Bacterial resistance to silver nanoparticles and how to overcome it

Abstract

Silver nanoparticles have already been successfully applied in various biomedical and antimicrobial technologies and products used in everyday life. Although bacterial resistance to antibiotics has been extensively discussed in the literature, the possible development of resistance to silver nanoparticles has not been fully explored. We report that the Gram-negative bacteria Escherichia coli 013, Pseudomonas aeruginosa CCM 3955 and E. coli CCM 3954 can develop resistance to silver nanoparticles after repeated exposure. The resistance stems from the production of the adhesive flagellum protein flagellin, which triggers the aggregation of the nanoparticles. This resistance evolves without any genetic changes; only phenotypic change is needed to reduce the nanoparticles’ colloidal stability and thus eliminate their antibacterial activity. The resistance mechanism cannot be overcome by additional stabilization of silver nanoparticles using surfactants or polymers. It is, however, strongly suppressed by inhibiting flagellin production with pomegranate rind extract.

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Fig. 1: Gradual aggregation and precipitation of silver NPs in microplates containing ‘Ag-resistant’ E. coli CCM 3954 after 0, 4, 8, 12, 16 and 24 hours of cultivation.
Fig. 2: High aggregation stability of silver NPs after culturing with ‘Ag-susceptible’ bacteria demonstrated by TEM images.
Fig. 3: UV/Vis absorption spectra.
Fig. 4: Effect of adding flagellin.
Fig. 5: Chemical mapping analysis of silver aggregates in presence of bacteria.

References

  1. 1.

    Wong, K. K. Y. & Liu, X. L. Silver nanoparticles—the real ‘silver bullet’ in clinical medicine? MedChemComm 1, 125–131 (2010).

    Article  Google Scholar 

  2. 2.

    Livermore, D. M. Fourteen years in resistance. Int. J. Antimicrob. Agents 39, 283–294 (2012).

    Article  Google Scholar 

  3. 3.

    May, M. Antibiotics. Nature 509, S1 (2014).

    Article  Google Scholar 

  4. 4.

    Hede, K. An infectious arms race. Nature 509, S2–S3 (2014).

    Article  Google Scholar 

  5. 5.

    Cui, L. et al. In situ study of the antibacterial activity and mechanism of action of silver nanoparticles by surface-enhanced Raman spectroscopy. Anal. Chem. 85, 5436–5443 (2013).

    Article  Google Scholar 

  6. 6.

    Lara, H. H., Ayala-Núñez, N. V., Ixtepan Turrent, L. del C. & Rodríguez Padilla, C. Bactericidal effect of silver nanoparticles against multidrug-resistant bacteria. World J. Microbiol. Biotechnol. 26, 615–621 (2010).

    Article  Google Scholar 

  7. 7.

    Li, W.-R. et al. Antibacterial effect of silver nanoparticles on Staphylococcus aureus. BioMetals 24, 135–141 (2011).

    Article  Google Scholar 

  8. 8.

    Morones, J. R. et al. The bactericidal effect of silver nanoparticles. Nanotechnology 16, 2346–2353 (2005).

    Article  Google Scholar 

  9. 9.

    Panáček, A. et al. Silver colloid nanoparticles: synthesis, characterization, and their antibacterial activity. J. Phys. Chem. B 110, 16248–16253 (2006).

    Article  Google Scholar 

  10. 10.

    Panáček, A. et al. Strong and nonspecific synergistic antibacterial efficiency of antibiotics combined with silver nanoparticles at very low concentrations showing no cytotoxic effect. Molecules 21, 26 (2016).

    Article  Google Scholar 

  11. 11.

    Panáček, A. et al. Silver nanoparticles strongly enhance and restore bactericidal activity of inactive antibiotics against multiresistant Enterobacteriaceae. Colloids Surf. B Biointerfaces 142, 392–399 (2016).

    Article  Google Scholar 

  12. 12.

    Brown, A. N. et al. Nanoparticles functionalized with ampicillin destroy multiple-antibiotic-resistant isolates of Pseudomonas aeruginosa and Enterobacter aerogenes and methicillin-resistant Staphylococcus aureus. Appl. Environ. Microbiol. 78, 2768–2774 (2012).

    Article  Google Scholar 

  13. 13.

    Sharma, V. K., Yngard, R. A. & Lin, Y. Silver nanoparticles: green synthesis and their antimicrobial activities. Adv. Colloid Interface Sci. 145, 83–96 (2009).

    Article  Google Scholar 

  14. 14.

    Morones-Ramirez, J. R., Winkler, J. A., Spina, C. S. & Collins, J. J. Silver enhances antibiotic activity against Gram-negative bacteria. Sci. Transl. Med. 5, 190ra81 (2013).

    Article  Google Scholar 

  15. 15.

    Lemire, J. A., Harrison, J. J. & Turner, R. J. Antimicrobial activity of metals: mechanisms, molecular targets and applications. Nat. Rev. Microbiol. 11, 371–384 (2013).

    Article  Google Scholar 

  16. 16.

    Haefeli, C., Franklin, C. & Hardy, K. Plasmid-determined silver resistance in Pseudomonas stutzeri isolated from a silver mine. J. Bacteriol. 158, 389–392 (1984).

    Google Scholar 

  17. 17.

    Li, X. Z., Nikaido, H. & Williams, K. E. Silver-resistant mutants of Escherichia coli display active efflux of Ag+ and are deficient in porins. J. Bacteriol. 179, 6127–6132 (1997).

