798 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



2. Egg. 



3. Glycerol. 



4. Bacillus phlei. 

 Preparation : 



(1) Grind up 1.0 g. of dried Bacillus phlei 

 in 12.0 cc. of glycerol and 22.0 cc. of 

 infusion broth (saline or peptone beef 

 broth may be used). 



(2) Autoclave (1). 



(3) Add (2) when cool to 66.0 cc. of mixed 

 yolk and white of fresh hen eggs. 



(4) Tube. 



Sterilization: Sterilize on each of three 

 successive days at 61 °C. and inspissate in 

 slopes. 



Use: Cultivation of Johns bacillus, human 

 lepra bacillus and a spirochaete. Bacil- 

 lus phlei was cultivated on 4.0% glycerol 

 infusion broth. 



Reference: Twort (1921 p. 798). 



2449. Harvey's Trypsinized Heart Egg 

 Medium 



Constituents : 



1. Water 1000.0 cc. 



2. Heart, horse 500.0 g. 



3. Egg 3000.0 cc. 



Preparation : 



(1) Beat up the contents of several fresh 

 eggs. 



(2) Mix 3 parts (1) with 1 part trypsinized 

 heart prepared as given under the 

 variants of medium 1117. 



(3) Filter thru muslin. 



(4) Tube. 



(5) Coagulate. 

 Sterilization: Method not given. 

 Use: General culture medium. 

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



2450. Rosenthal and Schulz's Meat Infusion 

 Egg Medium (Zimmermann) 



Constituents : 



1. Egg white. 



2. Meat infusion. 

 Preparation : 



(1) Prepare meat infusion. 



(2) Filter egg albumin thru a double 

 layer of muslin by hand under the 

 application of pressure. 



(3) Place the clear filtrate in a measuring 

 cylinder fitted with a ground glass 

 stopper. 



(4) Add to 5.0 cc. of albumin, 2.2 cc.^ of a 



sterile 1.0% solution of NaOH or 

 KOH and 2.8 cc. of sterile meat 

 infusion. 



(5) Allow to stand one hour, and shake 

 back and forth gently to mix. 



(6) Distribute into sterile test tubes or 

 other sterile culture containers. 



(7) Place the containers in hot (95°-98°) 

 water for a short time to coagulate the 

 albumin. 



Sterilization: Method not given. 



Use: General culture medium. 



Variants: The author reported that salts 

 as NaCl, NajCOs, K2CO.,, Na.SOi, 

 Na2HP04, etc., may be added if desired. 



Reference: Rosenthal and Schulz (1888 p. 

 307), Zimmermann (1888 p. 314). 



2451. Thoinot and Masselin's Egg Albumin 

 Medium 



Constituents: 



1. Egg albumin. 

 Preparation : 



(1) Remove the albumin of an egg under 

 aseptic conditions by means of a 

 pipette. 



(2) Distribute directly into sterile test 

 tubes. 



(3) Coagulate as serum or at 100°C. 

 Sterilization: Method not given. 



Use: Cultivation of B. pyocyaneus. Other 

 investigators cultivated a large number 

 of organisms on similar media. 



Variants: The following investigators pre- 

 pared the medimn as indicated : 



(a) Smith cultivated plant parasites on 

 the following medium: 



(1) Thoroly flame the end of the egg 

 from which the egg albumin is to be 

 poured. 



(2) Distribute the albumin in sterile 

 test tubes. 



(3) Coagulate in the same manner as 

 blood serum. 



(b) Duval isolated B. leprae on the 

 following medium. He reported that 

 the colonies at first were greyish 

 white but after several days they 

 assumed a distinct orange yellow tint. 

 Transfers may be made from this 

 medium to slants or other plates. 



(1) Pour egg albumin (human blood 

 serum), (or nutrient agar with or 

 without 1.0% tryptophane may be 



