Microbial Ecology

, Volume 71, Issue 3, pp 660–671 | Cite as

Seasonal Changes in Bacterial Communities Cause Foaming in a Wastewater Treatment Plant

  • Ping Wang
  • Zhisheng YuEmail author
  • Jihong Zhao
  • Hongxun Zhang
Environmental Microbiology


Bio-foaming is a major problem in solid separation in activated sludge (AS) wastewater treatment systems. Understanding the changes in bacterial communities during sludge foaming is vital for explaining foam formation. Changes in bacterial communities in the foam, corresponding foaming AS, and non-foaming AS in a seasonal foaming wastewater treatment plant (WWTP) in Northern China were investigated by high-throughput pyrosequencing and molecular quantification-based approaches. We found that bacterial communities of the foam and the corresponding foaming AS were similar but markedly different from those of the non-foaming AS. Actinobacteria was the predominant phylum in the foam and the corresponding foaming AS, whereas Proteobacteria was predominant in the non-foaming AS. Similar to the results of most previous studies, our results showed that CandidatusMicrothrix parvicella” was the predominant filamentous bacteria in the foam and the corresponding foaming AS and was significantly enriched in the foam compared to the corresponding foaming AS. Its abundance decreased gradually with a slow disappearance of sludge foaming, indicating that its overgrowth had a direct relationship with sludge foaming. In addition to Candidatus M. parvicella, Tetrasphaera and Trichococcus might play a role in sludge foaming, because they supported the changes in AS microbial ecology for foam formation. The effluent water quality of the surveyed plant remained stable during the period of sludge foaming, but the microbial consortia responsible for nitrogen and phosphorus transformation and removal markedly changed compared to that in the non-foaming AS. This study adds to the previous understanding of bacterial communities causing foaming in WWTPs.


Sludge foaming Bacterial community Filamentous bacteria Pyrosequencing qPCR 



This work was supported by the Main Direction Program of Knowledge Innovation of Chinese Academy of Sciences (No. KZCX2-YW-JC407-2) and the Strategic Priority Research Program (B) of the Chinese Academy of Sciences (XDB15010200). We would like to thank the staffs in the investigated WWTP for sampling.

Supplementary material

248_2015_700_MOESM1_ESM.doc (136 kb)
ESM 1 (DOC 136 kb)
248_2015_700_MOESM2_ESM.doc (50 kb)
ESM 2 (DOC 49.5 kb)
248_2015_700_MOESM3_ESM.doc (32 kb)
ESM 3 (DOC 32 kb)
248_2015_700_MOESM4_ESM.doc (37 kb)
ESM 4 (DOC 37 kb)
248_2015_700_MOESM5_ESM.doc (52 kb)
ESM 5 (DOC 51 kb)
248_2015_700_MOESM6_ESM.doc (41 kb)
ESM 6 (DOC 41 kb)
248_2015_700_MOESM7_ESM.doc (68 kb)
ESM 7 (DOC 68 kb)


