Applied Microbiology and Biotechnology

, Volume 102, Issue 21, pp 9351–9361 | Cite as

Long-term effects of multi-walled carbon nanotubes on the performance and microbial community structures of an anaerobic granular sludge system

  • Xiaohui Wang
  • Minghan Zhu
  • Feifei Li
  • Congxuan Zhang
  • Xiaobiao ZhuEmail author
Environmental biotechnology


Multi-walled carbon nanotubes (MWCNTs) released into the sewage may cause negative and/or positive effects on the treatment system. The objective of this study was to explore over 110 days’ effect of MWCNTs on the performance of anaerobic granular sludge and microbial community structures in an upflow anaerobic sludge blanket (UASB) reactor. The results showed that MWCNTs had no significant effect on the removal of chemical oxidation demand (COD) and ammonia in UASB reactor, but the total phosphorus (TP) removal efficiency increased by 29.34%. The biogas production of the reactor did not change. The anaerobic granular sludge tended to excrete more EPS to resist the effects of MWCNTs during the long-term impact. Illumina MiSeq sequencing of 16S rRNA gene revealed that MWCNTs did not affect the microbial diversity, but altered the composition and structure of microbial community in the reactor. In this process, Saccharibacteria replaced Proteobacteria as the highest abundant bacterial phylum. MWCNTs promoted the differentiation of methanogen structure, resulting in increase of Methanomassiliicoccus, Methanoculleus, and the uncultured WCHA1–57. These results indicated that MWCNTs impacted the performance of UASB reactor and the structures of the microbial community in anaerobic granular sludge.


Multi-walled carbon nanotubes Upflow anaerobic sludge blanket Anaerobic granular sludge Microbial community structure Methanogens 



This work was supported by the National Key Research and Development Program of China (2016YFC0401105), NSFC (51408020 and 51308319), and the Fundamental Research Funds for the Central Universities (BUCTRC201606).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical statement

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

Supplementary material

253_2018_9273_MOESM1_ESM.pdf (317 kb)
ESM 1 (PDF 316 kb)


