Short-term effects of excessive anaerobic reaction time on anaerobic metabolism of denitrifying polyphosphate- accumulating organisms linked to phosphorus removal and N₂O production

Abstract

The short-term effect of anaerobic reaction time (AnRT) (i.e., 90, 120 and 150 min) on the denitrifying phosphorus (P) removal performance and N₂O production was examined using a denitrifying enhanced biologic phosphorus removal (EBPR) sludge acclimatized with mixed acetate (HAc) and propionate (Pro) (in the molar ratio 3∶1) as carbon sources. The results showed that when the AnRT was prolonged from 90 to 150 min, the anaerobic polyhydroxyalkanoate (PHA) synthesis was decreased by 15.3%. Moreover, the ineffective PHA consumption occurred in anaerobic phases and contributed to an increased \mathrm{N}{\mathrm{O}}_{\mathrm{2}}^{-}-\mathrm{N} accumulation and higher free nitrous acid (FNA) concentrations (≥0.001–0.0011 mg HNO₂-N/L) in the subsequent anoxic phases, causing a severe inhibition on anoxic P-uptake and denitrification. Accordingly, the total nitrogen (TN) and total phosphorus (TP) removal efficiencies dropped by approximately 6.3% and 85.5%, respectively; and the ratio of anoxic N₂O-N production to TN removal increased by approximately 3.8%. The fluorescence in situ hybridization (FISH) analysis revealed that the sludge was mainly dominated by Accumulibacter (62.0% (SE_mean = 1.5%)). In conclusion, the short-term excessive anaerobic reaction time negatively impacted denitrifying P removal performance and stimulated more N₂O production, and its effect on P removal was more obvious than that on nitrogen removal.

Keywords

Denitrifying phosphorus removal / anaerobic reaction time / nitrous oxide / polyhydroxyalkanoate / free nitrous acid / fluorescence in-situ hybridization

References

[1] Kuba T, van Loosdrecht M C M, Heijnen J J. Phosphorus and nitrogen removal with minimal cod requirement by integration of denitrifying dephosphatation and nitrification in a two-sludge system. Water Research, 1996, 30(7): 1702–1710

[2] Osada T, Kuroda K, Yonaga M. Reducing nitrous oxide gas emissions from fill-and-draw type activated sludge process. Water Research, 1995, 29(6): 1607–1608

[3] Zeng R J, Lemaire R, Yuan Z, Keller J. Simultaneous nitrification, denitrification, and phosphorus removal in a lab-scale sequencing batch reactor. Biotechnology and Bioengineering, 2003, 84(2): 170–178

[4] Kampschreur M J, van der Star W R L, Wielders H A, Mulder J W, Jetten M S M, van Loosdrecht M C M. Dynamics of nitric oxide and nitrous oxide emission during full-scale reject water treatment. Water Research, 2008, 42(3): 812–826

[5] IPCC. Climate Change: The Scientific Basis. Cambridge: Cambridge University, 2001

[6] Barnard J L, Fothergill S. Secondary Phosphorus Release in Biological Phosphorus Removal Systems. Proceedings of the 71st Annual Water Environment Federation Technical Exposition and Conference; Orlando, Florida, Oct 3-7; Water Environment Federation: Alexandria, Virginia, 1998

[7] Coats E R, Watkins D L, Brinkman C K, Loge F J. Effect of anaerobic HRT on biological phosphorus removal and the enrichment of phosphorus accumulating organisms. Water Environ Res, 2011, 83(5): 461–469

[8] Barnard J L, Abraham K. Key features of successful BNR operation. Water Science and Technology, 2006, 53(12): 1–9

[9] Lopez C, Pons M N, Morgenroth E. Endogenous processes during long-term starvation in activated sludge performing enhanced biological phosphorus removal. Water Research, 2006, 40(8): 1519–1530

[10] Wang Y Y, Geng J J, Ren Z J, He W T, Xing M Y, Wu M, Chen S W. Effect of anaerobic reaction time on denitrifying phosphorus removal and N₂O production. Bioresource Technology, 2011, 102(10): 5674–5684

[11] Kuba T, Smolders G, Vanloosdrecht M C M, Heijnen J J. Biological phosphorus removal from waste-water by anaerobic-anoxic sequencing batch reactor. Environmental Science and Technology, 1993, 27(5-6): 241–252

[12] Pijuan M, Ye L, Yuan Z G. Free nitrous acid inhibition on the aerobic metabolism of poly-phosphate accumulating organisms. Water Research, 2010, 44(20): 6063–6072

[13] Zhu X Y, Chen Y G. Reduction of N₂O and NO generation in anaerobic-aerobic (low dissolved oxygen) biological wastewater treatment process by using sludge alkaline fermentation liquid. Environmental Science and Technology, 2011, 45(6): 2137–2143

[14] Chinese SEPA. Water and Wastewater Monitoring Methods. 4th ed. Beijing: Chinese Environmental Science Publishing House, 2002 (in Chinese)

