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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



954. Kahn's Casein Digest Egg Albumin 

 Bouillon 



Constituents: 



1. Water 1000.0 cc, 



2. Bouillon. 



3. Casein 200.0 g. 



Preparation : 



(1) Boil a few eggs for 15 minutes. 



(2) Carefully separate the white from 

 the yolk and wash in distilled water. 



(3) Cut the washed coagulated albumin in 

 cubes of about 2 mm. 



(4) Place a cube into each of several test 

 tubes. 



(5) Prepare casein digest according to the 

 method given by Kahn (see 648). 



(6) Add 10.0 cc. of (5) to each tube. 



(7) Seal the tube with a vaseline cap . 

 Sterilization: Autoclave at 15 pounds pres- 

 sure for 20 minutes. 



Use: To study proteolysis by spore forming 

 anaerobes. 



Variants : 



(a) Torrey and Kahn added 1.0 cc. of 

 casein digest containing 1.0% glu- 

 cose or lactose to each tube, or they 

 added 20.0% lactose, or whey broth 

 was added to the casein digest bouil- 

 lon. The authors reported that in a 

 proper medium Bacterium acid- 

 ophilus inhibited proteolysis by sev- 

 eral spore bearing proteolytic anaer- 

 obes including B. sporogenes, B. 

 histolyticus and B. botulinus for a 

 period of 10 days or more. 



Reference: Kahn (1922 p. 174), Torrey and 

 Kahn (1923 p. 483). 



955. Besredka and Jupille's Egg Bouillon 

 (Besson) 

 Constituents : 



1. Bouillon 500.0 cc. 



2. Egg white (10.0% soln.) 400.0 cc. 



3. Egg yolk (10.0% soln.) 100.0 cc. 



Preparation: (1) Mix 5 parts of bouillon 



with 4 parts of a 10.0% egg white solution 



and 1 part of 10.0% egg yolk emulsion in 



sterile water. 

 Sterilization: If the egg white and yolk are 



not obtained under aseptic conditions, 



sterilize by heating at 55°C. 

 Use: Cultivation of pneumococci, gono- 



cocci, meningococci, various other highly 



parasitic forms as well as saprophytic 



organisms. 



Variants : 



(a) Stitt prepared the medium as follows: 



(1) Prepare a 10.0% egg white solution 

 by constantly heating the egg 

 white while distilled water is 

 added, little by little. 



(2) Filter (1) thru absorbent cotton. 



(3) Heat to 100°C. 



(4) Filter thru filter paper. 



(5) Tube or flask. 



(6) Sterilize at 115° for 20 minutes. 



(7) Add 1.0 cc. of normal NaOH, to 

 100.0 cc. of a 10.0% emulsion of 

 egg yolk in water. Add sufficient 

 NaOH to slightly clarify, but still 

 be opaque when in rather thick 

 layers. 



(8) HeattolOO°C. 



(9) Filter. 



(10) Sterilize at 115°C. for 20 minutes. 



(11) Mix 100.0 cc. sterile bouillon (con- 

 taining no NaCl) with 80.0 cc. of 

 (6) and 20.0 cc. of (10) under 

 aseptic conditions. 



(12) Distribute in sterile tubes or 



Stitt reported that tubercle bacilli 

 grew better if the peptone be omitted 

 from the bouillon, 

 (b) Klimmer prepared the medium as 

 follows : 

 (1) Thoroly mix 1 part egg white with 

 10 parts distilled water. 



(2) Boil. 



(3) Filter. 



(4) Sterilize (method not given). 



(5) Thoroly mix 1 part egg yolk with 

 10 parts distilled water. 



(6) Add 1.0 cc. normal soda solution 

 to 100.0 cc. (5). 



(7) Boil. 



(8) Filter. 



(9) Sterilize (method not given). 

 (10) Mix 4 parts (4) with 1 part (9) 



and 5 parts nutrient bouillon. 

 References: Besson (1920 p. 55), Stitt (1923 

 p. 34), Klimmer (1923 p. 202). 



956. Robertson's Alkaline Egg Bouillon 



Constituents : 



1. Water 1000.0 cc. 



2. Eggs 4 



3. NaOH (normal) 12.0 cc. 



4. Bouillon 



