| Highly chlorinated Escherichia coli cannot be stained by propidium iodide. | |
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MedLine Citation:
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PMID: 17668025 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Several studies have shown that the staining by fluorochromes (DAPI, SYBR Green II, and TOTO-1) of bacteria is altered by chlorination. To evaluate the effect of chlorine (bleach solution) on propidium iodide (PI) staining, we studied Escherichia coli in suspension and biomolecules in solution (DNA, RNA, BSA, palmitic acid, and dextran) first subjected to chlorine and then neutralized by sodium thiosulphate. The suspensions and solutions were subsequently stained with PI. The fluorescence intensity of the PI-stained DNA and RNA in solution dramatically decreased with an increase in the chlorine concentration applied. These results explain the fact that for chlorine concentrations higher than 3 micromol/L Cl2, the E. coli cells were too damaged to be properly stained by PI. In the case of highly chlorinated bacteria, it was impossible to distinguish healthy cells (with a PI-impermeable membrane and undamaged nucleic acids), which were nonfluorescent after PI staining, from cells severely injured by chlorine (with a PI-permeable membrane and damaged nucleic acids) that were also nonfluorescent, as PI penetrated but did not stain chlorinated nucleic acids. Our results suggest that it would be prudent to be cautious in interpreting the results of PI staining, as PI false-negative cells (cells with compromised membranes but not stained by PI because of nucleic acid damage caused by chlorine) are obtained as a result of nucleic acid damage, leading to an underestimation of truly dead bacteria. |
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Authors:
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M-H Phe; M Dossot; H Guilloteau; J-C Block |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Canadian journal of microbiology Volume: 53 ISSN: 0008-4166 ISO Abbreviation: Can. J. Microbiol. Publication Date: 2007 May |
Date Detail:
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Created Date: 2007-08-01 Completed Date: 2007-11-30 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0372707 Medline TA: Can J Microbiol Country: Canada |
Other Details:
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Languages: eng Pagination: 664-70 Citation Subset: IM |
Affiliation:
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Laboratoire de Chimie Physique et Microbiologie pour l'Environnement, LCPME UMR 7564 CNRS - Nancy-Université, 15, avenue du Charmois, 54500 Vandoeuvre-lès-Nancy, France. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Cell Membrane Permeability
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physiology Chlorine / chemistry* Disinfection Escherichia coli / chemistry*, metabolism Propidium / chemistry* Staining and Labeling Water Microbiology |
| Chemical | |
Reg. No./Substance:
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36015-30-2/Propidium; 7782-50-5/Chlorine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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