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Fluctuation of microbial activities after influent load variations in a full-scale SBR: recovery of the biomass after starvation

Abstract

Due to variations in the production levels, a full-scale sequencing batch reactor (SBR) for post-treatment of tannery wastewater was exposed to low and high ammonia load periods. In order to study how these changes affected the N-removal capacity, the microbiology of the reactor was studied by a diverse set of techniques including molecular tools, activity tests, and microbial counts in samples taken along 3 years. The recover capacity of the biomass was also studied in a lab-scale reactor operated with intermittent aeration without feeding for 36 days. The results showed that changes in the feeding negatively affected the nitrifying community, but the nitrogen removal efficiencies could be restored after the concentration stress. Species substitution was observed within the nitrifying bacteria, Nitrosomonas europaea and Nitrobacter predominated initially, and after an ammonia overload period, Nitrosomonas nitrosa and Nitrospira became dominant. Some denitrifiers, with nirS related to Alicycliphilus, Azospirillum, and Marinobacter nirS, persisted during long-term reactor operation, but the community fluctuated both in composition and in abundance. This fluctuating community may better resist the continuous changes in the feeding regime. Our results showed that a nitrifying–denitrifying SBR could be operated with low loads or even without feeding during production shut down periods.

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References

  1. Amann RI (1995) In situ identification of microorganisms by whole cell hybridization with rRNA-targeted nucleic acid probes. In: Akkermans ADL, van Elsas JD, de Bruijn F (eds) Molecular microbial ecology manual. 3.3.6. Dluwer Academic Publishers, The Netherlands, pp 1–15

  2. APHA (1985) Standard methods for the examination of water and wastewater, 16th edn. American Public Health Association, Washington, DC

  3. Braker G, Zhou J, Wu L, Devol AH, Tiedje JM (2000) Nitrite reductase genes (nirK and nirS) as functional markers to investigate diversity of denitrifying bacteria in pacific northwest marine sediment communities. Appl Environ Microbiol 66:2096–2104

  4. Braker G, Ayala-del-Río HL, Devol AH, Fesefeldt A, Tiedje JM (2001) Community structure of denitrifiers, Bacteria, and Archaea along redox gradients in Pacific Northwest marine sediments by terminal restriction fragment length polymorphism analysis of amplified nitrite reductase (nirS) and 16S rRNA genes. Appl Environ Microbiol 67:1893–1901

  5. Cabezas A, Draper P, Muxí L, Etchebehere C (2006) Post-treatment of a slaughterhouse wastewater: stability of the microbial community of a sequencing batch reactor operated under oxygen limited conditions. Water Sci Technol 54:215–221

  6. Campos JL, Garrido JM, Mosquera-Corral A, Méndez R (2007) Stability of a nitrifying activated sludge reactor. Biochem Eng J 35:87–92

  7. Daims H, Nielsen JL, Nielsen PH, Schleifer K-H, Wagner M (2001) In situ characterization of Nitrospira-like nitrite-oxidizing bacteria active in wastewater treatment plants. Appl Environ Microbiol 67:5273–5284

  8. Dowing LS, Nerenberg R (2008) Effect of bulk liquid BOD concentration on activity and microbial community structure of a nitrifying, membrane-aerated biofilm. Appl Microbiol Biotechnol 8:153–162

  9. Dunbar J, Ticknor LO, Kuske CR (2001) Phylogenetic specificity and reproducibility and new method for analysis of terminal restriction fragment profiles of 16S rRNA genes from bacterial communities. Appl Environ Microbiol 67:190–197

  10. Egli K, Langer C, Siegrist HR, Zehnder AJ, Wagner M, van der Meer JR (2003) Community analysis of ammonia and nitrite oxidizers during start-up of nitritation reactors. Appl Environ Microbiol 69:3213–3222

  11. Etchebehere C, Errazquin MI, Dabert P, Moletta R, Muxí L (2001) Evaluation of the denitrifying microbiota of anoxic reactors. Fems Microbiol Ecol 35:259–265

