262 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



given under variants, all media not con- 

 taining peptone gave poor growth. 

 Variants: The author gave the following 

 method of preparing different media: 



(1) Soak one pound of meat with 500.0 cc. 

 of cold water, heat to 55° for a few 

 minutes and strain. Boil this extract 

 to remove coagulable materials and 

 filter. This is Extract I. 



(2) Take meat from extract I, add 

 500.0 cc. H2O, boil 5 minutes, strain 

 and filter. This is Extract II. 



(3) Take meat from extract II, add 

 500.0 cc. of H2O, boil 5 minutes, 

 strain and filter. This is Extact III. 



(4) Divide each e.xtract into two equal 

 parts. 



(5) Mix each of these two parts with an 

 equal amount of glucose salt solution 

 (see step (1) in the medium). 



(6) Add 2.0% peptone to one of these 

 parts, and make no addition to the 

 other. 



(7) Adjust to pH = 7.8 when containing 

 peptone and pH = 8.0 when not con- 

 taining peptone. 



Reference: Mueller (1922 pp. 316, 320, 321). 



843. Robinson and Rettger's Glucose 

 Infusion Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Lean beef 1.0 lb. 



3. Peptone (Witte) 20.0 g. 



4. Glucose 1.0 g. 



Preparation : 



(1) Extract 1.0 lb. of chopped lean beef 

 in one liter of water in the cold for 

 24 hours. 



(2) Coagulate on a boiling water bath. 



(3) Filter and add 2.0% Witte's peptone 

 to the filtrate. 



(4) Heat on the water bath for about 20 

 minutes. 



(5) Adjust the reaction to +0.6 to phenol- 

 phthalein. 



(6) Filter and distribute into 150.0 cc. 

 Erlenmeyer flasks in 20.0 cc. lots. 



(7) After sterilization add enough sterile 

 glucose to each flask to make 0.1% 

 glucose. 



Sterilization: Sterilize at 12 to 14 pounds 



pressure for 15 minutes. 

 Use: Cultivation of B. diphtheriae and 



toxin production. 



Variants: Authors adjusted the reaction to 

 + 1.0 to phenolphthalein instead of +0.6, 

 M.L.D. = 0.005 cc. 



Reference: Robinson and Rettger (1917 

 p. 363) 



844. Bronstein and Griinblatt's Indicator 



Glucose Infusion Broth 



Constituents : 



1. Meat infusion broth 1000.0 cc. 



2. Glucose (0.5%) 5.0 g. 



3. Indigo carmine solution. 



4. Fuchsin solution (acid). 

 Preparation : 



(1) Prepare a neutral 0.5% glucose meat 

 infusion broth. 



(2) Tube in 5.0 cc. lots. 



(3) Prepare a 2.0% watery solution of 

 indigo carmine. 



(4) Dissolve 10.0 g. of acid fuchsin in 

 100.0 cc. of 1.0% KOH solution. 



(5) Mix 2 parts (3) with 1 part (4) and 

 add to 22 parts distilled water. 



Sterilization: Not specified. 



Use: Differentiation of diphtheria organ- 

 isms. After 24 hours incubation add 3 

 drops of (5) to each tube. The authors 

 reported that the control tube was blue; 

 diphtheria cultures were ruby-red; pseudo 

 diphtheria cultures were green after 

 several minutes. 



Reference: Kolle and Wasserman (1912 

 p. 414). 



845. Ayers, Rupp and Johnson's Glucose 



Infusion Broth 



Constituents : 



1. Meat infusion 1000.0 cc. 



2. Peptone (Park Davis & Co.) 10.0 g. 



3. Glucose 5.0 g. 



4. Sodium ricinolate (Castor oil soap). 

 Preparation : 



(1) Prepare 1000.0 cc. of meat infusion. 



(2) Add 2 and 3 to (1). 



(3) Adjust reaction to pH = 7.5. 



(4) Add castor oil soap (sodium ricino- 

 late) or other surface reducing ma- 

 terial in sufficient amounts to lower 

 the surface tension to about 50 and 

 43 dynes. 



Sterilization: Method not given. 



Use: To study effect of surface tension on 

 growth of streptococci. Authors re- 

 ported that in general when the surface 



