240 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(13) Bring the volume up to 1000.0 cc. 



(14) Make the reaction 1.2% acid to 

 phenolphthalein. 



(15) Steam 30 minutes. 



(16) Filter while hot, thru well wetted, 

 thick filter paper. 



(17) Distribute the filtrate into test 

 tubes or flasks. 



(18) Sterilize. 



(d) Brown did not specify the method of 

 preparation of the veal infusion, but 

 specified the use of 10.0 g. of Fair- 

 child's, Witte's, Bacto peptone or 

 aminoids. The initial pH was be- 

 tween 6, 7 and 7. . 

 References: Warden, Connell and Holly 

 (1921 p. 104), Dernby and David (1921 

 p. 156), Dernby and Blanc (1921 p. 420), 

 Harvey (1921-22 p. 113), Brown (1921 

 p. 561). 



787. Schoenholz and Meyer's Veal Infusion 

 Broth 



Constituents : 



1. Water 1000.0 cc. 



2. Lean veal 500.0 g. 



3. Difco peptone 10.0 g. 



4. Na2HP04 



5. KH2PO4 

 Preparation : 



(1) Infuse 500.0 g. finely chopped veal in 

 a liter of water on ice over night. 



(2) Boil 20 minutes and strain thru cheese 

 cloth and allow to cool. 



(3) Filter to remove fat. 



(4) Add 3 to (3). 



(5) Adjust reaction to pH = 7.4. 



(6) Stabilize in Arnold for 30 minutes. 



(7) Filter and tube in 10.0 cc. lots. 



(8) To each 10.0 cc. tube HCl or NaOH is 

 added to obtain the desired reaction 

 (varying from pH 4.3 to pH 9.0). 



(9) Prepare 0.5M solutions of KH2PO4 

 and NajHPO^ and mix in equal 

 volumes. 



(10) Add 2.0 cc. of sterile (9) to each tube 

 of sterile (7). 



Sterilization: Sterilize (7) in steam on each 

 of 3 consecutive days. Method of steri- 

 lization of NaOH, HCl or phosphate 

 solutions not given. 



Use: To determine optimum pH values for 

 typhoid bacilli. Optimum growth oc- 

 curred between pH = 6.8 and 7.0 in a salt- 

 free medium. 



Variants : Omit the sodium and potassium 



phosphate. 

 Reference: Schoenholz and Meyer (1921 



p. 387). 



788. Bunker's Veal Infusion Polypeptide 

 Solution 



Constituents : 



1. Tap water 1000.0 cc. 



2. Lean veal 1.0 lb. 



3. NaCl 5.0 g. 



4. Polypeptides 2.0 g. 



Preparation: 



(1) Mix chopped veal and water in after- 

 noon and warm to 40°C. in large open 

 kettle. 



(2) Inoculate with B. coli. 



(3) Allow to stand over night. 



(4) Heat to coagulate proteins. 



(5) Strain and place in flasks. 



(6) Heat in the autoclave at 20 pounds 

 for I hour. 



(7) Add NaCl and polypeptides and 

 NaOH to give pH = 7.8. 



(8) Heat in autoclave at 15 pounds for 

 15 minutes. 



(9) Filter thru bibulous filter or thick 

 cloth. 



(10) Distribute in amber glass bottles. 

 Sterilization : Sterilize with steam in usual 



manner. 

 Use: Study of production of diphtheria 



toxin. 

 Reference: Bunker (1919 pp. 404, 405). 



789. Krasnow's et al. Sugar Free Veal Broth 



Constituents: 



1. Tap water 1000.0 cc. 



2. Bacto veal 75.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



Preparation : 



(1) Place 75.0 g. Bacto veal in 1000.0 cc. 

 tap water and heat in the Arnold 

 sterilizer for two hours. 



(2) Allow the coagulum thus formed to 

 settle to the bottom of the container. 

 Allow to cool very slowly. 



(3) Strain thru a wire sieve. 



(4) Dissolve 3 and 4 in (3). 



(5) Adjust to pH = 7.9. 



(6) Steam in the Arnold for 15 minutes. 



(7) Filter. 



Sterilization: Sterilize in the Arnold for 



