Summary. Serological variation in 71 oral isolates and three reference strains of Streptococcus milleri was examined. Antisera were raised by immunising rabbits with cells of 10 selected strains, followed by absorption of non-specific antibodies. Double diffusion of the typing sera and the Rantz and Randall extracts of the strains in agar gel demonstrated that 70 strains were divided into 10 serotypes (a-j) on the basis of cell-surface carbohydrate antigens. Only four strains were untypable. The typing scheme proposed depends on type antigens other than the Lancefield group antigens
Oral isolates of Streptococcus milleri were examined for their ability to coaggregate with actinomyces. Of the 68 S. mileri strains tested, including 3 reference strains, 40 strains coaggregated with Actinomyces naeslundii WVU45 (actinomyces coaggregation group B) and 36 strains coaggregated with Actinomyces viscosus T14V (actinomyces coaggregation group A). AU S. milleri strains of serotypes b (4 strains), e (2 strains), and f ( 24strains) coaggregated with both of the actinomyces. The coaggregation reactions between the S. milleri cells and A. naeslundii WVU45 cells were optimal at about pH 7.0 and were Ca2+ or Mg2+ dependent, but they were not inhibited by the presence of simple sugars or amino sugars, including lactose (up to 0.5 M). Treatment of the S. milleri cells with heat (100°C, 3 min) or proteases (trypsin, 1.0 mg/ml; pronase, 0.25 mg/ml; 37°C; 3 h) and of the actinomyces cells with periodate (0.01 M, 4°C, 16 h) destroyed their coaggregating abilities. The coaggregations between cells of the S. milleri strains, as well as cells of the Streptococcus sanguis Hi (reference strain for streptococcus coaggregation group 2) and the actinomyces strains (WVU45 and T14V), were inhibited by AFH1 (a carbohydrate receptor on T14V cells for a lectin on Hl cells). These interactions were also inhibited by anti-AFHl immunoglobulin G (IgG) and by anti-b, anti-e, and anti-f S. milleri IgG or anti-f IgG Fab fragments. These results suggest that S. milleri, at least strains of serotypes b, e, and f, belongs to streptococcus coaggregation group 2.
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