CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



29 



(3) Add a small quantity of sterile 

 CaCOs to each tube of sterile (2). 

 Sterilization: Sterilize (2) and the CaCOa 



into autoclave. 

 Use: To study oxidation of nitrites and 



nitrates. 

 Reference: Cunningham (1924 p. 151). 



88. Omelianski's Nitrite Solution 



Constituents: 



1. Distilled water 1000.0 cc. 



2. NaNOs (Merck) 1.0 g. 



3. NaaCOs (ustum) 1.0 g. 



4. Potassium phosphate 0.5 g. 



5. NaCl 0.5 g. 



6. FeS04 0-4 g. 



7. MgS04 0.3 g. 



8. MgCOa 

 Preparation: 



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



(2) Flask in flat bottomed flasks in 50.0 

 cc. lots. 



(3) Add 0.5 g. MgCOa to each flask. 

 Sterilization: Not specified. 



Use: Isolation of nitrate forming organ- 

 isms. The author found that repeated 

 cultivation on this medium yielded a pure 

 culture. 



Variants: Boullanger and Massol used the 

 above solution with the addition of slag. 



Reference: Omeliansky (1899 p. 548), Boul- 

 langer and Massol (1903 p. 494). 



89. Winogradsky and Omeliansky's Sodium 

 Nitrite Solution 



Constituents : 



1. Distilled water 1000.0 cc. 



2. NaNOa 1-0 g- 



3. Potassium phosphate 0.5 g. 



4. MgS04 0-3 g. 



5. NaCl 0.5 g. 



6. Na.COa 1-0 g. 



7. FeS04 0.4 g. 



Preparation : 



(1) Distill water with permanganate. 



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



(3) Distribute in 50.0 cc. lots. 



(4) Add 0.0, 0.1, 0.2, 0.4, 0.5 or 0.6 g. of 

 NaaCOa to each flask (1.0% gives best 

 results). 



(5) Iron may be added. 

 Sterilization: Not specified. 



Uses: To study nitrate production from 

 nitrites. The author found that the 



addition of 0.1% Na2C03 gave the best 

 results. The addition of FeS04 had no 

 effect on nitrate production. 

 Added nutrients : The authors added one of 

 the following materials to determine the 

 effect of organic material on nitrification. 

 glycerolO.l, 0.2, 0.4, 0.8, 1.0, 1.4 or 2.0% 

 glucose 0.05, 0.1, 0.2, 0.4, 0.8, 1.0 or 2.0% 

 urea 0.1, 1.0 or 1.6% 

 asparagin 0.1, 0.6, 0.8, 1.0 or 2.0% 

 hay infusion 4.0, 8.0, 16.0 or 32.0% 

 leaf infusion 4.0, 8.0, 16.0 or 32.0% 

 dirt infusion 4.0, 8.0, 16.0 or 32.0% 

 meat infusion 2.0, 4.0, 8.0 or 10.0% 

 urine 2.0, 4.0, 10.0 or 20.0% 

 (NH4)2S04 (1.0% solution) 0.2, 1.0, 1.6, 



2.0, 3.0, 4.0 or 6.0% 

 sodium acetate 1.0, 2.0, 3.0, 4.0 or 6.0% 

 sodium butyrate 0.1, 1.0, 2.0, 3.0 or 4.0% 

 Generally organic material slows up 

 oxidation unless it be present in very small 

 quantities. The more organic materials 

 present the slower the oxidation. 

 Variants : 



(a) Wimmer used the above solution 

 specifyingO.5 g. KH2PO4 and suggest- 

 ing the addition of 0.28 g. KCl. 



(b) Harvey used this solution employ- 

 ing 0.5 g. K2HPO4 and 0.3 g. anhy- 

 drous Na2C03 instead of 1.0 g. 



(c) Gibbs (Gowda) used 0.5 g. K2HPO4 

 and a trace of Fe2(S04)3 instead of 

 0.4 g. FeS04. 



Reference: Winogradsky and Omeliansky 

 (1899 p. 333), Wimmer (1904 p. 140), 

 Smith (1905 p. 199), Harvey (1921-22 p. 

 106), Gowda (1924 p. 258), Percival (1920 

 p. 149). 



90. Winogradsky's Basal Sodium Nitrite 



Solution 

 Constituents: 



1. Distilled water 1000.0 cc. 



2. NaN02 (Merck) 1.0 g 



3. Potassium phosphate 0.5 g 



4. MgS04 0.3 g 



5. NaoCOs 0.5 g 



6. NaCl 0.5 g 



Preparation : 



(1) Distill water two times with "chama- 

 leon." 



(2) Dis-solve 2, 3, 4 and 5 in 1. The 

 Na2C03 should be water free, heated 

 to a weak glowing. 



