Applied Microbiology and Biotechnology

, Volume 102, Issue 10, pp 4381–4392 | Cite as

Defining microbial community composition and seasonal variation in a sewage treatment plant in India using a down-flow hanging sponge reactor

  • Naoki Nomoto
  • Masashi Hatamoto
  • Yuga Hirakata
  • Muntjeer Ali
  • Komal Jayaswal
  • Akinori Iguchi
  • Tsutomu Okubo
  • Masanobu Takahashi
  • Kengo Kubota
  • Tadashi Tagawa
  • Shigeki Uemura
  • Takashi Yamaguchi
  • Hideki Harada
Environmental biotechnology


The characteristics of the microbial community in a practical-scale down-flow hanging sponge (DHS) reactor, high in organic matter and sulfate ion concentration, and the seasonal variation of the microbial community composition were investigated. Microorganisms related to sulfur oxidation and reduction (2–27%), as well as Leucobacter (7.50%), were abundant in the reactor. Anaerobic bacteria (27–38% in the first layer) were also in abundance and were found to contribute to the removal of organic matter from the sewage in the reactor. By comparing the Simpson index, the abundance-based coverage estimator (ACE) index, and the species composition of the microbial community across seasons (summer/dry, summer/rainy, autumn/dry, and winter/dry), the microbial community was found to change in composition only during the winter season. In addition to the estimation of seasonal variation, the difference in the microbial community composition along the axes of the DHS reactor was investigated for the first time. Although the abundance of each bacterial species differed along both axes of the reactor, the change of the community composition in the reactor was found to be greater along the vertical axis than the horizontal axis of the DHS reactor.


DHS (down-flow hanging sponge) Microbial community composition Sewage treatment Developing country 



We are grateful to the National River Conservation Directorate (NRCD); Ministry of Environment, Forest and Climate Change (MOEF); the Indian government; and Uttar Pradesh Jal Nigam (UPJN) for extending various forms of assistance. We also thank Mr. A. Noori and Mr. P. Sjati for contributing to the experiments in India.


This study was funded by the Science and Technology Research Partnership for Sustainable Development (SATREPS) in Japan.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with animals performed by any of the authors.

Supplementary material

253_2018_8864_MOESM1_ESM.pdf (289 kb)
ESM 1 (PDF 289 kb)