  1. Ambuchi JJ, Zhang ZH, Shan LL, Liang DD, Zhang P, Feng YJ (2017) Response of anaerobic granular sludge to iron oxide nanoparticles and multi-wall carbon nanotubes during beet sugar industrial wastewater treatment. Water Res 117:87–94. CrossRefPubMedGoogle Scholar
  2. APHA, AWWA, WEF (1998) Standard methods for the examination of water and wastewater, 20th edn. American Public Health Association, WashingtonGoogle Scholar
  3. Campos-Garcia J, Martinez DS, Alves OL, Leonardo AF, Barbieri E (2015) Ecotoxicological effects of carbofuran and oxidised multiwalled carbon nanotubes on the freshwater fish Nile tilapia: nanotubes enhance pesticide ecotoxicity. Ecotoxicol Environ Saf 111:131–137. CrossRefPubMedGoogle Scholar
  4. Campos-Garcia J, Martinez DS, Rezende KF, da Silva JR, Alves OL, Barbieri E (2016) Histopathological alterations in the gills of Nile tilapia exposed to carbofuran and multiwalled carbon nanotubes. Ecotoxicol Environ Saf 133:481–488. CrossRefPubMedGoogle Scholar
  5. Chao A (1984) Nonparametric estimation of the number of classes in a population. Scand J Stat 11(4):265–270Google Scholar
  6. Choi J, Liu Y (2014) Power generation and oil sands process-affected water treatment in microbial fuel cells. Bioresour Technol 169:581–587. CrossRefPubMedGoogle Scholar
  7. Frock AD, Notey JS, Kelly RM (2010) The genus Thermotoga: recent developments. Environ Technol 31(10):1169–1181. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Goyal D, Zhang XJ, Rooney-Varga JN (2010) Impacts of single-walled carbon nanotubes on microbial community structure in activated sludge. Lett Appl Microbiol 51(4):428–435. CrossRefPubMedGoogle Scholar
  9. Hai R, Wang Y, Wang X, Du Z, Li Y (2014) Impacts of multiwalled carbon nanotubes on nutrient removal from wastewater and bacterial community structure in activated sludge. PLoS One 9(9):e107345. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Iino T, Tamaki H, Tamazawa S, Ueno Y, Ohkuma M, Suzuki K, Igarashi Y, Haruta S (2013) Candidatus Methanogranum caenicola: a novel methanogen from the anaerobic digested sludge, and proposal of Methanomassiliicoccaceae fam. nov. and Methanomassiliicoccales ord. nov., for a methanogenic lineage of the class Thermoplasmata. Microbes Environ 28(2):244–250. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Jia G, Wang HF, Yan L, Wang X, Pei RJ, Yan T, Zhao YL, Guo XB (2005) Cytotoxicity of carbon nanomaterials: single-wall nanotube, multi-wall nanotube, and fullerene. Environ Sci Technol 39(5):1378–1383. CrossRefPubMedGoogle Scholar
  12. Kang S, Herzberg M, Rodrigues DF, Elimelech M (2008) Antibacterial effects of carbon nanotubes: size does matter. Langmuir 24(13):6409–6413. CrossRefPubMedGoogle Scholar
  13. Kindaichi T, Yamaoka S, Uehara R, Ozaki N, Ohashi A, Albertsen M, Nielsen PH, Nielsen JL (2016)Phylogenetic diversity and ecophysiology of candidate phylum Saccharibacteria in activated sludge. Fems Microbiol Ecol 92(6):fiw078 10.1093/femsec/fiw078CrossRefGoogle Scholar
  14. Lettinga G (1995) Anaerobic digestion and wastewater treatment systems. Antonie Van Leeuwenhoek 67(1):3–28. CrossRefPubMedGoogle Scholar
  15. Li LL, Tong ZH, Fang CY, Chu J, Yu HQ (2015) Response of anaerobic granular sludge to single-wall carbon nanotube exposure. Water Res 70:1–8. CrossRefPubMedGoogle Scholar
  16. Liu T, Zhang AN, Wang J, Liu S, Jiang X, Dang C, Ma T, Liu S, Chen Q, Xie S (2018) Integrated biogeography of planktonic and sedimentary bacterial communities in the Yangtze River. Microbiome 6(1):16. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Luongo LA, Zhang XQ (2010) Toxicity of carbon nanotubes to the activated sludge process. J Hazard Mater 178(1–3):356–362. CrossRefPubMedGoogle Scholar
  18. Mei XJ, Wang ZW, Miao Y, Wu ZC (2018) A pilot-scale anaerobic membrane bioreactor under short hydraulic retention time for municipal wastewater treatment: performance and microbial community identification. J Water Reuse Desalin 8(1):58–67. CrossRefGoogle Scholar
  19. Metenier K, Bonnamy S, Beguin F, Journet C, Bernier P, de La Chapelle ML, Chauvet O, Lefrant S (2002) Coalescence of single-walled carbon nanotubes and formation of multi-walled carbon nanotubes under high-temperature treatments. Carbon 40(10):1765–1773. CrossRefGoogle Scholar
  20. Ntim SA, Mitra S (2012) Adsorption of arsenic on multiwall carbon nanotube-zirconia nanohybrid for potential drinking water purification. J Colloid Interface Sci 375(1):154–159. CrossRefPubMedGoogle Scholar
  21. Owen WF, Stuckey DC, Jr JBH, Young LY, Mccarty PL (1979) Bioassay for monitoring biochemical methane potential and anaerobic toxicity. Water Res 13(6):485–492. CrossRefGoogle Scholar
