Enumeration of viable E. coli in rivers and wastewaters by fluorescent in situ hybridization

Journal of Microbiological Methods
T Garcia-Armisen, P Servais

Abstract

A combination of direct viable count (DVC) and fluorescent in situ hybridization (FISH) procedures was used to enumerate viable Escherichia coli in river waters and wastewaters. A probe specific for the 16S rRNA of E. coli labeled with the CY3 dye was used; enumeration of hybridized cells was performed by epifluorescence microscopy. Data showed that the method was able to accurately enumerate a minimum of 3000 viable E. coli among a large number of non-fecal bacteria. When applied to river water and wastewater samples, the DVC-FISH method gave systematically higher E. coli counts than a reference culture-based method (miniaturized MPN method). The ratio between both counts (DVC-FISH/MPN) increased with decreasing abundance of culturable E. coli indicating that the proportion of viable but non-culturable (VBNC) E. coli (detectable by the DVC-FISH procedure and not by a culture-based method) was higher in low contaminated environments. We hypothesized that the more stressing conditions, i.e. nutritional stress and sunlight effect, met in low contaminated environments were responsible for the larger fraction of VBNC E. coli. A survival experiment, in which sterile mineral water was inoculated with a pure E. coli strain and incubat...Continue Reading

References

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Citations

Jun 2, 2005·Biosensors & Bioelectronics·Wenting ZhaoI-Ming Hsing
Jun 7, 2005·Marine Pollution Bulletin·Philippe LebaronP Servais
Dec 6, 2011·Nano Letters·Susana CampuzanoJoseph Wang
Jul 6, 2010·Applied and Environmental Microbiology·G M LunaF Biavasco
Feb 12, 2009·Future Microbiology·Claire Cenciarini-BordeBernard La Scola
Mar 31, 2010·Annals of Microbiology·Myriam Ben SaidAbdennaceur Hassen
Dec 26, 2015·Environmental Monitoring and Assessment·Ryan A BlausteinYakov A Pachepsky
May 27, 2015·Pathogens·Flor Yazmín Ramírez-CastilloAlma Lilián Guerrero-Barrera
Jan 24, 2007·The Science of the Total Environment·Pierre ServaisGilles Billen
Mar 9, 2005·Letters in Applied Microbiology·T Garcia-ArmisenP Servais
Apr 25, 2006·Journal of Applied Microbiology·S DutilC Duchaine
May 26, 2010·Ground Water·Peter S K KnappettAlexander van Geen
May 20, 2008·Current Opinion in Biotechnology·Liv Fiksdal, Ingun Tryland
Aug 1, 2006·International Journal of Food Microbiology·A Jolivet-GougeonM Cormier
Apr 15, 2015·Environmental Science and Pollution Research International·Anna Gotkowska-PłachtaKamil Solarski
Jul 3, 2013·Journal of Microbiological Methods·Tarja Pitkänen
Dec 2, 2014·The Science of the Total Environment·Maya BenamiAmit Gross
Nov 28, 2012·Water Research·R A BlausteinG Whelan
Jul 26, 2019·Environmental Microbiology Reports·Lisa GorskiMichael B Cooley

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