Sensing of oxygen in microtiter plates: a novel tool for screening drugs against pathogenic yeasts |
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Authors: | Janine Wesolowski Rabeay Y A Hassan Stephan Hodde Christian Bardroff Ursula Bilitewski |
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Institution: | (1) Helmholtz Centre for Infection Research (HZI), Inhoffenstr. 7, 38124 Braunschweig, Germany |
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Abstract: | Most antibiotics were discovered via their inhibition of growth of target organisms. However, yeasts in particular have the
capability to adapt metabolic pathways to the availability of nutrients e.g. yeasts can easily switch between respiratory
and fermentative pathways in response to oxygen concentration, or can even use both simultaneously. Thus, we cultivated S. cerevisiae BY4741 and C. albicans 1386 in microtiter plates with integrated oxygen sensors to characterize the availability of oxygen for the organisms and
to detect influences of fungicides on the oxygen consumption rates. The relevance of the respiratory pathway was indicated
by the almost total consumption of oxygen during the first 1–3 h of the cultivation in the microtiter plates, when an increase
in turbidity could hardly be seen. Moreover, the sensitivity of S. cerevisiae to inhibitors of the respiratory chain, such as myxothiazol, could be detected via a reduced oxygen consumption rate, whereas
no inhibition of growth was observed. Thus, not only was the sensitivity of the test organism for the test compound detectable,
but the affected pathway was also highlighted. Other compounds, such as pyrrolnitrin and ambruticin VS-3, inhibited growth
of C. albicans 1386 and of S. cerevisiae (only pyrrolnitrin), which was additionally observed as reduced oxygen consumption rates. Thus, the determination of oxygen
in microtiter plates via fluorescent dyes is a versatile supplement to standard growth inhibition tests. |
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Keywords: | Myxothiazol Ambruticin VS-3 Pyrrolnitrin Candida albicans 1386 Saccharomyces cerevisiae Oxygen consumption Fluorescent oxygen-sensitive dye |
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