1887

Abstract

A sample of human dental plaque was homogenized in transport fluid and inoculated simultaneously into a glucose-limited and a glucose-excess chemostat maintained at pH 7·0 and a dilution rate () of 0·05 h. In an attempt to ensure the establishment of slow-growing bacterial populations, two further inoculations of each chemostat with fresh samples of dental plaque took place before a steady-state was attained at this dilution rate. The dilution rate was increased step-wise to = 0·6 h, and then returned directly to = 0·05 h. Contrary to chemostat theory, microbial communities with a high species diversity were maintained under all of the experimental conditions employed, although not all of the bacterial populations present in the inocula established successfully in the chemostat. At each steady-state the bacteriological composition and biochemical properties (fermentation products, enzyme assays and acid production) of the communities of each chemostat was determined. Higher cell yields and a slightly more diverse community were obtained from the glucose-limited chemostat at all dilution rates. A complex mixture of end products of metabolism was obtained from the glucose-limited chemostat, suggesting amino acid catabolism, while lactate was the predominant acid of the glucose-excess culture. In washed-cell experiments, communities from the glucose-excess chemostat produced the lower terminal pH values following a pulse of glucose, with the lowest pH values occurring at the higher dilution rates. A film of micro-organisms, which accumulated around the neck of the chemostat, was sampled at the end of the experiment. The microbial composition of the films from each chemostat differed markedly, and both were different to the community of the bulk fluid of the respective chemostat. Spirochaetes and a population of yeasts were detected in the films from the glucose-limited and glucose-excess chemostats, respectively. No invertase or glucosyltransferase activity, and little glucoamylase-specific glycogen was detected in the communities from either chemostat, although significant endogenous activity, particularly at high dilution rates, was obtained with washed-cells from the glucose-excess chemostat. The results suggest that the chemostat could make a valuable contribution to the study of the ecology of dental plaque.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-129-3-755
1983-03-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/129/3/mic-129-3-755.html?itemId=/content/journal/micro/10.1099/00221287-129-3-755&mimeType=html&fmt=ahah

References

  1. Alexander M. 1971 Microbial Ecology. New York: Wiley;
    [Google Scholar]
  2. Atkinson B., Fowler H. W. 1974; The significance of microbial film in fermenters. Advances in Biochemical Engineering 3:221–277
    [Google Scholar]
  3. Barker S. B., Summerson W. H. 1941; The colorimetric determination of lactic acid in biological material. Journal of Biological Chemistry 138:535–554
    [Google Scholar]
  4. Bowden G. H., Hardie J. M., Slack G. L. 1975; Microbial variations in approximal dental plaque. Caries Research 9:253–277
    [Google Scholar]
  5. Bowden G. H., Hardie J. M., Fillery E. D. 1976; Antigens from Actinomyces species and their value in identification. Journal of Dental Research 55: (Special Issue A) 192–204
    [Google Scholar]
  6. Bowden G. H., Ellwood D. C., Hamilton I. R. 1979; Microbial ecology of the oral cavity. Advances in Microbial Ecology 3:135–217
    [Google Scholar]
  7. Brown C. M., Ellwood D. C., Hunter J. R. 1977; Growth of bacteria at surfaces: influence of nutrient limitation. FEMS Microbiology Letters 1:163–166
    [Google Scholar]
  8. Brown C. M., Ellwood D. C., Hunter J. R. 1978; Enrichments in a chemostat. In Techniques for the Study of Mixed Populations pp. 213–222 Edited by Lovelock D. W., Davies R. London: Academic Press;
    [Google Scholar]
  9. Bull A. T. 1980; Biodegradation: some attitudes and strategies of microorganisms and microbiologists. In Contemporary Microbial Ecology pp. 107–136 Edited by Ellwood D. C., Hedger J. N., Latham M. J., Lynch J. M., Slater J. H. London: Academic Press;
    [Google Scholar]
  10. Carlsson J., Griffith C. J. 1974; Fermentation products and bacterial yields in glucose-limited and nitrogen-limited cultures of streptococci. Archives of Oral Biology 19:1105–1109
    [Google Scholar]
  11. Cole M. F., Bowden G. H., Korts D. C., Bowen W. H. 1978; The effect of pyridoxine, phytate and invert sugar on production of plaque acids in situ in the monkey (M. fascicularis). Caries Research 12:190–201
    [Google Scholar]
  12. Cowan S. T., Steel K. J. 1974 Manual for the Identification of Medical Bacteria, 2nd Edn. Cambridge: Cambridge University Press;
