TYPHOID TOXIN 377 



1. Toxin of Sanarelli. (a) Method of preparation. Sanarelli uses a virus 

 the virulence of which has been raised by passing it through the peritoneal 

 cavities of guinea-pigs (p. 368). The bacillus is sown in 2 per cent, glycerin- 

 broth and after incubating at 37 C. for a month is sterilized by heat and 

 allowed to remain at room temperature for 8 months. The flask containing 

 the culture is then sealed in the flame and heated to 60 C. for a few days. 

 During this long period of maceration the intra-cellular toxin diffuses into 

 the culture fluid and this, carefully decanted, constitutes the toxin of 

 Sanarelli. 



Gauthier and Balthazard justifiably point out that Sanarelli' s toxin is a complex 

 mixture containing substances foreign to the typhoid bacillus, and derived partly 

 from the albuminoid substances present in the medium, partly from the dead bodies 

 of the bacilli, etc. which have slowly undergone disintegration. It is nevertheless 

 true that animals inoculated with the different toxins prepared by SanarelH, Chante- 

 messe, and Balthazard exhibit identical symptoms. 



(/?) Action on laboratory animals. On rabbits. The toxin given sub- 

 cutaneously in doses of 10 c.c. per kg. of body weight kills rabbits weighing 

 700-1000 grams. 



Soon after inoculation the animal is seen to breathe more rapidly and to become 

 unsteady on its legs ; a general paralysis gradually comes on and about 10 hours 

 after inoculation convulsions occur ending in death. The temperature is at first 

 a little raised (about r V C.) but soon falls below normal and death takes place while 

 the temperature is still sub-normal. The effects of the toxin vary in different 

 animals : death is not infrequently delayed for some days and in that case is preceded 

 by a period of cachexia of which the characteristic signs are wasting, diarrhoea, etc. 

 Post mortem, the abdominal organs are found to be anaemic, and it is noticeable that 

 there is neither congestion of the intestinal mucous membrane nor swelling of the 

 Peyer's patches. 



On mice. 1 c.c. of toxin inoculated sub-cutaneously or P 2 c.c. intra- 

 peritoneally is generally fatal, death taking place in a few hours. Post mortem 

 the spleen is enlarged and there is a small amount of a sterile effusion in the 

 peritoneal cavity. 



On guinea-pigs. Guinea-pig inoculation is an excellent means of testing 

 typhoid toxin sub-cutaneously the minimal fatal dose is 1'5 c.c. per 100 



frams of body weight. Intra-peritoneally the results are less constant, 

 ub-cutaneous inoculation of 4 or 5 c.c. of toxin per 100 grams of body weight 

 leads to death in 15-20 hours. 



Prom the moment of inoculation the temperature falls and continues to do so until 

 death. About an hour after inoculation there is marked abdominal distension 

 accompanied by extreme tenderness, the animal does not move, but sits huddled up 

 and cries if touched : after 4 or 5 hours it is extremely dejected, its eyes are half- 

 closed and it is seen to be in an almost uninterrupted state of tremor ; a profuse 

 sometimes haemorrhagic diarrhoea is often present, and finally paralysis appears, 

 the meteorism vanishes and death takes place. Post mortem a variable quantity 

 of exudate rich in leucocytes and sometimes turbid is found in the peritoneal cavity ; 

 the spleen is enlarged, congested and friable : the walls of the small intestine 

 are distended and completely infiltrated with blood, the mucous membrane is red 

 and the lymphatic patches are infiltrated and congested ; the stomach and 

 suprarenal capsules are intensely congested and ecchymosed. The intestine is full 

 of liquid matter and contains a very virulent colon bacillus in pure culture. 



On monkeys. Monkeys are very susceptible to the toxin of the typhoid 

 bacillus : the course of the disease and the lesions are the same as in the 

 guinea-pig. 



2. Toxin of Chantemesse. (a) Method of preparation. Chantemesse at 

 first recommended growing an organism of increased virulence in a maceration 

 of spleen and bone marrow. He now, however, prefers to use a bacillus 



