514 



BUIXKTIN l67 



cc. portions of these various dilutions were added with sterile 

 pipettes to tubes containing 9 cc. of sterile broth, thus making 

 dilutions of 1-100, 1-500, 1-1000, 1-1500, and 1-2000 of liquid 

 phenol in broth. As in the case of the formalin 5 tubes were 

 inoculated and 3 retained as checks. Growth was observed at 

 the end of a day in all tubes containing 1 part liquid phenol to 

 2000 parts of broth, and in those containing 1 part to 1500, while 

 four of the five tubes containing 1 part of liquid phenol to 1000 

 parts of broth, showed slight clouding. Good growth eventually de- 

 veloped in all tubes containing 1 part to 1000. No further change 

 was observed in these tubes during the 2 weeks they were held 

 under observation. 



Nitrogen requirements. — The water used in the study of the 

 nitrogen requirements of the organism was re-distilled with potas- 

 sium permanganate, after which a third distillation employing 

 sulphuric acid was carried out. Only chemicals of known purity 

 were employed, except in the case of glycerin which, while of a 

 high grade of purity, was not free from traces of nitrogen. 



The following formulas were used : 



Sodium chlorid, 

 grams. 



Calcium chlorid. 

 .10 gram. 



Magnesium sulphate, 

 .35 gram. 



Di-potassium phos- 

 phate, 2.2 grams. 



Glycerin (Merck's blue 

 label), 35 grams. 



Water, 1000 cc. 



Sodium chlorid, 

 6 grams. 



Calcium chlorid, 

 .10 gram. 



Magnesium sulphate. 

 .35 gram. 



Di-potassium phos- 

 phate, 2.2 grams. 



Mannit, 35 grams. 



Water, 1000 cc. 



C. 



Sodium chlorid, 

 G grams. 



Calcium chlorid, 

 .10 gram. 



Magnesium sulphate, 

 .35 gram. 



Di-potassium phos- 

 phate, 2.2 grams. 



Dextrose, 35 grams. 



Water, 1000 cc. 



Each of these solutions, A., B., and C., were divided into five 

 portions, one of which in each case was reserved as a control. 

 To the other four were added 1% of asparagin, urea, ammonium 

 chlorid, and potassium nitrate respectively. The media were 

 tubed and sterilized in the autoclave fur 15 minutes under 5 

 pounds of steam. 



