56 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



5. NaCl 0.2 g. 



6. CaS04 0-1 g- 



7. CaCOs 5.0 g. 



Preparation : 



(1) Dissolve 4 in a little distilled water. 



(2) Neutralize (1) to phenolphthalein. 



(3) Add 2, 3, 5, 6 and 7 to (2) and make 

 up to 1000.0 cc. 



Sterilization: Not specified. 



Use : Cultivation of aerobic nitrogen fixing 



bacteria and protozoa. 

 Reference: Harvey (1921-22 p. 104). 



174. Sackett's Modified Lipman's Manni- 

 tol Salt Solution 



Constituents : 



1. Water (surface) 1000.0 cc. 



2. Mannitol 15.0 g. 



3. K2PO4 0-5 g- 



4. MgS04 0-2 g- 



5. CaCl2 002 g. 



6. 10.0% FeCU solution 1 drop 



Preparation : 



(1) Dissolve 2, 3, 4, 5 and 6 in 1000.0 cc. 

 of surface water. 



(2) Neutralize with N/1 NaOH to phenol- 

 phthalein. 



(3) Distribute in 100.0 cc. lots in 500.0 cc. 

 Erlenmeyer flasks. 



Sterilization: Autoclave for 5 minutes at 

 120°C. 



Use: Study nitrogen fixation by bacteria 

 found in soil. 



Variants: Heinemann used 0.2 g. K2HPO4 

 instead of 0.5 g. K3HPO4 and 0.01 g. 

 FeoCle. 



Reference: Sackett (1912 p. 87), Heine- 

 mann (1922 p. 39). 



175. Winogradsky's Mannitol Salt Solution 

 (Lantzsch) 



Constituents : 



1. Water 1000.0 cc. 



2. Mannitol 20.0 g. 



3. K2HPO4 10 g. 



4. MgS04 0.2 g. 



5. FeS04 0.01 g. 



6. NaCl 0.01 g. 



7. CaCOa 5.0 g. 



Preparation : 



(1) Dissolve one of 2, 3, 4, 5, 6 and 7 in 1. 



(2) To prepare anaerobic culture after 

 inoculation, stopper the flasks with 

 2 holed stoppers, seal with paraffin 



and pass N2 that has been washed in 

 potassium pyrogallic acid solution 

 three times, then in H2SO4 and 

 NaOH, thru the flasks. 

 Sterilization: Not specified. 

 Use: To study nitrogen fixation by Bacil- 

 lus amylobacter, Bad. pneumoniae and 

 torula under anaerobic and aerobic con- 

 ditions. 

 Variants: The author suggests the^use of 

 glucose instead of mannitol and specifies 

 that 10.0 or 20.0 g. of calcium humate 

 may or may not be added to the glucose 

 medium. 

 Reference: Lantzsch (1921 p. 2). 



176. Koser and Rettger's Basal Glycerol 

 Salt Solution 



Constituents : 



1. Distilled NH3 free water. . . 1000.0 cc. 



2. NaCl 5.0 g. 



3. MgS04 0.2 g. 



4. CaCl2 0.1 g. 



5. KH2PO4 1-0 g. 



6. K2HPO4 10 g. 



7. Glycerol 30.0 g. 



Preparation : 



(1) Dissolve 2, 3, 4, 5, 6 and 7 in 1. 



(2) Add one of the added nutrients to (1). 



(3) pH = 6.6. 



(4) Medium is clear and colorless and 

 requires no filtration. 



(5) Distribute into test tubes. 

 Sterilization : Autoclave at 10 to 12 pounds 



pressure for 15 minutes. 

 Use: To study bacterial nutrition. 

 Added nutrients: The authors used 1.0 g. 



of one of the following: 



valin 



glutamic acid 



tyrosin 



phenylalanin 



lysin picrate 



urea 



uric acid 



hypoxanthin 



allantoin 



creatin 



glycocoU 



aspartic acid 



asparagin 



tryptophan 



leucin 



histidin monochloride 



