1887

Abstract

The behaviour of in nutrient-poor filter-sterilized seawater was investigated at 23 and 5 °C with respect to its growth phase. At both temperatures, the culturable population declined below the detection level (0·1 c.f.u. ml) after 3–5 weeks, depending on the initial physiological state of the cells. During the first week, starved cells appeared more resistant to the seawater stress at 5 °C than cells initially in the exponential growth phase. This difference was not observed at 23 °C, where protein synthesis seemed to be required for long-term adaptation of cells from the exponential growth phase. Over the duration of the experiments, intact and total cell concentrations were not significantly affected, indicating that bacteria had entered a so-called viable but nonculturable state (VBNC). However, the incubated bacteria rapidly became heterogeneous with respect to their nucleic acid content, and their cell size decreased faster at 23 than at 5 °C. Resuscitation of VBNC cells was attempted by a temperature shift from 5 to 23 °C without exogenous nutrient addition. Comparison of the growth rates of the stressed population and of the untreated bacteria growing in the same autoclaved initial cell suspension showed significantly faster growth for the stressed cells, suggesting that in addition to growth of the few culturable stressed cells, a proportion of injured cells became culturable.

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2004-01-01
2024-12-02
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References

  1. Baumann P., Schubert R. H. W. 1984; Vibrionaceae. In Bergey's Manual of Systematic Bacteriology vol 1 pp. 516–560Edited by Krieg N. R., Holt J. C. Baltimore: Williams & Wilkins;
    [Google Scholar]
  2. Bogosian G., Bourneuf E. 2001; A matter of bacterial life and death. EMBO Rep 2:770–774 [CrossRef]
    [Google Scholar]
  3. Bogosian G., Aardema N. D., Bourneuf E. V., Morris P. J. L., O'Neil J. P. 2000; Recovery of hydrogen peroxide sensitive culturable cells of Vibrio vulnificus gives the appearence of resuscitation from a viable but nonculturable state. J Bacteriol 182:5070–5075 [CrossRef]
    [Google Scholar]
  4. Colwell R. R. 2000 In Nonculturable Microorganisms in the Environment pp. 325–342Edited by Colwell R. R., Grimes D. J. Washington: American Society for Microbiology;
    [Google Scholar]
  5. Desnues B., Cuny C., Grégori G., Dukan S., Aguilaniu H., Nystrom T. 2003; Differential oxidative damage and expression of stress defence regulons in culturable and nonculturable cells of Escherichia coli. EMBO Rep 4:400–404 [CrossRef]
    [Google Scholar]
  6. Dukan S., Nyström T. 1999; Oxidative stress defence and deterioration of growth-arrested Escherichia coli cells. J Biol Chem 274:26027–26032 [CrossRef]
    [Google Scholar]
  7. Dukan S., Touati D. 1996; Hypochlorous acid stress in Escherichia coli: resistance, DNA damage, and comparison with hydrogen peroxide stress. J Bacteriol 178:6145–6150
    [Google Scholar]
  8. Dukan S., Levi Y., Touati D. 1997; Recovery of culturability of an HOCl-stressed population of Escherichia coli after incubation in phosphate buffer: resuscitation or regrowth?. Appl Environ Microbiol 63:4204–4209
    [Google Scholar]
  9. Gasol J. M., Zweifel U. L., Peters F., Fuhrman J. A., Hagstrom A. 1999; Significance of size and nucleic acid content heterogeneity as measured by flow cytometry in natural planktonic bacteria. Appl Environ Microbiol 65:4475–4483
    [Google Scholar]
  10. Gavriel A. A., Landre J. P., Lamb A. J. 1998; Incidence of mesophilic Aeromonas within a public drinking water supply in north-east Scotland. J Appl Microbiol 84:383–392 [CrossRef]
    [Google Scholar]
  11. Grégori G., Citterio S., Ghiani A., Labra M., Scorbati S. B., Denis M. 2001; Resolution of viable and membrane-compromised bacteria in fresh water and marine waters based on analytical flow cytometry and nucleic acid double staining. Appl Environ Microbiol 67:4662–4670 [CrossRef]
    [Google Scholar]
  12. Hasan J. A. K., Huq A., Colwell R. R. 1991; Viable but nonculturable Aeromonas hydrophila in aquatic microcosms. Effect of temperature and salinity. (Abstract). . Am Soc Microbiol 91:299
    [Google Scholar]
  13. Hassani L., Imziln B., Boussaid A., Gauthier J. M. 1992; Seasonal incidence of and antibiotic resistance among Aeromonas species isolated from domestic wastewater before and after treatment in stabilization ponds. Microb Ecol 23:227–237 [CrossRef]
    [Google Scholar]
  14. Huq A., Rivera I. N. G., Colwell R. R. 2000; Epidemiological significance of viable but nonculturable microorganisms. In Nonculturable Microorganisms in the Environment pp. 301–323Edited by Colwell R. R., Grimes D. J. Washington, DC: American Society for Microbiolgy;
    [Google Scholar]
  15. Janda J. M., Abbott S. L. 1998; Evolving concepts regarding the genus Aeromonas: an expanding panorama of species, disease presentation, and unanswered questions. Clin Infect Dis 27:332–344 [CrossRef]