  1. Agrawal LK, Okui H, Ueki Y, Harada H, Ohashi A (1997) Treatment of raw sewage in a temperate climate using a UASB reactor and the hanging sponge cubes process. Water Sci Technol 36:433–440Google Scholar
  2. APHA (2005) Standard methods, 21st ed. Washington DC, pp 4–136–140Google Scholar
  3. Asano R, Nakai Y, Kawada W, Shimura Y, Inamoto T, Fukushima J (2013) Seawater inundation from the 2011 Tohoku tsunami continues to strongly affect soil bacterial communities 1 year later. Microb Ecol 66(3):639–646CrossRefPubMedGoogle Scholar
  4. Clarke KR, Warwick RM (2001) Change in marine communities: an approach to statistical analysis and interpretation, 2nd edn. Natural Environment Research Council, Plymouth Marine Laboratory, PlymouthGoogle Scholar
  5. Flaherty KE, Matheson RE Jr, McMichael RH Jr, Perry WB (2013) The influence of freshwater on nekton community structure in hydrologically distinct basins in Northeastern Florida Bay, FL, USA. Estuar Coasts 36:918–939CrossRefGoogle Scholar
  6. Garrity P (2009) Bergey’s manual of systematic bacteriology, vol 3: the Firmicutes. 737.
  7. Halpern M, Shakéd T, Pukall R, Schumann P (2009) Leucobacter chironomi sp. nov., a chromate resistant bacterium isolated from a chironomid egg mass. Int J Syst Evol Microbiol 59:665–670CrossRefPubMedGoogle Scholar
  8. Iguchi A, Okubo T, Tachibana M, Nagai H, Uemura S, Yamaguchi T, Kubota K, Harada H (2013) Bacterial community structure of practical-scale DHS reactor treating municipal wastewater by UASB-DHS system and quantification of denitrifying bacteria. J Jpn Soc Civil Eng Ser G (Environ Res) 69(7):III_539–III_546 (in Japanese)Google Scholar
  9. Ju F, Zhang T (2015) Bacterial assembly and temporal dynamics in activated sludge of a full-scale municipal wastewater treatment plant. ISME J 9(3):683–695CrossRefPubMedGoogle Scholar
  10. Ju F, Guo F, Ye L, Xia Y, Zhang T (2014) Metagenomic analysis on seasonal microbial variations of activated sludge from a full-scale wastewater treatment plant over 4 years. Environ Microbiol Rep 6(1):80–89CrossRefPubMedGoogle Scholar
  11. Kim N-K, Oh S, Liu W-T (2016) Enrichment and characterization of microbial consortia degrading soluble microbial products discharged from anaerobic methanogenic bioreactors. Water Res 90:395–404CrossRefPubMedGoogle Scholar
  12. Kubota K, Hayashi M, Matsunaga K, Iguchi A, Ohashi A, Li YY, Yamaguchi T, Harada H (2014) Microbial community composition of a down-flow hanging sponge (DHS) reactor combined with an up-flow anaerobic sludge blanket (UASB) reactor for the treatment of municipal sewage. Bioresour Technol 151:144–150CrossRefPubMedGoogle Scholar
  13. Kuhl M, Jorgensen BB (1992) Microsensor measurements of sulfate reduction and sulfide oxidation in compact microbial communities of aerobic biofilms. Am Soc Microbiol 58(4):1164–1174Google Scholar
  14. Kuroda K, Chosei T, Nakahara N, Hatamoto M, Wakabayashi T, Kawai T, Araki N, Syutsubo K, Yamaguchi T (2015) High organic loading treatment for industrial molasses wastewater and microbial community shifts corresponding to system development. Bioresour Technol 196:225–234CrossRefPubMedGoogle Scholar
  15. Lee SH, Kang HJ, Park HD (2015) Influence of influent wastewater communities on temporal variation of activated sludge communities. Water Res 73(15):132–144CrossRefPubMedGoogle Scholar
  16. Levén L, Eriksson ARB, Schnürer A (2006) Effect of process temperature on bacterial and archaeal communities in two methanogenic bioreactors treating organic household waste. Fed Eur Microbiol Soc 59:683–693Google Scholar
  17. Liao J, Fang C, Yu J, Sathyagal A, Willman E, Liu W-T (2017) Direct treatment of high-strength soft drink wastewater using a down-flow hanging sponge reactor: performance and microbial community dynamics. Appl Microbiol Biotechnol 101:5925–5936CrossRefPubMedGoogle Scholar
  18. Liu L, Liu M, Wilkinson DM, Chen H, Yu X, Yang J (2017) DNA metabarcoding reveals that 200-μm-size-fractionated filtering is unable to discriminate between planktonic microbial and large eukaryotes. Mol Ecol ResourGoogle Scholar
  19. Ma A, Wen X, Zhao F, Xi Y, Huang X, Waite D, Guan J (2013) Effect of temperature variation on membrane fouling and microbial community structure in membrane bioreactor. Bioresour Technol 133:462–468CrossRefPubMedGoogle Scholar
  20. Machdar I, Sekiguchi Y, Sumino H, Ohashi A, Harada H (2000) Combination of a UASB reactor and a curtain type DHS (downflow hanging sponge) reactor as a cost-effective sewage treatment system for developing countries. Water Sci Technol 42:83–88Google Scholar
  21. Meerbergen K, Geel MV, Waud M, Willems KA, Dewil R, Impe JV, Appels L, Lievens B (2017) Assessing the composition of microbial communities in textile wastewater treatment plants in comparison with municipal wastewater treatment plants. Microbiol Open 6(1):1–13CrossRefGoogle Scholar
  22. Nomoto N, Ali M, Jayaswal K, Iguchi A, Hatamoto M, Okubo T, Takahashi M, Kubota K, Tagawa T, Uemura S, Yamaguchi T, Harada H (2017a) Characteristics of DO, organic matter, and ammonium profile for practical-scale DHS reactor under various organic load and temperature conditions. Environ Technol, in pressGoogle Scholar
