Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay

Abstract Biocides such as quaternary ammonium compounds (QACs) are potentially important contributors towards bacterial antimicrobial resistance development, however, their contributions are unclear due to a lack of internationally recognized biocide testing standards. Methods to detect QAC toleranc...

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Autores principales:	Branden S. J. Gregorchuk, Shelby L. Reimer, Daniel R. Beniac, Shannon L. Hiebert, Timothy F. Booth, Michelle Wuzinski, Brielle E. Funk, Kieran A. Milner, Nicola H. Cartwright, Ali N. Doucet, Michael R. Mulvey, Mazdak Khajehpour, George G. Zhanel, Denice C. Bay
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fec3e9dcd54c4ed3ab03bc9c3e187c7a
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spelling	oai:doaj.org-article:fec3e9dcd54c4ed3ab03bc9c3e187c7a2021-12-02T16:08:37ZAntiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay10.1038/s41598-020-77446-82045-2322https://doaj.org/article/fec3e9dcd54c4ed3ab03bc9c3e187c7a2020-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-77446-8https://doaj.org/toc/2045-2322Abstract Biocides such as quaternary ammonium compounds (QACs) are potentially important contributors towards bacterial antimicrobial resistance development, however, their contributions are unclear due to a lack of internationally recognized biocide testing standards. Methods to detect QAC tolerance are limited to laborious traditional antimicrobial susceptibility testing (AST) methods. Here, we developed a rapid fluorescent dye-based membrane impermeant assay (RFDMIA) to discriminate QAC susceptibility among Gram-negative Enterobacterales and Pseudomonadales species. RFDMIA uses a membrane impermeant fluorescent dye, propidium iodide, in a 30-min 96-well fluorescent microplate-based assay where cell suspensions are exposed to increasing QAC concentrations. Our results demonstrate that RFDMIA can discriminate between QAC-susceptible and QAC-adapted Escherichia coli tolerant phenotypes and predict benzalkonium and cetrimide tolerance in all species tested except for intrinsically fluorescent Pseudomonas aeruginosa. RFDMIA identified a close association to minimum inhibitory concentration values determined by broth microdilution AST and increasing fluorescent dye emission values. RFDMIA emission values and scanning electron microscopy results also suggest that CET-adapted E. coli isolates have a CET dependence, where cells require sub-inhibitory CET concentrations to maintain bacilliform cell integrity. Overall, this study generates a new, rapid, sensitive fluorescent assay capable of detecting QAC-susceptible Gram-negative bacteria phenotypes and cell membrane perturbations.Branden S. J. GregorchukShelby L. ReimerDaniel R. BeniacShannon L. HiebertTimothy F. BoothMichelle WuzinskiBrielle E. FunkKieran A. MilnerNicola H. CartwrightAli N. DoucetMichael R. MulveyMazdak KhajehpourGeorge G. ZhanelDenice C. BayNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-16 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Branden S. J. Gregorchuk Shelby L. Reimer Daniel R. Beniac Shannon L. Hiebert Timothy F. Booth Michelle Wuzinski Brielle E. Funk Kieran A. Milner Nicola H. Cartwright Ali N. Doucet Michael R. Mulvey Mazdak Khajehpour George G. Zhanel Denice C. Bay Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay
description	Abstract Biocides such as quaternary ammonium compounds (QACs) are potentially important contributors towards bacterial antimicrobial resistance development, however, their contributions are unclear due to a lack of internationally recognized biocide testing standards. Methods to detect QAC tolerance are limited to laborious traditional antimicrobial susceptibility testing (AST) methods. Here, we developed a rapid fluorescent dye-based membrane impermeant assay (RFDMIA) to discriminate QAC susceptibility among Gram-negative Enterobacterales and Pseudomonadales species. RFDMIA uses a membrane impermeant fluorescent dye, propidium iodide, in a 30-min 96-well fluorescent microplate-based assay where cell suspensions are exposed to increasing QAC concentrations. Our results demonstrate that RFDMIA can discriminate between QAC-susceptible and QAC-adapted Escherichia coli tolerant phenotypes and predict benzalkonium and cetrimide tolerance in all species tested except for intrinsically fluorescent Pseudomonas aeruginosa. RFDMIA identified a close association to minimum inhibitory concentration values determined by broth microdilution AST and increasing fluorescent dye emission values. RFDMIA emission values and scanning electron microscopy results also suggest that CET-adapted E. coli isolates have a CET dependence, where cells require sub-inhibitory CET concentrations to maintain bacilliform cell integrity. Overall, this study generates a new, rapid, sensitive fluorescent assay capable of detecting QAC-susceptible Gram-negative bacteria phenotypes and cell membrane perturbations.
format	article
author	Branden S. J. Gregorchuk Shelby L. Reimer Daniel R. Beniac Shannon L. Hiebert Timothy F. Booth Michelle Wuzinski Brielle E. Funk Kieran A. Milner Nicola H. Cartwright Ali N. Doucet Michael R. Mulvey Mazdak Khajehpour George G. Zhanel Denice C. Bay
author_facet	Branden S. J. Gregorchuk Shelby L. Reimer Daniel R. Beniac Shannon L. Hiebert Timothy F. Booth Michelle Wuzinski Brielle E. Funk Kieran A. Milner Nicola H. Cartwright Ali N. Doucet Michael R. Mulvey Mazdak Khajehpour George G. Zhanel Denice C. Bay
author_sort	Branden S. J. Gregorchuk
title	Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay
title_short	Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay
title_full	Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay
title_fullStr	Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay
title_full_unstemmed	Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay
title_sort	antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/fec3e9dcd54c4ed3ab03bc9c3e187c7a
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Antiseptic quaternary ammonium compound tolerance by gram-negative bacteria can be rapidly detected using an impermeant fluorescent dye-based assay

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