    [Google Scholar]
  16. Jiang X., Chai T. 1996; Survival of Vibrio parahaemolyticus at low temperature under starvation conditions and subsequent resuscitation of viable, nonculturable cells. Appl Environ Microbiol 62:1300–1305
    [Google Scholar]
  17. Kell D. B., Kaprelyants A. S., Weichart D. H., Harwood C. R., Barer M. R. 1998; Viability and activity in readily culturable bacteria: a review and discussion of the practical issues. Antonie Van Leeuwenhoek 73:169–187 [CrossRef]
    [Google Scholar]
  18. Kersters I., Van Vooren L., Huys G., Janssen P., Kerster K., Verstraete W. 1995; Influence of temperature and process technology on the occurence of Aeromonas species and hygienic indicator organisms in drinking water production plants. Microb Ecol 30:203–218 [CrossRef]
    [Google Scholar]
  19. Kersters I., Huys G., Van Duffel H., Vancanneyt M., Kersters K., Verstraete W. 1996; Survival potential of Aeromonas hydrophila in freshwaters and nutrient-poor waters in comparison with other bacteria. J Appl Bacteriol 80:266–276 [CrossRef]
    [Google Scholar]
  20. Kjelleberg S., Albertson N., Flardh K., Holmquist L., Jouperjaan A., Marouga R., Ostling J., Svenblad B., Weichart D. 1993; How do non-differentiating bacteria adapt to starvation?. Antonie Van Leeuwenhoek 63:331–341
    [Google Scholar]
  21. Kolter R., Siegele D., Tormo A. 1993; The stationary phase of the bacterial life cycle. Annu Rev Microbiol 47:855–874 [CrossRef]
    [Google Scholar]
  22. Lange R., Hengge-Aronis R. 1991; Growth-phase regulated expression of bolA and morphology of stationary phase Escherichia coli cells are controlled by the novel sigma factorσs. J Bacteriol 196:4474–4481
    [Google Scholar]
  23. Lebaron P., Servais P., Aguogué H., Courties C., Joux F. 2001; Does the high nucleic acid content of individual bacteria cells allow us to discriminate between active cells and inactive cells in aquatic systems?. Appl Environ Microbiol 67:1775–1782 [CrossRef]
    [Google Scholar]
  24. Maalej S., Mahjoubi A., Elazri C., Dukan S. 2003; Simultaneous effects of environmental factors on motile Aeromonas dynamics in an urban effluent and in the natural seawater. Water Res 37:2865–2874 [CrossRef]
    [Google Scholar]
  25. Maniatis T., Fritsh E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  26. Mary P., Chihib N. E., Chrafeddine O., Defives C., Hornez J. P. 2002; Starvation survival and viable but nonculturable states in Aeromonas hydrophila. Microb Ecol 43:250–258 [CrossRef]
    [Google Scholar]
  27. Nioh I., Furusaka C. 1968; Growth of bacteria in the heat-killed cell suspensions of the same bacteria. J Gen Appl Microbiol 14:373–385 [CrossRef]
    [Google Scholar]
  28. Nyström T. 1994; The glucose starvation stimulant of Escherichia coli induced or repressed synthesis of enzyme of central metabolic pathways and role of acetyl phosphate in gene-expression and starvation survival. Mol Microbiol 12:833–843 [CrossRef]
    [Google Scholar]
  29. Oliver D. J. 2000; The public health significance of viable but nonculturable bacteria. In Nonculturable Microorganisms in the Environment pp. 277–300Edited by Colwell R. R., Grimes D. J. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  30. Oliver J. D., Nilsson L., Kjelleberg S. 1991; Formation of nonculturable Vibrio vulnificus cells and its relationship to the starvation state. Appl Environ Microbiol 57:2640–2644
    [Google Scholar]
  31. Oliver J. D., Hite F., McDougald D., Andon N. L., Simpson L. M. 1995; Entry into, and resuscitation from the viable but nonculturable state by Vibrio vulnificus in an estuarine environment. . Appl Environ Microbiol 61:2624–2630
    [Google Scholar]
  32. Rahman M. H., Suzuki S., Kawai K. 2001; The effect of temperature on Aeromonas hydrophila infection in goldfish,Carassius auratus. J Appl Ichthyol 17:282–285 [CrossRef]
    [Google Scholar]
  33. Rodriguez G. G., Phipps D., Ishiguro K., Ridgeway H. F. 1992; Use of a fluorescent probe for direct visualisation of actively respiring bacteria. Appl Environ Microbiol 58:1801–1808
    [Google Scholar]
  34. Roszak D. B., Colwell R. R. 1987; Survival strategies of bacteria in the natural environment. Microbiol Rev 51:365–379
    [Google Scholar]
  35. Schiavano G., Bruscolini F., Albano A., Brandi G. 1998; Virulence factors in Aeromonas spp. and their association with gastrointestinal disease. New Microbiol 21:23–28
    [Google Scholar]
  36. Wai S. N., Mizunoe Y., Takade A., Yoshida S. 2000; A comparison of solid and liquid media for resuscitation of starvation- and low-temperature-induced nonculturable cells of Aeromonas hydrophila. Arch Microbiol 173:307–310 [CrossRef]
    [Google Scholar]
  37. Whitesides M. D., Oliver J. D. 1997; Resuscitation of Vibrio vulnificus from the viable but nonculturable state. Appl Environ Microbiol 63:1002–1005
    [Google Scholar]
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