Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Nov;62(11):4114–4120. doi: 10.1128/aem.62.11.4114-4120.1996

Death of the Escherichia coli K-12 strain W3110 in soil and water.

G Bogosian 1, L E Sammons 1, P J Morris 1, J P O'Neil 1, M A Heitkamp 1, D B Weber 1
PMCID: PMC168233  PMID: 8900002

Abstract

Whether Escherichia coli K-12 strain W3110 can enter the "viable but nonculturable" state was studied with sterile and nonsterile water and soil at various temperatures. In nonsterile river water, the plate counts of added E. coli cells dropped to less than 10 CFU/ml in less than 10 days. Acridine orange direct counts, direct viable counts, most-probable-number estimates, and PCR analyses indicated that the added E. coli cells were disappearing from the water in parallel with the number of CFU. Similar results were obtained with nonsterile soil, although the decline of the added E. coli was slower. In sterile water or soil, the added E. coli persisted for much longer, often without any decline in the plate counts even after 50 days. In sterile river water at 37 degrees C and sterile artificial seawater at 20 and 37 degrees C, the plate counts declined by 3 to 5 orders of magnitude, while the acridine orange direct counts remained unchanged. However, direct viable counts and various resuscitation studies all indicated that the nonculturable cells were nonviable. Thus, in either sterile or nonsterile water and soil, the decline in plate counts of E. coli K-12 strain W3110 is not due to the cells entering the viable but nonculturable state, but is simply due to their death.

