1887

Abstract

SUMMARY:

cells were incubated at 4 °C in nutrient-limited artificial seawater (ASW) microcosms. Plate counts declined from 8 × 10 to less than 2 c.f.u. ml in about 23 d. When samples of microcosms were shifted to 30 °C, plate counts increased to 2·2×10 c.f.u. ml in 72 h. An experiment was performed to determine whether culturable cells obtained after temperature upshifts were the result of ‘resuscitation’, or outgrowth, of nonculturable cells or of cell division and growth of the few culturable cells that remained in samples. Prior to temperature upshift, samples from the microcosms were diluted 10− and 100−fold in filter-sterilized (0·1 μm) ASW from the microcosms. Undiluted, 1/10, and 1/100 diluted samples recovered culturability to about 2·2×10 c.f.u. ml within 72 h of temperature upshift. If resuscitation of nonculturable cells had occurred, the resultant number of culturable cells in diluted samples would have been 1/10 and 1/100 that of undiluted samples, respectively. In microcosms where plate counts had declined to less than 1 c.f.u. ml, 1/100 diluted samples did not regain culturability, i.e. no culturable cells remained from which growth could occur. Our conclusions are that in the experiments reported here, recovery of culturable cells on temperature upshifts resulted from growth and that there were no growthinhibiting factors in the spent growth medium, supported by the finding that about 10 recovered cells ml inoculated into filter-sterilized microcosm ASW grew to about 6·2 ×10 c.f.u. ml in 24 h, confirming that is capable of significant growth in ASW.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-2-377
1995-02-01
2021-10-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/2/mic-141-2-377.html?itemId=/content/journal/micro/10.1099/13500872-141-2-377&mimeType=html&fmt=ahah

