August 22, 1902.] 



SCIENCE. 



313 



acting as a ferment, or by forming some solu- 

 ble enzyme, may split up the proteids or other 

 constituents of the body or of the artiiicial cul- 

 ture medium, and among these split products 

 poisons may be produced. In this way the 

 formation of the specific toxins would be an 

 analytical process. In confirmation of this 

 theory, highly poisonous bodies have been 

 found in artificial culture media of some of 

 the pathogenic bacteria. Some of the first of 

 these poisonous bodies found were basic in 

 character and were known as ptomains, which 

 is a designation given to putrefactive alka- 

 loids. Many chemists have sought diligently 

 among the basic products of putrefaction for 

 the specific toxins of the infectious diseases, 

 and while a few highly poisonous ptomains 

 have been found, it is safe to say that no one 

 has yet been discovered to which all the sjonp- 

 toms and lesions of a disease could be 

 attributed. The researches of Roux and 

 Yersin, followed by those of Brieger and 

 Fraenkel, on the diphtheria toxin, led to the 

 conclusion that the specific poisons, instead of 

 being basic in character, are modified proteids, 

 and to these the term toxalbumins has been 

 given. However, competent workers have 

 failed to find anything like toxalbumins in 

 cultures of many of the most virulent patho- 

 genic bacteria. Anthrax is a disease to which 

 all of these theories concerning the modus 

 operandi of bacteria have been referred, and 

 although Brieger and Fraenkel thought at one 

 time that they had found a specific toxalbumin 

 in the bodies of animals dead from anthrax, 

 more careful investigations by other chemists 

 have failed to confirm their results, and it is 

 safe to say to-day that no specific bacterial 

 toxin has been found either in the body after 

 death from an infectious disease, or in artificial 

 culture media ; at least, no substance belonging 

 to this group of bodies to which the symptoms 

 and lesions of the disease could be attributed. 

 The other possible explanation of the produc- 

 tion of chemical poisons by pathogenic bac- 

 teria is that these substances are formed by 

 synthetical processes and are built up in the 

 cells of the microorganisms. The problem 

 which Dr. Vaughan and his students have at- 

 tempted to solve was that of determining 



whether or not the cells of pathogenic bacteria 

 contain specific toxins. By means of the large 

 incubating tanks, devised by Dr. Vaughan, and 

 which have been described in Science on page 

 378 of the issue of March 7, 1902, the cellular 

 substance of the pathogenic bacteria has been 

 obtained in large amount. The first experi- 

 ments were made witH the colon bacillus. A 

 virulent form of this germ was first grown in 

 ordinary beef tea cultures at 37° for periods 

 varying from fifteen to thirty days, and shown 

 to be toxic in both sterile and unsterilized con- 

 ditions, provided that the sterilization was 

 accomplished by means of heat, but when the 

 beef tea culture was deprived of germs by fil- 

 tration through porcelain, the germ-free fil- 

 trate could be injected into guinea-pigs in 

 amounts of from eight to ten cubic centime- 

 ters without any other harm than that which 

 would result from the introduction of an equal 

 amount of sterile beef tea into the abdominal 

 cavity of an animal. It follows from this that 

 in the colon bacillus at least there is no sol- 

 uble toxin formed. However, when the germ 

 substance was obtained in large amount, free 

 from constituents of the culture medium, ex- 

 tracted with alcohol and ether, dried and pul- 

 verized, and injected into animals in doses of 

 from one to five milligrams, death resulted. 

 This shows that the bacterial cell contains a 

 toxin. In case of the colon bacillus the bac- 

 terial cell suspended in water and placed in 

 a sealed tube may be heated in 18&° C, for 

 half an hour without loss of toxicity. Various 

 attempts were made to extract the toxin from 

 the cell by physical means. All kinds of sol- 

 vents, including various salts of different 

 strengths, were tried, without effect. These 

 investigations led to the belief that the spe- 

 cific toxin of the colon bacillus is an integral 

 part of the cell, and consists of a molecular 

 group of the cell proteid. It was then found 

 that when the dried colon cell substance was 

 heated at the temperature of the water bath 

 for a period not exceeding thirty minutes with 

 one-per-cent. aqueous solution of sulphuric 

 acid a certain constituent of the cell was split 

 off by this process and passed into solution. 

 The acid extract when filtered through porce- 

 lain or hard paper yields a voluminous pre- 



