Membrane potential dynamics unveil the promise of bioelectrical antimicrobial susceptibility Testing (BeAST) for anti-fungal screening.

Autor:	de Souza-Guerreiro TC; School of Life Sciences, University of Warwick, Coventry, United Kingdom.; Warwick Integrative Synthetic Biology Centre, University of Warwick, Coventry, United Kingdom., Huan Bacellar L; Institute of Chemistry, Biological Chemistry Laboratory, University of Campinas, Campinas, Brazil., da Costa TS; Institute of Chemistry, Biological Chemistry Laboratory, University of Campinas, Campinas, Brazil., Edwards CLA; Cytecom Ltd., Coventry, United Kingdom., Tasic L; Institute of Chemistry, Biological Chemistry Laboratory, University of Campinas, Campinas, Brazil., Asally M; School of Life Sciences, University of Warwick, Coventry, United Kingdom.; Warwick Integrative Synthetic Biology Centre, University of Warwick, Coventry, United Kingdom.
Jazyk:	angličtina
Zdroj:	MBio [mBio] 2024 Aug 14; Vol. 15 (8), pp. e0130224. Date of Electronic Publication: 2024 Jul 23.
DOI:	10.1128/mbio.01302-24
Abstrakt:	Membrane potential is a useful marker for antimicrobial susceptibility testing (AST) due to its fundamental roles in cell function. However, the difficulties associated with measuring the membrane potential in microbes restrict its broad application. In this study, we present bioelectrical AST (BeAST) using the model fungus Saccharomyces cerevisiae . Using fluorescent indicators [DiBAC4(3), ThT, and TMRM], we measured plasma and mitochondrial membrane-potential dynamics upon electric stimulation. We find that a 2.5 second electric stimulation induces hyperpolarization of plasma membrane lasting 20 minutes in vital S. cerevisiae , but depolarization in inhibited cells. The numerical simulation of FitzHugh-Nagumo model successfully recapitulates vitality-dependent dynamics. The model also suggests that the magnitude of plasma-membrane potential dynamics (PMD) correlates with the degree of inhibition. To test this prediction and to examine if BeAST can be used for assessing novel anti-fungal compounds, we treat cells with biogenic silver nanoparticles (bioAgNPs) synthesized using orange fruit flavonoids and Fusarium oxysporum. Comparing BeAST with optical density assay alongside various stressors, we show that PMD correlates with the magnitude of growth inhibitions. These results suggest that BeAST holds promise for screening anti-fungal compounds, offering a valuable approach to tackling antimicrobial resistance. Importance: Rapid assessment of the efficacy of antimicrobials is important for optimizing treatments, avoiding misuse and facilitating the screening of new antimicrobials. The need for rapid antimicrobial susceptibility testing (AST) is growing with the rise of antimicrobial resistance. Here, we present bioelectrical AST (BeAST). Combining time-lapse microscopy and mathematical modeling, we show that electrically induced membrane potential dynamics of yeast cells correspond to the strength of growth inhibition. Furthermore, we demonstrate the utility of BeAST for assessing antimicrobial activities of novel compounds using biogenic silver nanoparticles. Competing Interests: Munehiro Asally is a co-founder and a shareholder of Cytecom Ltd. Conor L. A. Edwards is an employee and a shareholder of Cytecom Ltd.
Databáze:	MEDLINE
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