CULTURE MEDIA FOR CULTIVATIOX OF MICROORGANISMS 



337 



(2) Make up to 1000.0 cc. by the addition 

 of water. 



(3) Filter. 



(4) Heat distilled water containing 0.2% 

 KH2PO4 and 0.01% MgS04 to boiling. 



(5) Cool (4) and filter. 



(6) To each 100.0 cc. of (5) add 0.1%, 

 1.0%, 2.0%, 5.0% or 10.0% of one of 

 (3). (Filtered extract solution.) 



(7) Incubate at room temperature for one 

 week to test sterility. 



Sterilization: Sterilize for 20 minutes on 

 each of 3 successive days in streaming 

 steam. 



Use: To study gum formation by Bacillus 

 radicicola. Author reported that tolu, 

 red clover blossoms, glycyrrhiza, kino, 

 baptisia and senna favored production. 

 Cathechu, physostigma, scoparius and 

 kino in high concentrations were un- 

 suitable. 



Variants: The author added 1.0% maltose. 



Reference: Buchanan (1909 pp. 72, 386). 



1158. Buchanan's Basal Clover Infusion 

 Solution 



Constituents: 



1. Water 4000.0 cc. 



2. Clover leaves and stems 

 {Trifolium pratense) 200.0 g. 



Preparation: 



(1) Extract 200.0 g. of the leaves and 

 stems of clover (Trifolium pratense) 

 with 4 liters of boiling tap water. 



(2) Filter until clear. 



(3) Add one of the added nutrients to (2). 



(4) Heat to boiling. 



(5) Cool and filter. 



(6) Incubate for one week at room tem- 

 perature to test sterility. 



Sterilization: Sterilize for 20 minutes on 

 each of 3 successive days in streaming 

 steam. 



Use: To study gum formation by Bacillus 

 radicicola, and cultivation of Bacillus 

 radicicola bacteroids. Author reported 

 that clover extracts alone favored the 

 production of gum. Peptone, and to a 

 less degree, asparagin and sodium aspara- 

 ginate inhibited the production of gum. 

 Nitrates except in large amounts did not 

 effect growth and gum production. 

 Clover extract did not constitute a 

 favorable medium for the development of 



the bacteroids, either unmodified or upon 

 the addition of asparagin or peptone. 

 Added nutrients: The author added one of 

 the following: 



KH2PO4 0.2% 



MgS04 0.1% 



Ammonium phosphate 0.5% 



Asparagin 1.0% 



Sodium asparaginate 1-0% 



Peptone 0.1, 0.5, 1.0, 



2.0 or 5.0% 



[Sucrose 2.0% 



\ [0.005, 0.01, 



[KNO3 i0.05, 0.1, 0.2 



[0.5 or 1.0% 



Glucose 2.0% 



Reference: Buchanan (1909 pp. 62, 386, 

 391). 



1159. Buchanan's Vetch Infusion Solution 



Constituents: 



1. Water 2000.0 cc. 



2. Vetch stems and leaves 200.0 g. 



Preparation: 



(1) Boil 200.0 g. of dry vetch stems and 

 leaves in 2 liters of water. 



(2) Filter. 



(3) Concentrate to ^ its volume by heat. 



(4) Add several times its volume of 

 alcohol. 



(5) Filter. 



(6) Evaporate the filtrate to dryness. 



(7) Wash the precipitate with absolute 

 alcohol, several times and evaporate 

 to dryness. 



(8) Dissolve each of these extracts in 

 100.0 cc. of distilled water. 



(9) Filter. 



(10) The first of these is called alcoholic 

 extract and the second the aqueous 

 extract. 



(11) Prepare 0.01, 0.1, 0.2 or 1.0% solu- 

 tions of the aqueous extract in dis- 

 tilled water and 0.1, 0.5, 1.0, 2.0 or 

 5.0% solutions of the alcoholic ex- 

 tract in distilled water. 



(12) Tube. 



(13) Incubate for one week at room tem- 

 peratures to test sterility. 



Sterilization: Sterilize in streaming steam 

 for 20 minutes on each of 3 successive 

 days. 



Use: To study gum formation by Bacillus 

 radicicola and cultivation of Bacillus 



