CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



253 



(11) Distribute 800.0 cc. in 2 liter flasks. 



(12) Autoclave for 30 minutes at 15 

 pounds pressure. 



(13) Add 8.0 cc. of a 10.0% glucose solu- 

 tion, sterilized in the Arnold on 

 each of 3 successive days, for 

 30 minutes to each flask. 



References: Wilcox (1916 p. 334), Walbum 

 (1922 p. 25) taken from (1922 p. 109), 

 Park, Williams and Krumwiede (1924 

 pp. 132, 133). 



824. Park, Williams and Krumwiede's 

 Glycerol Veal Infusion Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Veal 1.0 lb. 



3. Peptone (Fairchild's (1.0%). 10.0 g. 



4. NaCl (0.5%) 5.0 g. 



5. Glycerin (5.0%) 50.0 g. 



Preparation : 



(1) Mince 1.0 pound of lean veal free 

 from fascia with 1 liter of water. 



(2) Macerate over night at room tem- 

 perature. 



(3) Heat to 45 to 48° for one hour. 



(4) Boil up strongly. 



(5) Strain thru cheese cloth. 



(6) Add 1.0% Fairchild's peptone and 

 0.5% NaCl. 



(7) Boil to dissolve the peptone. 



(8) Titrate and adjust the reaction to 



+2.5 to phenolphthalein (room 

 temperature) (about pH = 6.0 

 to 6.4). 



(9) Autoclave for 15 minutes at 15 

 pounds pressure, to clear. 



(10) Filter thru cotton and paper. 



(11,' Measure the filtrate and add 5.0% 



glycerol. 

 (12) Distribute in 250.0 cc. quantities in 

 each quart Blake bottle or liter flask. 



Sterilization: Sterilize in the autoclave for 

 30 minutes at 15 pounds pressure. 



Use: Cultivation of B. mallei and malein 

 production for diagnosis of glanders. 

 Cultivation of tubercle bacilli and tuber- 

 culin production. 



Variants: The authors used the medium 

 with a different reaction to cultivate 

 tubercle bacilli and for the production of 

 tuberculin. This medium was adjusted 

 to +1.3 to phenolphthalein (pH about 

 7.1) and the final reaction +1.5 to phenol- 

 phthalein or pH about 6.9. 



Reference: Park, Williams and Krum- 

 wiede (1924 p. 133). 



825. Reeser's Potato Veal Infusion Broth 

 Constituents : 



1. Distilled water 3000.0 cc. 



2. Potato 



3. Peptone 30.0 g. 



4. NaCl 15.0 g. 



5. Glycerin 150.0 g. 



6. Veal 1500.0 g. 



Preparation : 



(1) Peel potatoes and carefully remove 

 all eyes. 



(2) Cut into thin slices. 



(3) Allow to stand in water for a day, 

 renewing the water occasionally. 



(4) Soak in distilled water over night. 



(5) Pour off the water and heat the 

 pieces of potato for f hour in auto- 

 clave at 1.5 atmospheres pressure. 



(6) Add 3.0 liters of distilled water. 



(7) After sterilization allow the po- 

 tatoes to stand in water for one day. 



(8) Filter the well extracted potato 

 slices and make the filtrate up to 

 3 liters. 



(9) Grind 1500.0 g. of lean veal in a meat 

 chopping machine. 



(10) Add (8) to (9) and place in the ice 

 box over night. 



(11) Filter thru a linen towel. 



(12) Add water to make 3 liters. 



(13) Boil 10 minutes. 



(14) Filter thru a flannel cloth. 



(15) Add 1%, peptone and 0.5% NaCl 

 which one has prepared a paste of 

 with a little of the hot liquid. 



(16) Add 5.0% glycerin. 



(17) Neutralize with N/1 Na2C03 using 

 litmus as an indicator. 



(18) Add 3.0 cc. N/1 NajCOa per liter to 

 give a slight alkaline reaction or 

 adjust the reaction so that 5.0 cc. of 

 the bouillon to which phenolphtha- 

 lein has been added, will be colored 

 weakly red when 0.4 cc. N/1 NaOH 

 or NaoCOs be added. 



(19) Sterilize for 2 hours, method not 

 given. 



(20) If not transparent after sterilization 

 filter thru paper or flannel. 



(21) Distribute in sterile Roux flasks. 

 Sterilization: Sterilize (6) by autoclaving 



for 45 minutes at 1.5 atmospheres of 



