1887

Abstract

The permeability of dormant spores of to malachite green (MG) and crystal violet (CV) was examined by using potassium trichloro(η-ethylene)platinum(II) (KTPt) as an electron-opaque marker for the dyes. The spores were treated with the dyes and other substances at 30 °C for 30 min or at 80 °C for 5 min. When the spores were incubated in 50 mM-MG solution at 30 °C and in 50 mM-CV solution at 30 °C or 80 °C, many small electron-dense precipitates, which were chemical complexes of dyes and platinum, were seen, mainly around the boundary between the inner and outer coat regions. The spores treated under the above conditions were not stained. Treatment with 50 mM-MG alone or a mixture of 25 mM-oxalic acid and 50 mM-CV at 80 °C made the spores stainable and dye-TPt precipitates were observed mainly in the outer pericortex region. Pretreatment with 25 mM-oxalic acid and 5% (v/v) phenol at 80 °C followed by 50 mM-CV treatment at 30 °C gave the same results as above. It was considered from these results that the inner coat itself might function as the primary permeability barrier to MG and CV, and that a secondary barrier to the dyes might exist around the cortex region.

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1991-03-01
2021-07-26
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References

  1. Beaman T. C., Pankratz H. S., Gerhardt P. 1988; Heat shock affects permeability and resistance of Bacillus stearothermophilus spores. Applied and Environmental Microbiology 54:2515–2520
    [Google Scholar]
  2. Beveridge T. J., Davies J. A. 1983; Cellular responses of Bacillus subtilis and Escherichia coli to the gram stain. Journal of Bacteriology 156:846–858
    [Google Scholar]
  3. Crafts-Lighty A., Ellar D. J. 1980; The structure and function of the spore outer membrane in dormant and germinating spores of Bacillus megaterium. Journal of Applied Bacteriology 48:135–145
    [Google Scholar]
  4. Davies J. A., Anderson G. K., Beveridge T. J., Clark H. C. 1983; Chemical mechanism of the gram stain and synthesis of a new electron-opaque marker for electron microscopy which replaces the iodine mordant of the stain. Journal of Bacteriology 156:837–845
    [Google Scholar]
  5. Dorner W. 1922; Ein neues Verfahren für isolierte Sporenfärbung. Landwirtschaftliches Jahrbuch der Schweiz 36:595–597
    [Google Scholar]
  6. Dutton L. O. 1928; Wright’s as a differential spore stain. Stain Technology 3:140–142
    [Google Scholar]
  7. Fitz-James P. C., Robinow C. F., Bergold G. H. 1954; Acid hydrolysis of the spores of B. cereus a correlation of chemical and cytological findings. Biochimica et Biophysica Acta 14:346–355
    [Google Scholar]
  8. Fujita Y., Yasuda Y., Kozuka S., Tochikubo K. 1989; Presence of proteins derived from the vegetative cell membrane in the dormant spore coat of Bacillus subtilis. Microbiology and Immunology 33:391–401
    [Google Scholar]
  9. Gould G. W., Hitchins A. D. 1963; Sensitization of bacterial spores to lysozyme and to hydrogen peroxide with agents which rupture disulphide bonds. Journal of General Microbiology 33:413–423
    [Google Scholar]
  10. Gould G. W., Stubbs J. M., King W. L. 1970; Structure and composition of resistant layers in bacterial spore coats. Journal of General Microbiology 60:347–355
    [Google Scholar]
  11. Koshikawa T., Beaman T. C., Pankratz H. S., Nakashio S., Corner T. R., Gerhardt P. 1984; Resistance, germination, and permeability correlates of Bacillus megaterium spores successively divested of integument layers. Journal of Bacteriology 159:624–632
