CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



739 



Sterilization: Sterilize intermittently on 3 



successive days. 

 Use: Cultivation of tubercle bacilli. 

 Variants : Dittrich solidified medium 1737 



with gelatin instead of agar. 

 Reference: Kowalski (1890 p. 266). 



2290. Uffelmann's Methyl Violet Infusion 



Gelatin 



Constituents: 



1. Infusion gelatin 1000.0 cc. 



2. Citric acid 



3. Methyl violet 

 Preparation : 



(1) Prepare meat infusion peptone gela- 

 tin in usual way, making slightly 

 alkaline and fiter. 



(2) Add solution of citric acid in such 

 amount that 10.0 cc. of medium re- 

 quires 14.0 cc. of a solution of 5.3 g. 

 sodium carbonate in 1000.0 cc. water 

 to neutralize exactly. 



(3) Filter. 



(4) Add to each 100.0 cc. of the filtrate, 

 2.5 mg. of methyl violet that has been 

 rubbed up with 1 drop absolute alco- 

 hol and 1.0 cc. distilled water. 



Sterilization: Sterilize in streaming steam 

 15 minutes. 



Use: Isolation of typhoid bacillus. The 

 author reported that the medium elim- 

 inated many non-typhoid organisms. 

 Good growth of typhoid bacilli. 



Reference: Uffelmann (1891 p. 855). 



2291. Vincent's Glucose Glycerol Gelatin 



Constituents : 



1. Infusion broth 500.0 cc. 



2. Gelatin (extra) 50.0 to 75.0 g. 



3. Glucose 5.0 g. 



4. Glycerol 5.0 g. 



Preparation : 



(1) Prepare infusion broth from beef 

 and peptone. (Method not given.) 



(2) Dissolve 2, 3 and 4 in (1). 



(3) Neutralize. (Indicator not specified.) 



(4) Tube. 



Sterilization: Method not given. 



Use: Cultivation of anaerobes in water 



analysis. 

 Reference: Vincent (1907 p. 65). 



2292. Huntoon's Hormone Gelatin 



Constituents : 



1. Water 1000.0 cc. 



2. Heart, fresh beef (or steak) 500.0 g. 



3. Peptone (Bacto) 10.0 g. 



4. Gelatin 10.0 g. 



5. Salt 5.0 g. 



6. Egg, whole 1 



Preparation : 



(1) Chop 2 and mix 1, 2, 3, 4, 5 and 6. 

 Place in an enamel ware vessel or a 

 large coffee pot. 



(2) Heat over a free flame with constant 

 stirring until the red color of the 

 meat infusion changes to browm at a 

 temperature of about 68°C. Do not 

 go beyond this temperature. 



(3) Adjust to slightly alkaline to litmus 

 with N/1 NaOH and then add 1.0 

 cc. per liter of medium. 



(4) Cover the vessel and place in an 

 Arnold sterilizer or in a water bath 

 at 100° for 1 hour. 



(5) Remove the vessel from the steri- 

 lizer and separate with a glass rod, 

 the firm clot which has formed from 

 the side of the vessel. 



(6) Return to the Arnold sterilizer at 

 100° for 90 minutes. 



(7) Remove the vessel and allow to 

 stand at room temperature for about 

 10 minutes in a slightly inclined 

 position. 



(8) Pipette off the fluid portion or 

 decant. If it is poured thru a fine 

 wire sieve, many of the fine pieces of 

 meat clot may be caught. (Avoid 

 filtering thru cheese cloth, cotton 

 or other adsorptive materials.) 



(9) Allow (8) to stand in tall cylinders 

 for 15 to 20 minutes until the fat 

 present has risen to the surface and 

 removed. 



(10) Tube in 10.0 cc. lots. 



(11) The medium may be further cleared 

 by filtering thru glass wool, asbestos 

 wool, sedimentation or centrifuga- 

 tion. 



Sterilization : Sterilize in the steamer using 

 the intermittent method. 



Use: Substitute for media containing ser- 

 ous fluids. The author reported the me- 



