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Free nitrous acid (FNA) inhibition on denitrifying poly-phosphate accumulating organisms (DPAOs)

  • Environmental Biotechnology
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Abstract

Free nitrous acid (FNA) has been identified to be a ubiquitous inhibitor of a wide range of microorganisms, including bacteria involved in wastewater treatment. The FNA-induced inhibition on the anoxic (nitrite as electron acceptor) metabolism of denitrifying poly-phosphate accumulating organisms (DPAOs) was investigated using sludge from a sequencing batch reactor performing carbon, nitrogen, and phosphorus removal from synthetic wastewater. We found that FNA had a much stronger inhibitory effect on phosphorus (P) uptake and glycogen production than on poly-β-hydroxyalkanoate degradation and nitrite reduction. The intracellular adenosine triphosphate levels decreased sharply during the FNA incubation, and the decreasing rates were positively correlated with increasing FNA concentrations. The electron transport activity of DPAOs when exposed to FNA displayed a similar trend. Further, at FNA concentrations above 0.044 mg HNO2-N/L, the anaerobic metabolism of DPAOs was initiated despite of the presence of nitrite, as evidenced by the release of phosphorus and the consumption of glycogen. DPAO metabolism did not recover completely from FNA inhibition in the subsequent FNA-free environment. The recovery rate depended on the concentration of FNA applied in the previous anoxic period. These results suggest that the inhibitory effects are diverse and may be attributable to different mechanisms operating simultaneously.

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References

  • Ahn JT, Daidou S, Tsuneda S, Hirata A (2001) Metabolic behavior of denitrifying phosphate-accumulating organisms under nitrate and nitrite electron acceptor conditions. J Biosci Bioeng 92:442–446

    Article  CAS  Google Scholar 

  • Almeida JS, Julio SM, Reis MAM, Carrondo MJT (1995) Nitrite inhibition of denitrification by Pseudomonas fluorescens. Biotechnol Bioeng 46:194–201

    Article  CAS  Google Scholar 

  • Amann R (1995) Fluorescently labeled, ribosomal-RNA-targeted oligonucleotide probes in the study of microbial ecology. Mol Ecol 4:543–553

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • APHA (1995) Standard methods for the examination of water and wastewater, 19th edn. American Public Health Association, Washiongton

    Google Scholar 

  • Brul S, Coote P (1999) Preservative agents in foods—mode of action and microbial resistance mechanisms. Intern J Food Microbiol 50:1–17

    Article  CAS  Google Scholar 

  • Burge BW, Pfefferkorn ER (1964) Conditional lethal mutants of RNA animal virus. Virol 24:126–128

    Article  CAS  Google Scholar 

  • Castellani AG, Niven CF (1955) Factors affecting the bacterostatic action of sodium nitrite. Appl Microbiol 3:154–159

    CAS  Google Scholar 

  • Crocetti GR, Hugenholtz P, Bond PL, Schuler A, Keller J, Jenkins D, Blackall LL (2000) Identification of polyphosphate-accumulating organisms and design of 16S rRNA-directed probes for their detection and quantitation. Appl Environ Microbiol 66:1175–1182

    Article  CAS  Google Scholar 

  • Crocetti GR, Banfield JF, Keller J, Bond PL, Blackall LL (2002) Glycogen accumulating organisms in laboratory-scale and full-scale activated sludge process. Microbiol 148:3353–3364

    CAS  Google Scholar 

  • 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 22:434–444

    CAS  Google Scholar 

  • Dainty RH, Meredith GC (1972) Mechanisms of inhibition of growth of bacteria by nitrite. Meat Research Institute Annual Report 1972–73. HMSO. London, UK. pp 80–81

  • Freitas F, Temudo MF, Carvalho G, Oehmen A, Reis MAM (2009) Robustness of sludge enriched with short SBR cycles for biological nutrient removal. Biores Technol 100:1969–1976

    Article  CAS  Google Scholar 

  • Hellinga C, Van Loosdrecht MCM, Heijnen JJ (1999) Model based design of a novel process for nitrogen removal from concentrated flows. Math Comput Model Dyn Syst 5:351–371

    Google Scholar 

  • Hiller RG, Bassham JA (1965) Inhibition of CO2 fixation by nitrous acid. Biochimica Et Biophysica Acta 109:607–610