  12. Etchebehere C, Errazquin MI, Cabezas A, Planzzola MJ, Mallo M, Lombardi P, Ottonello G, Borzacconi L, Muxi L (2002) Sludge bed development in denitrifying reactors using different inocula-performance and microbiological aspects. Water Sci Technol 45:365–370

  13. Fernández A, Huang S, Seston S, Xing J, Hickey R, Criddle C, Tiedje J (1999) How stable is stable? Function versus community composition. Appl Environ Microbiol 65:3697–3704

  14. Geets J, Boon N, Verstraete W (2006) Strategies of aerobic ammonia-oxidizing bacteria for coping with nutrient and oxygen fluctuations. Fems Microbiol Ecol 58:1–13

  15. Hammer Ø, Harper DAT, Ryan PD (2001) PAST: Paleontological Statistics Software Package for Education and Data Analysis. Palaeontologia Electronica 4:1–9. http://palaeo-electronica.org/2001_1/past/issue1_01.htm

  16. Heylen K, Gevers D, Vanparys B, Wittebolle L, Geets J, Boon N, De Vos P (2006) The incidence of nirS and nirK and their genetic heterogeneity in cultivated denitrifiers. Environ Microbiol 8:2012–2021

  17. Hiorns WD, Hastings RC, Head IM, McCarthy AJ, Saunders JR, Pickup RW, Hall GH (1995) Amplification of 16S ribosomal RNA genes of autotrophic ammonia-oxidizing bacteria demonstrates the ubiquity of nitrosospiras in the environment. Microbiology 141:2793–2800

  18. Horz H-P, Rotthauwe J-H, Lukow T, Liesack W (2000) Identification of major subgroups of ammonia-oxidizing bacteria in environmental samples by T-RFLP analysis of amoA PCR products. J Microbiol Methods 39:197–204

  19. Hoshino T, Terahara T, Tsuneda S, Hirata A, Inamori Y (2005) Molecular analysis of microbial population transition associated with the start of denitrification in a wastewater treatment process. J Appl Microbiol 99:1165–1175

  20. Hoshino T, Terahara T, Yamada K, Okuda H, Suzuki I, Tsuneda S, Hirata A, Inamori Y (2006) Long-term monitoring of the succession of a microbial community in activated sludge from a circulation flush toilet as a closed system. Fems Microbiol Ecol 55:459–470

  21. Hu J, Li D, Liu Q, Tao Y, He X, Wang X, Li X, Gao P (2009) Effect of organic carbon on nitrification efficiency and community composition of nitrifying biofilms. J Environ Sci 21(3):387–394

  22. Juretschko S, Timmermann G, Schmid M, Schleifer KH, Pommerening-Roser A, Koops HP, Wagner M (1998) Combined molecular and conventional analyses of nitrifying bacterium diversity in activated sludge: Nitrosococcus mobilis and Nitrospira-like bacteria as dominant populations. Appl Environ Microbiol 64:3042–3051

  23. Juretschko S, Loy A, Lehner A, Wagner M (2002) The microbial community composition of a nitrifying-denitrifying activated sludge from an industrial sewage treatment plant analyzed by the full-cycle rRNA approach. Syst Appl Microbiol 25:84–99

  24. Koops HP, Pommerening-Roser A (2001) Distribution and ecophysiology of the nitrifying bacteria emphasizing cultured species. Fems Microbiol Ecol 37:1–9

  25. Kuai LP, Verstraete W (1998) Ammonium removal by the oxygen limited autotrophic nitrification–denitrification system. Appl Environ Microbiol 64:4500–4506

  26. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163

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

  28. Lydmark P, Lind M, Sörensson F, Hermansson M (2006) Vertical distribution of nitrifying populations in bacterial biofilms from a full-scale nitrifying trickling filter. Environ Microbiol 8:2036–2049

  29. Miura Y, Hiraiwa MN, Ito T, Itonaga T, Watanabe Y, Okabe S (2007) Bacterial community structures in MBRs treating municipal wastewater: relationship between community stability and reactor performance. Water Res 41:627–637

