362 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(5) Inoculate thru the hole with a plati- 

 num wire or loop. 



(6) Shake well to mix. 



(7) Cover the opening with a small piece 

 of sterile paper. 



(8) Seal completely with a colloidion 

 membrane. 



Sterilization: Given under preparation. 



Use: General culture medium. 



Variants: Investigators have used slightly- 

 different methods in sterilization of the 

 shell, and several have specified that the 

 egg be thoroly shaken before use. The 

 differences are not of sufficient impor- 

 tance to warrant a separate discussion for 

 each investigator. 



References: Hueppe (1888 p. 80), Heim 

 (1891 p. 430), Hammerl (1894 p. 155), 

 Abel and Draer (1895 p. 65), Roux and 

 Rochaix (1911 p. 129), Ball (1919 p. 83), 

 Besson (1920 p. 54), Harvey (1921-22 

 p. 85). 



1242. Robertson's Egg Solution (Kahn) 



Constituents : 



1. Water 500.0 cc. 



2. Egg 

 Preparation : 



(1) Beat the yolk of one and the whites of 

 2 eggs in a beaker. 



(2) Add 6.0 cc. of N/ NaOH to (1). 



(3) Gradually add 500.0 cc. tap water 

 to (2). 



(4) Heat very slowly to 95°C. keeping the 

 mixture at this temperature for 

 about an hour. 



(5) Filter thru cotton, wool and muslin. 



(6) The reaction at this point is about 

 pH = 8.2. 



(7) Tube. 



Sterilization: Autoclave at 15 pounds pres- 

 sure for 20 minutes. 



Use: Cultivation of spore forming ana- 

 erobes. The tubes were sealed with a 



line cap. 



Variants: The author reported better 

 growth in a more acid medium (pH = 

 7.2) prepared as follows: 



(1) Add gradually the yolk of one and the 

 whites of 2 eggs to 500.0 cc. of tap 

 water. 



(2) Titrate the mixture so that the re- 

 action is pH = 7.2. 



(3) Heat very slowly to 95° and keep at 

 this temperature for an hour. 



(4) Autoclave at 15 pounds pressure for 

 20 minutes. 



(5) Some of the albumin will settle to the 

 bottom of the tubes. This does not 

 detract from the value of the medium. 



(6) The tubes are sealed with a vaseline 

 cap. 



Reference: Kahn (1922 pp. 175, 192). 



1243. Harvey's Egg Solution 



Constituents : 



1. Water 300.0 cc. 



2. Egg 

 Preparation : 



(1) Add the contents of one egg to 300.0 

 cc. water. 



(2) Shake to mix. 



(3) Raise the temperature slowly to 

 boiling point with frequent shaking. 



(4) Distribute into test tubes. 

 Sterilization: Sterilize in the autoclave or 



steamer. 

 Use: Cultivation of wound organisms and 



other anaerobes. Also used to cultivate 



other microorganisms. 

 Variants : Klimmer prepared the medium as 



follows for the cultivation of amoeba: 



(1) Shake the contents of one egg in a 

 flask containing glass beads. 



(2) Add 300.0 cc. of distilled water to (1). 



(3) Shake thoroly. 



(4) Boil in a water bath for 30 minutes 

 shaking occasionally. 



(5) Distribute in 5.0 cc. quantities in 

 tubes. 



(6) Sterilize in the autoclave. 

 Reference: Harvey (1921-22 p. 85), Klim- 

 mer (1923 p. 229), Stitt (1923 p. 35), 

 Park, Williams and Krumwiede (1924 

 p. 126). 



1244. Nastiukoff's Egg Yolk Solution. 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Egg yolk 100.0 cc. 



Preparation : 



(1) Separate the yolk of egg from the 

 white by Bunge's method (allow the 

 whites to roll from a blotting paper 

 containing the egg). 



(2) Add 0.5 cc. of a 10% NaOH solution 

 and 100.0 cc. of egg yolk to 1000.0 cc. 

 distilled water. 



(3) Place in a fiask and steam for 2 hours 

 in a steamer. 



