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Characterization of the Microbial Community in a Partial Nitrifying Sequencing Batch Biofilm Reactor

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Abstract

A lab-scale partial nitrifying sequencing batch biofilm reactor was a successful start-up. Denaturing gradient gel electrophoresis (DGGE) was used to investigate the bacterial community dynamics in three periods together with inocula sludge at ambient temperature. The DGGE profiles of bacteria and Shannon–Wiener index (H′) results showed that high free ammonia (FA) concentration referred to lower diversity in the bioreactor system. Cluster analysis indicated that microorganism in period III was similar with inocula sludge and was different from that in periods I and II. Similar results also appeared in ammonia-oxidizing bacteria (AOB) community structure and nitrite-oxidizing bacteria (NOB) community structure, and at least four AOB species and two NOB species were present in period III, respectively. Phylogenetic analysis of amoA gene sequences showed that Nitrosomonas eutropha cluster was predominant in all the three periods. With lower ammonium loads, three new operational taxonomic units formed and consisted Nitrosomonas sp. Cluster. This article demonstrated that microbial community, AOB, and NOB diversity were related with FA concentration closely at ambient temperature.

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References

  1. Anthonisen AC, Loehr RC, Prakasam TB, Srinath EG (1976) Inhibition of nitrification by ammonia and nitrous acid. J Water Pollut Control Fed 48:835–852

    PubMed  CAS  Google Scholar 

  2. Attard E, Poly F, Commeaux C, Laurent F, Terada A, Smets BF, Recous S, Le Roux X (2010) Shifts between Nitrospira- and Nitrobacter-like nitrite oxidizers underlie the response of soil potential nitrite oxidation to changes in tillage practices. Environ Microbiol 12:315–326

    Article  PubMed  CAS  Google Scholar 

  3. Bassam BJ, Caetano-Anolles G, Gresshoff PM (1991) Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal Biochem 196:80–83

    Article  PubMed  CAS  Google Scholar 

  4. Chuang HP, Imachi H, Tandukar M, Kawakami S, Harada H, Ohashi A (2007) Microbial community that catalyzes partial nitrification at low oxygen atmosphere as revealed by 16S rRNA and amoA genes. J Biosci Bioeng 104:525–528

    Article  PubMed  CAS  Google Scholar 

  5. Cui YW, Pang YZ, Peng CY, Gan XQ, Ye L (2006) Achieving biological nitrogen removal via nitrite by salt inhibition. Water Sci Technol 53:115–122

    PubMed  CAS  Google Scholar 

  6. Erguder TH, Boon N, Vlaeminck SE, Verstraete W (2008) Partial nitrification achieved by pulse sulfide doses in a sequential batch reactor. Environ Sci Technol 42:8715–8720

    Article  PubMed  CAS  Google Scholar 

  7. Gilbride KA, Lee DY, Beaudette LA (2006) Molecular techniques in wastewater: understanding microbial communities, detecting pathogens, and real-time process control. J Microbiol Methods 66:1–20

    Article  PubMed  CAS  Google Scholar 

  8. Guo J, Peng Y, Huang H, Wang S, Ge S, Zhang J, Wang Z (2010) Short- and long-term effects of temperature on partial nitrification in a sequencing batch reactor treating domestic wastewater. J Hazard Mater 179:471–479

    Article  PubMed  CAS  Google Scholar 

  9. Hellinga C, Schellen A, Mulder JW, van Loosdrecht MCM, Heijnen JJ (1998) The SHARON process: an innovative method for nitrogen removal from ammonium-rich waste water. Water Sci Technol 37:135–142

    CAS  Google Scholar 

  10. Hollocher TC, Tate ME, Nicholas DJ (1981) Oxidation of ammonia by Nitrosomonas europaea. Definite 18O-tracer evidence that hydroxylamine formation involves a monooxygenase. J Biol Chem 256:10834–10836

    PubMed  CAS  Google Scholar 

  11. Limpiyakorn T, Kurisu F, Sakamoto Y, Yagi O (2007) Effects of ammonium and nitrite on communities and populations of ammonia-oxidizing bacteria in laboratory-scale continuous-flow reactors. FEMS Microbiol Ecol 60:501–512

    Article  PubMed  CAS  Google Scholar 

  12. Lydmark P, Almstrand R, Samuelsson K, Mattsson A, Sorensson F, Lindgren PE, Hermansson M (2007) Effects of environmental conditions on the nitrifying population dynamics in a pilot wastewater treatment plant. Environ Microbiol 9:2220–2233

    Article  PubMed  CAS  Google Scholar 

  13. Nicolaisen MH, Ramsing NB (2002) Denaturing gradient gel electrophoresis (DGGE) approaches to study the diversity of ammonia-oxidizing bacteria. J Microbiol Methods 50:189–203

    Article  PubMed  CAS  Google Scholar 

  14. Otawa K, Asano R, Ohba Y, Sasaki T, Kawamura E, Koyama F, Nakamura S, Nakai Y (2006) Molecular analysis of ammonia-oxidizing bacteria community in intermittent aeration sequencing batch reactors used for animal wastewater treatment. Environ Microbiol 8:1985–1996

    Article  PubMed  CAS  Google Scholar 

  15. Ovreas L, Forney L, Daae FL, Torsvik V (1997) Distribution of bacterioplankton in meromictic lake saelenvannet, as determined by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA. Appl Environ Microbiol 63:3367–3373

    PubMed  CAS  Google Scholar 

  16. Park HD, Noguera DR (2004) Evaluating the effect of dissolved oxygen on ammonia-oxidizing bacterial communities in activated sludge. Water Res 38:3275–3286

