228 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(10) Dry in hot air oven or any other 

 convenient way. 



(11) Store powder in sterile glass stop- 

 pered bottle. 



(12) Add 6.0% by weight (11) to cold 

 water. 



(13) Extract (12) at room temperature 

 for 2 hours. 



(14) Filter. 



Sterilization: Method of sterilization of the 



filtrate not given. 

 Use: General culture medium. 

 Reference: Harvey and Iyengar (1921-22 



p. 366). 



779. Dunham's Infusion Broth 



Constituents : 



1. Water 1000.0 cc. 



2. Beef 500.0 g. 



3. NaCl (0.5%) 5.0 g. 



4. Peptone (1.0%,) 10.0 g. 



Preparation : 



(1 ) Boil finely chopped beef with a double 

 weight of water for 2 hours. 



(2) Filter. 



(3) Make slightly alkaline. 



(4) Dissolve 0.5% NaCl, and 1.0% pep- 

 tone in (3). 



Sterilization: Not specified. 



Use: Detection of cholera vibrio. Author 

 reported that cholera vibrio gave a red 

 ring where the H2SO4 and medium met, 

 when concentrated H2SO4 was poured 

 down the side of a culture tube. The 

 medium is used as a general culture 

 medium. 



Variants : Numerous authors have prepared 

 media from these constituents. The 

 method of preparation varied in a num- 

 ber of respects as indicated below, 

 (a) Schultz (1891). 



(1) Place 500.0 g. of the best quality 

 meat, without fat or tendons, in a 

 glass container fitted with a lid. 



(2) Pour 1300.0 cc. of distilled water 

 over the meat. 



(3) Store in a cool place until the 

 next day. 



(4) Filter thru 4 thicknesses of 

 "Morly" and press the remaining 

 meat to obtain as much fluid as 

 possible. 



(5) Pour the filtrate into a kettle, 

 and add 10.0 g. peptone (siccum), 



5.0 g. NaCl and the whites of two 

 eggs, beaten up in 2 or 3 volumes 

 of water. 



(6) Boil under a gas flame for 15 

 minutes. 



(7) Adjust to faint alkalinity, using 

 phenolphthalein as an indicator, 

 and the end point being a faint 

 red color. 



(8) Pour into an iron kettle, add 

 100.0 cc. distilled water, boil 

 strongly for 5 minutes and filter. 



(9) Distribute. 



(10) Sterilization not specified. 



(b) Frothingham (1895). 



(1) Add one pound of finely chopped 

 lean meat to 1000.0 cc. of water 

 and allow to stand from 12 to 24 

 hours in a cool place. 



(2) Strain thru a cheese cloth or a 

 coarse towel and squeeze in a 

 meat press or by twisting the ends 

 of the cloth until 1000.0 cc. of the 

 meat juice is obtained. Makeup 

 to 1000.0 cc. by the addition of 

 water if necessary. 



(3) Dissolve 5.0 g. NaCl and 10.0 g. 

 dried peptone in (2). 



(4) Boil 15 minutes, either in steam 

 or over a flame. 



(5) Make the reaction slightly alka- 

 line by the addition of a satu- 

 rated solution of Na2C03. 



(6) Boil from 45 minutes to an hour. 



(7) Filter. The filtrate should be 

 clear. If not, add the yolks of 

 2 eggs, mix rapidly and boil for 

 a quarter to a half hour. Filter. 



(8) Sterilization not specified. 



(c) Nicolle (1896). 



(1) Infuse 500.0 g. of fresh finely 

 chopped beef with 1 liter of water 

 overnight at 10°C. to 12°C. 



(2) Add 2.0% peptone and 0.5%, NaCl 

 to (1). 



(3) Boil, filter and make rather 

 strongly alkaline. 



(4) Heat for 10 minutes at 120°C. 



(5) Filter and distribute in flasks in 

 the ratio of one liter medium to a 

 two liter flask. 



(6) Sterilize for 15 minutes at 115°C. 



(d) Stutzer and Hartieb (1897). 



