ORIGINAL PAPER
Flow cytometric assessment of the antimicrobial properties of an essential oil mixture against Escherichia coli
1
Phytosynthese Innovation Centre, 63200 Mozac, France
2
Université Clermont-Auvergne, VetAgro Sup, INRA, UMR–Fromage UMRF, 15000 Aurillac, France
Publication date: 2019-06-13
Corresponding author
F. Leriche
Université Clermont-Auvergne, VetAgro Sup, INRA, UMR–Fromage UMRF, 15000 Aurillac, France
Université Clermont-Auvergne, VetAgro Sup, INRA, UMR–Fromage UMRF, 15000 Aurillac, France
J. Anim. Feed Sci. 2019;28(2):187-198
KEYWORDS
antimicrobial propertiesessential oilsflow cytometrycolistinmembrane integritymembrane depolarizationmembrane polarity
TOPICS
ABSTRACT
Essential oils are increasingly being used in human health and
animal farming as alternatives to antibiotics. The aim of this study was to
better understand the mode of antimicrobial action of a natural essential oil mix
(EO mix) by comparison with the colistin, as an antibiotic. The growth inhibitory
concentration (GIC) of the EO mix and colistin was determined by turbidimetry.
Escherichia coli exposed to EO mix and colistin were analysed by flow cytometry using the fluorescent dyes 3,3-diethyloxacarbocyanine iodide(DiOC2
(3)) to
assess membrane potential, and propidium iodide (PI) and SYTO9 to assess
membrane integrity following treatment at the GIC and ½-GIC of the EO mix
every h for 4 h. At 1 h, treatment with EO mix and colistin resulted in significant
cell membrane alteration and depolarization. Membrane integrity measurements identified four sub-populations that were not distributed in the same way
between EO mix and antibiotic treated cells. Colistin at GIC and ½-GIC drastically disintegrated the cells that appeared as debris (69.8% of cells were lysed
after 1 h of treatment) whereas the EO mix at GIC altered membrane of a majority of cells (67.4 ± 1.3% of cells were partially altered). Contact with ½-GIC
EO mix led to sub-populations that persisted or recovered a physiological state
with intact membrane (from 1 h to 4 h of treatment, intact cells increased from
23 to 33%). So, it was demonstrated that the EO mix presented antibacterial
action against E. coli. It altered membrane properties by decreasing its polarity
and integrity which were reversible phenomena here.
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