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Specificity of coaggregation reactions between human oral streptococci and strains of Actinomyces viscosus or Actinomyces naeslundii.

Coaggregation reactions between actinomycete and streptococcal cells occurred frequently when human strains of Actinomyces viscosus or A. naeslundii were mixed with human isolates of Streptococcus sanguis or S. mitis, but were infrequent with other oral actinomycetes and streptococci. Two groups of actinomycetes and four groups of streptococci were defined by the patterns of their coaggregation reactions and by the ability of beta-linked galactosides (i.e., lactose) to reverse these reactions. Coaggregations occurred by one of the following three kinds to cell-cell interactions: (i) coaggregation that was blocked by heating the streptococcus but not the actinomycete and was not reversed by lactose; (ii) coaggregation that was blocked by heating the actinomycete but not the streptococcus and was reversed by lactose; and (iii) coaggregation that was blocked only by heating both cell types. The latter reaction was a combination of the first two since lactose reversed coaggregation between heated streptococci and unheated actinomycetes but did not reverse coaggregations between unheated streptococci and heated actinomycetes. Cells that could be heat inactivated also were inactivated by amino group acetylation or protease digestion, whereas cells that were unaffected by heat were not inactivated by these treatments. Coaggregation reactions of each kind were Ca2+ dependent and insensitive to dextranase treatment. These findings are consistent with the hypothesis that human strains of A. viscosus and A. naeslundii coaggregate with strains of S. sanguis and S. mitis by a system of specific cell surface interactions between protein or glycoprotein receptors on one cell type and carbohydrates on the other type.

Acetylation↗

Immune labeling of certain strains of Actinomyces naeslundii and Actinomyces viscosus by fluorescence and electron microscopy.

A total of 12 well-characterized strains of Actinomyces viscosus and A. naeslundii grown on Trypticase soy agar plates supplemented with sheep erythrocytes were examined by light microscopy and transmission electron microscopy after treatment with appropriately labeled antisera to homologous and heterologous strains. Cells incubated with homologous rabbit antisera followed by fluorescein-isothiocyanate (FITC)-conjugated goat anti-rabbit immunoglobulin G (IgG) exhibited a completely smooth fluorescent outline in the case of A. naeslundii and and interrupted, irregular fluorescent outline in the case of human strains of A. viscosus. The different labeling patterns appeared to be related to the presence at the ultrastructural level of long, unevenly distributed strands of "fuzz" on the surface of human A. viscosus cells, whereas A. naeslundii cells had a narrower layer of fuzz, or more even thickness. The immunocoating reaction revealed homologous antibody binding to the irregular strands of fuzz on the surface of human A. viscosus cells, whereas homologous antisera to A. naeslundii coated A. naeslundii cells with a moderately electron-dense coating of antibody of even thickness. Human strains of A. viscosus incubated with heterologous antiserum to A. naeslundii followed by FITC-labeled goat anti-rabbit IgG exhibited a segmented fluorescent outline, which differed from that produced with homologous antisera. A. naeslundii incubated with heterologous rabbit antisera to human A. viscosus strains and FITC-labeled anti-rabbit IgG exhibited a completely smooth fluorescent outline similar to that produced with homologous antiserum. A. viscosus strains of hamster origin differed from A. viscosus strains of human origin by the absence of a surface fuzz and the comparatively smooth, even fluorescence produced by incubating these cells with homologous rabbit antiserum followed by FITC-labeled goat anti-rabbit IgG. Antiserum to a hamster strain did not cross-react with A. naeslundii or human strains of A. viscosus. Under the growth conditions of this experiment, ultrastructural features and labeling patterns with the indirect fluorescent technique may be useful in differentiating these serotypes from one another.

Actinomyces↗

Neuraminidase-dependent hamagglutination of human erythrocytes by human strains of Actinomyces viscosus and Actinomyces naeslundii.

Human A, B, and O erythrocytes (RBC) were agglutinated by many human strains of Actinomyces viscosus and A. naeslundii. At 37 degrees C, these bacterium-mediated hemagglutination reactions required the action of bacterial neuraminidase upon the RBC; however, at 4 degrees C, the requirement for neuraminidase was not as striking. Bacterial cell suspensions which caused hemagglutination at 37 degrees C contained both soluble extracellular and cell-associated neuraminidase activities as shown by enzyme assays using a soluble substrate (i.e., alpha 1-acid glycoprotein). Bacterium-mediated hemagglutination occurred only in the presence of soluble neuraminidase activity, and the rate of hemagglutination could be inhibited by 2-deoxy-2,3-dehydro-N-acetylneuraminic acid, a competitive inhibitor of purified soluble neuraminidase from A. viscosus T14V. Suspensions of bacteria which contained only cell-associated neuraminidase activity were unable to initiate hemagglutination, but they caused immediate hemagglutination when mixed with neuraminidase-treated RBC. All hemagglutination reactions were reversible in the presence of 0.02 M lactose and were abolished by heating (85 degrees C for 30 min) the actinomycete cells but not the RBC. The proposed mechanism of hemagglutination involves two sequential steps: (i) the action of neuraminidase to unmask galactose-containing receptors on the RBC and (ii) the multivalent binding of these receptors by many low-affinity lection sites on the bacterial surface.

Actinomyces↗

Identification of Actinomyces israelii and Actinomyces naeslundii by fluorescent-antibody and agar-gel diffusion techniques.

This study was an attempt to develop a fluorescent-antibody (FA) test to differentiate Actinomyces israelii and A. naeslundii as an aid in their laboratory identification. Two strains of A. israelii (X522 and A601) and two strains of A. naeslundii (X454 and X600), which had received intensive study by several investigators, were used for the immunization of rabbits. Working titers, based on tests with antigens prepared from the homologous strains and from well-established heterologous strains, were determined for each labeled antibody preparation. These conjugates and their normal serum control conjugates were used separately to stain 85 cultures of Actinomyes species and 23 strains of other species that might be confused with them. Acetone-precipitated soluble antigens from these same strains were tested with different antisera in the agar-gel diffusion test. Results showed that A. israelii (X522 and A601) and A. naeslundii (X454 and X600) labeled antiglobulins, when used at their working titers, stained most strains of their homologous species. Agar-gel diffusion results showed general agreement with those of the FA tests. The two tests appear to be equal in sensitivity, but the FA test is more specific, since several cross-reactions were noted with the agar-gel diffusion test whereas no cross-reactions were obtained with the FA reagents. Agar-gel and FA studies suggest that at least two serotypes of A. israelii may be associated with human disease. Although the majority of strains tested in this study appear to belong to a common serotype, "serotype 1," two strains of an apparent second serotype, "serotype 2," were encountered. FA staining of tissue impression smears from experimentally infected mice was successful when a counterstain, Evans Blue dye, was used.

Actinomyces↗

Incidence and characterization of anti-microbial effects produced by Actinomyces viscosus and Actinomyces naeslundii.

Sixty-two facultative Actinomyces strains isolated from dental plaque were tested for the production of bacteriocin-like inhibitory effects by a deferred antagonism method. When incubated anaerobically, all isolates produced identical inhibitory patterns against 15 indicator organisms, but under microaerophilic conditions, little inhibitory activity was observed. Activity was not evident after anaerobic incubation on a medium buffered by 0.5% (w/v) calcium carbonate. Gas-liquid chromatographic analyses of agar blocks removed from the inhibitory zones indicated that, compared with microaerophilic conditions, anaerobic incubation encouraged production of high concentrations of lactic and succinic acids, and the concomitant fall in the pH was probably responsible for the inhibitory effects.

Acids↗