266 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(2) Neutralize to phenolphthalein and 

 then add 5.5 cc. of normal acid per 

 liter. 



(3) Distribute (1) in 9.0 cc. lots in test 

 tubes. 



(4) Add 1.0 cc. of a 1.0% aqueous solution 

 of carbolic acid. 



Sterilization: Sterilize in the Arnold steri- 

 lizer (time not given). 

 Use: Enrichment medium in water analysis. 

 Variants : 



(a) Harvey added 1.0 to 5.0 g. of phenol 

 to infusion broth prepared according 

 to Harvey (see variant (bb) 665). 



(b) Harvey added 4.0 cc. of strong HCl 

 to 100.0 cc. of 5.0% phenol solution 

 and added 0.1, 0.2 or 0.3 cc. of this 

 mixture to 10.0 cc. of sterile meat 

 infusion (see 665). 



Reference: Jordan (1901 p. 298), Harvey 

 (1921-22 p. 90). 



858. Harvey's Egg Infusion Broth 



Constituents : 



1. Water 2000.0 cc. 



2. Beef 500.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



5. Whites of 4 eggs 



6. Yolks of 2 eggs 



7. NaOH (normal) ' 12.0 cc. 



Preparation : 



(1) Prepare meat infusion broth accord- 

 ing to Harvey's method (see variant 

 (bb) 665). 



(2) Prepare an egg stock solution by 

 mixing the whites of 4 eggs, the yolks 

 of 2 eggs and 12.0 cc. normal NaOH. 



(3) Add 1000.0 cc. of water to (2). 



(4) Heat very slowly to 90 °C. 



(5) Distribute in flasks. 



(6) Mix equal parts of (5) and sterile (1) 

 when ready for use. 



Sterilization: Sterilization of (1) in the 

 autoclave or steamer. Sterilize (5) in the 

 autoclave. 



Use: General culture medium. 



Variant: Harvey used a similar egg medium 

 by preparing the egg stock solution as 

 indicated above, or by mixing the whites 

 of 4 eggs and 8.0 cc. of normal NaOH with 

 660.0 cc. of water and then diluting to a 

 liter. When ready for use add 1 part of 

 the egg stock solution, prepared as indi- 



cated above, to 5 parts meat infusion 

 broth. 

 Reference: Harvey (1921-22 p. 86). 



859. Olitsky and Kligler's Egg White 

 Infusion Broth 



Constituents : 



1. Distilled water 90.0 cc. 



2. Meat infusion broth 300.0 cc. 



3. Egg white 10.0 cc. 



Preparation: 



(1) Prepare plain meat infusion broth 

 (method not given). 



(2) Add one volume of egg white to 9 

 volumes of distilled water. 



(3) Mix 300.0 cc. of (1) and 100.0 cc. of 

 (2), (^of (2) by volume). 



(4) Adjust to pH" from 7.6 to 7.8. 



(5) Distribute in 500.0 cc. lots into 2 liter 

 flasks to permit sufficient aeration. 



Sterilization: Autoclave for 45 minutes at 

 15 pounds pressure. 



Use: Cultivation of Bacillus dysenteriae 

 Shiga, and study of toxin production. 

 After 5 days incubation at 37° C. filter thru 

 Berkefeld N candle. This bacteria free 

 filtrate constitutes the exotoxin. 



Variants: Dopter and Sacquepee gave the 

 following method of preparation: 



(1) Place the whites of two eggs in a glass 

 graduate. 



(2) Shake constantly, adding little by 

 little three times their volume of dis- 

 tilled water. 



(3) Add 2.0 cc. of 10.0% soda solution 

 (0.5 cc. per 100.0 cc). Mix well. 



(4) Heat in the autoclave at 115°C. 



(5) Prepare meat infusion peptone solu- 

 tion (500.0 g. beef, 5.0 g. NaCl and 

 20.0 g. peptone per liter). 



(6) Mix one part (4) with 5 parts (5) . 



(7) Distribute. 



(8) Sterilize for 15 minutes at 112°C. 



860. Weiss and Wilkes-Weiss Egg 

 "Hormone" Broth 



Constituents 



1. Hormone broth 400.0 cc. 



2. Whole egg 100.0 cc. 



Preparation: (1) Mix whole egg and "hor- 

 mone" broth in the ratio of 1:4. 



Sterilization: Heat for 30 minutes in the 

 Arnold sterilizer. 



