Improving Substrate Consumption and Decrease of Growth Yield in Aerobic Cultures of Pseudomonas denitrificans By Applying Low Voltages in Bioelectric Systems

  • Luis F. Cházaro-RuizEmail author
  • María Irene López-Cázares
  • Ignacio González
  • Yanet Toriz
  • Felipe Alatriste-Mondragon
  • Marcela Santana
  • Lourdes B. CelisEmail author


It is well known that activated sludge treatment systems generate a lot of surplus sludge having environmental and economic impacts. Although several approaches have been proposed for the treatment/reuse of the excess of sludge, there are few studies focused on decreasing the biomass yield without affecting the metabolic activity. This work reports the effect of low magnitude electrical fields (0.07 to 0.2 V/cm) on the growth yield of a pure strain of Pseudomonas denitrificans (used as model microorganism). Cell potentials between 0.2 and 0.57 V were measured during 24 h to the aerobic culture; biomass production and substrate consumption were evaluated at regular intervals. Results indicated that the substrate (lactate) consumption efficiency increased with the applied potential, up to 100%, while the yield diminished 31% (0.34 g biomass/g lactate consumed) at 0.7 V vs. NHE. Bioenergetics showed that the fraction of electron equivalents toward biomass synthesis decreased from 0.68 (when no potential was applied) to 0.47 at 0.57 V, pointing out the redirection of the energy flow toward maintenance to cope with the stress caused by the imposed voltage. Therefore, the electrical stimulus could be used as control of biomass growth in aerobic wastewater treatment lines.


Activated sludge Bioelectric reactor Bioenergetics Biomass yield Biomass control Electric field 



The authors acknowledge the technical assistance of Elizabeth Cortés and Guillermo Vidriales. Y. Toriz acknowledges the scholarship provided by CONACYT to perform master studies. We also thank Alessandro Carmona for useful comments.


This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they do not have conflict of interest.

