, Volume 29, Issue 6, pp 567–577 | Cite as

Inhibition of benzene, toluene, phenol and benzoate in single and combination on Anammox activity: implication to the denitrification–Anammox synergy

  • Shuchan Peng
  • Xinkuan Han
  • Fuzhong SongEmail author
  • Lilan Zhang
  • Caiying Wei
  • Peili Lu
  • Daijun ZhangEmail author
Original Paper


A previous study demonstrated that denitrification synergized with Anammox could accelerate the anaerobic degradation of benzene. The inhibitory effects of benzene, toluene, phenol and benzoate in single and combination on Anammox activity were investigated by short-term batch tests. The results indicated that the inhibition of single compounds on Anammox could be well fitted with the extended non-competitive and Luong inhibition kinetic models. The inhibitions of the individual compound were in order as follows: benzene > toluene > phenol > benzoate. The joint inhibitions of bi-component mixtures of benzene with toluene, benzene with phenol and benzene with benzoate on Anammox activity were additive; the joint inhibition of a tri-component mixture (benzene, toluene and phenol) was partly additive; and the joint inhibition of a multicomponent mixture (benzene, toluene, phenol and benzoate) was synergistic. The effect of benzoate on the denitrification–Anammox synergy for benzene degradation was evaluated using a long-term test. Although the average rate of benzene degradation decreased by 13% with the addition of 10 mg L−1 benzoate, the average rate of NO3 and NH4+ increased by approximately 1- and 0.56-fold, respectively, suggesting that benzoate favors the stability of the denitrification–Anammox synergy. The carboxylation of benzene would be a more favorable pathway for the anaerobic degradation of benzene under denitrification synergized with Anammox.


Benzene Toluene Phenol Benzoate Inhibition Denitrification–Anammox 



The partial financial support from the State Fundamental Research Funds for the Central Universities in China (0222005202053), the research of foundational science and advanced technology of Chongqing (cstc2017jcyjBX0042), the Natural Science Foundation of China (NSF 51078365), the Scientific Research Foundation (2011DA105287-ZD201505) of the State Key Laboratory of Coal Mine Disaster Dynamics and Control and the National Science and Technology Major Project for Water Pollution Control and Remediation (2012ZX07307-001) is gratefully acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10532_2018_9853_MOESM1_ESM.docx (301 kb)
Supplementary material 1 (DOCX 300 kb)