    Article  PubMed  CAS  Google Scholar 

  17. Peng YZ, Li YZ, Peng CY, Wang SY (2004) Nitrogen removal from pharmaceutical manufacturing wastewater with high concentration of ammonia and free ammonia via partial nitrification and denitrification. Water Sci Technol 50:31–36

    PubMed  CAS  Google Scholar 

  18. Qiao S, Kanda R, Nishiyama T, Fujii T, Bhatti Z, Furukawa K (2010) Partial nitrification treatment for high ammonium wastewater from magnesium ammonium phosphate process of methane fermentation digester liquor. J Biosci Bioeng 109:124–129

    Article  PubMed  CAS  Google Scholar 

  19. Rotthauwe JH, Witzel KP, Liesack W (1997) The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations. Appl Environ Microbiol 63:4704–4712

    PubMed  CAS  Google Scholar 

  20. Sanchez O, Bernet N, Delgenes JP (2007) Effect of dissolved oxygen concentration on nitrite accumulation in nitrifying sequencing batch reactor. Water Environ Res 79:845–850

    Article  PubMed  CAS  Google Scholar 

  21. Sanz JL, Kochling T (2007) Molecular biology techniques used in wastewater treatment: an overview. Process Biochem 42:119–133

    Article  CAS  Google Scholar 

  22. Sofia A, Liu WT, Ong SL, Ng WJ (2004) In situ characterization of microbial community in an A/O submerged membrane bioreactor with nitrogen removal. Water Sci Technol 50:41–48

    PubMed  CAS  Google Scholar 

  23. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  PubMed  CAS  Google Scholar 

  24. Vadivelu VM, Keller J, Yuan ZG (2006) Effect of free ammonia and free nitrous acid concentration on the anabolic and catabolic processes of an enriched Nitrosomonas culture. Biotechnol Bioeng 95:830–839

    Article  PubMed  CAS  Google Scholar 

  25. van Dongen U, Jetten MS, van Loosdrecht MC (2001) The SHARON-Anammox process for treatment of ammonium rich wastewater. Water Sci Technol 44:153–160

    PubMed  Google Scholar 

  26. Van Hulle SWH, Volcke EIP, Teruel JL, Donckels B, van Loosdrecht MCM, Vanrolleghem PA (2007) Influence of temperature and pH on the kinetics of the Sharon nitritation process. J Chem Technol Biotechnol 82:471–480

    Article  Google Scholar 

  27. Wan C-Y, De Wever H, Diels L, Thoeye C, Liang J-B, Huang L-N (2011) Biodiversity and population dynamics of microorganisms in a full-scale membrane bioreactor for municipal wastewater treatment. Water Res 45:1129–1138

    Article  PubMed  CAS  Google Scholar 

  28. Wertz S, Poly F, Le Roux X, Degrange E (2008) Development and application of a PCR-denaturing gradient gel electrophoresis tool to study the diversity of Nitrobacter-like nxrA sequences in soil. FEMS Microbiol Ecol 63:261–271

    Article  PubMed  CAS  Google Scholar 

  29. Wojnowska-Baryla I, Cydzik-Kwiatkowska A, Zielinska M (2010) The application of molecular techniques to the study of wastewater treatment systems. Methods Mol Biol 599:157–183

    Article  PubMed  CAS  Google Scholar 

  30. Xia S, Li J, Wang R (2008) Nitrogen removal performance and microbial community structure dynamics response to carbon nitrogen ratio in a compact suspended carrier biofilm reactor. Ecol Eng 32:256–262

    Article  Google Scholar 

  31. Yamamoto T, Takaki K, Koyama T, Furukawa K (2008) Long-term stability of partial nitritation of swine wastewater digester liquor and its subsequent treatment by Anammox. Bioresour Technol 99:6419–6425

    Article  PubMed  CAS  Google Scholar 

  32. Ye L, Zhang T (2010) Estimation of nitrifier abundances in a partial nitrification reactor treating ammonium-rich saline wastewater using DGGE, T-RFLP and mathematical modeling. Appl Microbiol Biotechnol 88:1403–1412

    Article  PubMed  CAS  Google Scholar 

  33. Yu T, Qi R, Li D, Zhang Y, Yang M (2010) Nitrifier characteristics in submerged membrane bioreactors under different sludge retention times. Water Res 44:2823–2830

    Article  PubMed  CAS  Google Scholar 

  34. Zeng T, Li D, Zhang J (2011) Efficient star-up of partial nitrification process in sequencing batch biofilm reactor with domestic wastewater at room temperature. Adv Mater Res 255–260:2815–2818

    Article  Google Scholar 

  35. Zhang YX, Xu YL, Jia M, Zhou JT, Yuan SZ, Zhang JS, Zhang ZP (2009) Stability of partial nitrification and microbial population dynamics in a bioaugmented membrane bioreactor. J Microbiol Biotechnol 19:1656–1664

    PubMed  CAS  Google Scholar 

  36. Zhou J, Bruns MA, Tiedje JM (1996) DNA recovery from soils of diverse composition. Appl Environ Microbiol 62:316–322

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the National Natural Science Foundation of China (50878003), Beijing Natural Science Foundation (8092006), the State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology (QAK201005), and Major Science and Technology Program for Water Pollution Control and Treatment (2008ZX07208-003-003, 2008ZX07420-004-BJUT001, 2008ZX07208-005-003, 2009ZX07424-001, 2009ZX07424-002-003, 2009ZX07424-005).

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Correspondence to Jie Zhang.

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Zeng, T., Li, D. & Zhang, J. Characterization of the Microbial Community in a Partial Nitrifying Sequencing Batch Biofilm Reactor. Curr Microbiol 63, 543 (2011). https://doi.org/10.1007/s00284-011-0019-x

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