338 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



radicicola bacteroids. The author re- 

 ported that the aqueous and alcoholic 

 extracts in low dilutions tended to favor 

 the production of gum. Both the alco- 

 holic and aqueous extracts of the vetch 

 served as suitable nutrients for growth of 

 B. radicicola when in proper concentra- 

 tion. There was little bacteroid pro- 

 duction. 



Variants: The author specified the use of 

 V. Faba stems or leaves. 



Reference: Buchanan (1909 pp. 71, 386). 



1160. Wilhelmi's Clover Flower Infusion 

 Solution 



Constituents: 



1. Distilled water 1500.0 cc. 



2. Clover flowers 



3. Glucose 75.0 g. 



Preparation : 



(1) Fill a 2 liter flask with clover flowers 

 and add 1500.0 cc. distilled water 

 and 75.0 g. glucose. 



(2) Sterilize for 20 minutes in the auto- 

 clave at 120 °C. 



(3) Add sterile distilled water to restore 

 to 1500.0 cc. 



(4) Filter. 



(5) Add 25.0% HCl until litmus paper is 

 turned red (about 5.0 cc). 



(6) Allow to stand a day and again adjust 

 (about 1 or 2 cc . acid will be required) . 



(7) A slight acidity will be reached after 

 4 or 5 days, then observe the reaction 

 of the extract for 2 or 3 days. 



(8) Add the required amount of HCl (best 

 growth takes place when there is 

 about 5.5 or 5.8% pure hydrochloric 

 acid material present. 



Sterilization: Sterilization is effected in 



step (3) above. 

 Use: Isolation of Saccharovnjces guttulatus 



from intestines and stomach. Author 



reported that bacteria did not develop in 



this acid medium. 

 Reference: Wilhelmi (1898 p. 358). 



1161. Reed and Cooley's Bean Pod Infusion 

 Solution 



Constituents: 



1. Water. 



2. Bean pods (Phaseolus vulgaris). 

 Preparation: (1) Cook green pods of Pha- 

 seolus vulgaris in water (amount not 

 given). 



Sterilization: Method not given. 



Use: Cultivation of Heterosporium vari- 



abile. 

 Reference: Reed and Cooley (1911-12 



p. 50). 



1162. Maze's Sucrose Bean Infusion 

 Solution 



Constituents: 



1. Water 1000.0 cc. 



2. Beans (white) 



3. Sucrose (2.0%) 20.0 g. 



4. NaCl (1.0%) 10.0 g. 



Preparation : 



(1) Infuse white beans in water for 

 30 minutes at 100 °C. 



(2) Add 2.0%, sucrose, 1.0% NaCl and a 

 trace of sodium bicarbonate to (1). 

 (Filtering of (1) not specified.) 



Sterilization: Method not given. 

 Use: Cultivation of nitrogen fixing bac- 

 teria. 

 Variants : 



(a) Tanner, Besson, and Dopter and 

 Sacquep^e prepared the medium as 

 follows: 



(1) Macerate 50.0 to 60.0 g. haricot 

 beans in a liter of water for several 

 hours in the cold. 



(2) Boil for 30 minutes. 



(3) Pour on a coarse sieve. 



(4) Collect the liquid. 



(5) Add 1.0% NaCl, 2.0% sucrose and 

 a pinch of sodium bicarbonate. 



(6) Boil and filter thru paper. 



(7) Tube. 



(8) Sterilize at 115°C. 



(b) Harvey prepared the medium as 

 follows: 



(1) Mince finely haricot beans. 



(2) Add 250.0 g. to 1000.0 cc. distilled 

 water or clear tap water. 



(3) Heat the mixture 20 minutes at a 

 temperature not exceeding 50 °C. 



(4) Raise the temperature to boiling 

 point. 



(5) Boil 10 minutes. 



(6) Add 1.0 cc. 1.0% sodium bi- 

 carbonate. 



(7) Add 10.0 g. sodium chloride. 



(8) Steam 45 minutes. 



(9) Bring the volume up to 1000.0 cc. 

 by the addition of distilled water. 



(10) Filter while hot thru thick filter 

 paper. 



