & Holloway, K. (2012). Irrational use of antibiotics and role of the pharmacist: An insight from a qualitative study in New Delhi, India. J Clin Pharm Ther, 37, 308–312. https://doi.org/10.1111/j.1365-2710.2011.01293.x.

    Article  CAS  Google Scholar 

  • Kumar, A., & Pal, D. (2018). Antibiotic resistance and wastewater: Correlation, impact and critical human health challenges. J Environ chem eng, 6, 52–58.

    Article  Google Scholar 

  • Kümmerer, K. (2009). Antibiotics in the aquatic environment–A review–part I. Chemospere, 75, 417–434.

    Article  Google Scholar 

  • Lamba, M., & Ahammad, S. Z. (2017a). Sewage treatment effluents in Delhi: A key contributor of β-lactam resistant bacteria and genes to the environment. Chemosphere, 188, 249–256.

    Article  CAS  Google Scholar 

  • Lamba, M., & Ahammad, S. Z. (2017b). Performance comparison of secondary and tertiary treatment systems for treating antibiotic resistance. Water res, 127, 172–182.

    Article  CAS  Google Scholar 

  • Langergraber, G., Pressl, A., Leroch, K., et al. (2010). Comparison of single-stage and a two-stage vertical flow constructed wetland systems for different load scenarios. Water Sci Technol, 61, 1341–1348.

    Article  CAS  Google Scholar 

  • Liao, X., Chen, C., Wang, Z., et al. (2013). Changes of biomass and bacterial communities in biological activated carbon filters for drinking water treatment. Process Biochem, 48, 312–316. https://doi.org/10.1016/j.procbio.2012.12.016.

    Article  CAS  Google Scholar 

  • Liu, L., Liu, C., Zheng, J., et al. (2013). Elimination of veterinary antibiotics and antibiotic resistance genes from swine wastewater in the vertical flow constructed wetlands. Chemosphere, 91, 1088–1093. https://doi.org/10.1016/j.chemosphere.2013.01.007.

    Article  CAS  Google Scholar 

  • Liu, W., He, Z., Yang, C., et al. (2016). Microbial network for waste activated sludge cascade utilization in an integrated system of microbial electrolysis and anaerobic fermentation. Biotechnol Biofuels, 9, 1–15. https://doi.org/10.1186/s13068-016-0493-2.

    Article  CAS  Google Scholar 

  • Łuczkiewicz, A., Jankowska, K., Fudala-Książek, S., et al. (2010). Antimicrobial resistance of fecal indicators in municipal wastewater treatment plant. Water res, 44, 5089–5097.

    Article  Google Scholar 

  • Ma, X, Song, X, Li, X, et al. (2018) Characterization of microbial communities in pilot-scale constructed wetlands with Salicornia for treatment of marine aquaculture effluents. Archaea 2018.

  • Mai, X., Tao, R., Yang, Y., et al. (2015). Investigation of antibiotic resistance of indigenous bacteria and abundance of class I integron in matrix of constructed wetlands of different configurations. J Environ Sci, 36, 1776–1784.

    Google Scholar 

  • Marano RB, Cytryn E (2017) The mobile resistome in wastewater treatment facilities and downstream environments. In: Keen PL & Fugere R (eds) Antimicrobial resistance in wastewater treatment processes. John Wiley & Sons Inc, pp.129-155.

  • McLain, J. E., Cytryn, E., Durso, L. M., & Young, S. (2016). Culture-based methods for detection of antibiotic resistance in agroecosystems: Advantages, challenges, and gaps in knowledge. J Environ Qual, 45, 432–440. https://doi.org/10.2134/jeq2015.06.0317.

    Article  CAS  Google Scholar 

  • Mcleod, M. P., Warren, R. L., Hsiao, W. W. L., et al. (2006). The complete genome of Rhodococcus sp. RHA1 provides insights into a catabolic powerhouse. Proc Natl Acad Sci, 103, 15582–15587.

