724 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Reference: Boekhout and Ott de Vries 

 (1918 p. 130). 



2258. Matzuschita's Glucose Glycerol 



Peptone Gelatin 



Constituents : 



1. Water 1000.0 cc. 



2. Gelatin (10.0%) 100.0 g 



3. NaCl (0.5%) 5.0 g 



4. Peptone (1.0%) 10.0 g 



5. Glycerol (6.0%) 60.0 g 



6. Glucose (2.0%) 20.0 g 



Preparation : 



(1) Dissolve gelatin in water. 



(2) Neutralize (indicator not specified). 



(3) Add 0.5% NaCl, 10.0% peptone, 6.0% 

 glycerol and 2.0% glucose. 



(4) Filter. 



Sterilization: Method not given. 



Use: Cultivation of mammalian and 

 chicken tubercle bacilli. The author 

 reported that mammalian type gave gray 

 white flakes which sank to the bottom 

 of the liquid at 37°C. In gelatin a gray 

 white membrane formed on top. Chicken 

 type gave large flakes and a gray white 

 precipitate at the bottom. A membrane 

 also formed on the surface. 



Reference: Matzuschita (1899 p. 128). 



2259. Dombrowski's Glucose Peptone 



Gelatin 



Same as medium 631 but solidified by the 

 addition of 10.0% gelatin. 



2260. Beijerinck's Asparagin Peptone 



Gelatin 



Constituents : 



1. Water 900.0 cc. 



2. Sucrose 10.0 g. 



3. Asparagin 2.0 g. 



4. Peptone 8.0 g. 



5. Gelatin 80.0 g. 



Preparation : 



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



(2) Adjust (1) to neutral or slightly acid. 

 Indicator not specified. 



Sterilization: Method not specified. 



Use: To study liquefaction of gelatin and 



pigment production by Chlorococcum. 



Chlorococcum did not liquefy the gelatin. 

 Reference: Beijerinck (1890 p. 461). 



2261. Remy's Phenolated Lactose Peptone 



Gelatin 

 Constituents : 



1. Distilled water. .. 1000.0 cc. 



2. Asparagin 6.0 g. 



3. Oxalic acid 0.5 g. 



4. Lactic acid 0.15 g. 



5. Citric acid 0.15 g. 



6. Bisodium phos- 

 phate 5.0 g. 



7. MgS04 2.5 g. 



8. K0SO4 1.25 g. 



9. NaCl 2.0 g. 



10. Gelatin 120.0 to 150.0 g. 



11. Peptone (Witte's). 30.0 g. 



12. Lactose 



13. Phenol 

 Preparation : 



(1) Grind 6, 8 and 9 in a mortar. 



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



(3) Add 30.0 g. peptone (Witte or Griib- 

 ler's) to (2) and autoclave under 

 pressure for 15 minutes. 



(4) Pour the hot solution into a flask 

 containing 120.0 to 150.0 g. of gela- 

 tin. Shake thoroly until solution is 

 complete. 



(5) Make slightly alkaline by the addi- 

 tion of soda. 



(6) Heat in the autoclave under pressure 

 for 15 minutes at 110°C. 



(7) Acidify by the addition of 0.5N 

 H2SO4 so that the acidity of 10.0 cc. 

 of gelatin will be neutralized by 

 0.2 cc. of a 0.5 N soda solution. 



(8) Steam again for 8 or 10 minutes. 



(9) Filter. 



(10) Retest the reaction by adding 10.0 cc. 

 of gelatin to 100.0 cc. water and 

 several drops of phenolphthalein. 

 Add 0.5 N soda solution drop by 

 drop until a red coloration appears. 

 0.2 cc. of the soda solution should 

 be required. 



(11) Add 2.5 g. MgS04. 



(12) Distribute in 10.0 cc. lots. 



(13) Just before use, add 1.0 cc. of a 

 35.0% lactose solution and 0.1 cc. of 

 a 2.5% phenol solution to each tube 

 of sterile (12). 



Sterilization: Sterilize by heating on 3 



successive days. 

 Use: Isolation of typhoid bacillus. 

 Reference: Remy (1900 p. 561). 



