CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



581 



(6) Add 100. cc. of (5) to each flask. 



(7) Solidify the agar by immersing the 

 flask in cold water for 15 minutes. 



(8) Mix one part infusion broth (2) and 

 two parts sterile ascitic fluid. 



(9) Fill each flask (7) to the neck with 

 the mixture of (8). 



(10) Seal with a layer of sterile vaseline 

 1.0 cm. deep. 

 Sterilization: Methods of sterilization not 



specified. 

 Use: Mass cultivation of anaerobic 



organisms found in influenzae. 

 Reference: Olitsky and Gates (1921 p. 716). 



1857. Pelouze and Viteri's Brain Veal 

 Infusion Agar 



Constituents : 



1. Dist. water 500.0 cc. 



2. Brain, calf 500.0 g. 



3. KH2PO4 (0.5%) 2.5 g. 



4. Peptone (1.0%) 5.0 g. 



5. Veal infusion agar (2.5%) 

 Preparation : 



(1) Force 500.0 g. of calf brain thru a 

 wide meshed gauze into 500.0 cc. 

 distilled water and place in the ice 

 box for 24 hours. 



(2) Filter thru cotton of varying degrees 

 of compactness. The filtrate will 

 not be clear. 



(3) Add 0.5%, KH2PO4 and 1.0% peptone. 



(4) Prepare standard 2.5% agar from veal 

 infusion with the addition of 0.5% 

 NaCl and 1.0% peptone. 



(5) Mix one part (2) with three parts (4). 



(6) Adjust to pH 7.8. 



(7) Tube. 



(8) Following sterilization and solidify- 

 ing replace the cotton plug with a 

 sterile rubber cork. 



Sterilization: Sterilize in the autoclave. 



Use: Cultivation of gonococci. 



Variants: Following the mixing of the 

 brain and agar the medium may be 

 placed in the autoclave, brought to 15 

 pounds pressure quickly and then filtered 

 before tubing and sterilizing. This proc- 

 ess gives a clear medium but gives also a 

 scanty growth. The author does not 

 recommend the clear medium. 



Reference: Pelouze and Viteri (1926 p. 

 685). 



1858. Avery's Oleate Blood Cell Agar 

 (Harvey) 

 Constituents : 



1. Infusion agar 950.0 cc. 



2. Sodium oleate (2.0% neutral 

 solution) 50.0 cc. 



3. Erythrocytes, human 10.0 cc. 



Preparation : 



(1) Prepare infusion agar according to 

 Harvey's method (see variant (v) 

 medium 1661). 



(2) Adjust (1) to 0.4% acid to phenol- 

 phthalein and heat to 45 °C. 



(3) Mix 950.0 cc. (2) with 50.0 cc. of a 

 2.0% neutral sodium oleate solution, 

 and 10.0 cc. of sterile washed human 

 erythrocytes, both heated to 45°C. 



Sterilization: Not specified. 

 Use: Cultivation of B. influenzae. 

 Variants: Park, Williams and Krumwiede 

 prepared a similar medium as follows: 



(1) Remove the corpuscles from sterile 

 defibrinated human or rabbit blood 

 by centrifugation. 



(2) Discard the supernatant fluid by 

 pipetting off. 



(3) Make up the original volume of (1) 

 by the addition of sterile infusion 

 broth. 



(4) Prepare a 2.0% solution of neutral 

 sodium oleate. (Kahlbaum's pre- 

 ferred.) 



(5) Sterilize (4) in the autoclave. 



(6) To 94.0 cc. of hot 2.0%, nutrient agar 

 (vitamin agar, see variant (e) medium 

 1863 is recommended) add 5.0 cc. of 

 (5) and add 1.0 cc. of (3) under 

 aseptic conditions. 



References: Harvey (1921-22 p. 87), Park, 

 Williams and Krumwiede (1924 p. 130). 



1859. Brown and Orcutt's Blood Cell Agar 

 Constituents: 



1. Veal infusion agar. 



2. Blood corpuscles. 

 Preparation : 



(1) Prepare veal infusion agar. 



(2) Defibrinate blood and wash re- 

 peatedly with sterile physiological 

 salt solution. 



(3) Add some of the washed blood cor- 

 puscles to melted (1), (amounts not 

 given) and pour into plates. 



Sterilization: Not specified. 