    Article  Google Scholar 

  • Morató, J., Codony, F., Sánchez, O., et al. (2014). Science of the total environment key design factors affecting microbial community composition and pathogenic organism removal in horizontal subsurface flow constructed wetlands. Sci Total Environ, 481, 81–89. https://doi.org/10.1016/j.scitotenv.2014.01.068.

    Article  CAS  Google Scholar 

  • Nascimento, A. L., Souza, A. J., Andrade, P. A. M., et al. (2018). Sewage sludge microbial structures and relations to their sources, treatments, and chemical attributes. Front Microbiol, 9. https://doi.org/10.3389/fmicb.2018.01462.

  • Negreanu, Y., Pasternak, Z., Jurkevitch, E., & Cytryn, E. (2012). Impact of treated wastewater irrigation on antibiotic resistance in agricultural soils. Environ Sci Technol, 46, 4800–4808. https://doi.org/10.1021/es204665b.

    Article  CAS  Google Scholar 

  • Nõlvak, H., Truu, M., Tiirik, K., et al. (2013). Dynamics of antibiotic resistance genes and their relationships with system treatment efficiency in a horizontal subsurface flow constructed wetland. Sci Total Environ, 461–462, 636–644. https://doi.org/10.1016/j.scitotenv.2013.05.052.

    Article  CAS  Google Scholar 

  • Olson, M. R., Axler, R. P., & Hicks, R. E. (2004). Effects of freezing and storage temperature on MS2 viability. J Virol Methods, 122, 147–152. https://doi.org/10.1016/j.jviromet.2004.08.010.

    Article  CAS  Google Scholar 

  • Pandey, N., & Bhatt, R. (2016). Role of soil associated Exiguobacterium in reducing arsenic toxicity and promoting plant growth in Vigna radiata. Eur J Soil Biol, 75, 142–150. https://doi.org/10.1016/j.ejsobi.2016.05.007.

    Article  CAS  Google Scholar 

  • Pruden, A., Pei, R., Storteboom, H., & Carlson, K. H. (2006). Antibiotic resistance genes as emerging contaminants: Studies in northern Colorado. Environ Sci Technol, 40, 7445–7450. https://doi.org/10.1021/es060413l.

    Article  CAS  Google Scholar 

  • Rampuria, A., Gupta, A. B., & Brighu, U. (2020). Nitrogen transformation processes and mass balance in deep constructed wetlands treating sewage, exploring the anammox contribution. Bioresour Technol, 314, 123737. https://doi.org/10.1016/j.biortech.2020.123737.

    Article  CAS  Google Scholar 

  • Sidrach-Cardona, R., & Bécares, E. (2013). Fecal indicator bacteria resistance to antibiotics in experimental constructed wetlands. Ecol Eng, 50, 107–111. https://doi.org/10.1016/j.ecoleng.2012.01.001.

    Article  Google Scholar 

  • Sinton, L. W., Donnison, A. M., & Hastie, C. M. (1993). Faecal streptococci as faecal pollution indicators: A review part I: Taxonomy and enumeration. New Zeal J Mar Freshw Res, 27, 101–115. https://doi.org/10.1080/00288330.1993.9516549.

    Article  Google Scholar 

  • Solyanikova, I., & Golovleva, L. A. (2011). Biochemical features of the degradation of pollutants by Rhodococcus as a basis for contaminated wastewater and soil cleanup. Microbiology, 80, 591.

    Article  CAS  Google Scholar 

  • Stevik, T. K., Aa, K., Ausland, G., & Hanssen, J. F. (2004). Retention and removal of pathogenic bacteria in wastewater percolating through porous media: A review. Water Res, 38, 1355–1367. https://doi.org/10.1016/j.watres.2003.12.024.

    Article  CAS  Google Scholar 

  • Tunçsiper, B., Ayaz, S. Ç., & Akça, L. (2012). Coliform bacteria removal from septic wastewater in a pilot-scale combined constructed wetland system. Environ Eng Manag J, 11, 1873–1879. https://doi.org/10.30638/eemj.2012.233.

