[Vol. 6 



116 ANNALS OF THE MISSOURI BOTANICAL GARDEN 



nomenclature used is that of Migula ( '00). The following or- 

 ganisms were employed: 



(1) Alkaline on litmus milk. 



Bacillus v id gat us (Fliigge) Mig. 



Bacterium mycoides (Sclioll) Mig. 



(2) Acid on litmus milk. 



Bacillus vulgaris (Hauser) Mig. 



Bacillus coli (Escherlich) Mig. 



Bacillus prodigiosus (Ehrenberg) Fliigge. 



(3) Nitrogen-fixing form. 



Azotobacter chroococcum Beijerinck. 



All forms were plated in the usual manner, to make certain 

 that pure cultures were being considered, and then transferred 

 to standard beef broth. 



Method of inoculation of culture bottles. — Transfers from 

 the sawdust cultures of the fungi were made to sterile agar 

 plates. When the entire surface of the plate was overgrown 



with the mycelium it was cut into small squares of approxi- 

 mately one square centimeter size by means of a sterile scalpel. 

 A single square was added to each culture bottle. In most 

 cases the mycelium could be seen growing through the sawdust 

 about one week after inoculation. 



When the fungus mycelium had become well established in 

 the sawdust the culture bottles were inoculated a second 

 time either with another fungus or with bacteria. Previous to 

 this second inoculation all culture bottles were carefully 

 inspected and those in which the infection of the sawdust did 

 not occur were thrown out. The second fungus inoculation 

 was made in a similar manner to the first. The bacteria were 

 added by transferring a considerable number of the organisms 

 from hard agar slants to 250-cc. Erlenmeyer flasks containing 

 distilled water. Where more than one bacterium was added 

 to the culture bottles a mixture of the organisms was made in 

 the same way. Approximately one cc. of bacterium suspension 

 was added to each culture bottle. After all of the culture bot- 

 tles were inoculated with the bacterial suspension transfers 

 were made to agar slants to make certain that the action of the 



