1887

Abstract

Summary: The ability of flow cytometry to detect and enumerate viable bacteria during survival in a lakewater microcosm was assessed using as a model organism. Counts of colony-forming units (c.f.u.) on nutrient agar were not significantly different from those obtained by flow cytometric detection of rhodamine 123 stained bacteria and there was no evidence for a viable but nonculturable state using these methods. However c.f.u. were significantly lower when estimated using mannitol salts agar compared with nutrient agar. was also enumerated immunofluorescently after staining with FITC-lgG. There was no significant difference between the population estimated immunofluorescently and by acridine orange direct counting, and unlike estimations of viability, only slight reductions in total cell numbers were observed. Changes in the protein and nucleic acid content of during survival were also measured by flow cytometry to investigate any potential heterogeneity arising within the starved population. Flow cytometric determinations were found to correlate significantly with their respective chemical determinations. These results demonstrate the ability of flow cytometry to detect viable bacteria during starvation and to study changes in macromolecular content. They also illustrate the importance of using appropriate methods for the detection of viable bacteria in environmental samples.

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1994-01-01
2021-10-21
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References

  1. Allman R., Hann A.C., Phillips A.P., Martin K. L., Lloyd D. 1990; Growth of Azotobacter vinelandii with correlation of coulter cell size, flow cytometric parameters and ultrastructure. Cytometry 11:822–831
    [Google Scholar]
  2. Allman R., Hann A. C., Manchee R., Lloyd D. 1992; Characterization of bacteria by multiparameter flow cytometry. J Appl Bacteriol 73:438–444
    [Google Scholar]
  3. Amann R. I., Binder B. J., Olson R. J., Chisholm S. W., Devereux R., Stahl D.A. 1990; Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl Environ Microbiol 56:1919–1925
    [Google Scholar]
  4. Bohlool B.B., Schmidt E.L. 1980; The immunofluorescence approach in microbial ecology. Adv Microb Ecol 4:204–241
    [Google Scholar]
  5. Burkill P.H. 1987; Analytical flow cytometry and its application to marine microbial ecology. In Microbes in the Sea139–166 Edited by Sleigh M. A. Chichester: Ellis Horwood;
    [Google Scholar]
  6. Diaper J. P., Tither K., Edwards C. 1992; Rapid assessment of bacterial viability by flow cytometry. Appl Microbiol Biotechnol 38:268–272
    [Google Scholar]
  7. van Dilla M. A., Langlois R. G., Pinkel D., Yajko D., Hadley W.K. 1982; Bacterial characterisation by flow cytometry. Science 220:620–622
    [Google Scholar]
  8. Donnelly C.W., Baigent G.J. 1986; Method for flow cytometric detection of Listeria monocytogenes in milk. Appl Environ Microbiol 52:689–695
    [Google Scholar]
  9. Edwards C. 1993; The significance of in situ activity on the efficiency of monitoring methods. In Monitoring Genetically Manipulated Microorganisms in the Environment pp 1–25 Edited by Edwards C. Chichester: John Wiley;
    [Google Scholar]
  10. Edwards C., Diaper J. P., Porter J., Pickup R. 1992a; Applications of flow cytometry in bacterial ecology. In Flow Cytometry in Microbiology pp 121–129 Edited by Lloyd D. London: Springer-Verlag;
    [Google Scholar]
  11. Edwards C., Porter J., Saunders J. R., Diaper J., Morgan J. A. W., Pickup R.W. 1992b; Flow cytometry and microbiology. Soc Gen Microbiol Quarterly 19:105–108
    [Google Scholar]
  12. Kaprelyants A.S., Kell D.B. 1992; Rapid assessment of bacterial viability and vitality using rhodamine 123 and flow cytometry. J Appl Bacteriol 72:410–422
    [Google Scholar]
  13. Kell D. B., Ryder H. M., Kaprelyants A.S., Westerhoff H.V. 1991; Quantifying heterogeneity: flow cytometry of bacterial culture. Antonie Leeuwenboek 60:145–422
    [Google Scholar]
  14. Kjelleberg S., Hermansson M., Marden P., Jones G.W. 1987; The transient phase between growth and nongrowth of heterotrophic bacteria, with emphasis on the marine environment. Annu Rev Microbiol 41:25–49
    [Google Scholar]
  15. Fry J.G. 1990; Direct methods and biomass estimation. Methods Microbiol 22:41–85
    [Google Scholar]
  16. Herbert D., Phillips P.J., Strange R.E. 1971; Chemical analysis of microbial cells. Methods Microbiol 5B:210–344
    [Google Scholar]
  17. Hutter K.-J., Eipel H.E. 1979; Microbial determinations by flow cytometry. J Gen Microbiol 113:369–375
    [Google Scholar]
  18. Mackensie N.M., Pinder A.G. 1987; Flow cytometry and its applications in veterinary medicine. Res Vet Sci 42:131–139
    [Google Scholar]
  19. Martin A. 1992; Physiology, molecular biology and applications of the bacterial starvation response. J Appl Bacteriol Symp49S–57S
    [Google Scholar]
  20. Miller J.S., Quales J.M. 1980; Flow cytometric identification of microorganisms by dual staining with FITC. Cytometry 11:667–675
    [Google Scholar]
  21. Muirhead K. A., Horan P.K., Poste G. 1985; Flow cytometry: present and future. Biotechnology 3:337–356
    [Google Scholar]
  22. O’Shea M.L., Field R. 1992; An evaluation of bacterial standards and disinfection practices used for the assessment and treatment of stormwater. Adv Appl Microbiol 37:21–40
    [Google Scholar]
  23. Pinder A. C., Purdy P. W., Poulter S. A. G., Clark D.C. 1990; Validation of flow cytometry for rapid enumeration of bacterial concentrations in pure cultures. J Appl Bacteriol 69:92–100
    [Google Scholar]
  24. Roszak D.B., Colwell R.R. 1987; Survival strategies of bacteria in the natural environment. Microbiol Rev 51:365–379
    [Google Scholar]
  25. Ryan H. B., Jointer B.I., Ryan B.I. 1985 MINIT AB Student Handbook, 2nd end. Boston: PWS-Kent;
    [Google Scholar]
  26. Skarstad K., Steen H.B., Boye E. 1983; Cell cycle parameters of slowly growing Escherichia coli B/r studied by flow cytometry. J Bacteriol 154:656–662
    [Google Scholar]
  27. Sokai R.R., Rohlf F.J. 1981 Biometry, 2nd edn. San Francisco: W. H. Freeman;
    [Google Scholar]
  28. Steen H. B., Skarsted K., Boye E. 1990; DNA measurements of bacteria. Methods Cell Biol 33:519–526
    [Google Scholar]
  29. Thorsen B. K., Enger O., Norland S., Hoff K.A. 1992; Longterm starvation survival of Yersinia ruckeri at different salinities studied by microscopical and flow cytometric methods. Appl Environ Microbiol 58:1624–1628
    [Google Scholar]
  30. Tyndall R. L., Hand R. E., Mann R. C., Evans C., Jernigan R. 1985; Application of flow cytometry to detection and characterization of Legionella spp. Appl Environ Microbiol 49:852–857
    [Google Scholar]
  31. Vessy G., Slade J.S., Fricker C.R. 1991; Taking the eye strain out of environmental Cryptosporidium analysis. Lett Appl Microbiol 13:62–65
    [Google Scholar]
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