    Article  CAS  Google Scholar 

  • Hinze H, Holzer H (1985) Accumulation of nitrite and sulfite in yeast-cells and synergistic depletion of the intracellular ATP content. Zeitschrift Fur Lebensmittel-Untersuchung Und-Forschung 180:117–120

    Article  CAS  Google Scholar 

  • Hu JY, Ong SL, Ng WJ, Lu F, Fan XJ (2003) A new method for characterizing denitrifying phosphorus removal bacteria by using three different types of electron acceptors. Water Res 37:3463–3471

    Article  CAS  Google Scholar 

  • Martin HG, Ivanova N, Kunin V, Warnecke F, Barry KW, McHardy AC, Yeates C, He SM, Salamov AA, Szeto E, Dalin E, Putnam NH, Shapiro HJ, Pangilinan JL, Rigoutsos I, Kyrpides NC, Blackall LL, McMahon KD, Hugenholtz P (2006) Metagenomic analysis of two enhanced biological phosphorus removal (EBPR) sludge communities. Nat Biotechnol 24:1263–1269

    Article  CAS  Google Scholar 

  • Meinhold J, Arnold E, Isaacs S (1999) Effect of nitrite on anoxic phosphate uptake in biological phosphorus removal activated sludge. Water Res 33:1871–1883

    Article  CAS  Google Scholar 

  • Meyer RL, Saunders AM, Blackall LL (2006) Putative glycogen-accumulating organisms belonging to the Alphaproteobacteria identified through rRNA-based stable isotope probing. Microbiology-Sgm 152:419–429

    Article  CAS  Google Scholar 

  • Mortensen HD, Jacobsen T, Koch AG, Arneborg N (2008) Intracellular pH homeostasis plays a role in the tolerance of Debaryomyces hansenii and Candida zeylanoides to acidified nitrite. Appl Environ Microbiol 74:4835–4840

    Article  CAS  Google Scholar 

  • Oehmen A, Keller-Lehmann B, Zeng RJ, Yuan ZG, Keller J (2005) Optimisation of poly-β-hydroxyalkanoate analysis using gas chromatography for enhanced biological phosphorus removal systems. J Chromatogr A 1070:131–136

    Article  CAS  Google Scholar 

  • Oehmen A, Carvalho G, Freitas F, Reis MAM (2010) Assessing the abundance and activity of denitrifying polyphosphate accumulating organisms through molecular and chemical techniques. Water Sci Technol 61:2061–2068

    Article  CAS  Google Scholar 

  • O’Leary V, Solberg M (1976) Effect of sodium nitrite inhibition on intracellular thiol groups and on the activity of certain glycolytic enzymes in Clostridium perfringens. Appl Environ Microbiol 31:208–212

    Google Scholar 

  • Pijuan M, Yuan Z (2010) Development and optimization of a sequencing batch reactor for nitrogen and phosphorus removal from abattoir wastewater to meet irrigation standards. Water Sci Technol 61:2105–2112

    Article  CAS  Google Scholar 

  • Saito T, Brdjanovic D, van Loosdrecht MCM (2004) Effect of nitrite on phosphate uptake by phosphate accumulating organisms. Water Res 38:3760–3768

    Article  CAS  Google Scholar 

  • Sijbesma WFH, Almeida JS, Reis MAM, Santos H (1996) Uncoupling effect of nitrite during denitrification by Pseudomonas fluorescens: an in vivo 31P-NMR study. Biotechnol Bioeng 52:176–182

    Article  CAS  Google Scholar 

  • Smolders GJF, Van der Meij J, Van Loosdrecht MCM, Heijnen JJ (1994) Model of the anaerobic metabolism of the biological phosphorus removal process: stoichiometry and pH influence. Biotechnol Bioeng 43:461–470

    Article  CAS  Google Scholar 

  • Tchobanoglous G, Burton FL, Stensel HD (2002) Wastewater engineering: treatment and reuse, 4th edn. Metcalf & Eddy Inc., McGraw-Hill Science Engineering, New York

    Google Scholar 

  • Turk O, Mavinic DS (1986) Preliminary assessment of a shortcut in nitrogen removal from wastewater. Can J Civ Eng 13:600–605