Full Text

The Full Text of this article is available as a PDF (227.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Balbas P., Soberon X., Bolivar F., Rodriguez R. L. The plasmid, pBR322. Biotechnology. 1988;10:5–41. doi: 10.1016/b978-0-409-90042-2.50007-6. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bogosian G., Morris P. J., Hale M. D., Kane J. F. Fate in water of a recombinant Escherichia coli K-12 strain used in the commercial production of bovine somatotropin. J Ind Microbiol. 1992 Jan;9(1):27–36. doi: 10.1007/BF01576365. [DOI] [PubMed] [Google Scholar]
  4. Byrd J. J., Colwell R. R. Maintenance of plasmids pBR322 and pUC8 in nonculturable Escherichia coli in the marine environment. Appl Environ Microbiol. 1990 Jul;56(7):2104–2107. doi: 10.1128/aem.56.7.2104-2107.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caldwell B. A., Ye C., Griffiths R. P., Moyer C. L., Morita R. Y. Plasmid expression and maintenance during long-term starvation-survival of bacteria in well water. Appl Environ Microbiol. 1989 Aug;55(8):1860–1864. doi: 10.1128/aem.55.8.1860-1864.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Colwell R. R. Nonculturable but still viable and potentially pathogenic. Zentralbl Bakteriol. 1993 Jun;279(2):154–156. doi: 10.1016/s0934-8840(11)80392-0. [DOI] [PubMed] [Google Scholar]
  7. Davies C. M., Long J. A., Donald M., Ashbolt N. J. Survival of fecal microorganisms in marine and freshwater sediments. Appl Environ Microbiol. 1995 May;61(5):1888–1896. doi: 10.1128/aem.61.5.1888-1896.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Duncan S., Glover L. A., Killham K., Prosser J. I. Luminescence-based detection of activity of starved and viable but nonculturable bacteria. Appl Environ Microbiol. 1994 Apr;60(4):1308–1316. doi: 10.1128/aem.60.4.1308-1316.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fish J. T., Pettibone G. W. Influence of freshwater sediment on the survival of Escherichia coli and Salmonella sp. as measured by three methods of enumeration. Lett Appl Microbiol. 1995 May;20(5):277–281. doi: 10.1111/j.1472-765x.1995.tb00445.x. [DOI] [PubMed] [Google Scholar]
  10. Flint K. P. The long-term survival of Escherichia coli in river water. J Appl Bacteriol. 1987 Sep;63(3):261–270. doi: 10.1111/j.1365-2672.1987.tb04945.x. [DOI] [PubMed] [Google Scholar]
  11. Gauthier M. J., Le Rudulier D. Survival in seawater of Escherichia coli cells grown in marine sediments containing glycine betaine. Appl Environ Microbiol. 1990 Sep;56(9):2915–2918. doi: 10.1128/aem.56.9.2915-2918.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gonzalez J. M., Sherr E. B., Sherr B. F. Size-selective grazing on bacteria by natural assemblages of estuarine flagellates and ciliates. Appl Environ Microbiol. 1990 Mar;56(3):583–589. doi: 10.1128/aem.56.3.583-589.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. González J. M., Iriberri J., Egea L., Barcina I. Characterization of culturability, protistan grazing, and death of enteric bacteria in aquatic ecosystems. Appl Environ Microbiol. 1992 Mar;58(3):998–1004. doi: 10.1128/aem.58.3.998-1004.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Halvorson H. O., Ziegler N. R. Application of Statistics to Problems in Bacteriology: I. A Means of Determining Bacterial Population by the Dilution Method. J Bacteriol. 1933 Feb;25(2):101–121. doi: 10.1128/jb.25.2.101-121.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hobbie J. E., Daley R. J., Jasper S. Use of nuclepore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol. 1977 May;33(5):1225–1228. doi: 10.1128/aem.33.5.1225-1228.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Klein T. M., Alexander M. Bacterial inhibitors in lake water. Appl Environ Microbiol. 1986 Jul;52(1):114–118. doi: 10.1128/aem.52.1.114-118.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kogure K., Simidu U., Taga N. A tentative direct microscopic method for counting living marine bacteria. Can J Microbiol. 1979 Mar;25(3):415–420. doi: 10.1139/m79-063. [DOI] [PubMed] [Google Scholar]
  18. Linder K., Oliver J. D. Membrane fatty acid and virulence changes in the viable but nonculturable state of Vibrio vulnificus. Appl Environ Microbiol. 1989 Nov;55(11):2837–2842. doi: 10.1128/aem.55.11.2837-2842.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Porter J., Edwards C., Pickup R. W. Rapid assessment of physiological status in Escherichia coli using fluorescent probes. J Appl Bacteriol. 1995 Oct;79(4):399–408. doi: 10.1111/j.1365-2672.1995.tb03154.x. [DOI] [PubMed] [Google Scholar]
  20. Quinn J. P. The modification and evaluation of some cytochemical techniques for the enumeration of metabolically active heterotrophic bacteria in the aquatic environment. J Appl Bacteriol. 1984 Aug;57(1):51–57. doi: 10.1111/j.1365-2672.1984.tb02355.x. [DOI] [PubMed] [Google Scholar]
  21. Roszak D. B., Colwell R. R. Metabolic activity of bacterial cells enumerated by direct viable count. Appl Environ Microbiol. 1987 Dec;53(12):2889–2893. doi: 10.1128/aem.53.12.2889-2893.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roszak D. B., Colwell R. R. Survival strategies of bacteria in the natural environment. Microbiol Rev. 1987 Sep;51(3):365–379. doi: 10.1128/mr.51.3.365-379.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Roszak D. B., Grimes D. J., Colwell R. R. Viable but nonrecoverable stage of Salmonella enteritidis in aquatic systems. Can J Microbiol. 1984 Mar;30(3):334–338. doi: 10.1139/m84-049. [DOI] [PubMed] [Google Scholar]
  24. Roth W. G., Leckie M. P., Dietzler D. N. Restoration of colony-forming activity in osmotically stressed Escherichia coli by betaine. Appl Environ Microbiol. 1988 Dec;54(12):3142–3146. doi: 10.1128/aem.54.12.3142-3146.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scheuerman P. R., Schmidt J. P., Alexander M. Factors affecting the survival and growth of bacteria introduced into lake water. Arch Microbiol. 1988;150(4):320–325. doi: 10.1007/BF00408301. [DOI] [PubMed] [Google Scholar]
  26. Smith J. J., Howington J. P., McFeters G. A. Survival, physiological response and recovery of enteric bacteria exposed to a polar marine environment. Appl Environ Microbiol. 1994 Aug;60(8):2977–2984. doi: 10.1128/aem.60.8.2977-2984.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  28. Wilson M., Lindow S. E. Relationship of total viable and culturable cells in epiphytic populations of Pseudomonas syringae. Appl Environ Microbiol. 1992 Dec;58(12):3908–3913. doi: 10.1128/aem.58.12.3908-3913.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES