Resazurin assay for high throughput analysis of antimicrobial properties of nanofibre membranes

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F70883521%3A28610%2F18%3A63522136" target="_blank" >RIV/70883521:28610/18:63522136 - isvavai.cz</a>

Výsledek na webu

—

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

Resazurin assay for high throughput analysis of antimicrobial properties of nanofibre membranes

Popis výsledku v původním jazyce

Adhesion of microorganisms and organic matter to membrane surfaces triggers the development of biofilms that can decrease transmembrane flux and affect filtration efficiency, therefore, biofouling control is challenging. High throughput method using resazurin dye (known also as alamarBlueTM) as metabolic indicator, was used to assess an antimicrobial (antifouling, anti-biofilm) efficiency of electrospun nanofibre filtration membranes functionalized by quarternary ammonium salts and nanosilver particles. Membrane cuts were placed in 12-wells microplate, inoculated by Escherichia coli suspension and incubated at 37 °C for 4 and 24 hours for resazurin assay and cultivation, respectively. Then, resazurin was added, incubated for another 30 minutes. Membranes were removed, and the fluorescence was measured each 30 minutes for 6 hours. To express amount of bacteria in colony forming units per milliliter (CFU/mL), relative fluorescence units were converted through the calibration curve that formulates relation between growth rate as OD600 and respective fluorescence of metabolized resazurin. Based on the calibration curve, limit of detection was set to 4.5 logCFU/mL. Cultivation method was also used to relate the results with resazurin assay. The results showed good relation between resazurin and cultivation methods. Membranes with nanosilver resulted in &gt;5 logCFU/mL removal compared to control membranes. Membranes with quarternary ammonium salts were far less efficient with bacterial removal only about 1 logCFU/mL. Compared to standard cultivation method, resazurin assay does not require sonication or vortexing even when working with porous and complex matrices, and results are delivered in shorter time period. Moreover, microplate design is much less laborious compared to work with tubes and agar plates.

Název v anglickém jazyce

Resazurin assay for high throughput analysis of antimicrobial properties of nanofibre membranes

Popis výsledku anglicky

Adhesion of microorganisms and organic matter to membrane surfaces triggers the development of biofilms that can decrease transmembrane flux and affect filtration efficiency, therefore, biofouling control is challenging. High throughput method using resazurin dye (known also as alamarBlueTM) as metabolic indicator, was used to assess an antimicrobial (antifouling, anti-biofilm) efficiency of electrospun nanofibre filtration membranes functionalized by quarternary ammonium salts and nanosilver particles. Membrane cuts were placed in 12-wells microplate, inoculated by Escherichia coli suspension and incubated at 37 °C for 4 and 24 hours for resazurin assay and cultivation, respectively. Then, resazurin was added, incubated for another 30 minutes. Membranes were removed, and the fluorescence was measured each 30 minutes for 6 hours. To express amount of bacteria in colony forming units per milliliter (CFU/mL), relative fluorescence units were converted through the calibration curve that formulates relation between growth rate as OD600 and respective fluorescence of metabolized resazurin. Based on the calibration curve, limit of detection was set to 4.5 logCFU/mL. Cultivation method was also used to relate the results with resazurin assay. The results showed good relation between resazurin and cultivation methods. Membranes with nanosilver resulted in &gt;5 logCFU/mL removal compared to control membranes. Membranes with quarternary ammonium salts were far less efficient with bacterial removal only about 1 logCFU/mL. Compared to standard cultivation method, resazurin assay does not require sonication or vortexing even when working with porous and complex matrices, and results are delivered in shorter time period. Moreover, microplate design is much less laborious compared to work with tubes and agar plates.

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

10404 - Polymer science

Projekt

—

Návaznosti

V - Vyzkumna aktivita podporovana z jinych verejnych zdroju

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů