76 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Preparation : 



(1) Dilute HCl so that 1.0 cc. is not quite 

 neutralized by 1.0 cc. of silicate solu- 

 tion made by dissolving 24.0 g. 

 KsSiOs and 8.4 g. NasSiOa in 500.0 g. 

 distilled water. (Phenolphthalein as 

 indicator.) 



(2) Add to the HCl the following salts: 



MgS04 0.5 g. 



CaO 0.01 g. 



Fe2(S04)3 0.01 g. 



MnS04 0.01 g. 



(NH4)2S04 1.0 g. 



(3) Standardize (2) against the silicate 

 solution so that 1.0 cc. silicate equiva- 

 lent to 1.0 cc. (2) using methyl orange 

 as indicator. 



(4) Standardize a solution of H2SO4 in 

 same way as HCl omitting the salts. 



(5) Standardize H3PO4 in similar way as 

 HCl omitting salt and using phenol- 

 phthalein as indicator. 



(6) Mix the acids in the following ratio: 



HCl 153.5 cc. 



H2SO4 77.0 cc. 



H3PO4 116.0 cc. 



(7) Mix equal quantities of N/0.6205 

 KOH and N/0.2578 NaOH. 



(8) 1.0 cc. of (7) should neutralize 1.0 cc. 

 of (6), using phenolphthalein as 

 indicator. 



(9) Draw acid and base into separate 

 plugged burettes, allow to stand 

 several hours to sterilize. 



(10) Add enough sterile glucose solution 

 to equal amounts of (6) and (7) to give 

 concentration of glucose of 10.0 grams 

 per liter to a mixture of equal amounts 

 of (6) and (7). 



Use: General synthetic culture medium. 



Reference: Doryland (1916 pp. 146-148). 



252. Winogradsky's Glucose Ammonium 

 Sulphate Solution 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Dextrose 20.0 g. 



3. Potassium phosphate 1.0 g. 



4. MgS04 0.2 g. 



5. NaCl ] 



6. FeS04 '""^^^ , 



7. MnS04 J ^°'°"°*' 



8. (NH4)2S04 0.01 g. 



9. Chalk 



Preparation : 



(1) Dissolve 2, 3, 4, 5, 6, 7 and 8 in 1. 



(2) Adjustment of reaction not specified. 



(3) Distribute into Drechsel's flasks with 

 a ground cover. 



(4) Add an excess of chalk to each flask. 



(5) After sterilization connect the flask 

 with a nitrogen generator and after 

 inoculation replace the air with 

 nitrogen. 



Sterilization: Sterilize in an autoclave. 

 Use: Cultivation of Clostridium Pastori- 



anum. 

 Reference: Winogradsky (1902 p. 49). 



253. Gage's Glucose Ammonium Sulphate 

 Solution 



Constituents: 



1. Water 1000.0 cc. 



2. (NH4)2S04 4.0 g. 



3. Glucose 1.0 g. 



4. KH2PO4 2.0 g. 



5. MgS04 1.0 g. 



6. Iron sulphate 0.8 g. 



7. NaCl 4.0 g. 



8. MgCOa 

 Preparation : 



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



(2) Distribute in 100.0 cc. lots in 150.0 cc. 

 flasks. 



(3) To each flask add 1.0 cc. of a heavy 

 suspension of MgCOs. 



Sterilization: Not specified. 



Use: To study nitrification by Nitroso- 



hacter and Pseudomonas radicicola. 

 Reference: Gage (1910 p. 33). 



254. Stoklasa's Glucose Ammonium 

 Sulphate Solution 



Constituents: 



1. Distilled water 1000.0 cc. 



2. d-glucose 25.0 g. 



3. K2SO4 1.0 g. 



4. MgCl2 0.5 g. 



5. Iron sulphate 0.1 g. 



6. (NH4)2S04 



7. One of the following: 1.0 g. 

 Monodiferric-phosphate Fe203(P20s) 3 

 Monodialuminum-phosphate 



A1203P205-8H20 



Preparation : 



(1) Dissolve 2, 3, 4, 5 and 2.0 g. nitrogen 

 in the form of (NH4)2S04 in 1. 



