326 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(3) Autoclave. (Time not specified.) 



(4) Add asparagin, ammonium lactate, 

 K0HPO4 and MgS04. 



(5) Make up to 1 liter and adjust to +1 

 phenolphthalein. 



Sterilization: Final sterilization not speci- 

 fied. 



Use: To show production of indol by 

 bacteria. 



Reference: Norton and Sawyer (1921 

 p. 473). 



1134. Duval and Harris' Tryptic Digest 

 Solution 



Constituents: 

 1. Serum, egg albumin, liver or 



placenta 200.0 cc. 



Preparation : 



(1) 200.0 cc. of fresh sterile human serum, 

 egg albumin, or the equivalent in 

 grams of liver or placenta tissue are 

 placed in a flask and 20.0 cc. of a 

 1.0% physiological salt solution of 

 sterile trypsin added. 



(2) Digest for 5 days at 37 °C., changing 

 the reaction to neutral as occasion 

 demands. 



(3) Heat at 70°C. for one hour. 

 Sterilization: Filter thru a Berkefeld filter. 

 Use: Cultivation of leprosy bacilli. The 



author stated that the same materials 

 might be autolyzed under sterile condi- 

 tions at 40°C. for 2 weeks or hydrolyzed 

 with a culture of Finkler and Prior's 

 vibrio or some other bacterial proteoly- 

 zer for 24 to 36 hours, and then neu- 

 tralized with normal NaOH. Sterilize as 

 indicated above. 

 Reference: Duval and Harris' (1911 p. 168). 



1135. Harvey's Tryptic Digest Solution 



Constituents : 



1. Water 1000.0 cc. 



2. Heart (ox), or placenta 

 (human) 500.0 g. 



3. K2HPO4 2.0 g. 



Preparation : 



(1) Free fresh hog pancreas from fat as 

 far as possible. 



(2) Mince finely and weigh. 



(3) Add 3.0 cc. 0.5% hydrochloric acid 

 for every gram of the minced pan- 

 creas. 



(4) Stir the mixture at intervals for 

 30 minutes. 



(5) Add 6.4 cc. 5.0% sodium hydroxide 

 for every 100.0 cc. of 0.5% hydro- 

 chloric acid used. 



(6) Stir well and filter thru folded filter 

 paper. 



(7) Shake up with a little toluol. 



(8) Make the reaction less acid by the 

 cautious addition of 10.0% sodium 

 hydroxide. 



(9) Store in a stoppered bottle in a cool, 

 dark place. 



(10) Mince finely ox heart or human 

 placenta. 



(11) Add 500.0 g. to 1000.0 cc. tap water. 



(12) Make faintly alkaline to litmus. 



(13) Heat slowly to 75°C. and maintain 

 at this temperature 10 minutes. 



(14) Cool to 37°C. 



(15) Add 1.0 or 2.0% of (9) to (14). 



(16) Keep at 37°C. for 4 hours. If the 

 digestion is extended to 6 hours or 

 longer it is necessary to add chloro- 

 form or toluene. Control the prog- 

 ress of digestion by Biuret and 

 tryptophane tests. 



(17) Make faintly acid to litmus with 

 glacial acetic acid. 



(18) Raise slowly to the boiling point. 



(19) Boil gently 15 minutes. 



(20) Filter. 



(21) Adjust the reaction. 



(22) Add 0.2% di-potassium phosphate. 



(23) Steam 20 minutes. 



(24) Clarify and filter. 

 Sterilization: Sterilize in the autoclave or 



steamer. 

 Use: General culture medium. 

 Reference: Harvey (1921-22 p. 114). 



1136. Stickel and Meyer's Tryptic Digest 

 Solution 



Constituents: 



1. Distilled water 1000.0 cc. 



2. NaCl 5.0 g. 



3. CaCl2 0.1 g. 



4. MgS04 0.2 g. 



5. CaHP04 2.0 g. 



6. K2HPO4 2.0 g. 



7. Heart, beef or placenta, 

 human 500.0 g. 



Preparation : 



(1) Remove fat from beef heart and chop 

 finely, or rinse human placenta in 

 water and pass thru a meat chopper. 



