1 89(5.] rapidly producing Diphtheria Antitoxines. 291 



for more than a year. The toxines described by Continental observers 

 have been those obtained from broth cnltivations of the organism, 

 where we cannot expect the products, even if identical in nature, to 

 be present in the same proportions as when vegetating in the living 

 body of an animal. More than eight years ago Hueppe* and I pointed 

 out that we must grow the pathogenic organism on a natural 

 albumen if we wished to obtain the toxines proper of the microbe 

 and working on this principle by inoculating hens' eggs with Koch's 

 cholera bacillus we succeeded in producing much more powerful 

 poisons than had been previously obtained. The toxines produced by 

 the action of the diphtheria bacillus on albumen have been minutely 

 investigated by Sidney Martin, and the occurrence of the same sub- 

 stances in the bodies of patients who have succumbed to the disease 

 has proved the correctness of this view. In this communication I 

 shall not enter into the precise nature of the products in question, as 

 my present purpose is to describe a practical method of rapidly pro- 

 ducing powerful diphtheria antitoxines. 



In these experiments two species of diphtheria toxine were made 

 use of : firstly, the ordinary toxine produced by the organism in 

 peptone broth ; secondly, the substances present in serum-broth cul- 

 tivations which had been filtered and heated up to 65° C. [In the 

 former the active principle consists almost entirely of the so-called 

 ferment toxine, while in the latter this has been destroyed by heating 

 up to 65° C, so that its action must depend on the presence of other 

 substances. ]f The medium employed for the production of the serum 

 toxine was ordinary peptone broth, to which an addition of 10 or 20 

 per cent, blood serum or plasma had been added ; where the latter 

 was used the broth was previously decalcified to prevent coagulation. 

 As a rule the broth was inoculated with a virulent diphtheria culture 

 some three or four days previous to the addition of the serum or plasma, 

 and then incubated at a temperature of 37° C. for at least three or four 

 weeks. Before being used for injection it was subjected to a tem- 

 perature of 65° C. for about an hour and then filtered through a 

 sterilised Chamberland candle to remove the bodies of the bacilli. 

 This fluid will be spoken of subsequently as " serum " toxine in contra- 

 distinction to the ordinary poison, which will be spoken of as 

 " broth " toxine. The serum toxine is characterised chiefly by giving 

 rise to little local irritation but marked febrile reaction which is 

 still more pronounced when the injection is repeated. As the 

 diphtheria albumose described by Sidney Martin was characterised 

 by precisely these properties, in all probability the potency of the 

 serum toxine depends on its presence. In addition to these properties, 

 however, it was found that animals which had been subjected to its 

 * ' Cent. f. Bakt.,' No. 5, 1884. 



f The passages in brackets were added after the reading of the paper. 



