PSEUDOMONAS MRUGINOSA 369 



the enzymes encountered in the animal body? Which? and 

 Why? 



Extract with chloroform a six days' old bouillon culture 

 of this organism. In which portion of the liquid so extracted 

 is the proteolytic ferment contained, the chloroform extract 

 or the supernatant fluid? 



Mix slowly a two weeks' old bouillon culture of this 

 organism, grown at body temperature, with six times its 

 volume of absolute alcohol. Allow to stand over night. 

 Filter. Redissolye the precipitate in a few c.c. (5 or 6), of 

 physiological salt solution. In the meantime evaporate the 

 alcohol filtrate to dryness at a temperature not exceeding 40° 

 C, and redissolve the sedement in 5 or 6 c.c. of physio- 

 logical salt solution. Test both of these solutions on car- 

 bolized gelatin for proteolytic activity. What are the 

 results and how are they explained? 



Inoculation into Animals. — As a rule, cultures of this 

 organism obtained directly from the discharges of the wound 

 are capable, when introduced into animals, of producing 

 diseased conditions; but cultures kept on artificial media 

 for a long time may in part, or completely, lose this power. 



When guinea-pigs or rabbits are inoculated subcutaneously 

 with 1 c.c. of virulent fluid cultures of this organism, death 

 usually results in from eighteen to thirty-six hours. At 

 the seat of inoculation there are found an extensive purulent 

 infiltration of the tissues and a marked zone of inflammatory 

 edema. 



When introduced directly into the peritoneal cavity the 

 results are also fatal, and at autopsy a genuine fibrinous 

 peritonitis is found. There is usually an accumulation of 

 serum in both the peritoneal and pleural cavities. At 

 autopsies after both methods of inoculation the organisms 

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