    [Google Scholar]
  12. Kushida H. 1979; Ultrathin sectioning (5). Saibo 11:350–352 (in Japanese)
    [Google Scholar]
  13. Lagerberg I. 1917; Eine neue Methode der Sporenfärbung nebst Bemerkung über säurefeste Granula in sporenhaltigen Bakterien. Zentralblatt für Bakteriologie, Parasitenkunde und Infektionskrankheiten. Abteilung 1, Originale 79:191–192
    [Google Scholar]
  14. Lechtman M. D., Bartholomew J. W., Philips A., Russo M. 1965; Rapid methods of staining bacterial spores at room temperature. Journal of Bacteriology 89:848–854
    [Google Scholar]
  15. Maneval W. E. 1941; Staining bacteria and yeasts with acid dyes. Stain Technology 16:13–19
    [Google Scholar]
  16. May H. G. 1926; A safe spore stain for class use. Stain Technology 1:105–106
    [Google Scholar]
  17. Möller H. 1891; Über eine neue Methode der Sporenfärbung. Zentralblatt fur Bakteriologie, Parasitenkunde und Infektionskrankheiten. Abteilung 1, Originale 10:273–277
    [Google Scholar]
  18. Ohye D. F., Murrell W. G. 1962; Formation and structure of the spore of Bacillus coagulans. Journal of Cell Biolology 14:111–123
    [Google Scholar]
  19. Robinow C. F. 1951; Observations on the structure of Bacillus spores. Journal of General Microbiology 5:439–457
    [Google Scholar]
  20. Rode L. J., Foster J. W. 1960; Induced release of dipicolinic acid from spores of Bacillus megaterium. Journal of Bacteriology 79:650–656
    [Google Scholar]
  21. Rode L. J., Lewis C. W., Foster J. W. 1962; Electron microscopy of spores of Bacillus megaterium with special reference to the effects of fixation and thin sectioning. Journal of Cell Biology 13:423–435
    [Google Scholar]
  22. Schaeffer A. B., Fulton M. D. 1933; A simplified method of staining endospores. Science 77:194
    [Google Scholar]
  23. Scott I. R., Ellar D. J. 1978; Study of calcium dipicolinate release during bacterial spore germination by using a new, sensitive assay for dipicolinate. Journal of Bacteriology 135:133–137
    [Google Scholar]
  24. Stäubli W. 1960; Nouvelle matière d’inclusion hydrosoluble pour la cytologie électronique. Comptes Rendus Hebdomadaires des Séances de l’Académie des Sciences 250:1137–1139
    [Google Scholar]
  25. Sykes G. 1970; The sporicidal properties of chemical disinfectants. Journal of Applied Bacteriology 33:147–156
    [Google Scholar]
  26. Thomas S., Russell A. D. 1974; Studies on the mechanism of the sporicidal action of glutaraldehyde. Journal of Applied Bacteriology 37:83–92
    [Google Scholar]
  27. Tochikubo K., Yasuda Y., Kozuka S. 1986; Decreased particulate NADH oxidase activity in Bacillus subtilis spores after polymyxin B treatment. Journal of General Microbiology 132:277–287
    [Google Scholar]
  28. Tochikubo K., Kozuka S., Yasuda Y. 1988; Permeability of gentamicin into the inside of Bacillus subtilis spores. FEMS Microbiology Letters 50:137–140
    [Google Scholar]
  29. Vary J. C. 1973; Germination of Bacillus megaterium spores after various extraction procedures. Journal of Bacteriology 116:797–802
    [Google Scholar]
  30. Watabe K., Ichikawa T., Kondo M. 1974; Biochemical studies on germination of bacterial spores. I. Incorporation of 14C-L-alanine into spores of Bacillus thiaminolyticus during germination. Japanese Journal of Microbiology 18:173–180
    [Google Scholar]
  31. Wirtz R. 1908; Eine einfache Art der Sporenfärbung. Zentralblatt für Bakteriologie, Parasitenkunde und Infektionskrankheiten. Abteilung 1, Originale 46:727–728
    [Google Scholar]
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