MICHIGAN ACADEMY OF SCIENCE. 301 



added. By the addition of sterile normal NaOH solution the reaction 

 of portions of the original uninociilated medium was adjusted to 5° and 

 12° alkaline respectively. Twenty-five cc. of each of these was inocu- 

 lated from the same culture as the other two. The following table gives 

 the results : 



Table 9. 



Effect of Reaction upon Growth, etc. Test with Filtered Extracts. 



7 days 12 days 21 days 



Culture liquid plus 2% glucose - - 



Culture liquid no glucose - - 



Sterile clover juice medium, 5° alkaline -|- -f- + H — h 



Same— 12° alkaline 4- ++ + + 



-, no growth; -|-, fair growth; -f- -|-, good growth. 



From the above results it is evident that neither the alkalinity nor the 

 absence of food are the causes of the discontinuance of growth. The 

 accumulation of metabolic by-products is the })robable cause of the re- 

 tardation of growth. 



Changes in Carbohydrate Media: 



Neither acid or gas is formed in fermentation tubes when the fungus 

 grown in 2% glucose, 2% maltose, or 2% saccharose. The quantity of 

 growth in these solutions is fair, and if any acid is formed, the amount 

 is so small that its presence cannot be detected with litmus paper. 



Anaerobic Growth: 



A Lavaran tube was used for growing the fungus in the absence of 

 oxygen. In one arm of the tube clover juice agar which had been boiled 

 for several minutes to drive out the air was poured and slanted. When 

 solidified this was at once inoculated. In the other arm of the tube 

 pyrogallic acid and KOH solution were placed, and the mouths of both 

 tubes quickly sealed with rubber stoppers which were then heavily coated 

 with paraffin. Within two weeks the first signs of growth usually 

 ai^peared. From that time on growth proceeded slowly with light spore 

 formation between the fourth and fifth week. This experiment was 

 repeated several times with the same results. It is extremely unlikely 

 that a perfect anerobic condition was present inside the tube, since it is 

 jjossible that the agar upon cooling absorbed sufficient air for the growth 

 of the fungus. At all events the fungus can grow, though slowly with as 

 meager supply of free oxygen as the average anaerobic bacterium. 



TEMPERATURE RELATIONS. 



This series of temperature studies was made by inoculating two tubes 

 each of corn meal agar, clover agar, and clover juice and keeping incuba- 

 tors at the various temperatures. 



