A Novel Method to Reveal a Ureolytic Biofilm Attachment and In Situ Growth Monitoring by Electrochemical Impedance Spectroscopy

Abstract

The formation of biofilms capable of efficiently carrying out ureolysis is of fundamental importance in several biotechnological systems such as urinary tract infections, building materials and municipal wastewater treatment. This work proposes a straightforward method for the formation of a ureolytic biofilm attached to graphite. The proposed strategy reduced the time needed to complete ureolysis to 3 days instead of 16 days required in suspension culture. To confirm the formation of a ureolytic biofilm, scanning electron microscopy and confocal laser scanning microscopy studies were employed ex situ. However, it is imperative to analyse the biofilm by direct non-invasive techniques. Accordingly, open circuit potential (OCP) and electrochemical impedance spectroscopy (EIS) were used as in situ monitoring techniques. The reduction in OCP from − 0.01 to − 0.2 V vs. Ag/AgCl and the increase in capacitance from 200 to 260 μF cm−2 were related to biofilm attachment. To the best of our knowledge, this is the first time in which a ureolytic biofilm attachment has been analysed by EIS. The increase in the biomass from 0.04 to 2.81 μm3 μm−2 and in average thickness from 10.19 to 32.78 μm was related to biofilm maturation.

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Acknowledgements

The authors would like to thank David Sepúlveda-Sánchez from the Laboratory of Electron Microscopy (UAM-I). They also acknowledge María de Jesús Perea Flores from the Multidisciplinary Laboratory of Characterisation of Nanostructures and Materials (IPN) for her support in confocal laser scanning microscopy. Additionally, the authors would like to thank Guillermo Huerta-Miranda from IBT-IER-UNAM for confocal image analysis.

Funding

This research was supported by the National Council for Science and Technology (CONACyT), Mexico, and scholarship 328072.

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Correspondence to Florina Ramírez.

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Romero, M.C., Ramos, G., González, I. et al. A Novel Method to Reveal a Ureolytic Biofilm Attachment and In Situ Growth Monitoring by Electrochemical Impedance Spectroscopy. Appl Biochem Biotechnol 193, 1379–1396 (2021). https://doi.org/10.1007/s12010-020-03386-8

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Keywords

  • Ureolytic biofilm
  • Ureolysis
  • Ammonium recovery
  • Enrichment culture
  • Bacterial attachment
  • Electrochemical impedance spectroscopy