INTRACELLULAR TOXIN OP BACILLUS PYOCYANEUS. 163 



THE INTRACELLULAR TOXIN OF BACILLUS TYOCYANEUS.* 



DONALD R. MACINTYRB. 



The object of this research was to ascertain whether or not Bacillus 

 pyocyaneus contains an intracelluhir toxin. Some years ago Buchner 

 obtained from this organism a large quantity of a pyogenetic proteid. 

 He took the germ substance from potato cultures, rubbed it up with 

 water and then treated it with about fifty volumes of a 0.5 per cent, 

 solution of caustic potash. This formed in the cold a mucilaginous 

 mass, which dissolved at the temperature of the water bath and after 

 being heated for several hours was filtered and acidified with acetic or 

 hydrochloric acid. This acid threw down a precipitate, which was col- 

 lected, washed with water, and was injected subcutaneously into ani- 

 mals. It was also placed in sterilized spindle-shaped glass tubes, and 

 introduced under aseptic precautions under the skin of rabbits. After 

 from two to three days the tubes were removed and found to contain, 

 in addition to some of the proteid, several cubic millimeters of fibrinous 

 pus, which proved to be sterile. 



In 1889, Charrin and Roger, working with Bacillus pyocyaneus, made 

 the experiments which may be said to be the starting point of all subse- 

 quent work on agglutination. These facts made it desirable to ascertain 

 whether or not this organism contains an intracellular toxin. 



My own work may be summarized as follows: Pure cultures were 

 grown in the large tanks used in this laboratory, and after fourteen days, 

 the germ substance was removed from the agar, precipitated with alcohol, 

 collected on a hard filter and extracted with alcohol and ether. The 

 toxicity of the germ substance for guinea-pigs is shown in Table 1. 



♦This paper and the two which follow are from a series of studies of bacterial cells, research in 

 the Hygienic Laboratory of the University of Michigan, 1903. 