    [Google Scholar]
  13. Cybulska J., Pakula R. 1963; Streptococcal polyglucosidase. I. A medium suitable for polyglucosidase production. Experimental Medical Microbiology 15:187–198
    [Google Scholar]
  14. Dennis D. A., Gawronski T. H., Sudo S. Z., Harris R. S., Folke L. E. A. 1975; Variations in microbial and biochemical components of four-day plaque during a four-week controlled diet period. Journal of Dental Research 54:716–722
    [Google Scholar]
  15. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F. 1956; Colorimetric method for determination of sugar and related substances. Analytical Chemistry 28:350–356
    [Google Scholar]
  16. Edwardsson S. 1974; Bacteriological studies on deep areas of carious dentine. Odontologiskrevy 25: supplement 32
    [Google Scholar]
  17. Ellwood D. C. 1976; Chemostat studies of oral bacteria. In Microbial Aspects of Dental Caries 3 pp. 785–798 Edited by Stiles H. M., Loesche W. J., O’Brien T. C. Washington, D.C.: Information Retrieval;
    [Google Scholar]
  18. Ellwood D. C., Hunter J. R., Longyear V. M. C. 1974; Growth of Streptococcus mutans in a chemostat. Archives of Oral Biology 19:659–665
    [Google Scholar]
  19. Ellwood D. C., Baird J. K., Hunter J. R., Longyear V. M. C. 1976; Variations in surface polymers of Streptococcus mutans. Journal of Dental Research 55: (Special Issue C) 42–49
    [Google Scholar]
  20. Ellwood D. C., Phipps P. J., Hamilton I. R. 1979; Effect of growth rate and glucose concentration on the activity of the phosphoenolpyruvate phosphotransferase system in Streptococcus mutansgrown in continuous culture. Infection and Immunity 23:224–231
    [Google Scholar]
  21. Ellwood D. C., Keevil C. W., Marsh P. D., Brown C. W., Wardell J. N. 1982; Surface-associated growth. In New Dimensions in Microbiology: Mixed Substrates, Mixed Cultures and Microbial Communities pp. 71–86 Edited by Quayle J. R., Bull A. T. London: The Royal Society;
    [Google Scholar]
  22. Geddes D. A. M. 1972; The production of l(+) and D(−) lactic acid and volatile acids by human dental plaque and the effect of plaque buffering and acidic strength on pH. Archives of Oral Biology 17:537–545
    [Google Scholar]
  23. Geddes D. A. M. 1975; Acids produced by human dental plaque metabolism in situ. Caries Research 9:98–109
    [Google Scholar]
  24. Gibbons R. J., Van Houte J. 1975; Bacterial adherence in oral microbial ecology. Annual Review of Medicine 26:121–136
    [Google Scholar]
  25. Gilmour M. N., Green G. C., Zahn L. M., Sparmann C. D., Pearlman J. 1976; The C1-C4 monocarboxylic and lactic acids in dental plaques before and after exposure to sucrose in vivo. In Microbial Aspects of Dental Caries 2 pp. 539–556 Edited by Stiles H. M., Loesche W. J., O’Brien T. C. Washington, D.C.: Information Retrieval;
    [Google Scholar]
  26. Hamilton I. R. 1976; Intracellular polysaccharide synthesis by cariogenic microorganisms. In Microbial Aspects of Dental Caries 3 pp. 683–701 Edited by Stiles H. M., Loesche W. J., O’Brien T. C. Washington, D.C.: Information Retrieval;
    [Google Scholar]
  27. Hamilton I. R., Ellwood D. C. 1978; Effects of fluoride on carbohydrate metabolism by washed cells of Streptococcus mutans grown at various pH values in a chemostat. Infection and Immunity 19:434–442
    [Google Scholar]
  28. Hamilton I. R., Phipps P. J., Ellwood D. C. 1979; Effect of growth rate and glucose concentration on the biochemical properties of Streptococcus mutansIngbritt in continuous culture. Infection and Immunity 26:861–869
    [Google Scholar]
  29. Harder W., Veldkamp H. 1971; Competition of marine psychrophilic bacteria at low temperatures. Antonie van Leeuwenhoek 37:51–63
    [Google Scholar]
  30. Hardie J. M., Bowden G. H. 1976; Physiological classification of oral viridans streptococci. Journal of Dental Research 55: (Special Issue A) 166–176
    [Google Scholar]
  31. Herbert D., Kornberg H. L. 1976; Glucose transport as a rate-limiting step in the growth of Escherichia coli on glucose. Biochemical Journal 156:477–480
    [Google Scholar]
  32. Herbert D., Phipps P. J., Tempest D. W. 1965; The chemostat: design and instrumentation. Laboratory Practice 14:1150–1161
    [Google Scholar]
  33. Van Der Hoeven J. S., Franken H. C. M., Camp P. J. M., Dellebarre C. W. 1978; Analysis of bacterial fermentation products by isotachophoresis. Applied and Environmental Microbiology 35:17–23
    [Google Scholar]
  34. Hoshino E., Araya A. 1980; Lactate degradation by polysaccharide-producing Neisseria isolated from human dental plaque. Archives of Oral Biology 25:211–212
    [Google Scholar]
  35. Jannasch H. W. 1965; Continuous culture in microbial ecology. Laboratory Practice 14:1162–1167
    [Google Scholar]
