506 Bulletin 167 



trols in a Now jar, the cover of which was sealed with Darwin's 

 wax mixture. Carbon dioxid prepared in a Kipp generator 

 from boiled marble chips and chemically pure hydrochloric acid 

 was passed successively through wash bottles containing 10% 

 sodium carbonate, 10% potassium permanganate and boiled dis- 

 tilled water respectively, and then through the Novy jar. The 

 cotton plugs employed were as loose as was consistent with the 

 safety of the cultures and the jar was exhausted with the vacuum 

 apparatus several times to aid in removing all oxygen. After 

 carbon dioxid had passed through the jar two hours and a half 

 it was sealed and incubated at room temperature. After 20 

 days the jar was opened and the tubes examined for growth 

 with negative results. Within 24 hours after air was admitted 

 to the culture all except the controls developed characteristic 

 growth. 



A second series of cultures was prepared in the same man- 

 ner as the first except that a 2.' < dextrose nutrient broth was 

 employed. Five days after inoculation fermentation became ap- 

 parent in several tubes, as could be seen from the gas collecting 

 in bubbles at the surface. After two weeks all inoculated tul>es 

 showed fair growth. Pellicle and ring formation were absent. 

 The growth was characterized by a tendency to granular forma- 

 tion throughout the medium and the evolution of appreciable 

 amounts of gas on agitation. When air was admitted to the 

 cultures moderate clouding promptly developed and the usual 

 aerobic characters appeared. 



Desiccation. — For desiccation tests small portions of 24 

 hour old broth cultures were transferred to sterile cover slips 

 in sterile Petri dishes. Only a very small amount of material 

 w as transferred and this was spread in a thin film on the cover 

 slip. The preparations were allowed to dry at room tempera- 

 ture, 20 to 24 . Slips were transferred to tubes of nutrient 

 broth with sterile forceps as soon as dry. and at the end of 3 

 hours. 17 hours. 24 hours. 4T hours, 50 hours, 6 days, 9 days, and 

 20 days. In no case was the organism killed by less than 