  22. Parks AN, Chandler GT, Ho KT, Burgess RM, Ferguson PL (2015) Environmental biodegradability of C-14 single-walled carbon nanotubes by Trametes versicolor and natural microbial cultures found in New Bedford Harbor sediment and aerated wastewater treatment plant sludge. Environ Toxicol Chem 34(2):247–251. CrossRefPubMedGoogle Scholar
  23. Petersen EJ, Zhang LW, Mattison NT, O'Carroll DM, Whelton AJ, Uddin N, Nguyen T, Huang QG, Henry TB, Holbrook RD, Chen KL (2011) Potential release pathways, environmental fate, and ecological risks of carbon nanotubes. Environ Sci Technol 45(23):9837–9856. CrossRefPubMedGoogle Scholar
  24. Pettes MT, Shi L (2009) Thermal and structural characterizations of individual single-, double-, and multi-walled carbon nanotubes. Adv Funct Mater 19(24):3918–3925. CrossRefGoogle Scholar
  25. Pires ACC, Cleary DFR, Almeida A, Cunha A, Dealtry S, Mendonca-Hagler LCS, Smalla K, Gomes NCM (2012) Denaturing gradient gel electrophoresis and barcoded pyrosequencing reveal unprecedented archaeal diversity in mangrove sediment and rhizosphere samples. Appl Environ Microbiol 78(16):5520–5528CrossRefGoogle Scholar
  26. Qu YY, Ma Q, Deng J, Shen WL, Zhang XW, He ZL, Van Nostrand JD, Zhou JT, Zhou JZ (2015) Responses of microbial communities to single-walled carbon nanotubes in phenol wastewater treatment systems. Environ Sci Technol 49(7):4627–4635. CrossRefPubMedGoogle Scholar
  27. Qu YY, Zhang XW, Shen WL, Ma Q, You SN, Pei XF, Li SZ, Ma F, Zhou JT (2016) Illumina MiSeq sequencing reveals long-term impacts of single-walled carbon nanotubes on microbial communities of wastewater treatment systems. Bioresour Technol 211:209–215. CrossRefPubMedGoogle Scholar
  28. Rotaru AE, Shrestha PM, Liu F, Shrestha M, Shrestha D, Embree M, Zengler K, Wardman C, Nevin KP, Lovley DR (2014) A new model for electron flow during anaerobic digestion: direct interspecies electron transfer to Methanosaeta for the reduction of carbon dioxide to methane. Energy Environ Sci 7(1):408–415. CrossRefGoogle Scholar
  29. Shannon CE, Weaver W (1949) The mathematical theory of communication (Urbana, IL). M.D. computing computers in medical practice 60(3)Google Scholar
  30. Shin HS, Han SK, Song YC, Lee CY (2001) Performance of UASB reactor treating leachate from acidogenic fermenter in the two-phase anaerobic digestion of food waste. Water Res 35(14):3441–3447. CrossRefPubMedGoogle Scholar
  31. Simpson EH (1949) The measurement of diversity. Nature 163(4148):688CrossRefGoogle Scholar
  32. Stegen JC, Lin X, Konopka AE, Fredrickson JK (2012) Stochastic and deterministic assembly processes in subsurface microbial communities. ISME J 6(9):1653–1664CrossRefGoogle Scholar
  33. Wang X, Li J, Liu R, Hai R, Zou D, Zhu X, Luo N (2017) Responses of bacterial communities to CuO nanoparticles in activated sludge system. Environ Sci Technol 51(10):5368–5376. CrossRefPubMedGoogle Scholar
  34. Yadav T, Mungray AA, Mungray AK (2016) Effect of multiwalled carbon nanotubes on UASB microbial consortium. Environ Sci Pollut Res 23(5):4063–4072. CrossRefGoogle Scholar
  35. Yin Y, Zhang X (2008) Evaluation of the impact of single-walled carbon nanotubes in an activated sludge wastewater reactor. Water Sci Technol 58(3):623–628. CrossRefPubMedGoogle Scholar
  36. Zhang HL, Fang W, Wan YP, Sheng GP, Zeng RJ, Li WW, Yu HQ (2013) Phosphorus removal in an enhanced biological phosphorus removal process: roles of extracellular polymeric substances. Environ Sci Technol 47(20):11482–11489. CrossRefPubMedGoogle Scholar
  37. Zhang M, Wang W, Zhang Y, Teng Y, Xu Z (2017) Effects of fungicide iprodione and nitrification inhibitor 3, 4-dimethylpyrazole phosphate on soil enzyme and bacterial properties. Sci Total Environ 599:254–263. CrossRefPubMedGoogle Scholar
  38. Zhou JZ, Ning DL (2017) Stochastic community assembly: does it matter in microbial ecology? Microbiol Mol Biol R 81(4):e00002–e00017CrossRefGoogle Scholar
  39. Brar SK, Verma M, Tyagi RD, Surampalli RY (2010) Engineered nanoparticles in wastewater and wastewater sludge—evidence and impacts. Waste Manag 30(3):504–520. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Xiaohui Wang
    • 1
  • Minghan Zhu
    • 1
  • Feifei Li
    • 2
  • Congxuan Zhang
    • 1
    • 3
  • Xiaobiao Zhu
    • 1
    Email author
  1. 1.Department of Environmental Science and Engineering, College of Chemical EngineeringBeijing University of Chemical TechnologyBeijingChina
  2. 2.School of EnvironmentTsinghua UniversityBeijingChina
  3. 3.Beijing Sander Environmental Engineering Co. Ltd.BeijingChina

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