CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



259 



(11) Add 0.15% dextrose and enough 

 laked blood to give a slight pink 

 tint. 



(12) Tube in 10.0 cc. lots. 

 Sterilization: Sterilize by the intermittent 



method. 



Use: To cultivate highly pathogenic organ- 

 isms. Author reported that if spinal 

 fluid be taken from a meningitis case in 

 which no organism can be found in 

 smears, a distinct growth appeared in 

 this medium after 9 hours rendering a 

 positive diagnosis possible. Organisms 

 multiplied in the leukocytes transferred. 



Variants : 



(a) Author used fresh beef steak instead 

 of heart. 



(b) Bailey prepared a similar medium as 

 follows: 



(1) Dissolve 10.0 g. gelatin in 1000.0 cc. 

 distilled water and heat to 50 to 

 60°C. 



(2) Add 500.0 g. of moderately fine 

 chopped beef to (1). 



(3) Bring to a boil and cook slowly for 

 15 to 20 minutes. 



(4) Filter thru a 16 mesh (cullender 

 type) until clear. 



(5) Add 10.0 g. peptone and 5.0 g. 

 NaCl. 



(6) Boil 5 minutes. 



(7) Adjust to the desired reaction (pH 

 = 7.5). 



(8) Allow to stand several minutes 

 and decant the supernatant fluid. 



(9) Tube. 



(10) Sterilize fractionally or at 5 

 pounds pressure for 5 minutes. 

 Reference: Huntoon (1918 p. 172). Bailey 

 (1925 p. 341). 



838. Macnoughton's Blood Infusion Broth 



Constituents : 



1. Heart infusion broth. 



2. Blood. 

 Preparation : 



(1 ) Prepare infusion broth using ox heart, 

 and adjust to pH between 7.4 and 7.5. 



(2) Tube in 10.0 cc. lots. 



(3) The evening before the medium is re- 

 quired, add 1.0 cc. of sterile human 

 blood to each 10.0 cc. of (2). 



(4) Thoroughly mix by rolling the tubes 

 between the hands. 



(5) Allow to stand at room temperature 

 over night. 

 Sterilization: Not specified. 

 Use: Isolation of gonococci. 

 Reference: Macnoughton (1923 p. 297). 



839. Ficker and Hoffmann's Caffeine 

 Infusion Broth 



Constituents: 



1. Distilled water 4000.0 cc. 



2. Beef 1000.0 g. 



3. Peptone (Witte) (6.2%) .... 120.0 g. 



4. NaCl (0.5%) 10.0 g. 



5. Caffein (0.6%) 24.0 g. 



6. Crystal violet (0.1%) 

 (solution) 28.0 cc. 



Preparation : 



(1) Pour 2 liters of distilled water over 

 1000.0 g. finely chopped lean beef 

 in an enamel kettle. 



(2) Weigh the kettle and contents. 



(3) Heat at 50 to 55°C. for 30 minutes. 



(4) Stir with a glass rod and heat to 

 boiling. 



(5) Weigh and add distilled water to 

 make up the loss in weight. 



(6) Filter thru filter gauze. 



(7) Measure and add 6.0% (not given as 

 0.6%) Witte peptone and 0.5% NaCl. 



(8) Heat until the peptone is dissolved. 



(9) Filter. 



(10) Distribute in Erlenmeyer or beer 

 flasks. 



(11) Provide each flask with an absorbent 

 paper cap. 



(12) Prepare a 1.2% caffeine solution in 

 sterile distilled water. Shake, but 

 heating is not necessary to obtain 

 complete solution. Weigh the caf- 

 feine on a chemical balance. 



(13) Measure 100.0 cc. of sterile (11) into 

 a sterile Erlenmeyer flask and adjust 

 to —2.7 to phenolphthalein. 



(14) Measure 105 cc. of (12) in a sterile 

 measuring flask and add to each 

 100.0 cc. of cool (13) under aseptic 

 conditions. Do not heat after the 

 addition of caffeine. 



(15) Prepare a 0.1% crystal violet solu- 

 tion (use chemical balance) in sterile 

 distilled cold water. 



(16) Add 1.4 cc. of (15) by means of a 

 sterile pipette to each flask (14). 



Sterilization: Sterilize (1) in streaming 