  1. 1.
    Ye L, Zhang T (2013) Bacterial communities in different sections of a municipal wastewater treatment plant revealed by 16S rDNA 454 pyrosequencing. Appl Microbiol Biotechnol 97:2681–2690CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Frigon D, Michael Guthrie R, Timothy Bachman G, Royer J, Bailey B, Raskin L (2006) Long-term analysis of a full-scale activated sludge wastewater treatment system exhibiting seasonal biological foaming. Water Res 40:990–1008CrossRefPubMedGoogle Scholar
  3. 3.
    Pujol R, Duchene P, Schetrite S, Canler J (1991) Biological foams in activated sludge plants: characterization and situation. Water Res 25:1399–1404CrossRefGoogle Scholar
  4. 4.
    Seviour E, Williams C, Seviour R, Soddell J, Lindrea K (1990) A survey of filamentous bacterial populations from foaming activated sludge plants in eastern states of Australia. Water Res 24:493–498CrossRefGoogle Scholar
  5. 5.
    Guo F, Wang ZP, Yu K, Zhang T (2015) Detailed investigation of the microbial community in foaming activated sludge reveals novel foam formers. Sci Rep 5Google Scholar
  6. 6.
    Slijkhuis H (1983) Microthrix parvicella, a filamentous bacterium isolated from activated sludge: cultivation in a chemically defined medium. Appl Environ Microbiol 46:832–839PubMedPubMedCentralGoogle Scholar
  7. 7.
    Shen FT, Huang HR, Arun A, Lu HL, Lin TC, Rekha P, Young CC (2007) Detection of filamentous genus Gordonia in foam samples using genus-specific primers combined with PCR-denaturing gradient gel electrophoresis analysis. Can J Microbiol 53:768–774CrossRefPubMedGoogle Scholar
  8. 8.
    Marsh TL, Liu WT, Forney LJ, Cheng H (1998) Beginning a molecular analysis of the eukaryal community in activated sludge. Water Sci Technol 37:455–460CrossRefGoogle Scholar
  9. 9.
    Blackall LL, Burrell PC, Gwilliam H, Bradford D, Bond PL, Hugenholtz P (1998) The use of 16S rDNA clone libraries to describe the microbial diversity of activated sludge communities. Water Sci Technol 37:451–454CrossRefGoogle Scholar
  10. 10.
    Erhart R, Bradford D, Seviour RJ, Amann R, Blackall LL (1997) Development and use of fluorescent in situ hybridization probes for the detection and identification of ‘Microthrix parvicella’ in activated sludge. Syst Appl Microbiol 20:310–318Google Scholar
  11. 11.
    Seviour RJ, Kragelund C, Kong Y, Eales K, Nielsen JL, Nielsen PH (2008) Ecophysiology of the Actinobacteria in activated sludge systems. Antonie Van Leeuwenhoek 94:21–33Google Scholar
  12. 12.
    Lechevlier MP, Lechevalier H (1974) Nocardia amarae sp. nov., an actinomycete common in foaming activated sludge. Int J Syst Bacteriol 24:278–288Google Scholar
  13. 13.
    Wang J, Li Q, Qi R, Tandoi V, Yang M (2014) Sludge bulking impact on relevant bacterial populations in a full-scale municipal wastewater treatment plant. Process Biochem 49:2258–2265CrossRefGoogle Scholar
  14. 14.
    Eikelboom DH (2000) Process control of activated sludge plants by microscopic investigation. IWA publishing, London, pp 85–102Google Scholar
  15. 15.
    Khan MA, Mohsin J, Faheem SM (2013) Monitoring microbial diversity of a full-scale municipal wastewater treatment plant in Dubai. APCBEE Proc 5:102–106CrossRefGoogle Scholar
  16. 16.
    State Environmental Protection Administration of China (2002) Standard methods of water and wastewater monitoring. 4th edGoogle Scholar
  17. 17.
    Amann RI, Krumholz L, Stahl DA (1990) Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology. J Bacteriol 172:762PubMedPubMedCentralGoogle Scholar
  18. 18.
    Daims H, Bruhl A, Amann R, Schleifer KH, Wagner M (1999) The domain-specific probe EUB338 is insufficient for the detection of all bacteria: development and evaluation of a more comprehensive probe set. Syst Appl Microbiol 22:434–444CrossRefPubMedGoogle Scholar
  19. 19.
    Kragelund C, Müller E, Schade M, Nguyen H, Lemmer H, Seviour RJ, Nielsen PH (2009) FISH handbook for biological wastewater treatment. IWA publishing, London, pp 73–84Google Scholar
  20. 20.
    Lane D (1991) 16S/23S rRNA sequencing. Nucleic acid techniques in bacterial systematics. 125–175Google Scholar
  21. 21.
