STUDIES ONSTREPTOCOCCUS PYOGENESII

Abstract

S. pyogenes (S. hae-molyticus) type 6 strain S43 was disintegrated and solubilized by shaking with glass beads. All cellular components not sedimented after centrifugation for 1/2 hr. at 20,000 x G were considered soluble. Fifty-eight % of cellular dry weight was solubilized in water in 1/2 hr., and the viable count was reduced to less than 1%. Maximum cell material solubilization (68%) was reached in approximately 3 hr. Bicarbonate extracted an additional 1.5%; acetate was negligible. The cell wall constituent rhamnose was found in all soluble fractions; the concentration in each fraction increased gradually until a concentration of 6% was present in the fraction obtained between the 4th and 5th hour. The largest quantity of rhamnose was solubilized in the first 15 minutes of shaking; 81% remained in the final insoluble residue. Positive precipi-tin tests for the group-specific C-carbohydrate and type-specific M-protein were obtained on all extracts; the strongest on fractions released in the later stages of shaking. Electron microscopic observations of disintegrated cells at various stages of shaking show extensive destruction of cell structures occurred after several hours. Granular particles of about 100 m[mu] diameter are seen upon rupture of the cell. The disintegration and solubilization of streptococci by shaking with "ballotini" can be divided into 2 phases rupture of the cell wall and release of cytoplasmic constituents, and partial solubilization of structures associated with or a part of the cell wall. Differences between the sonic oscillation and shaking methods of solubilizing streptococci are discussed.