  1. Ali M, Okabe S (2015) Anammox-based technologies for nitrogen removal: advances in process start-up and remaining issues. Chemosphere 141:144–153. CrossRefPubMedGoogle Scholar
  2. APHA (2006) American Public Health Association. Standard Methods for the Examination of Water and Wastewater. American Public Health Association, American Water Works Association, Water Environment Federation 1085Google Scholar
  3. Carrera J, Torrijos M, Baeza JA, Lafuente J, Vicent T (2003) Inhibition of nitrification by fluoride in high-strength ammonium wastewater in activated sludge. Process Biochem 39:73–79. CrossRefGoogle Scholar
  4. Dapena-Mora A, Fernández I, Campos JL, Mosquera-Corral A, Méndez R, Jetten MSM (2007) Evaluation of activity and inhibition effects on Anammox process by batch tests based on the nitrogen gas production. Enzyme Microb Technol 40:859–865. CrossRefGoogle Scholar
  5. Ding S, Wu J, Zhang M, Lu H, Mahmood Q, Zheng P (2015) Acute toxicity assessment of ANAMMOX substrates and antibiotics by luminescent bacteria test. Chemosphere 140:174–183. CrossRefPubMedGoogle Scholar
  6. Evans WC, Fuchs G (1988) Anaerobic degradation of aromatic compounds. Annu Rev Microbiol 42:289–317CrossRefGoogle Scholar
  7. Gitiafroz R (2012) Microorganisms and metabolic pathways involved in anaerobic benzene biodegradation under nitrate-reducing conditions. Dissertation, University of TorontoGoogle Scholar
  8. González-Blanco G, Beristain-Cardoso R, Cuervo-López F, Cervantes FJ, Gómez J (2012) Simultaneous oxidation of ammonium and p-cresol linked to nitrite reduction by denitrifying sludge. Bioresour Technol 103:48–55. CrossRefPubMedGoogle Scholar
  9. Henze MCM, van Loosdrecht GA, Ekama DB (2010) Biological wastewater treatment: principles, modelling and design. Copyright @ IWA PUBLISHINGGoogle Scholar
  10. Hernández SM, Sun W, Sierra-Alvarez R, Field JA (2013) Toluene-nitrite inhibition synergy of anaerobic ammonium oxidizing (anammox) activity. Process Biochem 48:926–930. CrossRefGoogle Scholar
  11. Huang XL, Gao DW, Tao Y, Wang XL (2014) C2/C3 fatty acid stress on anammox consortia dominated by Candidatus Jettenia asiatica. Chem Eng J 253:402–407. CrossRefGoogle Scholar
  12. Jin R, Yang G, Yu J, Zheng P (2012) The inhibition of the Anammox process: a review. Chem Eng J 197:67–79. CrossRefGoogle Scholar
  13. Kartal B, De Almeida NM, Maalcke WJ, Op den Camp HJM, Jetten MSM, Keltjens JT (2013) How to make a living from anaerobic ammonium oxidation. FEMS Microbiol Rev 37:428–461. CrossRefPubMedGoogle Scholar
  14. Kasai Y, Kodama Y, Takahata Y, Hoaki T, Watanabe K (2007) Degradative capacities and bioaugmentation potential of an anaerobic benzene-degrading bacterium strain DN11. Environ Sci Technol 41:6222–6227. CrossRefPubMedGoogle Scholar
  15. Krebs HA, Wigginst D, Stubbs M (1983) Studies on the mechanism of the antifungal action of benzoate. Biochem J 07:657–663CrossRefGoogle Scholar
  16. Kumar M, Lin JG (2010) Co-existence of anammox and denitrification for simultaneous nitrogen and carbon removal-Strategies and issues. J Hazard Mater 178:1–9. CrossRefPubMedGoogle Scholar
  17. Lennerz BS, Vafai SB, Delaney NF, Clish CB, Deik AA, Pierce KA, Ludwig DS, Mootha VK (2015) Effects of sodium benzoate, a widely used food preservative, on glucose homeostasis and metabolic profiles in humans. Bioresour Technol 114:73–79. CrossRefGoogle Scholar
  18. Londry KL, Fedorak PM, Suflita JM (1997) Anaerobic degradation of m-Cresol by a sulfate-reducing bacterium. Appl Environ Microbiol 63:3170–3175PubMedPubMedCentralGoogle Scholar
  19. Lotti T, Van Der Star WRL, Kleerebezem R, Lubello C, Van Loosdrecht MCM (2012) The effect of nitrite inhibition on the anammox process. Water Res 46:2559–2569. CrossRefPubMedGoogle Scholar
  20. Luo F, Gitiafroz R, Devine CE, Gong Y, Hung LA, Raskin L, Edwards EA (2014) Metatranscriptome of an anaerobic benzene-degrading, nitrate-reducing enrichment culture reveals involvement of carboxylation in benzene ring activation. Appl Environ Microbiol 80:4095–4107. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Luong JHT (1985) Kinetics of ethanol inhibition in alcohol fermentation. Biotechnol Bioeng 27:280–285. CrossRefPubMedGoogle Scholar
  22. Mou Y (2012) The toxicity evaluation of sodium benzoate to organism at the molecular level by spectroscopies. Dissertation, Shandong UniversityGoogle Scholar
  23. Mulder A, Van De Graaf AA, Robertson LA, Kuenen JG (1995) Anaerobic ammonium oxidation discovered in a denitrifying fluidized bed reactor. FEMS Microbiol Ecol 16:177–184CrossRefGoogle Scholar
  24. Ni SQ, Ni JY, Hu DL, Sung S (2012) Effect of organic matter on the performance of granular anammox process. Bioresour Technol 110:701–705. CrossRefPubMedGoogle Scholar
  25. Peng S, Zhang L, Zhang D, Lu P, Zhang X (2017) Denitrification synergized with ANAMMOX for the anaerobic degradation of benzene: performance and microbial community structure. Appl Microbiol Biotechnol 101:4315–4325. CrossRefPubMedGoogle Scholar
  26. Ramos C, Fernández I, Suárez-Ojeda ME, Carrera J (2015) Inhibition of the anammox activity by aromatic compounds. Chem Eng J 279:681–688. CrossRefGoogle Scholar
  27. Schweighofer P, Frey W, Matsche N (1992) Nitrification inhibition-a source identification method for combined municipal combined municipal and/or industrial wastewater treatment plants. Water Sci Technol 26:1135–1146CrossRefGoogle Scholar
  28. Sierra-Alvarez R, Lettinga G (1991) The effect of aromatic structure on the inhibition of acetoclastic methanogenesis in granular sludge. Appl Microbiol Biotechnol 34:544–550. CrossRefGoogle Scholar
  29. Sikkema J, de Bont JA, Poolman B (1995) Mechanisms of membrane toxicity of hydrocarbons. Microbiol Rev 59:201–222PubMedPubMedCentralGoogle Scholar
  30. Yang GF, Guo XL, Chen SX, Liu JH, Guo LX, Jin RC (2013) The evolution of Anammox performance and granular sludge characteristics under the stress of phenol. Bioresour Technol 137:332–339. CrossRefPubMedGoogle Scholar
  31. Yao Z, Lu P, Zhang D, Wan X, Li Y, Peng S (2015) Stoichiometry and kinetics of the anaerobic ammonium oxidation (Anammox) with trace hydrazine addition. Bioresour Technol 198:70–76. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Coal Mine Disaster Dynamics and ControlChongqing UniversityChongqingPeople’s Republic of China
  2. 2.Department of Environmental ScienceChongqing UniversityChongqingPeople’s Republic of China

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