    Haas BJ, Gevers D, Earl AM, Feldgarden M, Ward DV, Giannoukos G, Ciulla D, Tabbaa D, Highlander SK, Sodergren E (2011) Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons. Genome Res 21:494–504CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Pruesse E, Quast C, Knittel K, Fuchs BM, Ludwig W, Peplies J, Glöckner FO (2007) SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res 35:7188–7196CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Guo F, Zhang T (2012) Profiling bulking and foaming bacteria in activated sludge by high throughput sequencing. Water Res 46:2772–2782CrossRefPubMedGoogle Scholar
  24. 24.
    Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ (2009) Introducing mothur: open-source, platform-independent, community supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537–7541CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27:2194–2200CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Klammer S, Knapp B, Insam H, Dell’Abate MT, Ros M (2008) Bacterial community patterns and thermal analyses of composts of various origins. Waste Manag Res 26:173–187CrossRefPubMedGoogle Scholar
  27. 27.
    De Gregoris TB, Aldred N, Clare AS, Burgess JG (2011) Improvement of phylum- and class-specific primers for real-time PCR quantification of bacterial taxa. J Microbiol Methods 86:351–356CrossRefGoogle Scholar
  28. 28.
    Ritalahti KM, Amos BK, Sung Y, Wu Q, Koenigsberg SS, Löffler FE (2006) Quantitative PCR targeting 16S rRNA and reductive dehalogenase genes simultaneously monitors multiple dehalococcoides strains. Appl Environ Microbiol 72:2765–2774CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Bjornsson L, Hugenholtz P, Tyson GW, Blackall LL (2002) Filamentous Chloroflexi (green non-sulfur bacteria) are abundant in wastewater treatment processes with biological nutrient removal. Microbiology 148:2309–2318CrossRefPubMedGoogle Scholar
  30. 30.
    Weber CF, King GM (2010) Quantification of Burkholderia coxL genes in Hawaiian volcanic deposits. Appl Environ Microbiol 76:2212–2217CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Kaetzke A, Jentzsch D, Eschrich K (2005) Quantification of Microthrix parvicella in activated sludge bacterial communities by realtime PCR. Lett Appl Microbiol 40:207–211CrossRefPubMedGoogle Scholar
  32. 32.
    VandeWalle J, Goetz G, Huse S, Morrison H, Sogin M, Hoffmann R, Yan K, McLellan S (2012) Acinetobacter, Aeromonas and Trichococcus populations dominate the microbial community within urban sewer infrastructure. Environ Microbiol 14:2538–2552CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Klein AN, Frigon D, Raskin L (2007) Populations related to Alkanindiges, a novel genus containing obligate alkane degraders, are implicated in biological foaming in activated sludge systems. Environ Microbiol 9:1898–1912CrossRefPubMedGoogle Scholar
  34. 34.
    Sokal RR, Rohlf FJ (1995) Biometry. The principles and practice of statistics in biology research. Biometry the principles & practice of statistics in biological researchGoogle Scholar
  35. 35.
    Legendre P, Legendre L (2012) Developments in numerical ecology. Springer, BerlinGoogle Scholar
  36. 36.
    Xu M, Wu WM, Wu L, He Z, Nostrand JDV, Deng Y, Luo J, Carley J, Ginder-Vogel M, Gentry TJ (2010) Responses of microbial community functional structures to pilot-scale uranium in situ bioremediation. ISME J 4:1060–1070CrossRefPubMedGoogle Scholar
  37. 37.
    Van Etten E (2005) Multivariate analysis of ecological data using canoco. Aust Ecol 30:486–487CrossRefGoogle Scholar
  38. 38.
    Levantesi C, Rossetti S, Thelen K, Kragelund C, Krooneman J, Eikelboom D, Nielsen PH, Tandoi V (2006) Phylogeny, physiology and distribution of Candidatus ‘Microthrix calida’, calida’, a new microthrix species isolated from industrial activated sludge wastewater treatment plants. Environ Microbiol 8:1552–1563CrossRefPubMedGoogle Scholar
  39. 39.
    Petrovski S, Tillett D, Seviour RJ (2012) Isolation and complete genome sequence of a bacteriophage lysing Tetrasphaera jenkinsii, a filamentous bacteria responsible for bulking in activated sludge. Virus Genes 45:380–388CrossRefPubMedGoogle Scholar
  40. 40.
