Abstract
Biofilm formation by the pathogenic bacteria generates a serious threat to the public health as it can increase the virulence potential, resistance to drugs, and escape from the host immune response mechanisms. Among the environmental factors that influence the biofilm formation, there are only limited reports available on the role of antimicrobial agents. During the antimicrobial drug administration or application for any purpose, the microbial population can expect to get exposed to the sub-minimum inhibitory concentration (sub-MIC) of the drug which will have an unprecedented impact on microbial responses. Hence, the study has been conducted to investigate the effects of sub-MIC levels of zinc oxide nanoparticles (ZnO NPs) on the biofilm formation of Klebsiella pneumoniae and Staphylococcus aureus. Here, the selected bacteria were primarily screened for the biofilm formation by using the Congo red agar method, and their susceptibility to ZnO NPs was also evaluated. Quantitative difference in biofilm formation by the selected organisms in the presence of ZnO NPs at the sub-MIC level was further carried out by using the microtiter plate-crystal violet assay. Further, the samples were subjected to atomic force microscopy (AFM) analysis to evaluate the properties and pattern of the biofilm modulated under the experimental conditions used. From these, the organisms treated with sub-MIC levels of ZnO NPs were found to have enhanced biofilm formation when compared with the untreated sample. Also, no microbial growth could be observed for the samples treated with the minimum inhibitory concentration (MIC) of ZnO NPs. The results observed in the study provide key insights into the impact of nanomaterials on clinically important microorganisms which demands critical thinking on the antimicrobial use of nanomaterials.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author upon reasonable request.
References
Allahverdiyev AM, Abamor ES, Bagirova M, Rafailovich M (2011) Antimicrobial effects of TiO2 and Ag2O nanoparticles against drug-resistant bacteria and leishmania parasites. Future Microbiol 6:933–940. https://doi.org/10.2217/FMB.11.78
Ashitha A, Radhakrishnan EK, Mathew J (2020) Characterization of biosurfactant produced by the endophyte Burkholderia sp. WYAT7 and evaluation of its antibacterial and antibiofilm potentials. J Biotechnol 313:1–10. https://doi.org/10.1016/J.JBIOTEC.2020.03.005
Ballah FM, Islam MS, Rana ML (2022) Phenotypic and genotypic detection of biofilm-forming Staphylococcus aureus from different food sources in Bangladesh. Biology 11:949. https://doi.org/10.3390/BIOLOGY11070949
Bernardi S, Anderson A, Macchiarelli G (2021) Subinhibitory antibiotic concentrations enhance biofilm formation of clinical Enterococcus faecalis isolates. Antibiotics 10:874. https://doi.org/10.3390/ANTIBIOTICS10070874
Chu L, Zhou X, Shen Y, Yu Y (2020) Inhibitory effect of trisodium citrate on biofilms formed by Klebsiella pneumoniae. J Glob Antimicrob Resist 22:452–456. https://doi.org/10.1016/J.JGAR.2020.04.025
Fadwa AO, Alkoblan DK, Mateen A, Albarag AM (2021) Synergistic effects of zinc oxide nanoparticles and various antibiotics combination against Pseudomonas aeruginosa clinically isolated bacterial strains. Saudi J Biol Sci 28:928. https://doi.org/10.1016/J.SJBS.2020.09.064
Harika K, Shenoy V, Narasimhaswamy N, Chawla K (2020) Detection of biofilm production and its impact on antibiotic resistance profile of bacterial isolates from chronic wound infections. J Glob Infect Dis 12:129–134. https://doi.org/10.4103/JGID.JGID_150_19
Huang R, Zhang S, Zhang W, Yang X (2021) Progress of zinc oxide-based nanocomposites in the textile industry. IET Collaborative Intelligent Manufacturing 3:281–289. https://doi.org/10.1049/CIM2.12029
Ivanova A, Ivanova K, Perelshtein I et al (2021) Sonochemically engineered nano-enabled zinc oxide/amylase coatings prevent the occurrence of catheter-associated urinary tract infections. Mater Sci Eng C Mater Biol Appl. https://doi.org/10.1016/J.MSEC.2021.112518
Jayakumar A, Heera KV, Sumi TS et al (2019) Starch-PVA composite films with zinc-oxide nanoparticles and phytochemicals as intelligent pH sensing wraps for food packaging application. Int J Biol Macromol 136:395–403. https://doi.org/10.1016/J.IJBIOMAC.2019.06.018
Jin S-E, Eon Jin J, Hwang W et al (2019) Photocatalytic antibacterial application of zinc oxide nanoparticles and self-assembled networks under dual UV irradiation for enhanced disinfection. Int J Nanomedicine 14:1737–1751. https://doi.org/10.2147/IJN.S192277
