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. 1986 Dec;54(3):846–854. doi: 10.1128/iai.54.3.846-854.1986

Potential role of lysozyme in bactericidal activity of in vitro-acquired salivary pellicle against Streptococcus faecium 9790.

G R Germaine, L M Tellefson
PMCID: PMC260248  PMID: 3023239

Abstract

The adherence of Streptococcus faecium 9790 to hydroxyapatite (HA) coated with whole saliva supernatant proteins (S-HA) or parotid fluid proteins was studied. The organism was labeled with [3H]thymidine, and adherence was estimated as the radioactivity remaining associated with the variously coated HA preparations after incubation and removal of unbound microbes by washing the adherence substratum. Adherence was time dependent and saturable, characteristics typical of oral streptococci in this in vitro adherence model system. However, adherence to S-HA, but not bare HA, was decreased 20-fold at 4 degrees C compared with room temperature. Furthermore, adherence at 4 degrees C to S-HA was decreased 20-fold relative to bare HA at 4 degrees C. Adherence to HA coated with parotid fluid proteins also was reduced at 4 degrees C. The magnitude of the temperature dependence and the inhibitory effect at 4 degrees C of whole saliva or parotid fluid pellicles on HA was unexpected. Of several sugars and amino sugars tested, the chitin saccharides, chitotriose, chitobiose, and N-acetylglucosamine caused greater than 90% inhibition of adherence to S-HA. These same saccharides were previously shown to inhibit lysozyme, polylysine, or autolytic lysis of the organism (N. J. Laible and G. R. Germaine, Infect. Immun. 48:720-728, 1985). Examination of unbound and adherent microbes revealed that lysis of the organism occurred during the adherence assays. A strong association (r = 0.83) between the extent of lysis and the extent of adherence was found under a variety of conditions. Depletion of lysozyme from saliva specimens used to coat HA resulted in a greater than 90% decrease in both cell lysis and adherence. Lysis of the microbe appeared dependent upon the presence of the saliva pellicle (coating) on HA, since solutions containing proteins desorbed from HA during mock-adherence incubations possessed lytic activity that was 2- to 10-fold too low to account for the extents of lysis observed with greater than or equal to 10(8) input cells. These results demonstrate the potential antibacterial activity of acquired salivary pellicle on enamel in vivo and the likely role of lysozyme in this activity. The data also serve to caution that this widely used in vitro adherence model will not distinguish whole-cell adherence from the adsorption of radiolabeled DNA released from lysing cells. Several additional controls are suggested that will indicate whether test microbes remain intact or lyse during adherence trials.