    Article  Google Scholar 

  18. 18.

    Gupta, A., Matsui, K., Lo, J.-F. & Silver, S. Molecular basis for resistance to silver cations in Salmonella. Nat. Med. 5, 183–188 (1999).

    Article  Google Scholar 

  19. 19.

    Nies, D. H. Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol. Rev. 27, 313–339 (2003).

    Article  Google Scholar 

  20. 20.

    Silver, S. Bacterial silver resistance: molecular biology and uses and misuses of silver compounds. FEMS Microbiol. Rev. 27, 341–353 (2003).

    Article  Google Scholar 

  21. 21.

    Silver, S., Phung, L. T. & Silver, G. Silver as biocides in burn and wound dressings and bacterial resistance to silver compounds. J. Ind. Microbiol. Biotechnol. 33, 627–634 (2006).

    Article  Google Scholar 

  22. 22.

    Graves, J. L. Jr et al. Rapid evolution of silver nanoparticle resistance in Escherichia coli. Front. Genet. 6, 42 (2015).

    Article  Google Scholar 

  23. 23.

    Losasso, C. et al. Antibacterial activity of silver nanoparticles: sensitivity of different Salmonella serovars. Front. Microbiol. 5, 227 (2014).

    Article  Google Scholar 

  24. 24.

    Gunawan, C., Teoh, W. Y., Marquis, C. P. & Amal, R. Induced adaptation of Bacillus sp. to antimicrobial nanosilver. Small 9, 3554–3560 (2013).

    Article  Google Scholar 

  25. 25.

    Kvítek, L. et al. Effect of surfactants and polymers on stability and antibacterial activity of silver nanoparticles (NPs). J. Phys. Chem. C 112, 5825–5834 (2008).

    Article  Google Scholar 

  26. 26.

    Sivera, M. et al. Silver nanoparticles modified by gelatin with extraordinary pH stability and long-term antibacterial activity. PLoS One 9, e103675 (2014).

    Article  Google Scholar 

  27. 27.

    Hunter, R. J. in Foundations of Colloid Science 2nd edn, 601–603 (Oxford Univ. Press, New York, 2001).

  28. 28.

    Bardy, S. L., Ng, S. Y. M. & Jarrell, K. F. Prokaryotic motility structures. Microbiology 149, 295–304 (2003).

    Article  Google Scholar 

  29. 29.

    Metlina, A. L. Bacterial and archaeal flagella as prokaryotic motility organelles. Biochemistry (Moscow) 69, 1203–1212 (2004).

    Article  Google Scholar 

  30. 30.

    Hayashi, F. et al. The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410, 1099–1103 (2001).

    Article  Google Scholar 

  31. 31.

    Friedlander, R. S. et al. Bacterial flagella explore microscale hummocks and hollows to increase adhesion. Proc. Natl Acad. Sci. USA 110, 5624–5629 (2013).

    Article  Google Scholar 

  32. 32.

    Haiko, J. & Westerlund-Wikström, B. The role of the bacterial flagellum in adhesion and virulence. Biology 2, 1242–1267 (2013).

    Article  Google Scholar 

  33. 33.

    Pratt, L. A. & Kolter, R. Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Mol. Microbiol. 30, 285–293 (1998).

    Article  Google Scholar 

  34. 34.

    Asadishad, B., Hidalgo, G. & Tufenkji, N. Pomegranate materials inhibit flagellin gene expression and flagellar-propelled motility of uropathogenic Escherichia coli strain CFT073. FEMS Microbiol. Lett. 334, 87–94 (2012).

    Article  Google Scholar 

  35. 35.

    Brauner, A., Fridman, O., Gefen, O. & Balaban, N. O. Distinguishing between resistance, tolerance and persistence to antibiotic treatment. Nat. Rev. Microbiol. 14, 320–330 (2016).

    Article  Google Scholar 

  36. 36.

    Petrovská, B. et al. Proteomic analysis of barley cell nuclei purified by flow sorting. Cytogenet. Genome Res. 143, 78–86 (2014).

    Article  Google Scholar 

  37. 37.

    Koboldt, D. C. et al. VarScan: variant detection in massively parallel sequencing of individual and pooled samples. Bioinformatics 25, 2283–2285 (2009).

    Article  Google Scholar 

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Acknowledgements

We acknowledge the support provided by projects LO1305 and LM2015073 of the Ministry of Education, Youth and Sports of the Czech Republic, the Czech Science Foundation (project no.15–22248S), the Ministry of Health of the Czech Republic (AZV VES 15-27726A) and the Internal Grants of Palacký University in Olomouc (IGA_PrF_2015_022 and IGA_LF_2016_022).

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A.P. and L.K. conceived the project and designed the experiments dealing with synthesis of silver NPs. M.S. and R.P. carried out the experiments dealing with synthesis and stabilizations of silver NPs including their characterizations. R.V. and M.K. designed and carried out the microbiological experiments. M.R. carried out the experiments dealing with genomic analysis. F.D. and M.S. carried out the experiments dealing with proteomic analysis and viability tests. O.T. was responsible for HRTEM characterizations. R.Z. contributed clarifications and guidance on the manuscript. A.P. and R.Z. wrote the paper with input from all co-authors.

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Correspondence to Libor Kvítek or Radek Zbořil.

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Panáček, A., Kvítek, L., Smékalová, M. et al. Bacterial resistance to silver nanoparticles and how to overcome it. Nature Nanotech 13, 65–71 (2018). https://doi.org/10.1038/s41565-017-0013-y

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