    Article  Google Scholar 

  • Tzouvelekis, L. S., Markogiannakis, A., Psichogiou, M., et al. (2012). Carbapenemases in Klebsiella pneumoniae and other Enterobacteriaceae: An evolving crisis of global dimensions. Clin Microbiol Rev, 25, 682–707. https://doi.org/10.1128/CMR.05035-11.

    Article  CAS  Google Scholar 

  • Ulrich, H., Klaus, D., Irmgard, F., et al. (2005). Microbiological investigations for sanitary assessment of wastewater treated in constructed wetlands. Water Res, 39, 4849–4858.

    Article  CAS  Google Scholar 

  • Van Boeckel, T. P., Gandra, S., Ashok, A., et al. (2014). Articles Global antibiotic consumption 2000 to 2010: An analysis of national pharmaceutical sales data. Lancet Infect Dis, 14, 742–750. https://doi.org/10.1016/S1473-3099(14)70780-7.

    Article  Google Scholar 

  • WHO (2017) Prioritization of pathogens to guide discovery, research and development of new antibiotics for drug-resistant bacterial infections, including tuberculosis. World Health Organization. www.who.int/medicines/areas/rational_use/PPLreport_2017_09_19.pdf?ua=1.

  • Winward, G. P., Avery, L. M., Frazer-Williams, R., et al. (2008). A study of the microbial quality of grey water and an evaluation of treatment technologies for reuse. Ecol Eng, 32, 187–197. https://doi.org/10.1016/j.ecoleng.2007.11.001.

    Article  Google Scholar 

  • Wu, S., Carvalho, P. N., Müller, J. A., et al. (2016). Science of the total environment sanitation in constructed wetlands : A review on the removal of human pathogens and fecal indicators. Sci Total Environ, 541, 8–22. https://doi.org/10.1016/j.scitotenv.2015.09.047.

    Article  CAS  Google Scholar 

  • Zhang, T., & Li, B. (2011). Occurrence, transformation, and fate of antibiotics in municipal wastewater treatment plants. Crit Rev Environ Sci Technol, 41, 951–998. https://doi.org/10.1080/10643380903392692.

    Article  CAS  Google Scholar 

  • Zhang B, Xu X, Zhu L (2018) Activated sludge bacterial communities of typical wastewater treatment plants: distinct genera identification and metabolic potential differential analysis. AMB Express 8. https://doi.org/10.1186/s13568-018-0714-0

  • Zhao, R., Feng, J., Liu, J., et al. (2019). Deciphering of microbial community and antibiotic resistance genes in activated sludge reactors under high selective pressure of different antibiotics. Water Res, 151, 388–402.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Special thanks to Dr Dinesh Kumar from M/s. Rebound Enviro Tech Pvt. Ltd. for providing access to the wetland facilities and permitting us to work even when the system overloaded and was performing suboptimally, which finally led to vast improvement in the process efficiencies.

Availability of Data and Materials

All data generated or analyzed during this study are included in this published article (and its supplementary information files).

Code Availability

Not applicable

Funding

The financial support was provided by the Department of Science and Technology, Govt of India (Grant no. DST/TM/WTI/WIC/2K17/83).

Author information

Authors and Affiliations

  1. Department of Civil Engineering, Malaviya National Institute of Technology, Raj, Jaipur, 302017, India

    Aakanksha Rampuria, Akhilendra Bhushan Gupta, Niha Mohan Kulshreshtha & Urmila Brighu

Authors
  1. Aakanksha Rampuria
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Akhilendra Bhushan Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Niha Mohan Kulshreshtha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Urmila Brighu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

ABG conceptualized and supervised the study and acquired the funding, AR and NMK performed the experiments, AR and NMK wrote the original draft, and ABG and UB revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Aakanksha Rampuria.

Ethics declarations

Conflict of Interest

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rampuria, A., Gupta, A.B., Kulshreshtha, N.M. et al. Microbiological Analysis of Two Deep Constructed Wetlands with Special Emphasis on the Removal of Pathogens and Antibiotic-Resistant Bacteria. Water Air Soil Pollut 232, 174 (2021). https://doi.org/10.1007/s11270-021-05121-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11270-021-05121-3

Keywords

Search

Navigation