Studies on Streptococcus pyogenes. II. Observations on the microscopical and biological aspects of the disintegration and solubilization of a type 6 strain by shaking with glass beads.

Slade and Vetter (1956) have shown that exposure to sonic oscillation of an aqueous suspension of a type 6 strain of Streptococcus pyogenes (Streptococcus haemolyticus) resulted in the solubilization of considerable quantities of the group-specific C-antigen, the type-specific M1antigen, and the cell wall constituent rhamnose. Several hours of oscillation resulted in the release into solution of 85 per cent of the dry weight of the cell. Electron microscopic observations at various times during the oscillation process demonstrated that extensive disintegration of the structural elements of the cell occurred. Hess and Slade (1955) found that shaking the same strain of S. pyogenes with glass beads in a Mickle disintegrator resulted in the solubilization of only 60 per cent as much of the cell material as had been obtained by the sonic process. This result, plus the fact that cell walls of group A streptococci prepared by shaking with glass beads (Salton, 1953; Barkulis and Ekstedt, 1955) were found to contain M-antigen, indicated that the disintegration and solubilization of the cell by the latter process was not as extensive as that caused by sonic energy. It was thus of considerable interest to investigate further the shaking procedure. In addition, the previous results made it desirable to examine the possible advantages of MIickle extracts as compared to sonic extracts as a starting material for the preparation of purified MI-antigen. It will be shown that the extracts obtained by the two procedures differed sharply in several instances. In addition, microscopical observations of the process of cell destruction are presented.