  23. Nomoto N, Ali M, Jayaswal K, Iguchi A, Hatamoto M, Okubo T, Takahashi M, Kubota K, Tagawa T, Uemura S, Yamaguchi T, Harada H (2017b) Removal and oxygen consumption of retained sludge for organic matter, ammonium, and sulfur in a practical-scale down-flow hanging sponge sewage treatment reactor. In Proceedings of 6th annual international conference on sustainable energy and environmental sciences (SEES 2017), pp 52-58Google Scholar
  24. Oksanen J, Blanchet FG, Kindt R, Legendre P, Minchin PR, O'Hara RB, Simpson GL, Solymos P, Stevens MHH, Wagner H (2016) Vegan: community ecology package. R package version 2.3–4Google Scholar
  25. Okubo T, Kubota K, Yamaguchi T, Uemura S, Harada H (2016) Development of a new non-aeration-based sewage treatment technology: performance evaluation of a full-scale down-flow hanging sponge reactor employing third-generation sponge carriers. Water Res 102:138–146CrossRefPubMedGoogle Scholar
  26. Onodera T, Tandukar M, Sugiyama D, Uemura S, Ohashi A, Harada H (2014) Development of a sixth-generation down-flow hanging sponge (DHS) reactor using rigid sponge media for post-treatment of UASB treating municipal sewage. Bioresour Technol 152:93–100CrossRefPubMedGoogle Scholar
  27. Sato N, Okubo T, Onodera T, Ohashi A, Harada H (2006) Prospects for a self-sustainable sewage treatment system: a case study on full-scale UASB system in India’s Yamuna River basin. J Environ Manag 80:198–207CrossRefGoogle Scholar
  28. Sturm G, Jacobs J, Sproer C, Schumann P, Gescher J (2011) Leucobacter chromiiresistens sp. nov., a chromate-resistant strain. Int J Syst Evol Microbiol 61:956–960CrossRefPubMedGoogle Scholar
  29. Tandukar M, Uemura S, Ohashi A, Harada H (2006) Combining UASB and the “fourth generation” down-flow hanging sponge reactor for municipal wastewater treatment. Water Sci Technol 53:209–218CrossRefPubMedGoogle Scholar
  30. Tandukar M, Ohashi A, Harada H (2007) Performance comparison of a pilot scale UASB and DHS system and activated sludge process for the treatment of municipal wastewater. Water Res 41:2697–2705CrossRefPubMedGoogle Scholar
  31. Tang JC, Shibata A, Zhou Q, Katayama A (2007) Effect of temperature on reaction rate and microbial community in composting of cattle manure with rice straw. J Biosci Bioeng 104(4):321–328CrossRefPubMedGoogle Scholar
  32. Tawfik A, Ohashi A, Harada H (2006) Sewage treatment in a combined up-flow anaerobic sludge blanket (UASB)—down-flow hanging sponge (DHS) system. Biochem Eng J 29:210–219CrossRefGoogle Scholar
  33. Wang X, Wen X, Yan H, Ding K, Zhao F, Hu M (2011) Bacterial community dynamics in a functionally stable pilot-scale wastewater treatment plant. Bioresour Technol 102:2352–2357CrossRefPubMedGoogle Scholar
  34. Wang X, Hu M, Xia Y, Wen X, Ding K (2012) Pyrosequencing analysis of bacterial diversity in 14 wastewater treatment systems in China. Appl Environ Microbiol 78(19):7042–7047CrossRefPubMedPubMedCentralGoogle Scholar
  35. Xin X, He J, Wang Y, Feng J, Qiu W (2016) Role of aeration intensity on performance and microbial community profiles in a sequencing batch reaction kettle (SBRK) for wastewater nutrients rapid removal. Bioresour Technol 201:140–147CrossRefPubMedGoogle Scholar
  36. Yergeau E, Lawrence JR, Sanschagrin S, Waiser MJ, Korber DR, Greera CW (2012) Next-generation sequencing of microbial communities in the Athabasca River and its tributaries in relation to Oil Sands mining activities. Appl Environ Microbiol 78(21):7626–7637CrossRefPubMedPubMedCentralGoogle Scholar
  37. Zhang T, Shao MF, Ye L (2012) 454 pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants. ISME J 6(6):1137–1147CrossRefPubMedGoogle Scholar
  38. Zheng D, Sun Y, Li H, Lu S, Shan M, Xu S (2016) Multistage A-O activated sludge process for paraformaldehyde wastewater treatment and microbial community structure analysis. J Chem 2016:7Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Naoki Nomoto
    • 1
    • 2
  • Masashi Hatamoto
    • 3
  • Yuga Hirakata
    • 4
  • Muntjeer Ali
    • 5
  • Komal Jayaswal
    • 6
  • Akinori Iguchi
    • 7
  • Tsutomu Okubo
    • 8
  • Masanobu Takahashi
    • 5
  • Kengo Kubota
    • 9
  • Tadashi Tagawa
    • 10
  • Shigeki Uemura
    • 8
  • Takashi Yamaguchi
    • 4
  • Hideki Harada
    • 5
  1. 1.Department of Energy and Environment ScienceNagaoka University of TechnologyNiigataJapan
  2. 2.Department of Chemical and Biological EngineeringNational Institute of Technology, Ube CollegeYamaguchiJapan
  3. 3.Department of Civil and Environmental EngineeringNagaoka University of TechnologyNiigataJapan
  4. 4.Department of Science of Technology InnovationNagaoka University of TechnologyNiigataJapan
  5. 5.New Industry Creation Hatchery CenterTohoku UniversitySendaiJapan
  6. 6.Department of Civil EngineeringIndian Institute of Technology RoorkeeRoorkeeIndia
  7. 7.Faculty of Applied Life SciencesNiigata University of Pharmacy and Applied Life SciencesNiigataJapan
  8. 8.Department of Civil EngineeringNational Institute of Technology, Kisarazu CollegeKisarazuJapan
  9. 9.Department of Civil and Environmental EngineeringTohoku UniversitySendaiJapan
  10. 10.Department of Civil EngineeringNational Institute of Technology, Kagawa CollegeTakamatsuJapan

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