CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



99 



352. Weissenberg's Glycerol Nitrate 

 Solution 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Sodium phosphate (neu- 

 tral) 2.5 g. 



3. Glycerol 40.0 g. 



4. NaCl 5.0 g. 



5. NH4NO3 3.75 g. 



Preparation : 



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



(2) Place 0.75 g. NH4NO3 in 175.0 cc. 

 Erlenmeyer flasks. 



(3) Fill each flask containing NH4NO3 

 with (1). 



Sterilization: Sterilize carefully (method 

 not given). 



Use: To study denitrification by B. pyo- 

 cyaneus. The author found that ammo- 

 nia was produced. 



Reference: Weissenberg (1897 p. 284). 



353. Lantzsch's Glycerol Nitrate Solution 



Constituents: 



1. Water 1000.0 cc. 



2. KNO3 0.5 to 1.5 g. 



3. Glycerin 20.0 g. 



4. Malic acid 7.0 g 



5. NaoHPOi 0.5 g 



6. NaCl 0.5 g 



7. NasCOa 0.5 g 



8. MgS04 0.1 g 



Preparation: (1) Dissolve 2, 3, 4 (neutralize 



acid with soda) 5, 6, 7 and 8 in 1. 



Sterilization: Not specified. 



Use: To determine nitrate reduction by 

 fluorescens group. Acidify the culture 

 with 5.0% H2SO4 and add a strong zinc 

 iodide solution. A blue color indicates 

 nitrate formation or presence. Test for 

 NH3 with Nessler's reagent. 



Reference: Lantzsch (1921 p. 82). 



354. Stoklasa's Mannitol Nitrate Solution 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Mannite 20.0 g 



3. NaNOa 2.0 g 



4. K2HPO4 0.5 g 



5. K2SO4 0.2 g 



6. CaCl2 0.05 g 



7. MgCl. 0.05 g 



8. FePOi 0.05 g 



Preparation: (1) Dissolve 2, 3, 4, 5, 6 and 8 



in 1. 

 Sterilization: Not specified. 

 Use: To study action of Azotobacter and 



Radiobacter on nitrates. Author found 



that nitrates were reduced. 

 Reference: Stoklasa (1908 p. 493). 



355. Doryland's Acetic Acid Nitrate Solution 



Constituents: 



1. Distilled water 500.0 cc. 



2. HCl dilute 



3. MgS04 0.5 g. 



4. CaO 0.01 g. 



5. re2(S04)3 0.01 g. 



6. MnS04 0.01 g. 



7. HNO3 (amount not given) 



8. H2SO4 



9. CH3COOH 



10. H3PO4 



11. N/0.2578 NaOH 



12. N/0.6205 KOH 

 Preparation : 



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

 quite neutralized by 1.0 cc. silicate 

 solution. Prepared by dissolving 

 24.0 g. KjSiOa and 8.4 g. Na2Si03 in 

 500.0 cc. distilled water (phenol- 

 phthalein). 



(2) Add the following salts to the HCl: 



MgS04 0.5 g. 



CaO 0.01 g. 



Fe2(S04)3 0.01 g. 



MnS04 0.01 g. 



HNO3 (amount not given) 



(3) Standardize (2) so 1.0 cc. equivalent 

 to 1.0 cc. silicate solution, using 

 methyl orange as indicator. 



(4) Standardize a solution of H2SO4 in 

 the same way as HCl omitting the 

 salts. 



(5) Standardize H3PO4 and CH3COOH 

 solution in the same way as HCl 

 omitting the salts and using phenol- 

 phthalein as indicator. 



(6) Mix the acids in the following ratio: 



HCl 153.5 cc. 



CH3COOH 153.5 cc. 



H2SO4 77.0 cc. 



H3PO4 116.0 cc. 



(7) Mi.x equal quantities of N/0.6205 

 KOH and N/0.2578 NaOH solutions. 



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



