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A New Approach to Detection of the Protective Effect of Escherichia coli on Gram-Positive Bacteria in Binary Biofilms in the Presence of Antibiotics

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Abstract—

Binary biofilms containing gram-positive bacteria Micrococcus thailandicus HB or Kocuria rhizophila strain 4A-2G and gram-negative bacteria Escherichia coli strain K-12 or strain ET12567 pRAG56 were obtained. Strain E. coli ET12567 contained a plasmid encoding aminoglycoside phosphotransferase, the enzyme which inactivates kanamycin. A new kinetic approach was developed for selective detection of metabolically active gram-positive bacteria in binary biofilms with E. coli, based on the differences in reduction rate of the electron acceptor, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). It was found that the studied E. coli strains did not exert a protective effect upon the action of antibiotics vancomycin and oxacillin on gram-positive bacteria in the binary biofilms under formation. However, a protective effect was found in the case of preformed binary biofilms: the growth of gram-positive bacteria in binary biofilms was observed at the concentrations of antibiotics oxacillin and kanamycin, which almost completely suppressed growth in the mono-species biofilms of these gram-positive bacteria. The presence of a protective effect was confirmed by several methods (dynamics of MTT reduction, densitometric evaluation of biofilm staining, measurement of the optical density of MTT extracts, and CFU enumeration).

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REFERENCES

  1. Burmølle, M., Webb, J.S., Rao, D., Hansen, L.H., Sørensen, S.J., and Kjelleberg, S., Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms, Appl. Environ. Microbiol., 2006, vol. 72, pp. 3916‒3923.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Gannesen, A.V., Zhurina, M.V., Veselova, M.A., Khmel’, I.A., and Plakunov, V.K., Regulation of biofilm formation by Pseudomonas chlororaphis in an in vitro system, Microbiology (Moscow), 2015, vol. 84, pp. 319–327.

    Article  CAS  Google Scholar 

  3. Lee, K.W., Periasamy, S., Mukherjee, M., Xie, C., Kjelleberg, S., and Rice, S.A., Biofilm development and enhanced stress resistance of a model, mixed-species community biofilm, ISME J., 2014, vol. 8, pp. 894‒907.

    Article  CAS  PubMed  Google Scholar 

  4. Leriche, V., Briandet, R., and Carpentier, B., Ecology of mixed biofilms subjected daily to a chlorinated alkaline solution: spatial distribution of bacterial species suggests a protective effect of one species to another, Environ. Microbiol., 2003, vol. 5, pp. 64‒71.

    Article  CAS  PubMed  Google Scholar 

  5. Lopes, S.P., Ceri, H., Azevedo, N.F., and Pereira, M.O., Antibiotic resistance of mixed biofilms in cystic fibrosis: impact of emerging microorganisms on treatment of infection, Int. J. Antimicrob. Agents, 2012, vol. 40, pp. 260–263.

    Article  CAS  PubMed  Google Scholar 

  6. Mart’yanov, S.V., Zhurina, M.V., El’-Registan, G.I., and Plakunov, V.K. Activation and prevention of formation of bacterial biofilms by azithromycin, Microbiology (Moscow), 2014, vol. 83, no. 6, pp. 723–731.

    Article  CAS  Google Scholar 

  7. Mazzariol, A., Cornaglia, G., and Nikaido, H., Contributions of the AmpC beta-lactamase and the AcrAB multidrug efflux system in intrinsic resistance of Escherichia coli K-12 to beta-lactams, Antimicrob. Agents Chemother., 2000, vol. 44, pp. 1387–1390.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Normark, S., Edlund, T., Grundström, T., Bergström, S., and Wolf-Watz, H., Escherichia coli K-12 mutants hyperproducing chromosomal beta-lactamase by gene repetitions, J. Bacteriol., 1977, vol. 132, pp. 912–922.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Nozhevnikova, A.N., Botchkova, E.A., and Plakunov, V.K., Multispecies biofilms in ecology, medicine, and biotechnology, Microbiology (Moscow), 2015, vol. 84, pp. 731–750.

    Article  CAS  Google Scholar 

  10. Paulson, D.S., Biostatistics and Microbiology, Springer Science + Business Media, 2008.

  11. Plakunov, V.K., Mart’yanov, S.V., Teteneva, N.A., and Zhurina, M.V., Controlling of microbial biofilms formation: anti- and probiofilm agents, Microbiology (Moscow), 2017, vol. 86, pp. 423–438.

    Article  CAS  Google Scholar 

  12. Sanchez-Vizuete, P., Orgaz, B., Aymerich, S., Le Coq, D., and Briandet, R., Pathogens protection against the action of disinfectants in multispecies biofilms, Front. Microbiol., 2015, vol. 6, p. 705. https://doi.org/10.3389/fmicb.2015.0070

    Article  PubMed  PubMed Central  Google Scholar 

  13. Si, T., Li, B., Zhang, K., Xu, Y., Zhao, H., and Sweedler, J.V., Characterization of Bacillus subtilis colony biofilms via mass spectrometry and fluorescence imaging, J. Proteome Res., 2016, vol. 15, pp. 1955–1962.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Strelkova, E.A., Zhurina, M.V., Plakunov, V.K., and Belyaev, S.S., Stimulation of biofilm formation by antibiotics, Microbiology (Moscow), 2012, vol. 81, pp. 259–262.

    Article  CAS  Google Scholar 

  15. Yang, L., Liu, Y., Wu, H., Hóiby, N., Molin, S., and Song, Z.J., Current understanding of multi-species biofilms, Int. J. Oral. Sci., 2011, vol. 3, pp. 74–81.

    Article  PubMed  PubMed Central  Google Scholar 

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FUNDING

This work was supported by the Ministry of Science and Higher Education of the Russian Federation.

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Authors and Affiliations

  1. Winogradsky Institute of Microbiology, Research Center of Biotechnology, Russian Academy of Sciences, 119071, Moscow, Russia

    V. K. Plakunov, Yu. A. Nikolaev, A. V. Gannesen, D. S. Chemaeva & M. V. Zhurina

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  1. V. K. Plakunov
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  2. Yu. A. Nikolaev
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  3. A. V. Gannesen
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  4. D. S. Chemaeva
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  5. M. V. Zhurina
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Correspondence to V. K. Plakunov.

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Statement of the welfare of animals. The present article does not contain any data obtained using animals as subjects.

Conflict of interests. The authors declare that they have no conflict of interests.

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Translated by A. Oleskin

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Plakunov, V.K., Nikolaev, Y.A., Gannesen, A.V. et al. A New Approach to Detection of the Protective Effect of Escherichia coli on Gram-Positive Bacteria in Binary Biofilms in the Presence of Antibiotics. Microbiology 88, 275–281 (2019). https://doi.org/10.1134/S0026261719030093

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  • DOI: https://doi.org/10.1134/S0026261719030093

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