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. 1972 Sep;111(3):674–681. doi: 10.1128/jb.111.3.674-681.1972

Interference-Contrast and Phase-Contrast Microscopy of Sporulation in Clostridium thermosaccharolyticum Grown Under Strict Anaerobiosis

Charles Eller 1, Z John Ordal 1
PMCID: PMC251339  PMID: 4559820

Abstract

Cells of Clostridium thermosaccharolyticum grown under strict anaerobiosis (modified Hungate technique) were examined during growth and sporulation by employing Nomarski interference-contrast and Zernike phase-contrast optics to delineate the sequence of morphological changes leading to the formation of free, mature spores. A 0.5% l-arabinose, liquid, complex medium was used to obtain a yield of 30 to 40% free, refractile spores (ca. 108/ml) by 48 hr of incubation. The mean doubling time for the glucose culture (vegetative cells) was found to be 80 min, and that for the l-arabinose culture (sporulating cells), 498 min. By 8 hr of incubation, beginning spore formation became evident in the arabinose culture by the development of a distinct arrowhead-shaped terminal swelling. By 32 hr of incubation or shortly thereafter, Nomarski optics showed the mature spore to be uniformly spherical, whereas the enlarged terminal swelling containing it was not. The use of phase-contrast and interference-contrast optics permitted the characterization of the distinctive morphological changes occurring during sporulation of C. thermosaccharolyticum.

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Selected References

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

  1. Balassa G. The genetic control of spore formation in bacilli. Curr Top Microbiol Immunol. 1971;56:99–192. doi: 10.1007/978-3-642-65241-7_4. [DOI] [PubMed] [Google Scholar]
  2. Eller C., Crabill M. R., Bryant M. P. Anaerobic roll tube media for nonselective enumeration and isolation of bacteria in human feces. Appl Microbiol. 1971 Oct;22(4):522–529. doi: 10.1128/am.22.4.522-529.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fitz-James P. C. MORPHOLOGY OF SPORE DEVELOPMENT IN CLOSTRIDIUM PECTINOVORUM. J Bacteriol. 1962 Jul;84(1):104–114. doi: 10.1128/jb.84.1.104-114.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. HUNGATE R. E. The anaerobic mesophilic cellulolytic bacteria. Bacteriol Rev. 1950 Mar;14(1):1–49. doi: 10.1128/br.14.1.1-49.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hitchins A. D., Kahn A. J., Slepecky R. A. Interference contrast and phase contrast microscopy of sporulation and germination of Bacillus megaterium. J Bacteriol. 1968 Nov;96(5):1811–1817. doi: 10.1128/jb.96.5.1811-1817.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hoeniger J. F., Stuart P. F., Holt S. C. Cytology of spore formation in Clostridium perfringens. J Bacteriol. 1968 Nov;96(5):1818–1834. doi: 10.1128/jb.96.5.1818-1834.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hsu E. J., Ordal Z. J. Comparative metabolism of vegetative and sporulating cultures of Clostridium thermosaccharolyticum. J Bacteriol. 1970 May;102(2):369–376. doi: 10.1128/jb.102.2.369-376.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hsu E. J., Ordal Z. J. Sporulation of Clostridium thermosaccharolyticum under conditions of restricted growth. J Bacteriol. 1969 Mar;97(3):1511–1512. doi: 10.1128/jb.97.3.1511-1512.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hsu E. J., Ordal Z. J. Sporulation of Clostridium thermosaccharolyticum. Appl Microbiol. 1969 Nov;18(5):958–960. doi: 10.1128/am.18.5.958-960.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McClung L. S. Studies on Anaerobic Bacteria: III. Historical Review and Technique of Culture of Certain Thermophilic Anaerobes. J Bacteriol. 1935 Feb;29(2):173–187. doi: 10.1128/jb.29.2.173-187.1935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McClung L. S. Studies on Anaerobic Bacteria: IV. Taxonomy of Cultures of a Thermophilic Species Causing "Swells" of Canned Foods. J Bacteriol. 1935 Feb;29(2):189–203. doi: 10.1128/jb.29.2.189-203.1935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pheil C. G., Ordal Z. J. Sporulation of the "thermophilic anaerobes". Appl Microbiol. 1967 Jul;15(4):893–898. doi: 10.1128/am.15.4.893-898.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Santo L. M., Hohl H. R., Frank H. A. Ultrastructure of putrefactive anaerobe 3679h during sporulation. J Bacteriol. 1969 Sep;99(3):824–833. doi: 10.1128/jb.99.3.824-833.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. TAKAGI A., KAWATA T., YAMAMOTO S., KUBO T., OKITA S. Electron microscopic studies on ultrathin sections of spores of Clostridium tetani and Clostridium histolyticum, with special reference to sporulation and spore germination process. Jpn J Microbiol. 1960 Apr;4:137–155. doi: 10.1111/j.1348-0421.1960.tb00162.x. [DOI] [PubMed] [Google Scholar]

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