CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



115 



Preparation : 



(1) Dissolve 2 and 3 in part of 1. 



(2) Dissolve 4, 5, 6, 7 and 8 in the rest of 1. 



(3) Neutralize (2) with KOH during the 

 heating. 



(4) Mix (3) and (1). 



Sterilization: Sterilize for 20 minutes on 

 each of two successive days. On the 

 third day add 1.0% of one of the added 

 nutrients and sterilize for 15 minutes. 



Use: To study decomposition of sugars by 

 Bacterium coli and \Bacterium Fitzi- 

 anus. The author reported the following 

 reactions: 



Variants: The author used 10.0 g. asparagin 

 instead of 5.0 g. in the basic solution. 



Added nutrients: The author employed 

 1.0% of one of the following as carbon 

 sources: 



glucose lactose 



maltose sucrose 



Reference: Mendel (1911 p. 297). 



395. Maassen's Basal Malic Acid Asparagin 



Solution (Smith) 

 Constituents: 



1. Distilled water 1000.0 cc. 



2. Malic acid 7.0 g. 



3. Asparagin 10.0 g. 



4. Sodium phosphate 



(secondary) 5.0 g 



5. MgS04 2.5 g. 



6. NaOH 2.5 g. 



7. CaCl2 0.01 g. 



Preparation : 



(1) Dissolve 2 in 1000.0 cc. of distilled 

 water. 



(2) Neutralize exactly to litmus, using 

 7.0% KOH. 



(3) Make up to 1000.0 cc. 



(4) Dissolve 4, 5 and 6 in (3). 



(5) Add 0.01 g. CaCl2 to (4). 



(6) Dissolve one of the added nutrients 

 in (5). 



Sterilization: Not specified. 



Use: As a general culture medium. 



Variants: Kolle and Wasserman used 0.4 g. 

 MgS04 in the basal solution and 2.5 g. 

 of pure crystalline soda instead of 

 NaOH. He specified the use of 0.5 to 

 1.0% of any desired carbohydrate, alco- 

 hol, etc. as added nutrients. 



Added nutrients: Smith added any desired 

 carbohydrate, alcohol, etc. 



Reference: Smith (1905 p. 198), Kolle and 

 Wasserman (1912 p. 394). 



396. Lockemann's Basal Citrate Glycerol 



Solution 

 Constituents: 

 1- Water lOOO.O cc. 



2. MgHP04-7H20 4.93 g. 



3. H3PO4 (1/3 molar) 20.0 cc. 



4. Citric acid (3 normal) 10.0 cc. 



5. Asparagin 5.29 g. 



6. Glycerol 20.0 g. 



Preparation : 



(1) Dissolve 2, 3, 4, 5 and 6 in 1. 



(2) Add one of the combinations given 

 under added nutrients. The reaction 

 of the various solutions will also be 

 given there. 



Sterilization: Not specified. 

 Use: Cultivation of tubercle bacilli. 

 Added nutrients and modifications: The 

 author added the following materials: 



(a) K2HPO4 0.35 g. and 10.0 cc. normal 

 H2SO4. The acid titer was +4.7. 

 Luxuriant growth was reported. 

 (No indicator specified.) 



(b) Used 5.0 g. asparagin, 12.0 cc. citric 

 acid, 25.0 cc. H3PO4, 4.0 g. MgHP04-- 

 7H2O and added 0.25 g. of NaH2P04- 

 12 H2O, and 0.25 g. of KH2PO4. The 

 acid titer was +4.0. Growth was 

 reported as being quite good. 



(c) Used 5.0 g. asparagin 12.0 cc. citric 

 acid, 25.0 cc. of H3PO4, 3.75 g. 

 MgHP04-7H20 and added 0.25 g. 

 CaHP04, 0.25 g. KH2PO4 and 0.25 g. 

 NaH2P04. The reaction was +4.2. 

 Growth was reported as slight. 



(d) Added 0.72 g. NaH2P04, and 10.0 cc. 

 normal H2SO4 to the basic solution. 

 The acid titer was +4.7. Growth 

 was reported as very slight. 



(e) Added 10.0 cc. of normal H2SO4 and 

 0.35 g. K2HPO4 to the basic solution. 

 The acid titer was +4.6. Growth 

 reported as luxuriant. 



(f) Added 10.0 cc. normal H2SO4, 0.18 g. 

 K2HPO4 and 0.36 g. KH2PO4 to the 

 basic solution. The acid titer was 

 +4.2. Growth reported as being 

 luxuriant. 



Reference: Lockemann (1919 p. 421). 



397. Mendel's Basal Citrate Glycerol 

 Solution 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Asparagin 5.0 g. 



