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Antibacterial effect of graphene oxide (GO) nano-particles against Pseudomonas putida biofilm of variable age

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Abstract

Graphene oxide (GO) has been reported to possess antibacterial activity; therefore, its accumulation in the environment could affect microbial communities such as biofilms. The susceptibility of biofilms to antimicrobials is known to depend on the stage of biofilm maturity. The aim of this study was to investigate the effect of GO nano-particles on Pseudomonas putida KT2440 biofilm of variable age. FT-IR, UV-vis, and Raman spectroscopy confirmed the oxidation of graphene while XPS confirmed the high purity of the synthesised GO over 6 months. Biofilms varying in maturity (24, 48, and 72 h) were formed using a CDC reactor and were treated with GO (85 μg/mL or 8.5 μg/mL). The viability of P. putida was monitored by culture on media and the bacterial membrane integrity was assessed using flow cytometry. P. putida cells were observed using confocal microscopy and SEM. The results showed that GO significantly reduced the viability of 48-h biofilm and detached biofilm cells associated with membrane damage while the viability was not affected in 24- and 72-h biofilms and detached biofilm cells. The results showed that susceptibility of P. putida biofilm to GO varied according to age which may be due to changes in the physiological state of cells during maturation.

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Acknowledgements

The authors would like to thank Dr. James Bowen for his technical support in AFM and Dr. Isaac Vizcaino for his assistance in confocal microscopy.

Conflict of interest

The authors declare that there are no conflicts of interest.

Funding

This study was financially supported by King Abdulaziz City of Science and Technology (KACST).

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Authors and Affiliations

  1. School of Chemical Engineering, University of Birmingham, B15 2TT, Birmingham, UK

    Hussam Fallatah, Tim W Overton, Hani El Kadri & Konstantinos Gkatzionis

  2. School of Pharmacy, University of Birmingham, B15 2TT, Birmingham, UK

    Mohamad Elhaneid & Hanene Ali-Boucetta

  3. Department of Food Science and Nutrition, School of the Environment, University of the Aegean, Metropolite Ioakeim 2, 81400, Myrina, Lemnos, Greece

    Konstantinos Gkatzionis

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  1. Hussam Fallatah
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  2. Mohamad Elhaneid
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Highlights

• First study showing antibacterial effects of GO colloidal suspension on biofilms.

• Effect of GO on P. putida was specific to 48-h mature biofilm.

• Susceptibility of 48-h mature biofilm-detached cells followed a similar pattern.

• Susceptibility of P. putida to GO was associated with membrane damage.

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Fallatah, H., Elhaneid, M., Ali-Boucetta, H. et al. Antibacterial effect of graphene oxide (GO) nano-particles against Pseudomonas putida biofilm of variable age. Environ Sci Pollut Res 26, 25057–25070 (2019). https://doi.org/10.1007/s11356-019-05688-9

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