542 Professor David James Hamilton [March 8, 



When the peritoneal or other serous effusion is inculjated at a 

 temperature of 88'^ C, nearly the whole of the rods will be found to 

 have vanished within forty-eight hours, while the number of free 

 spores has correspondingly increased. In giving birth to a spore, 

 under these circumstances, the mother bacillus seems to perish and is 

 bacteriolysed. 



Thus incul)ated, the spores may lie retained for a matter of years. 

 I cannot say for how many years, but I may affirm that the earliest 

 samples of peritoneal liquid in my possession will still reproduce 

 the disease with almost perfect certainty. The liquid does not 

 undergo ordinary putrefaction, and the vitality of the spores is not 

 influenced injuriously by the moisture. I generally keep the liquid 

 in sealed tubes, but it may be preserved in sterile bottles equally 

 well. 



The reaction of the peritoneal liquid is alkaline as a rule, and 

 abundant triple phosphate crystals are usually found in it. This 

 fact probal)ly accounts for the organism growing to such excess in 

 this liquid within the living body. 



Boiling of the spores for a matter of five minutes usually kills 

 them, but they are able to retain their vitality without fail at a 

 temperature of 80° C. continued for twenty minutes. Jensen* found 

 that after the stomach of a sheep had been kept in dilute spirit for 

 a period of seven weeks, he was still able to start a growth from it. 



The bacillus stains with gentian-violet, perhaps more intensely 

 than with any other of the aniline dyes, and for photographic pur- 

 poses this reagent is possibly the most serviceable ; but fuchsin and 

 Loeffler's methylene-blue give excellent results. The latter stains 

 the chromatic granules very delicately, and, like the others, leaves 

 the spore quite uncoloured. I would emphasise, however, that the 

 view obtained of the unstained organism in its living state and 

 suspended in the serous liquid in which it has been growing, is 

 incomparably truer to nature than that of any stained and clarified 

 preparation. 



The bacillus when taken fresh from the peritoneal liquid some- 

 times gives a reaction l)y Gram's method, l)ut a positive result is 

 uncertain, and reliance should not be placed on this reaction as a 

 point in the diagnosis. More frequently than otherwise the reaction 

 fails completely, and I have never obtained it in beef -tea cultures. 



Its Culture. — When cultivated on glucose-beef-tea under oil, all 

 germination should be over at the end of thirty-six hours' incubation, 

 and the organism in great part have settled down on the bottom and 

 the sides of the tube in fine particulate, sand-like masses. If the 

 tube has been retained in a sloping position, the deposit takes place 

 in a fine line all along its dependent aspect, and at the bottom. The 

 part of the deposit at the bottom of the tube has a slimy appearance 



* Deut. Ztschr. fur Tliiermcdiciu und vergl. Path., Bd. viii., 189G, p. 265. 



