1887

Abstract

SUMMARY: Steadily growing organisms were largely killed by slow freezing in buffer or by freeze drying. 100% survival was obtained after dropping bacteria suspended in 10% aqueous glycerol into liquid nitrogen and thawing. Ten per cent solutions of diethylene glycol, -erythritol, glucose, sucrose or polyethylene glycol (MW = 10,000) protected equally well; the last three substances did not penetrate the cell cytoplasm. The most lethal medium was dilute NaCl; broth, water or a dilute salt mixture were moderately lethal. These findings are incompatible with the view that the lethal effects of freezing are connected with osmotic shock or that protection from freezing damage requires ) penetration of the protective agent or ) osmotic dehydration of the cytoplasm. Cells frozen and thawed, even with a protective agent, showed a lowered rate of glycerol oxidation and a higher death rate when starved at the optimal temperature and pH value for growth. The storage life of frozen organisms at −20° depended on the protective agent used; only glycerol permitted extended storage.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-26-3-367
1961-11-01
2024-05-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/26/3/mic-26-3-367.html?itemId=/content/journal/micro/10.1099/00221287-26-3-367&mimeType=html&fmt=ahah

References

  1. Clement M. T. 1961; Effects of freezing, freeze-drying and storage in the freeze-dried and frozen state on viability of Escherichia coli cells. Canad. J. Microbiol 7:99
    [Google Scholar]
  2. Floodgate G. D., Hayes P. R. 1961; The preservation of marine bacteria. J. appl. Bact24 (in the Press)
    [Google Scholar]
  3. Fox M. S., Hotchkiss R. D. 1957; Initiation of bacterial transformation. Nature, Lond 179:1322
    [Google Scholar]
  4. Fry R. M., Greaves R. I. N. 1951; The survival of bacteria during freezing and after drying. J. Hyg., Camb 49:220
    [Google Scholar]
  5. Gorrill R. H., McNiel E. M. 1960; The effect of cold diluent on the viable count of Pseudomonas pyocyanea. J. gen. Microbiol 22:437
    [Google Scholar]
  6. Haines R. B. 1938; The effect of freezing on bacteria. Proc. roy. Soc B 124:451
    [Google Scholar]
  7. Harrison A. P. 1955; Survival of bacteria on repeated freezing and thawing. J. Bact 70:711
    [Google Scholar]
  8. Harrison A. P. 1956; Causes of death of bacteria in frozen suspensions. Antonie Van Leeuwenhoek J. Microbiol. Serol 22:407
    [Google Scholar]
  9. Harrison A. P., Cerroni R. E. 1956; Fallacy of ‘crushing death’ in frozen bacterial suspension. Proc. Soc. exp. Biol., N.Y 91:577
    [Google Scholar]
  10. Hegarty C. P., Weeks O. B. 1940; Sensitivity of Escherichia coli to cold-shock during the logarithmic growth phase. J. Bact 39:475
    [Google Scholar]
  11. Herbert D., Elsworth R., Telling R. C. 1956; The continuous culture of bacteria; a theoretical and experimental study. J. gen. Microbiol 14:601
    [Google Scholar]
  12. Hollander D. H., Nell E. E. 1954; Improved preservation of Treponema pallidum and other bacteria by freezing with glycerol. Appl. Microbiol 2:164
    [Google Scholar]
  13. Howard D. H. 1956; The preservation of bacteria by freezing in glycerol broth. J. Bact 71:625
    [Google Scholar]
  14. Lovelock J. E. 1953; The mechanism of the protective action of glycerol against haemolysis by freezing and thawing. Biochim. biophys. Acta 11:28
    [Google Scholar]
  15. Luyet B. J., Gehenio P. M. 1940 Life and Death at Law Temperatures Normandy, Mo., U.S.A.: Biodynamica;
    [Google Scholar]
  16. Mager J., Kuczynski M., Schatzberg G., Avidor Y. 1956; Turbidity changes in bacterial suspensions in relation to osmotic pressure. J. gen. Microbiol 14:69
    [Google Scholar]