[15] Ma J, Yang Q, Wang S Y, Wang L, Takigawa A, Peng Y Z. Effect of free nitrous acid as inhibitors on nitrate reduction by a biological nutrient removal sludge. Journal of Hazardous Materials, 2010, 175(1-3): 518–523

[16] Jenkins D, Richard M G, Daigger G T. Manual on the Causes and Control of Activated Sludge Bulking, Foaming and other Solids Separation Problems. 2nd ed. Colorado: Lewis Publishers, 2003

[17] Oehmen A, Keller-Lehmann B, Zeng R J, Yuan Z G, Keller J. Optimisation of poly-beta-hydroxyalkanoate analysis using gas chromatography for enhanced biological phosphorus removal systems. Journal of Chromatography. A, 2005, 1070(1-2): 131–136

[18] Amann R I. Partial and complete 16S rDNA sequences, their use in generation of 16S rDNA phylogenetic trees and their implications in molecular ecology studies. In: Akkermans, A D L, van Elsas, J D, de Bruijn F J, eds. Molecular Microbial Ecology Manual, vol.3.3.6. Holland: Kluwer Academic Publications Dordrecht, 1995, 1–15

[19] Daims H, Brühl A, Amann R, Schleifer K H, Wagner M. The domain-specific probe EUB338 is insufficient for the detection of all Bacteria: development and evaluation of a more comprehensive probe set. Systematic and Applied Microbiology, 1999, 22(3): 434–444

[20] Crocetti G R, Hugenholtz P, Bond P L, Schuler A, Keller J, Jenkins D, Blackall L L. Identification of polyphosphate-accumulating organisms and design of 16S rRNA-directed probes for their detection and quantitation. Applied Microbiology and Biotechnology, 2000, 66(3): 1175–1182

[21] Crocetti G R, Banfield J F, Keller J, Bond P L, Blackall L L. Glycogen-accumulating organisms in laboratory-scale and full-scale wastewater treatment processes. Microbiology (Reading, England), 2002, 148(Pt 11): 3353–3364

[22] Kong Y H, Ong S L, Ng W J, Liu W T. Diversity and distribution of a deeply branched novel proteobacterial group found in anaerobic-aerobic activated sludge processes. Environmental Microbiology, 2002, 4(11): 753–757

[23] Kuba T, Murnleitner E, van Loosdrecht M C M, Heijnen J J. A metabolic model for biological phosphorus removal by denitrifying organisms. Biotechnology and Bioengineering, 1996, 52(6): 685–695

[24] Carvalho G, Lemos P C, Oehmen A, Reis M A M. Denitrifying phosphorus removal: linking the process performance with the microbial community structure. Water Research, 2007, 41(19): 4383–4396

[25] Zeng R J, Yuan Z, Keller J. Enrichment of denitrifying glycogen-accumulating organisms in anaerobic/anoxic activated sludge system. Biotechnology and Bioengineering, 2003, 81(4): 397–404

[26] Kong Y H, Beer M, Rees G N, Seviour R J. Functional analysis of microbial communities in aerobic-anaerobic sequencing batch reactors fed with different phosphorus/carbon (P/C) ratios. Microbiology (Reading, England), 2002, 148(Pt 8): 2299–2307

[27] Lopez-Vazquez C M, Oehmen A, Hooijmans C M, Brdjanovic D, Gijzen H J, Yuan Z G, van Loosdrecht M C M. Modeling the PAO-GAO competition: effects of carbon source, pH and temperature. Water Research, 2009, 43(2): 450–462

[28] Lu H B, Keller J, Yuan Z G. Endogenous metabolism of Candidatus Accumulibacter phosphatis under various starvation conditions. Water Research, 2007, 41(20): 4646–4656

[29] Smolders G J F, van der Meij J, van Loosdrecht M C M, Heijnen J J. Model of the anaerobic metabolism of the biological phosphorus removal process: Stoichiometry and pH influence. Biotechnology and Bioengineering, 1994, 43(6): 461–470

[30] Zhou Y, Oehmen A, Lim M, Vadivelu V, Ng W J. The role of nitrite and free nitrous acid (FNA) in wastewater treatment plants. Water Research, 2011, 45(15): 4672–4682

[31] Yang Q, Liu X H, Peng C Y, Wang S Y, Sun H W, Peng Y Z. N₂O production during nitrogen removal via nitrite from domestic wastewater: main sources and control method. Environmental Science and Technology, 2009, 43(24): 9400–9406

[32] Kampschreur M J, Temmink H, Kleerebezem R, Jetten M S M, van Loosdrecht M C M. Nitrous oxide emission during wastewater treatment. Water Research, 2009, 43(17): 4093–4103

Acknowledgements

This study was supported by the National Natural Science Foundation of China (Grant Nos. 51078283 and 51178325), the Shanghai Science and Technology Committee Rising-Star Tracking Program (12QH1402400) and the Fundamental Research Funds for the Central University (Tongji University).

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2014 Higher Education Press and Springer-Verlag Berlin Heidelberg