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Selected References

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

  1. Appelbaum B., Golub E., Holt S. C., Rosan B. In vitro studies of dental plaque formation: adsorption of oral streptococci to hydroxyaptite. Infect Immun. 1979 Aug;25(2):717–728. doi: 10.1128/iai.25.2.717-728.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armstrong W. G. Characterisation studies on the specific human salivary proteins adsorbed in vitro by hydroxyapatite. Caries Res. 1971;5(3):215–227. doi: 10.1159/000259749. [DOI] [PubMed] [Google Scholar]
  3. Clark W. B., Bammann L. L., Gibbons R. J. Comparative estimates of bacterial affinities and adsorption sites on hydroxyapatite surfaces. Infect Immun. 1978 Mar;19(3):846–853. doi: 10.1128/iai.19.3.846-853.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Drasar B. S. Some factors associated with geographical variations in the intestinal microflora. Soc Appl Bacteriol Symp Ser. 1974;3(0):187–196. [PubMed] [Google Scholar]
  5. Germaine G. R., Tellefson L. M. Simple and rapid procedure for the selective removal of lysozyme from human saliva. Infect Immun. 1979 Dec;26(3):991–995. doi: 10.1128/iai.26.3.991-995.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gibbons R. J. Adherent interactions which may affect microbial ecology in the mouth. J Dent Res. 1984 Mar;63(3):378–385. doi: 10.1177/00220345840630030401. [DOI] [PubMed] [Google Scholar]
  7. Gibbons R. J., Houte J. V. Bacterial adherence in oral microbial ecology. Annu Rev Microbiol. 1975;29:19–44. doi: 10.1146/annurev.mi.29.100175.000315. [DOI] [PubMed] [Google Scholar]
  8. Gibbons R. J., Spinell D. M., Skobe Z. Selective adherence as a determinant of the host tropisms of certain indigenous and pathogenic bacteria. Infect Immun. 1976 Jan;13(1):238–246. doi: 10.1128/iai.13.1.238-246.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gibbons R. J., Van Houte J., Liljemark W. F. Parameters that effect the adherence of Streptococcus salivarius to oral epithelial surfaces. J Dent Res. 1972 Mar-Apr;51(2):424–435. doi: 10.1177/00220345720510023101. [DOI] [PubMed] [Google Scholar]
  10. Gold O. G., Jordan H. V., van Houte J. The prevalence of enterococci in the human mouth and their pathogenicity in animal models. Arch Oral Biol. 1975 Jul;20(7):473–477. doi: 10.1016/0003-9969(75)90236-8. [DOI] [PubMed] [Google Scholar]
  11. Hillman J. D., Van Houte J., Gibbons R. J. Sorption of bacteria to human enamel powder. Arch Oral Biol. 1970 Sep;15(9):899–903. doi: 10.1016/0003-9969(70)90163-9. [DOI] [PubMed] [Google Scholar]
  12. Kilian M., Roland K., Mestecky J. Interference of secretory immunoglobulin A with sorption of oral bacteria to hydroxyapatite. Infect Immun. 1981 Mar;31(3):942–951. doi: 10.1128/iai.31.3.942-951.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laible N. J., Germaine G. R. Adsorption of lysozyme from human whole saliva by Streptococcus sanguis 903 and other oral microorganisms. Infect Immun. 1982 Apr;36(1):148–159. doi: 10.1128/iai.36.1.148-159.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laible N. J., Germaine G. R. Bactericidal activity of human lysozyme, muramidase-inactive lysozyme, and cationic polypeptides against Streptococcus sanguis and Streptococcus faecalis: inhibition by chitin oligosaccharides. Infect Immun. 1985 Jun;48(3):720–728. doi: 10.1128/iai.48.3.720-728.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Levine M., Keller P. J. The isolation of some basic proline-rich proteins from human parotid saliva. Arch Oral Biol. 1977;22(1):37–41. doi: 10.1016/0003-9969(77)90137-6. [DOI] [PubMed] [Google Scholar]
  16. Liljemark W. F., Gibbons R. J. Proportional distribution and relative adherence of Streptococcus miteor (mitis) on various surfaces in the human oral cavity. Infect Immun. 1972 Nov;6(5):852–859. doi: 10.1128/iai.6.5.852-859.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liljemark W. F., Schauer S. V. Studies on the bacterial components which bind Streptococcus sanguis and Streptococcus mutans to hydroxyapatite. Arch Oral Biol. 1975 Sep;20(9):609–615. doi: 10.1016/0003-9969(75)90082-5. [DOI] [PubMed] [Google Scholar]
  18. MacKay B. J., Denepitiya L., Iacono V. J., Krost S. B., Pollock J. J. Growth-inhibitory and bactericidal effects of human parotid salivary histidine-rich polypeptides on Streptococcus mutans. Infect Immun. 1984 Jun;44(3):695–701. doi: 10.1128/iai.44.3.695-701.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. MacKay B. J., Pollock J. J., Iacono V. J., Baum B. J. Isolation of milligram quantities of a group of histidine-rich polypeptides from human parotid saliva. Infect Immun. 1984 Jun;44(3):688–694. doi: 10.1128/iai.44.3.688-694.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moore W. E., Holdeman L. V., Smibert R. M., Good I. J., Burmeister J. A., Palcanis K. G., Ranney R. R. Bacteriology of experimental gingivitis in young adult humans. Infect Immun. 1982 Nov;38(2):651–667. doi: 10.1128/iai.38.2.651-667.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Orstavik D. Sorption of Streptococci to glass: Effects of macromolecular solutes. Acta Pathol Microbiol Scand B. 1977 Feb;85B(1):47–53. [PubMed] [Google Scholar]
  22. Orstavik D. Sorption of Streptococcus faecium to glass. Acta Pathol Microbiol Scand B. 1977 Feb;85B(1):38–46. [PubMed] [Google Scholar]
  23. Pollock J. J., Denepitiya L., MacKay B. J., Iacono V. J. Fungistatic and fungicidal activity of human parotid salivary histidine-rich polypeptides on Candida albicans. Infect Immun. 1984 Jun;44(3):702–707. doi: 10.1128/iai.44.3.702-707.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shockman G. D., Conover M. J., Kolb J. J., Phillips P. M., Riley L. S., Toennies G. LYSIS OF STREPTOCOCCUS FAECALIS. J Bacteriol. 1961 Jan;81(1):36–43. doi: 10.1128/jb.81.1.36-43.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tellefson L. M., Germaine G. R. Adherence of Streptococcus sanguis to hydroxyapatite coated with lysozyme and lysozyme-supplemented saliva. Infect Immun. 1986 Mar;51(3):750–759. doi: 10.1128/iai.51.3.750-759.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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