114 CROWN-GALL OF PLANTS. 



uschinsky's solution. 



Grovjth scanty, not viscid. Tubes were inoculated from a beef- 

 broth culture 3 da3^s old, a l-mm. loop being used for each tube of 

 Uschinsk3^'s solution. In 2 days a scanty growth, which was slightly 

 flocculent, could be seen, together with a few white filamentous flaky 

 particles which were in suspension in the liquid. At the end of 6 days 

 no further change was perceptible and the fluid did not become viscid, 

 nor fluorescent. There was no pellicle. Under the microscope, at 

 the end of 2 months, the filamentous flakes consisted of numerous 

 short rods staining readily in carbol fuchsin. These rods appear to 

 lie in an unstained slime. No chains were detected. 



SODIUM CHLORIDE BOUILLON. 



Four per cent of salt inhibits growth, 3 per cent retards growth or 

 inhibits it. 



A l-mm. loop of a 3-day-old -M5 peptonized beef-broth culture 

 was placed in each tube of peptonized beef broth, containing 1, 2, 

 3, 4, 5, and 6 per cent c. p. sodium chloride — several tubes of each sort. 

 At the end of 6 days, growth was apparent in tubes containing 1, 2, 

 and 3 per cent, but no growth could be detected in tubes containing 



4 per cent or more of sodium chloride, indicating that 4 per cent will 

 inhibit the growth of the organism. The growth in the 3 per cent was 

 slight. 



In another experiment the daisy organism refused to grow in 3 per 

 cent salt bouillon (Table VI). 



In a repetition test it grew in 3.5 per cent. 



GROWTH IN BOUILLON OVER CHLOROFORM. 



Growth is unrestrained. Chloroform to the amount of 5 c. c. was 

 iim into 5 tubes of bouillon by means of a sterile pipette. Three tubes 

 were then inoculated with the organism from a 10-day-old bouillon 

 culture. In 2 days there was a good growth at the top of the bouillon ; 

 1 2 days after inoculating a heavy growth was present. The tubes were 

 not shaken. 



NITROGEN NUTRITION. 



Nitrogen is obtained from peptone, asparagin, etc. In filtered river 

 water containing 0.5 per cent dextrose and 0.5 per cent urea there 

 was no growth. The experiment was repeated some months later 

 with the same result In filtered river water containing 1 per cent 

 asparagin the organism made a slow initial growth, first visible after 



5 days. Strain B, which had been in the laboratory several years 

 and had lost its virulence, grew better than a recent isolation. This 



213 



