Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacin

Given the current need for predictive persisting model for Mycobacterium abscessus, we adopted a classical assay to study drug-tolerant bacterial persisters, focusing on the behavior of a small antibiotic-insensitive subpopulation during prolonged exposure to moxifloxacin. Our study showed a wide-ra...

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Main Authors: Elena G. Salina, Billy A. Martini, Vladimir V. Sorokin, Andrey L. Mulyukin
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-11-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2024.1494147/full
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author Elena G. Salina
Elena G. Salina
Billy A. Martini
Vladimir V. Sorokin
Andrey L. Mulyukin
author_facet Elena G. Salina
Elena G. Salina
Billy A. Martini
Vladimir V. Sorokin
Andrey L. Mulyukin
author_sort Elena G. Salina
collection DOAJ
description Given the current need for predictive persisting model for Mycobacterium abscessus, we adopted a classical assay to study drug-tolerant bacterial persisters, focusing on the behavior of a small antibiotic-insensitive subpopulation during prolonged exposure to moxifloxacin. Our study showed a wide-ranging response of M. abscessus, depending on antibiotic concentration, growth stage of mycobacterial cultures, and the availability of potassium ions in the medium. Mid-logarithmic cultures, initially grown in either balanced or K+-free medium, contained small sup-populations capable of prolonged and stable survival in the presence of moxifloxacin. The response of these mid-log cultures to antibiotic exposure involved initial killing, followed by regrowth at 1–2 MBCs of moxifloxacin or a substantial reduction of the antibiotic-insensitive subpopulation to fewer than 102 CFU/mL at 16 MBCs. In stationary-phase cultures grown in a complete medium, a consistent number of viable cells was observed when exposed to a high dose of moxifloxacin. In contrast, antibiotic-insensitive subpopulations in stationary-phase M. abscessus cultures under potassium-deficient conditions experienced gradual killing across a wide range of moxifloxacin concentrations (1–16 MBCs). Studies on electron microscopy demonstrated that singular cells were rapidly destroyed after relatively short-term exposure to moxifloxacin, while cells in aggregates or clumps persisted longer, explaining the delayed biocidal effect. The small subpopulation that survived under intense moxifloxacin pressure was notably heterogeneous in cell morphology and fine structure, consisting of ovoid forms and cell-wall-deficient cells with reduced size. These findings suggest that the same antibiotic dose may have varying effects on M. abscessus cells, depending on their physiological state and abundance within infected cells or tissues. Taken together, our study may contribute to the development of strategies to combat recalcitrant survivor subpopulations.
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spelling doaj-art-aed3ca5dab38498e919e7cea491bfe9c2024-11-28T14:26:04ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-11-011510.3389/fmicb.2024.14941471494147Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacinElena G. Salina0Elena G. Salina1Billy A. Martini2Vladimir V. Sorokin3Andrey L. Mulyukin4Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, RussiaShemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, RussiaBach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, RussiaWinogradsky Institute of Microbiology, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, RussiaWinogradsky Institute of Microbiology, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, RussiaGiven the current need for predictive persisting model for Mycobacterium abscessus, we adopted a classical assay to study drug-tolerant bacterial persisters, focusing on the behavior of a small antibiotic-insensitive subpopulation during prolonged exposure to moxifloxacin. Our study showed a wide-ranging response of M. abscessus, depending on antibiotic concentration, growth stage of mycobacterial cultures, and the availability of potassium ions in the medium. Mid-logarithmic cultures, initially grown in either balanced or K+-free medium, contained small sup-populations capable of prolonged and stable survival in the presence of moxifloxacin. The response of these mid-log cultures to antibiotic exposure involved initial killing, followed by regrowth at 1–2 MBCs of moxifloxacin or a substantial reduction of the antibiotic-insensitive subpopulation to fewer than 102 CFU/mL at 16 MBCs. In stationary-phase cultures grown in a complete medium, a consistent number of viable cells was observed when exposed to a high dose of moxifloxacin. In contrast, antibiotic-insensitive subpopulations in stationary-phase M. abscessus cultures under potassium-deficient conditions experienced gradual killing across a wide range of moxifloxacin concentrations (1–16 MBCs). Studies on electron microscopy demonstrated that singular cells were rapidly destroyed after relatively short-term exposure to moxifloxacin, while cells in aggregates or clumps persisted longer, explaining the delayed biocidal effect. The small subpopulation that survived under intense moxifloxacin pressure was notably heterogeneous in cell morphology and fine structure, consisting of ovoid forms and cell-wall-deficient cells with reduced size. These findings suggest that the same antibiotic dose may have varying effects on M. abscessus cells, depending on their physiological state and abundance within infected cells or tissues. Taken together, our study may contribute to the development of strategies to combat recalcitrant survivor subpopulations.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1494147/fullMycobacterium abscessusmoxifloxacinantimicrobialsbactericidal activitysurvivorsdrug insensitive subpopulation
spellingShingle Elena G. Salina
Elena G. Salina
Billy A. Martini
Vladimir V. Sorokin
Andrey L. Mulyukin
Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacin
Frontiers in Microbiology
Mycobacterium abscessus
moxifloxacin
antimicrobials
bactericidal activity
survivors
drug insensitive subpopulation
title Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacin
title_full Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacin
title_fullStr Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacin
title_full_unstemmed Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacin
title_short Fate of in vitro cultured Mycobacterium abscessus populations when exposed to moxifloxacin
title_sort fate of in vitro cultured mycobacterium abscessus populations when exposed to moxifloxacin
topic Mycobacterium abscessus
moxifloxacin
antimicrobials
bactericidal activity
survivors
drug insensitive subpopulation
url https://www.frontiersin.org/articles/10.3389/fmicb.2024.1494147/full
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