References

  1. Allen-Austin D., Austin B., Colwell R. R. 1984; Survival of Aeromonas salmonicida in river water. FEMS Microbiol Lett 21:143–146
    [Google Scholar]
  2. Baker R. M., Singleton F. L., Hood M. A. 1983; The effects of nutrient deprivation on Vibrio cholerae . Appl Environ Microbiol 46:930–940
    [Google Scholar]
  3. Brayton P. R., Colwell R. R. 1987; Fluorescent antibody staining method for enumeration of viable environmental Vibrio cholerae 01. J Microbiol Methods 6:309–314
    [Google Scholar]
  4. Byrd J. J., Colwell R. R. 1990; Maintenance of plasmids pBR322 and pUC8 in nonculturable Escherichia coli in the marine environment. Appl Environ Microbiol 56:2104–2107
    [Google Scholar]
  5. Chowdhury M.A.R., Miyoshi S.-I., Yamanaka H., Shinoda S. 1992; Ecology and distribution of toxigenic Vibrio cholerae in aquatic environment of a temperate region. Microbios 72:203–213
    [Google Scholar]
  6. Colwell R. R., Brayton P. R., Grimes D. J., Roszack D. R., Huq S. A., Palmer L. M. 1985; Viable, but non-culturable, Vibrio cholerae and related pathogens in the environment: implications for release of genetically engineered microorganisms. Bio/Technology 3:817–820
    [Google Scholar]
  7. Colwell R. R., Tamplin M.L., Brayton P. R., Gauzens A.L., Tall B. D., Harrington D., Levine M. M., Hall S., Huq A., Sack D.A. 1990 Environmental aspects of V. cholerae in transmission of cholera. In Advances in Research on Cholera and Related Areas pp 327–343 Edited by Sack R. B., Zinnaka Y. Tokyo: KTK Scientific Publishers;
    [Google Scholar]
  8. Epstein P. R., Ford T. E., Colwell R. R. 1993; Marine ecosystems. Lancet 342:1216–1219
    [Google Scholar]
  9. Hobbie J. E., Daleu R. J., Jasper S. 1977; Use of nucleopore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol 33:1225–1228
    [Google Scholar]
  10. Islam S., Hasan M. K., Miah M. A., Sur G. C., Felsenstein A., Venkatesan M., Sack R. B., Albert M. J. 1993; Use of the polymerase chain reaction and fluorescent-antibody methods for detecting viable but nonculturable Shigella dysenteriae type 1 in laboratory microcosms. Appl Environ Microbiol 59:536–540
    [Google Scholar]
  11. Kaper J. B., Lockman H., Baldini M. M., Levine M. M. 1984; A recombinant live oral cholera vaccine. Bio/Technology 2:345–349
    [Google Scholar]
  12. Kaprelyants A. S., Kell D. B. 1993; Dormancy in stationary- phase cultures of Micrococcus luteus: flow cytometric analysis of starvation and resuscitation. Appl Environ Microbiol 59:3187–3196
    [Google Scholar]
  13. Kaprelyants A. S., Mukamolova G. V., Kell D. B. 1994; Estimation of dormant Micrococcus luteus cells by penicillin lysis and by resuscitation in cell-free spent culture medium at high dilution. FEMS Microbiol Lett 115:347–352
    [Google Scholar]
  14. Kogure K., Simidu U., Taga N. 1979; A tentative direct microscopic method for counting living marine bacteria. Can J Microbiol 25:415–420
    [Google Scholar]
  15. Linder K., Oliver J. D. 1989; Membrane fatty acid and virulence changes in the viable but non culturable state of Vibrio vulnificus . Appl Environ Microbiol 55:2837–2842
    [Google Scholar]
  16. Mai U.E.H., Shahamat M., Colwell R. R. 1990 Survival of Helicobacter pylori in the aquatic environment. In Helicobacter pylori 1990 pp 90–96 Edited by Menge H., Gregor M., Tytgat G.N.J., Marshal B. J., McNulty C. A. M. Berlin: Springer-Verlag;
    [Google Scholar]
  17. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Nilsson L., Oliver J. D., Kjelleberg S. 1991; Resuscitation of Vibrio vulnificus from the viable but not culturable state. J Bacteriol 173:5054–5059
    [Google Scholar]
  19. Peele E. R., Colwell R. R. 1981; Application of a direct microscopic method for enumeration of substrate-responsive marine bacteria. Can J Microbiol 27:1071–1075
    [Google Scholar]
  20. Rahman I., Shahamat M., Kirchman P. A., Russek-Cohen E., Colwell R. R. 1994; Methionine uptake and cytopathogenicity of viable but nonculturable Shigella dynsenteriae type I. Appl Environ Microbiol 60:3573–3578
    [Google Scholar]
  21. Rodriguez G. G., Phipps D., Ishiguro K., Ridgway H. F. 1992; Use of fluorescent redox probe for direct visualization of actively respiring bacteria. Appl Environ Microbiol 58:1801–1808
    [Google Scholar]
  22. Rollins D. M., Colwell R. R. 1986; Viable but nonculturable stage of Campylobacter jejuni and its role in survival in the natural aquatic environment. Appl Environ Microbiol 52:531–538
    [Google Scholar]
  23. Roszak D. B., Colwell R. R. 1987; Survival strategies of bacteria in the natural environment. Microbiol Rev 51:365–379
    [Google Scholar]
  24. Roszak D. B., Grimes D. J., Colwell R. R. 1984; Viable but nonrecoverable stage of Salmonella enteritidis in aquatic systems. Can J Microbiol 30:334–338
    [Google Scholar]
  25. Shahamat M., Mai U., Paszko-Kolva C., Kessel M., Colwell R. R. 1993; Use of microautoradiography to assess viability of Helicobacter pylori in water. Appl Environ Microbiol 59:1231–1235
    [Google Scholar]
  26. Weichart D., Oliver J. D., Kjelleberg S. 1992; Low temperature induced non-culturability and killing of Vibrio vulnificus . FEMS Microbiol Lett 100:205–210
    [Google Scholar]
  27. Wolf P. W., Oliver J. D. 1992; Temperature effects on the viable but nonculturable state of Vibrio vulnificu . s. FEMS Microbiol Lett 101:33–39
    [Google Scholar]
  28. Xu H.-S, Roberts N., Singleton F. L., Attwell R. W., Grimes D. J., Colwell R. R. 1982; Survival and viability of nonculturable Escherichia coli and Vibrio cholerae in the estuarine and marine environment. Microb Ecol 8:313–323
    [Google Scholar]
  29. Zimmermann R., Iturriaga R., Becker-Birck J. 1978; Simultaneous determination of the total number of aquatic bacteria and the number thereof involved in respiration. Appl Environ Microbiol 36:926–935
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-2-377
Loading
/content/journal/micro/10.1099/13500872-141-2-377
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error