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Applied and Environmental Microbiology, October 2002, p. 5057-5063, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.5057-5063.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Rapid and Sensitive Enumeration of Viable Diluted Cells of Members of the Family Enterobacteriaceae in Freshwater and Drinking Water

Julia Baudart,1,2* Josée Coallier,1 Patrick Laurent,1 and Michèle Prévost2

NSERC Industrial Chair on Drinking Water, Civil, Geological and Mining Engineering, École Polytechnique de Montréal, Montréal, Québec, Canada H3C 3A7,1 US Filter-John Meunier, St. Laurent, Québec, Canada H4S 2B32

Received 19 February 2002/ Accepted 2 July 2002

Water quality assessment involves the specific, sensitive, and rapid detection of bacterial indicators and pathogens in water samples, including viable but nonculturable (VBNC) cells. This work evaluates the specificity and sensitivity of a new method which combines a fluorescent in situ hybridization (FISH) approach with a physiological assay (direct viable count [DVC]) for the direct enumeration, at the single-cell level, of highly diluted viable cells of members of the family Enterobacteriaceae in freshwater and drinking water after membrane filtration. The approach (DVC-FISH) uses a new direct detection device, the laser scanning cytometer (Scan RDI). Combining the DVC-FISH method on a membrane with Scan RDI detection makes it possible to detect as few as one targeted cell in approximately 108 nontargeted cells spread over the membrane. The ability of this new approach to detect and enumerate VBNC enterobacterial cells in freshwater and drinking water distribution systems was investigated and is discussed.

* Corresponding author. Present address: Observatoire Océanologique, Laboratoire Arago, Université Pierre et Marie Curie (Paris 6), Institut National des Sciences de l'Univers (INSU) CNRS-UMR-7621, BP 44, 66651 Banyuls-sur-Mer Cedex, France. Phone: (33) 4-6888-7347. Fax: (33) 4-6888-7398. E-mail: baudart{at}obs-banyuls.fr.

Applied and Environmental Microbiology, October 2002, p. 5057-5063, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.5057-5063.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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