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Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli

The use of nanoparticles in multiple industries has raised concerned voices about the assessment of their toxicity/antimicrobial activity and the development of standardized handling protocols. Issues emerge during the antimicrobial assaying of multiple cargo, colorimetric, colloidal nanoformulation...

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Autores principales: Marcelo, Gonçalo A., Galhano, Joana, Duarte, Maria Paula, Capelo-Martínez, José Luis, Lodeiro, Carlos, Oliveira, Elisabete
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9268724/
https://www.ncbi.nlm.nih.gov/pubmed/35807997
http://dx.doi.org/10.3390/nano12132164
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author Marcelo, Gonçalo A.
Galhano, Joana
Duarte, Maria Paula
Capelo-Martínez, José Luis
Lodeiro, Carlos
Oliveira, Elisabete
author_facet Marcelo, Gonçalo A.
Galhano, Joana
Duarte, Maria Paula
Capelo-Martínez, José Luis
Lodeiro, Carlos
Oliveira, Elisabete
author_sort Marcelo, Gonçalo A.
collection PubMed
description The use of nanoparticles in multiple industries has raised concerned voices about the assessment of their toxicity/antimicrobial activity and the development of standardized handling protocols. Issues emerge during the antimicrobial assaying of multiple cargo, colorimetric, colloidal nanoformulations, as standard protocols often rely on visual evaluations, or optical density (OD) measurements, leading to high variance inhibitory concentrations (MIC). Thus, a fast, luminescence-based assay for the effective assessment of the antimicrobial activity of nanoparticles is herein reported, using the bioluminescence of an in-house E. coli ATCC(®) 8739(TM) construct with the pMV306G13 + Lux plasmid (E. coli Lux). The new strain’s sensitivity to ofloxacin as a standard antibiotic was confirmed, and the methodology robustness verified against multiple nanoparticles and colorimetric drugs. The reduction of incubation from 24 to only 8 h, and the sole use of luminescence (LUX(490)) to accurately determine and distinguish MIC(50) and MIC(90), are two main advantages of the method. By discarding OD measurements, one can avoid turbidity and color interferences when calculating bacterial growth. This approach is an important tool that contributes to the standardization of methods, reducing samples’ background interference and focusing on luminescence as a direct probe for bacterial metabolic activity, growth and, most importantly, the correct assessment of nanomaterials’ antimicrobial activity.
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spelling pubmed-92687242022-07-09 Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli Marcelo, Gonçalo A. Galhano, Joana Duarte, Maria Paula Capelo-Martínez, José Luis Lodeiro, Carlos Oliveira, Elisabete Nanomaterials (Basel) Article The use of nanoparticles in multiple industries has raised concerned voices about the assessment of their toxicity/antimicrobial activity and the development of standardized handling protocols. Issues emerge during the antimicrobial assaying of multiple cargo, colorimetric, colloidal nanoformulations, as standard protocols often rely on visual evaluations, or optical density (OD) measurements, leading to high variance inhibitory concentrations (MIC). Thus, a fast, luminescence-based assay for the effective assessment of the antimicrobial activity of nanoparticles is herein reported, using the bioluminescence of an in-house E. coli ATCC(®) 8739(TM) construct with the pMV306G13 + Lux plasmid (E. coli Lux). The new strain’s sensitivity to ofloxacin as a standard antibiotic was confirmed, and the methodology robustness verified against multiple nanoparticles and colorimetric drugs. The reduction of incubation from 24 to only 8 h, and the sole use of luminescence (LUX(490)) to accurately determine and distinguish MIC(50) and MIC(90), are two main advantages of the method. By discarding OD measurements, one can avoid turbidity and color interferences when calculating bacterial growth. This approach is an important tool that contributes to the standardization of methods, reducing samples’ background interference and focusing on luminescence as a direct probe for bacterial metabolic activity, growth and, most importantly, the correct assessment of nanomaterials’ antimicrobial activity. MDPI 2022-06-23 /pmc/articles/PMC9268724/ /pubmed/35807997 http://dx.doi.org/10.3390/nano12132164 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Marcelo, Gonçalo A.
Galhano, Joana
Duarte, Maria Paula
Capelo-Martínez, José Luis
Lodeiro, Carlos
Oliveira, Elisabete
Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli
title Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli
title_full Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli
title_fullStr Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli
title_full_unstemmed Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli
title_short Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli
title_sort validation of a standard luminescence method for the fast determination of the antimicrobial activity of nanoparticles in escherichia coli
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9268724/
https://www.ncbi.nlm.nih.gov/pubmed/35807997
http://dx.doi.org/10.3390/nano12132164
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