Role of Colonization in the Virulence of Actinomyces viscosus Strains T14-Vi and T14-Av

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Germfree rats fed a high-sucrose diet were inoculated with Actinomyces viscosus strain T14-Vi (virulent) or T14-Av (avirulent). The mean recovery of strain T14-Vi from six extracted finely ground molars of rats sacrificed after 90 days was 1.1 × 108 colony-forming units (CFU). The mean recovery of strain T14-Av was 5.7 × 107 CFU, which was significantly less. Strain T14-Vi caused severe alveolar bone loss, but only minimal bone loss occurred in rats infected with strain T14-Av. Scanning electron microscopy of teeth of germfree rats revealed that strain T14-Vi colonized in the fissures as well as on tooth surface areas near the gingiva; strain T14-Av also colonized in fissures but was unable to colonize the teeth near the gingiva. In studies with conventional rats fed a high-sucrose diet, streptomycin-resistant strain T14-Vi colonized on the teeth of all rats inoculated with in the order of 108 or 107 CFU and on the teeth of about half of the rats inoculated with 106 or 105 CFU. In contrast, streptomycin-resistant strain T14-Av could not be detected on the teeth of any of the rats in groups similarly inoculated. In vitro “resting” cells of both strains suspended in conventional or germfree rat saliva survived to comparable degrees. [3H]thymidine-labeled T14-Vi cells adhered well to hydroxyapatite (HA) beads and to HA beads pretreated with saliva obtained from germfree or conventional rats. In contrast, T14-Av cells adhered less well than did T14-Vi cells to HA, whereas their adherence to saliva-coated HA was negligible. Transmission electron microscopy of negatively stained T14-Vi and T14-Av cells repeatedly passed in 1% phosphotungstic acid revealed fibrils on cells of both strains. T14-Av cells were covered by large amounts of extracellular material which was presumably heteropolysaccharide; little extracellular material was present on the surface of T14-Vi cells. T14-Vi cells had a relatively low affinity for the heteropolysaccharide synthesized by strain T14-Av. Other evidence also suggested that this polysaccharide had a relatively low affinity for saliva-coated HA. Collectively, the evidence indicates that the difference in periodontopathic potential between strains T14-Vi and T14-Av results from their different abilities to colonize teeth. This difference is probably due to the lower adherence of T14-Av cells to teeth rather than to their ability to grow in the mouth. The low affinity of T14-Av cells for tooth surfaces may be due, in part, to the presence of large amounts of cell-surface-associated polysaccharide.

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