Karami A, Xie Z, Zhang J et al (2020) Insights into the antimicrobial mechanism of Ag and I incorporated ZnO nanoparticle derivatives under visible light. Mater Sci Eng, C 107:110220. https://doi.org/10.1016/J.MSEC.2019.110220
Kelly S, Lanigan N, O’Neill I et al (2020) Bifidobacterial biofilm formation is a multifactorial adaptive phenomenon in response to bile exposure. Sci Rep. https://www.nature.com/articles/s41598-020-68179-9
Martins KB, Ferreira AM, Pereira VC et al (2019) In vitro effects of antimicrobial agents on planktonic and biofilm forms of Staphylococcus saprophyticus isolated from patients with urinary tract infections. Front Microbiol. https://doi.org/10.3389/FMICB.2019.00040
Mendes CR, Dilarri G, Forsan CF et al (2022) Antibacterial action and target mechanisms of zinc oxide nanoparticles against bacterial pathogens. Sci Rep 12:1–10. https://doi.org/10.1038/s41598-022-06657-y
Nakasone I, Kinjo T, Yamane N et al (2007) Laboratory-based evaluation of the colorimetric VITEK-2 compact system for species identification and of the advanced expert system for detection of antimicrobial resistances: VITEK-2 compact system identification and antimicrobial susceptibility testing. Diagn Microbiol Infect Dis 58:191–198. https://doi.org/10.1016/J.DIAGMICROBIO.2006.12.008
Neethu S, Midhun SJ, Radhakrishnan EK, Jyothis M (2020) Surface functionalization of central venous catheter with mycofabricated silver nanoparticles and its antibiofilm activity on multidrug resistant Acinetobacter baumannii. Microb Pathog 138:103832. https://doi.org/10.1016/J.MICPATH.2019.103832
Omer HS, Aka ST (2022) Effects of subminimal inhibitory concentrations of chlorhexidine on the chlorhexidine resistance and biofilm formation in clinical drug-resistant Acinetobacter baumannii isolates. Polytechnic J 12:85–91. https://doi.org/10.25156/PTJ.V12N2Y2022.PP85-91
Panichikkal J, Jose A, Sreekumaran S et al (2022) Biofilm and biocontrol modulation of Paenibacillus sp. CCB36 by supplementation with zinc oxide nanoparticles and chitosan nanoparticles. Appl Biochem Biotechnol 194:1606–1620. https://doi.org/10.1007/S12010-021-03710-W
Pinto H, Simões M, Borges A (2021) Prevalence and impact of biofilms on bloodstream and urinary tract infections: a systematic review and meta-analysis. Antibiotics (Basel). https://doi.org/10.3390/ANTIBIOTICS10070825
Rambabu K, Bharath G, Banat F, Show PL (2021) Green synthesis of zinc oxide nanoparticles using Phoenix dactylifera waste as bioreductant for effective dye degradation and antibacterial performance in wastewater treatment. J Hazard Mater. https://doi.org/10.1016/J.JHAZMAT.2020.123560
Saddik MS, Elsayed MMA, El-Mokhtar MA et al (2022) Tailoring of novel azithromycin-loaded zinc oxide nanoparticles for wound healing. Pharmaceutics. https://doi.org/10.3390/PHARMACEUTICS14010111
Singh R, Sahore S, Kaur P et al (2016) Penetration barrier contributes to bacterial biofilm-associated resistance against only select antibiotics, and exhibits genus-, strain- and antibiotic-specific differences. Pathog Dis. https://doi.org/10.1093/FEMSPD/FTW056
Sun Y, Jiang W, Zhang M et al (2021) The inhibitory effects of ficin on Streptococcus mutans biofilm formation. Biomed Res Int. https://doi.org/10.1155/2021/6692328
Yang B, Lei Z, Zhao Y et al (2017) Combination susceptibility testing of common antimicrobials in vitro and the effects of sub-MIC of antimicrobials on Staphylococcus aureus biofilm formation. Front Microbiol 8:2125. https://doi.org/10.3389/FMICB.2017.02125/BIBTEX
Yu J, Jiang F, Zhang F et al (2021) Thermonucleases contribute to Staphylococcus aureus biofilm formation in implant-associated infections–a redundant and complementary story. Front Microbiol. https://doi.org/10.3389/FMICB.2021.687888/FULL
Yuan L, Dai H, He G et al (2023) Multi-omics reveals the increased biofilm formation of Salmonella Typhimurium M3 by the induction of tetracycline at sub-inhibitory concentrations. SSRN Electron J. https://doi.org/10.2139/SSRN.4363616
Zhao X, Tang H, Jiang X (2022) Deploying gold nanomaterials in combating multi-drug-resistant bacteria. ACS Nano. https://doi.org/10.1021/ACSNANO.2C02269
Funding
The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
K., S., Nechikkadan, S., Theresa, M. et al. ZnO nanoparticles induced biofilm formation in Klebsiella pneumoniae and Staphylococcus aureus at sub-inhibitory concentrations. Folia Microbiol (2024). https://doi.org/10.1007/s12223-024-01158-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s12223-024-01158-z