Rapid assessment of physiological status in Escherichia coli using fluorescent probes.

Rapid and direct viability assessment of Escherichia coli in filtered, sterile lake water was possible using multiparameter flow cytometry. Fluorescent dyes were used as probes for different cellular functions (membrane potential, membrane integrity and intracellular enzyme activity), which were correlated with the ability of the cells to respond to nutrient addition while in a stressed state. Measurement of several criteria circumvented limitations imposed by other methods, and provided extensive evidence for the validity of the methods for monitoring cell viability during adoption of a viable-but-non-culturable state in starved E. coli. Macromolecular staining was concomitantly used to monitor changes in cellular protein, RNA and DNA as additional indicators of physiological status during starvation/stress.

[1]  C. Edwards,et al.  Rapid, automated separation of specific bacteria from lake water and sewage by flow cytometry and cell sorting , 1993, Applied and environmental microbiology.

[2]  H. Steen,et al.  Escherichia coli DNA distributions measured by flow cytometry and compared with theoretical computer simulations , 1985, Journal of bacteriology.

[3]  J. Saunders,et al.  Use of a xylE marker gene to monitor survival of recombinant Pseudomonas putida populations in lake water by culture on nonselective media , 1991, Applied and environmental microbiology.

[4]  M. Hood,et al.  Survival of Vibrio cholerae and Escherichia coli in estuarine waters and sediments , 1982, Applied and environmental microbiology.

[5]  R. Kolter,et al.  A novel DNA-binding protein with regulatory and protective roles in starved Escherichia coli. , 1992, Genes & development.

[6]  B. Monger,et al.  Flow Cytometric Analysis of Marine Bacteria with Hoechst 33342 , 1993, Applied and environmental microbiology.

[7]  C. Edwards,et al.  The use of fluorogenic esters to detect viable bacteria by flow cytometry , 1994 .

[8]  D. J. Mason,et al.  The ability of membrane potential dyes and calcafluor white to distinguish between viable and non-viable bacteria. , 1995, The Journal of applied bacteriology.

[9]  G. Rhodes,et al.  Survival of nonculturable Aeromonas salmonicida in lake water , 1993, Applied and environmental microbiology.

[10]  R. Pickup,et al.  Survival of Aeromonas salmonicida in lake water , 1991, Applied and environmental microbiology.

[11]  K P Flint,et al.  The long-term survival of Escherichia coli in river water. , 1987, The Journal of applied bacteriology.

[12]  H. Steen,et al.  Cell cycle parameters of slowly growing Escherichia coli B/r studied by flow cytometry , 1983, Journal of bacteriology.

[13]  K Kogure,et al.  A tentative direct microscopic method for counting living marine bacteria. , 1979, Canadian journal of microbiology.

[14]  D Lloyd,et al.  Characterization of bacteria by multiparameter flow cytometry. , 1992, The Journal of applied bacteriology.

[15]  John C. Fry,et al.  2 Direct Methods and Biomass Estimation , 1990 .

[16]  F. Singleton,et al.  Variations in rRNA content of marine Vibrio spp. during starvation-survival and recovery , 1992, Applied and environmental microbiology.

[17]  A. Matin,et al.  Role of protein degradation in the survival of carbon-starved Escherichia coli and Salmonella typhimurium , 1984, Journal of bacteriology.

[18]  D B Kell,et al.  Dormancy in non-sporulating bacteria. , 1993, FEMS microbiology reviews.

[19]  D Lloyd,et al.  Growth of Azotobacter vinelandii with correlation of Coulter cell size, flow cytometric parameters, and ultrastructure. , 1990, Cytometry.

[20]  K. A. Hoff,et al.  Long-term starvation survival of Yersinia ruckeri at different salinities studied by microscopical and flow cytometric methods , 1992, Applied and environmental microbiology.

[21]  D. Hedley,et al.  Measurement of intracellular pH. , 1994, Methods in cell biology.

[22]  M. Riley,et al.  Organization of the bacterial chromosome , 1990, Microbiological reviews.

[23]  B. D. Davis,et al.  Role of ribosome degradation in the death of starved Escherichia coli cells , 1986, Journal of bacteriology.

[24]  J Porter,et al.  Applications of flow cytometry to bacterial ecology. , 1993 .

[25]  P. Lebaron,et al.  Flow cytometric analysis of the cellular DNA content of Salmonella typhimurium and Alteromonas haloplanktis during starvation and recovery in seawater , 1994, Applied and environmental microbiology.

[26]  J. Miller,et al.  Flow cytometric identification of microorganisms by dual staining with FITC and PI. , 1990, Cytometry.

[27]  J. Gottschal Phenotypic response to environmental changes , 1990 .

[28]  J. Saunders,et al.  Differential regulation of lambda pL and pR promoters by a cI repressor in a broad-host-range thermoregulated plasmid marker system , 1989, Applied and environmental microbiology.

[29]  B. Robertson,et al.  Characterizing aquatic bacteria according to population, cell size, and apparent DNA content by flow cytometry. , 1989, Cytometry.

[30]  Sallie W. Chisholm,et al.  A novel free-living prochlorophyte abundant in the oceanic euphotic zone , 1988, Nature.

[31]  D. Kell,et al.  Dormancy in Stationary-Phase Cultures of Micrococcus luteus: Flow Cytometric Analysis of Starvation and Resuscitation , 1993, Applied and environmental microbiology.

[32]  A. Matin,et al.  Role of protein synthesis in the survival of carbon-starved Escherichia coli K-12 , 1984, Journal of bacteriology.

[33]  C. Edwards,et al.  Survival of Staphylococcus aureus in lakewater monitored by flow cytometry. , 1994, Microbiology.

[34]  R. R. Colwell,et al.  Viable but Non-Culturable Vibrio cholerae and Related Pathogens in the Environment: Implications for Release of Genetically Engineered Microorganisms , 1985, Bio/Technology.

[35]  D. Pinkel,et al.  Bacterial characterization by flow cytometry. , 1983, Science.

[36]  S. Kjelleberg,et al.  Formation of nonculturable Vibrio vulnificus cells and its relationship to the starvation state , 1991, Applied and environmental microbiology.