    Nielsen PH, Kragelund C, Seviour RJ, Nielsen JL (2009) Identity and ecophysiology of filamentous bacteria in activated sludge. FEMS Microbiol Rev 33:969–998CrossRefPubMedGoogle Scholar
  41. 41.
    Wang J, Qi R, Liu M, Li Q, Bao H, Li Y, Wang S, Tandoi V, Yang M (2014) The potential role of Candidatus ‘Microthrix parvicella’ in phosphorus removal during sludge bulking in two full-scale enhanced biological phosphorus removal plants. Water Sci Technol 70:367–375CrossRefPubMedGoogle Scholar
  42. 42.
    Heylen K (2006) Cultivation of denitrifying bacteria: optimization of isolation conditions and diversity study. Appl Environ Microbiol 72:2637–2643CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Martín HG, Ivanova N, Kunin V, Warnecke F, Barry KW, McHardy AC, Yeates C, He S, Salamov AA, Szeto E (2006) Metagenomic analysis of two enhanced biological phosphorus removal (EBPR) sludge communities. Nat Biotechnol 24:1263–1269CrossRefGoogle Scholar
  44. 44.
    Zhang T, Shao MF, Ye L (2012) 454 pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants. ISME J 6:1137–1147CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Ju F, Guo F, Ye L, Xia Y, Zhang T (2014) Metagenomic analysis on seasonal microbial variations of activated sludge from a fullscale wastewater treatment plant over 4 years. Environ Microbiol Rep 6:80–89CrossRefPubMedGoogle Scholar
  46. 46.
    Hu M, Wang X, Wen X, Xia Y (2012) Microbial community structures in different wastewater treatment plants as revealed by 454- pyrosequencing analysis. Bioresour Technol 117:72–79CrossRefPubMedGoogle Scholar
  47. 47.
    Hug T, Ziranke M, Siegrist H (2005) Dynamics of population and scumming on a full-scale wastewater treatment plant in Switzerland. Acta Hydrochim Hydrobiol 33:216–222CrossRefGoogle Scholar
  48. 48.
    Wong MT, Mino T, Seviour RJ, Onuki M, Liu WT (2005) In situ identification and characterization of the microbial community structure of full-scale enhanced biological phosphorous removal plants in Japan. Water Res 39:2901–2914CrossRefPubMedGoogle Scholar
  49. 49.
    Kong Y, Xia Y, Nielsen JL, Nielsen PH (2007) Structure and function of the microbial community in a full-scale enhanced biological phosphorus removal plant. Microbiology 153:4061–4073CrossRefPubMedGoogle Scholar
  50. 50.
    Jenkins D (1992) Towards a comprehensive model of activated sludge bulking and foaming. Water Sci Technol 25:215–230Google Scholar
  51. 51.
    Nielsen PH, Roslev P, Dueholm TE, Nielsen JL (2002) Microthrix parvicella, a functional lipid consumer in anaerobic-aerobic activated sludge plants. Water Sci Technol 46:73–80Google Scholar
  52. 52.
    de los Reyes FL III, Raskin L (2002) Role of filamentous microorganisms in activated sludge foaming: relationship of mycolata levels to foaming initiation and stability. Water Res 36:445–459CrossRefPubMedGoogle Scholar
  53. 53.
    Pitt P, Jenkins D (1990) Causes and control of Nocardia in activated sludge. Res J Water Pollut Control Fed 62:143–150Google Scholar
  54. 54.
    Rossetti S, Tomei MC, Nielsen PH, Tandoi V (2005) “Microthrix parvicella”, a filamentous bacterium causing bulking and foaming in activated sludge systems: a review of current knowledge. FEMS Microbiol Rev 29:49–64Google Scholar
  55. 55.
    Andreasen K, Nielsen PH (2000) Growth of Microthrix parvicella in nutrient removal activated sludge plants: studies of in situ physiology. Water Res 34:1559–1569Google Scholar
  56. 56.
    Mcilroy SJ (2013) Metabolic model for the filamentous Candidatus ‘Microthrix parvicella’ based on genomic and metagenomic analyses. ISME J 7:1161–1172Google Scholar
  57. 57.
    Mielczarek AT, Saunders AM, Larsen P, Albertsen M, Stevenson M, Nielsen JL, Nielsen PH (2013) The microbial database for Danish wastewater treatment plants with nutrient removal (MiDas-DK) a tool for understanding activated sludge population dynamics and community stability. IWA World Water Congress, London, pp 84–87Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.College of Resources and EnvironmentUniversity of Chinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.Henan Collaborative Innovation Center of Environmental Pollution Control and Ecological RestorationZhengzhouPeople’s Republic of China

Personalised recommendations