  36. Jannasch J. W. 1967; Enrichments of aquatic bacteria in continuous culture. Archiv für Mikro- biologie 59:165–173
    [Google Scholar]
  37. Konings W. N., Veldkamp H. 1980; Phenotypic responses to environmental change. In Contemporary Microbial Ecology pp. 161–191 Edited by Ellwood D. C., Hedger J. N., Latham M. J., Lynch J. M., Slater J. H. London: Academic Press;
    [Google Scholar]
  38. Marsh P. D. 1980 Oral Microbiology. Walton-on-Thames: Nelson;
    [Google Scholar]
  39. Marsh P. D., Williamson M. I., Keevil C. W., Mcdermid A. S., Ellwood D. C. 1982; The influence of sodium and potassium ions on acid production by washed cells of Streptococcus mutansIngbritt and Streptococcus sanguis NCTC 7865 grown in a chemostat. Infection and Immunity 36:476–483
    [Google Scholar]
  40. Michels P. A. M., Michels J. P. J., Boonstra J., Konings W. N. 1979; Generation of an electrochemical proton gradient in bacteria by the excretion of metabolic end-products. FEMS Microbiology Letters 5:357–364
    [Google Scholar]
  41. Mlkx F. H. M., Van Der Hoeven J. S. 1975; Symbiosis of Streptococcus mutans and Veillonella alcalescens in mixed continuous cultures. Archives of Oral Biology 20:407–410
    [Google Scholar]
  42. Mikx F. H. M., Van Der Hoeven J. S., König K. G., Plasschaert A. J. M., Guggenheim B. 1972; Establishment of defined microbial ecosystems in germ-free rats. Caries Research 6:211–223
    [Google Scholar]
  43. Mikx F. H. M., Van Der Hoeven J. S., Walker G. J. 1976; Microbial symbiosis in dental plaque studied in gnotobiotic rats and in the chemostat. In Microbial Aspects of Dental Caries 3 pp. 763–771 Edited by Stiles H. M., Loesche W. J., O’Brien T. C. Washington, D.C.: Information Retrieval;
    [Google Scholar]
  44. Otto R., Sonnenberg A. S. M., Veldkamp H., Konings W. N. 1980; Generation of an electrochemical proton gradient in Streptococcus cremoris by lactate efflux. Proceedings of the National Academy of Sciences of the United States of America 77:5502–5505
    [Google Scholar]
  45. Slater J. H., Bull A. T. 1978; Interactions between microbial populations. In Companion to Microbiology pp. 181–201 Edited by Bull A. T., Meadow P. M. London: Longmans;
    [Google Scholar]
  46. Slater J. H., Godwin D. 1980; Microbial adaptation and selection. In Contemporary Microbial Ecology pp. 137–160 Edited by Ellwood D. C., Hedger J. N., Latham M. J., Lynch J. M., Slater J. H. London: Academic Press;
    [Google Scholar]
  47. Slater J. H., Somerville H. J. 1979; Microbial aspects of waste treatment with particular attention to the degradation of organic compounds. Symposia of the Society for General Microbiology 29:221–261
    [Google Scholar]
  48. Staat R. H., Gawronski T. H., Cressey D. E., Harris R. S., Folke L. E. A. 1975; Effects of dietary sucrose levels on the quantity and microbial composition of human dental plaque. Journal of Dental Research 54:872–880
    [Google Scholar]
  49. Stouthamer A. H. 1976 Yield Studies in Microorganisms. Shildon: Meadowfield;
    [Google Scholar]
  50. Sumney D. L., Jordan H. V. 1974; Characterization of bacteria isolated from human root surface carious lesions. Journal of Dental Research 53:343–351
    [Google Scholar]
  51. Veilleux B. G., Rowland I. 1981; Simulation of the rat intestinal ecosystem using a two-stage continuous culture system. Journal of General Microbiology 123:103–115
    [Google Scholar]
  52. Veldkamp H. 1976; Mixed culture studies with the chemostat. In Sixth International Symposium on Continuous Culture pp. 315–328 Edited by Dean A. C. R., Ellwood D. C., Evans C. G. T., Melling J. Chichester: Ellis Horwood;
    [Google Scholar]
  53. Veldkamp H., Jannasch H. W. 1972; Mixed culture studies with the chemostat. Journal of Applied Chemistry and Biotechnology 22:105–123
    [Google Scholar]
  54. Wardell J. N., Brown C. M., Ellwood D. C. 1980; A continuous culture study of the attachment of bacteria to surfaces. In Microbial Adhesion to Surfaces pp. 221–230 Edited by Berkeley R. C. W., Lynch J. M., Melling J., Rutter P. R., Vincent B. Chichester: Ellis Horwood;
    [Google Scholar]
  55. Wimpenny J. W. T. 1982; Responses of microorganisms to physical and chemical gradients. In New Dimensions in Microbiology: Mixed Substrates, Mixed Cultures and Microbial Communities pp. 51–68 Edited by Quayle J. R., Bull A. T. London: The Royal Society;
    [Google Scholar]
  56. Wimpenny J. W. T., Coombs J. P., Lovitt R. W., Whittaker S. G. 1981; A gel-stabilized model ecosystem for investigating microbial growth in spatially ordered solute gradients. Journal of General Microbiology 127:277–287
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-129-3-755
Loading
/content/journal/micro/10.1099/00221287-129-3-755
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