Supplementary material

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  1. 1.
    Liu, Y. (2003). Chemically reduced excess sludge production in the activated sludge process. Chemosphere, 50(1), 1–7. Scholar
  2. 2.
    Liu, Y., & Tay, J. H. (2001). Strategy for minimization of excess sludge production from the activated sludge process. Biotechnology Advances, 19(2), 97–107. Scholar
  3. 3.
    Pan, S. C., & Tseng, D. H. (2001). Sewage sludge ash characteristics and its potential applications. Water Science and Technology, 44(10), 261–267. Scholar
  4. 4.
    Stolarek, P., & Ledakowicz, S. (2001). Thermal processing of sewage sludge by drying, pyrolysis, gasification and combustion. Water Science and Technology, 44(10), 333–340. Scholar
  5. 5.
    Guo, W. Q., Yang, S. S., Xiang, W. S., Wang, X. J., & Ren, N. Q. (2013). Minimization of excess sludge production by in-situ activated sludge treatment processes - A comprehensive review. Biotechnology Advances, 31(8), 1386–1396. Scholar
  6. 6.
    Wang, Q., Wei, W., Gong, Y., Yu, Q., Li, Q., Sun, J., & Yuan, Z. (2017). Technologies for reducing sludge production in wastewater treatment plants: State of the art. Science of the Total Environment, 587-588, 510–521. Scholar
  7. 7.
    Ray, S., & Peters, C. A. (2008). Changes in microbiological metabolism under chemical stress. Chemosphere, 71(3), 474–483. Scholar
  8. 8.
    Schimel, J., Balser, T. S., & Wallenstein, M. (2007). Microbial stress-response physiology and its implications. Ecology, 88(6), 1386–1394. Scholar
  9. 9.
    Schröder, U., Harnisch, F., & Angenent, L. T. (2015). Microbial electrochemistry and technology: Terminology and classification. Energy and Environmental Science, 8(2), 513–519. Scholar
  10. 10.
    Thrash, J. C., & Coates, J. D. (2008). Review : Direct and indirect electrical stimulation of microbial metabolism. Environmental Science and Technology, 42(11), 3921–3931. Scholar
  11. 11.
    Fleming, J. T. (2010). Electronic interfacing with living cells. Advances in Biochemical Engineering / Biotechnology, 117, 155178. Scholar
  12. 12.
    Bajracharya, S., Sharma, M., Mohanakrishna, G., Dominguez Benneton, X., Strik, D. P. B. T. B., Sarma, P. M., & Pant, D. (2016). An overview on emerging bioelectrochemical systems (BESs): Technology for sustainable electricity, waste remediation, resource recovery, chemical production and beyond. Renewable Energy, 98, 153–170. Scholar
  13. 13.
    Velasco-Alvarez, N., González, I., Damian-Matsumura, P., & Gutiérrez-Rojas, M. (2011). Enhanced hexadecane degradation and low biomass production by Aspergillus niger exposed to an electric current in a model system. Bioresource Technology, 102(2), 1509–1515. Scholar
  14. 14.
    Parvanova-Mancheva, T., & Beschkov, V. (2009). Microbial denitrification by immobilized bacteria Pseudomonas denitrificans stimulated by constant electric field. Biochemical Engineering Journal, 44(2–3), 208–213. Scholar
  15. 15.
    Beschkov, V., Velizarov, S., Agathos, S. N., & Lukova, V. (2004). Bacterial denitrification of waste water stimulated by constant electric field. Biochemical Engineering Journal, 17(2), 141–145. Scholar
  16. 16.
    Thrash, J. C., Van Trump, J. I., Weber, K. A., Miller, E., Achenbach, L. A., & Coates, J. D. (2007). Electrochemical stimulation of microbial perchlorate reduction. Environmental Science and Technology, 41(5), 1740–1746. Scholar
  17. 17.
    Yin, X., Qiao, S., & Zhou, J. (2015). Using electric field to enhance the activity of Anammox bacteria. Applied Microbiology and Biotechnology, 99(16), 6921–6930. Scholar
  18. 18.
    Rittmann, B. E., & McCarty, P. L. (2000). Environmental biotechnology: Principles and applications (First ed.). Boston: McGraw-Hill.Google Scholar
  19. 19.
    APHA/AWWA/WEF. (2012). Standard methods for the examination of water and wastewater. Standard Methods, 541 ISBN 9780875532356.Google Scholar
  20. 20.
    Soga, T., & Ross, G. A. (1999). Simultaneous determination of inorganic anions, organic acids and metal cations by capillary electrophoresis. Journal of Chromatography A, 834(1–2), 65–71. Scholar
  21. 21.
    Guo, K., Soeriyadi, A. H., Feng, H., Prévoteau, A., Patil, S. A., Gooding, J. J., & Rabaey, K. (2015). Heat-treated stainless steel felt as scalable anode material for bioelectrochemical systems. Bioresource Technology, 195, 46–50. Scholar
  22. 22.
    Cantu, J. C., Tarango, M., Beier, H. T., & Ibey, B. L. (2016). The biological response of cells to nanosecond pulsed electric fields is dependent on plasma membrane cholesterol. Biochimica et Biophysica Acta - Biomembranes, 1858(11), 2636–2646. Scholar
  23. 23.
    Ibey, B. L., Xiao, S., Schoenbach, K. H., Murphy, M. R., & Pakhomov, A. G. (2009). Plasma membrane permeabilization by 60- and 600-ns electric pulses is determined by the absorbed dose. Bioelectromagnetics, 30(2), 92–99. Scholar
  24. 24.
    Dehghani, S., Rezaee, A., & Hosseinkhani, S. (2018). Effect of alternating electrical current on denitrifying bacteria in a microbial electrochemical system: biofilm viability and ATP assessment. Environmental Science and Pollution Research, 25(33), 33591–33598. Scholar
  25. 25.
    Qiao, S., Yin, X., Zhou, J., & Furukawa, K. (2014). Inhibition and recovery of continuous electric field application on the activity of Anammox biomass. Biodegradation, 25(4), 505–513. Scholar
  26. 26.
    Shin, H., Zeikus, J., & Jain, M. (2002). Electrically enhanced ethanol fermentation by Clostridium thermocellum and Saccharomyces cerevisiae. Applied Microbiology and Biotechnology, 58(4), 476–481. Scholar
  27. 27.
    Baudler, A., Schmidt, I., Langner, M., Greiner, A., & Schröder, U. (2015). Does it have to be carbon? Metal anodes in microbial fuel cells and related bioelectrochemical systems. Energy and Environmental Science, 8(7), 2048–2055. Scholar
  28. 28.
    Valle, A., Zanardini, E., Abbruscato, P., Argenzio, P., Lustrato, G., Ranalli, G., & Sorlini, C. (2007). Effects of low electric current (LEC) treatment on pure bacterial cultures. Journal of Applied Microbiology, 103(5), 1376–1385. Scholar
  29. 29.
    Kumar, Y., Kumar Patel, K., & Kumar, V. (2015). Pulsed electric field processing in food technology. International Journal of Engineering Studies and Technical Approach, 1(2), 6–17 ISSN 2395-0900.Google Scholar
  30. 30.
    Smith, K. C., Son, R. S., Gowrishankar, T. R., & Weaver, J. C. (2014). Emergence of a large pore subpopulation during electroporating pulses. Bioelectrochemistry, 100, 3–10. Scholar
  31. 31.
    Van Loey, A., Verachtert, B., & Hendrickx, M. (2002). Effects of high electric field pulses on enzymes. Trends in Food Science & Technology, 12, 94–102. Scholar
  32. 32.
    Aragón, C., Quiroga, J. M., & Coello, M. D. (2009). Comparison of four chemical uncouplers for excess sludge reduction. Environmental Technology, 30(7), 707–714. Scholar
  33. 33.
    Gostomski, P. A., & De Vela, R. J. (2018). Metabolic uncouplers for controlling biomass accumulation in biological waste treatment systems. Reviews in Environmental Science and Biotechnology, 17(1), 1–18. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.División de Ciencias AmbientalesInstituto Potosino de Investigación Científica y TecnológicaMéxicoMexico
  2. 2.Departamento de QuímicaUniversidad Autónoma Metropolitana – IztapalapaMéxico CityMexico

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