    Article  Google Scholar 

  • Vadivelu VM, Yuan ZG, Fux C, Keller J (2006a) The inhibitory effects of free nitrous acid on the energy generation and growth processes of an enriched Nitrobacter culture. Environ Sci Technol 40:4442–4448

    Article  CAS  Google Scholar 

  • Vadivelu VM, Yuan ZG, Fux C, Keller J (2006b) Stoichiometric and kinetic characterisation of Nitrobacter in mixed culture by decoupling the growth and energy generation processes. Biotechnol Bioeng 94:1176–1188

    Article  CAS  Google Scholar 

  • Van Loosdrecht MCM, Jetten MSM (1998) Microbiological conversions in nitrogen removal. Water Sci Technol 38:1–7

    Google Scholar 

  • Weon SY, Lee CW, Lee SI, Koopman B (2002) Nitrite inhibition of aerobic growth of Acinetobacter sp. Water Res 36:4471–4476

    Article  CAS  Google Scholar 

  • Wong MT, Tan FM, Ng WJ, Liu WT (2004) Identification and occurrence of tetrad-forming Alphaproteobacteria in anaerobic–aerobic activated sludge process. Microbiol 150:3741–3748

    Article  CAS  Google Scholar 

  • Yarbrough JM, Rake JB, Eagon RG (1980) Bacterial inhibition effects of nitrite: inhibition of active transport, but not of group translocation, and of intracellular enzymes. Appl Environ Microbiol 39:831–834

    CAS  Google Scholar 

  • Zeng RJ, Saunders AM, Yuan Z, Blackall LL, Keller J (2003a) Identification and comparison of aerobic and denitrifying polyphosphate-accumulating organisms. Biotechnol Bioeng 83:140–148

    Article  CAS  Google Scholar 

  • Zeng RJ, Van Loosdrecht MCM, Yuan Z, Keller J (2003b) Metabolic model for glycogen-accumulating organisms in anaerobic/aerobic activated sludge systems. Biotechnol Bioeng 81:92–105

    Article  CAS  Google Scholar 

  • Zhou Y, Pijuan M, Yuan ZG (2007) Free nitrous acid inhibition on anoxic phosphorus uptake and denitrification by poly-phosphate accumulating organisms. Biotechnol Bioeng 98:903–912

    Article  CAS  Google Scholar 

  • Zhou Y, Pijuan M, Zeng RJ, Yuan Z (2008) Free nitrous acid inhibition on nitrous oxide reduction by a denitrifying-enhanced biological phosphorus removal sludge. Environ Sci Technol 42:8260–8265

    Article  CAS  Google Scholar 

Download references

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Authors and Affiliations

  1. Nanyang Environment and Water Research Institute (NEWRI), Nanyang Technological University, Block N1, Level B3b-29, 50 Nanyang Avenue, Singapore, 639798, Singapore

    Yan Zhou, Lily Ganda & Wun Jern Ng

  2. School of Civil and Environmental Engineering and Singapore Membrane Technology Center (SMTC-NEWRI), Nanyang Technological University, Block N2, Level 1c-101, 50 Nanyang Avenue, Singapore, 639798, Singapore

    Melvin Lim

  3. Advanced Water Management Centre (AWMC), The University of Queensland, St Lucia, Brisbane, QLD 4072, Australia

    Zhiguo Yuan

  4. Advanced Environmental Biotechnology Center (AEBC-NEWRI) Nanyang Technological University, SBS North wing, Level B2-01, 60 Nanyang Avenue, Singapore, 637551, Singapore

    Staffan Kjelleberg

  5. School of Biotechnology and Biomolecular Sciences and Centre for Marine Bio-Innovation, The University of New South Wales, UNSW, Sydney, 2052, Australia

    Staffan Kjelleberg

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  1. Yan Zhou
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  2. Lily Ganda
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  3. Melvin Lim
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  4. Zhiguo Yuan
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  5. Staffan Kjelleberg
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  6. Wun Jern Ng
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Correspondence to Wun Jern Ng.

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Zhou, Y., Ganda, L., Lim, M. et al. Free nitrous acid (FNA) inhibition on denitrifying poly-phosphate accumulating organisms (DPAOs). Appl Microbiol Biotechnol 88, 359–369 (2010). https://doi.org/10.1007/s00253-010-2780-3

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  • DOI: https://doi.org/10.1007/s00253-010-2780-3

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