BACTERIAL CELLULAR SUBSTANCE 39 



in vacuo, taken up in a small volume of water, and injected 

 into animals without effect. That the extraction of the 

 cellular substance of the colon bacillus with alcohol and 

 ether has no destructive action on the intracellular poison 

 is shown by the fact that this material after extraction with 

 these agents does not lose any of its toxicity. Furthermore, 

 it may be stated that prolonged boiling of the cellular sub- 

 stance of the colon bacillus with alcohol or ether, or with 

 the two successively, neither sets free nor destroys the 

 intracellular poison. These agents dissolve the fats, waxes, 

 and coloring matter from bacterial cells, but do not remove 

 or destroy the intracellular poison. As will be seen later, 

 this is also true of the intracellular poison of those bacteria 

 that produce a soluble toxin in their cultures. Moreover, 

 it will later appear that the poisonous group not only in 

 bacterial but in vegetable and animal proteins is soluble 

 in absolute alcohol after it has been well detached from 

 the other groups in the protein molecule, but it is not 

 removed from its place in the complex molecule by either 

 alcohol or ether. Indeed, the poisonous group in the protein 

 molecule is not removed from its attachment to other 

 groups by any purely physical solvents, and the molecule 

 must be disrupted by high temperature, chemical agents, 

 or enzymes before its poisonous constituent can be extracted 

 by physical solvents. 



3. The cellular substance of the colon bacillus may be 

 heated with water without destruction of its poisonous group. 



Two hundred milligrams of the cellular substance of the 

 colon bacillus was suspended in 10 c.c. of water in a tube, 

 which was then sealed and heated at 184 for thirty minutes. 

 On opening the tube, the milky content was found on 

 microscopic examination to contain granular debris with a 

 few unbroken cells. Portions of this heated substance 

 injected into guinea-pigs caused death, and autopsy revealed 

 the same lesions that are seen after death from either the 

 living bacillus or the unbroken cellular substance. 



Another portion of the content of this heated tube was 

 placed in a centrifuge and separated into a deposit and a 



