CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



745 



(3) Mix 10.0 g. of egg albumin in 

 100.0 cc. of water and add to (2). 



(4) Add 3.0 g. meat extract, 10.0 g. 

 peptone and 5.0 g. NaCl to (3). 



(5) Cover the container, place in the 

 autoclave and heat at about 15 

 pounds pressure for 45 minutes. 



(6) Adjust the reaction to +1.5% by 

 the addition of NaOH or HCl. 



(7) Filter while boiling hot thru 

 plaited filter paper, washed with 

 500.0 g. boiling water. 



(8) Distribute as desired. 



(9) Autoclave for 20 minutes at 

 10 pounds pressure. 



(10) Cool in a running water bath im- 

 mediately, 

 (j) Stitt prepared the medium as follows: 



(1) Place 3.0 g. Liebig's extract, 

 10.0 g. peptone and 5.0 g. NaCl in 

 a mortar. 



(2) Dissolve the white of one or two 

 eggs in 1000.0 cc. of water. 



(3) Add (2), little by little, to (1), 

 until a brownish color is obtained. 



(4) Heat (3) to 45° in a water bath. 



(5) Dissolve about 120.0 g. of Gold 

 Label or other good quality gelatin 

 in (4). 



(6) Add sufficient normal NaOH to 

 bring the reaction to +1. 



(7) Place in the inner compartment of 

 a rice cooker. 



(8) Boil for 15 minutes. 



(9) Filter thru filter paper in a hot 

 funnel. The paper must be very 

 thoroly wetted with very hot 

 water. 



(10) Tube. 



(11) Sterilize either in the Arnold on 

 3 successive days or in the auto- 

 clave at 8 to 10 pounds pressure 

 for 10 minutes. 



(12) Cool the tubes as quickly as 

 possible. 



(k) Klimmer gave the following method 

 of preparation: 



(1) Dissolve 10.0 g. Liebig's meat 

 extract, 10.0 g. Witte's peptone, 

 and 5.0 g. NaCl in 1000.0 cc. water 

 by heating for 30 minutes in the 

 steamer. 



(2) Allow to stand until cool. 



(3) Filter. 



(4) To 900.0 g. of (2) add 100.0 g. of 

 gelatin. 



(5) Steam until solution is complete 

 (not longer than 30 minutes). 



(6) Add 4.0% NaOH solution until 

 blue litmus paper is no longer 

 turned red. 



(7) Boil for 15 minutes in the steamer. 



(8) Add 1.5 g. of crystalline (not 

 effloresced) soda per liter. 



(9) Boil 30 minutes. 



(10) Filter. 



(11) Tube in 10.0 cc. quantities. 



(12) Sterilize by a single heating at 15 

 to 20 minutes in steam. 



References: Frost (1903 p. 6), Stoklasa 

 (1908 p. 489), Abel (1912 p. 19), Day and 

 Walker (1912-13 p. 435), Burger (1916-17 

 p. 477), Meier (1918 p. 435), Besson (1920 

 p. 40), Wolf and Shunk (1921 p. 325) 

 Giltner (1921 p. 35), Stitt (1923 p. 38), 

 Klimmer (1923 p. 193), Park, Williams 

 and Krumwiede (1924 p. 177). 



2300. Kohn's Peptone Extract Gelatin 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Meat extract 



(Liebig's) 6.0 g. 



3. Peptone (Witte's) . . 10.0 g. 



4. NaCl 5.0 g. 



5. K2HPO4 2.0 g. 



6. Gelatin 100.0 or 120.0 g. 



Preparation : 



(1) Dissolve 2, 3, 4 and 5 in 1 by heating 

 on the water bath. 



(2) Add and dissolve 100.0 to 120.0 g. 

 gelatin in (1). 



(3) Neutralize to litmus with NaOH and 

 then add 1.5 g. crystalline soda 

 (15.0 cc. of a 10.0% soda solution). 



Sterilization: Not specified. 



Use: Medium used for water analysis. 



Reference: Kohn (1909 p. 127). 



2301. Buchan's Neutral Red Peptone 



Extract Gelatin 



Constituents : 



1. Distilled water 850.0 cc. 



2. KOH (5.0% solution) 10.0 cc. 



3. Gelatin 100.0 g. 



4. Lemco 10.0 g. 



5. Peptone 20.0 g. 



6. Neutral red 