  30. Mobarry BK, Wagner M, Urbain V, Rittmann BE, Stahl DA (1996) Phylogenetic probes for analyzing abundance and spatial organization of nitrifying bacteria. Appl Environ Microbiol 62:2156–2162

  31. Moura A, Tacão M, Henriques I, Dias J, Ferreira P, Correia A (2007) Characterization of bacterial diversity in two aerated lagoons of a wastewater treatment plant using PCR–DGGE analysis. Microbiol Res. doi:https://doi.org/10.1016/j.micres.2007.06.005

  32. Orhon D, Ates Genceli E, Sözen S (2000) Experimental evaluation of the nitrification kinetics for tannery wastewaters. Water SA 26:43–50

  33. Osaka T, Yoshie S, Tsuneda S, Hirata A, Iwami N, Inamori Y (2006) Identification of acetate- or methanol-assimilating bacteria under nitrate-reducing conditions by stable-isotope probing. Microb Ecol 52:253–266

  34. Philippot L (2005) Use of functional genes to quantify denitrifiers in the environment. Biochem Soc Trans 34:101–103

  35. Quevedo M, Guynot E, Muxí L (1996) Denitrifying potential of methanogenic sludge. Biotechnol Lett 18:1363–1368

  36. Rowan AK, Snape JR, Fearnside D, Barer MR, Curtis TP, Head IM (2003) Composition and diversity of ammonia-oxidising bacterial communities in wastewater treatment reactors of different design treating identical wastewater. Fems Microbiol Ecol 43:195–206

  37. Satoh H, Yamakawa T, Kindaichi T, Ito T, Okabe S (2006) Community structures and activities of nitrifying and denitrifying bacteria in industrial wastewater-treating biofilms. Biotechnol Bioeng 94:762–772

  38. Schramm A, De Beer D, Wagner M, Aman R (1998) Identification and activities in situ of Nitrosospira and Nitrospira spp. as dominant populations in a nitrifying fluidized bed reactor. Appl Environ Microbiol 64:3480–3485

  39. Smith NR, Yul Z, Mohn WW (2003) Stability of the bacterial community in a pulp mill effluent treatment system during normal operation and a system shutdown. Water Res 37:4873–4884

  40. Surmacz-Gorska J, Garnaey K, Demuynck C, Vanrolleghem P, Verstraete W (1996) Nitrification monitoring in activated sludge by oxygen uptake rate (OUR) measurements. Water Res 30:1228–1236

  41. Vlaeminck SE, Geets J, Vervaeren H, Boon N, Verstraete W (2007) Reactivation of aerobic and anaerobic ammonium oxidizers in OLAND biomass after long-term storage. Appl Microbiol Biotechnol 74:1376–1384

  42. Wagner M, Loy A, Nogueira R, Purkhold U, Lee N, Daims H (2002) Microbial community composition and function in wastewater treatment plants. Antonie Van Leeuwenhoek 81:665–680

  43. Wittebolle L, Marzorati M, Clement L, Balloi A, Daffonchio D, Heylen K, De Vos P, Verstraete W, Boon N (2009) Initial community evenness favours functionality under selective stress. Nature 458:623–626

  44. Yoshie S, Noda N, Tsuneda S, Hirata A, Inamori Y (2004) Salinity decreases nitrite reductase gene diversity in denitrifying bacteria of wastewater treatment systems. Appl Environ Microbiol 70:3152–3157

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Acknowledgements

The authors are grateful to the Center for Microbial Ecology in the Michigan State University for assistance in the cloning libraries and sequences. The authors also are grateful to the personal in charge of the WWTP in the tannery in Uruguay for providing data and samples. Financial support for this study was provided by CSIC-SP from the Universidad de la República in Uruguay.

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Correspondence to Claudia Etchebehere.

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Cabezas, A., Draper, P. & Etchebehere, C. Fluctuation of microbial activities after influent load variations in a full-scale SBR: recovery of the biomass after starvation. Appl Microbiol Biotechnol 84, 1191–1202 (2009). https://doi.org/10.1007/s00253-009-2138-x

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Keywords

  • Nitrification
  • Denitrification
  • Full scale
  • Long term
  • Community analysis
  • SBR