  17. Major C. P., McDougal J. D., Harrison A. P. 1955; The effect of initial cell concentration upon survival of bacteria at — 22° C. J. Bact 69:244
    [Google Scholar]
  18. Mazur P. 1960; Physical factors implicated in the death of micro-organisms at sub-zero temperatures. Ann. N.Y. Acad. Sci 85:610
    [Google Scholar]
  19. Mazur P., Rhian M. A., Mahlandt B. G. 1957; Survival of Pasteurella tularensis in gelatin-saline after cooling and warming at sub-zero temperatures. Arch. Biochem 71:31
    [Google Scholar]
  20. Meynell G. G. 1958; Effect of sudden shilling on Escherichia coli. J. gen. Microbiol 19:380
    [Google Scholar]
  21. Mitchell P., Moyle J. 1956; Osmotic function and structure in bacteria. In Bacterial Anatomy. Symp. Soc. gen. Microbiol 6:150
    [Google Scholar]
  22. Moses V. 1955; Tricarboxylic acid cycle reactions in the fungus Zygorrhynchus moelleri. J. gen. Microbiol 13:235
    [Google Scholar]
  23. Norris K. P., Powell E. O. 1961; Improvements in determining total counts of bacteria. J. R. micr. Soc 80: (in the Press)
    [Google Scholar]
  24. Polge C., Smith A. U., Parkes A. S. 1949; Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, Lond 164:666
    [Google Scholar]
  25. Postgate J. R. 1960; Cytochrome c3 . Proceedings of the Haemaiin Enzyme Symposium, Canberra Ed Falk J. E., Lemberg M. R., Morton R. K. London: Pergamon Press;
    [Google Scholar]
  26. Postgate J. R., Crumpton J. E., Hunter J. R. 1961; Measurement of bacterial viabilities by slide culture. J. gen. Microbiol 24:15
    [Google Scholar]
  27. Quadling C. 1960; Preservation of Xanihomonas by freezing in glycerol broth. Canad. J. Microbiol 6:475
    [Google Scholar]
  28. Record B. R., Taylor R. 1953; Some factors influencing the survival of Bacteriumcoli on freeze drying. J. gen. Microbiol 9:475
    [Google Scholar]
  29. Record B. R., Taylor R. 1960; Survival of bacteria on drying in sugar/protein mixtures. Nature, Lond 185:944
    [Google Scholar]
  30. Sanderson E. S. 1925; Effect of freezing and thawing on the bacteriophage. Science 62:377
    [Google Scholar]
  31. Sherman J. M., Cameron G. M. 1934; Lethal environmental factors within the neutral range of growth. J. Bact 27:341
    [Google Scholar]
  32. Smith E. F., Swingle D. B. 1905; Der Einfluss des Gefrierens auf Bakterien. Zbl. Bakt 37:357
    [Google Scholar]
  33. Squires R. W., Hartsell S. E. 1955; Survival and growth initiation of defrosted Escherichia coli as affected by frozen storage menstrua. Appl. Microbiol 3:40
    [Google Scholar]
  34. Straka R. P., Stokes J. L. 1959; Metabolic injury to bacteria at low temperatures. J. Bact 78:181
    [Google Scholar]
  35. Tanguay A. E. 1959; Preservation of microbiological assay organisms by direct freezing. Appl. Microbiol 7:84
    [Google Scholar]
  36. Toyokama K., Hollander D. H. 1956; Variation in sensitivity of Escherichia coli to freezing damage during the growth cycle. Proc. Soc. exp. Biol., N.Y 92:499
    [Google Scholar]
  37. Wade H. E., Lovett S. 1961; Polynucleotide phosphorylase in ribosomes from Escherichia coli. Biochem. J 81:319
    [Google Scholar]
  38. Wieser R. S., Osterud C. M. 1945; Studies on the death of bacteria at low temperatures. 1. The influence of the intensity of the freezing temperature, repeated fluctuations of temperature, and the period of exposure to freezing temperatures on the mortality of Escherichia coli J. Bact 50:413
    [Google Scholar]
  39. Wood T. H., Rosenberg A. M. 1957; Freezing in yeast cells. Biochim, biophys. Acta 25:78
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-26-3-367
Loading
/content/journal/micro/10.1099/00221287-26-3-367
Loading

Data & Media loading...

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