Abstract
The long-term action of recommended (RC) and near-recommended concentrations of several commercial biocides (Lonzabac 12.100, Genamin CS302D, benzalkonium chloride and 2-phenoxyethanol) on cells ofS. cerevisiae wild-type strain DTXII was described using plating tests while short-term effects were determined using the potentiometric fluorescent probe diS-C3(3) that detects both changes in membrane potential and impairment of membrane integrity. A 2-d plating of cells exposed to 0.5×RC of benzalkonium chloride and Genamin CS302D for 15 min showed a complete long-term cell killing, with 2-phenoxyethanol the killing was complete only at 2×RC and Lonzabac caused complete killing at RC but not at 0.5×RC. The diS-C3(3) fluorescence assay performed immediately after a 10-min biocide exposure revealed several concentration-dependent modes of action: Lonzabac at 0.5×RC caused a mere depolarization, higher concentrations causing gradually increasing cell damage; benzalkonium chloride and Genamin CS302D rapidly damaged the membrane of some cells and depolarized the rest whereas 2-phenoxyethanol, which had the lowest effect in the plating test, produced a concentration-dependent fraction of cells with impaired membranes. Cell staining slightly increased during the diS-C3(3) assay; addition of a protonophore showed that part of the remaining undamaged cells retained their membrane potential. Comparison of short-term and long-term data implies that membrane depolarization alone is not sufficient for complete long-term killing of yeast cells under the action of a biocide unless it is accompanied by perceptible impairment of membrane integrity. The results show that the diS-C3(3) fluorescence assay, which reflects the short-term effects of a biocide on cell membranes, can be successfully used to assess the microbicidal efficiency of biocides.
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The work was supported by theGrant Agency of the Academy of Sciences of the Czech Republic (grant S5020202), by grants Kontakt CZE 01-032 and ME577 of the German and CzechMinistries of Education and by theInstitutional Research Concept AV0Z 5020 903. The support bySchülke & Mayr GmbH (Norderstedt, Germany) is gratefully acknowledge.
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Chládková, K., Hendrych, T., Gášková, D. et al. Effect of biocides onS. cerevisiae: Relationship between short-term membrane affliction and long-term cell killing. Folia Microbiol 49, 718–724 (2004). https://doi.org/10.1007/BF02931555
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DOI: https://doi.org/10.1007/BF02931555