CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



471 



Preparation : 



(1) Add2, 3, 4, 5and6to(l). 



(2) Heat slowly until the gelatin is 

 dissolved. Stir. 



(3) Neutralize the alkaline mixture by 

 the careful addition of HCl, drop by 

 drop, using litmus as an indicator. 



(4) Add 2% of a soda solution containing 

 two parts water to one part soda. 



(5) Boil two hours in the steamer. 



(6) Filter thru a thin layer of sterile 

 cotton. 



(7) Distribute into sterile petri dishes 

 and solidify. 



Sterilization: Sterilization given in the 

 preparation. 



Use: Diagnosis of cholera. Author re- 

 ported that cholera colonies after 4 or 5 

 hours appeared similar to colonies 15 to 20 

 hours old in gelatin, having the same 

 characteristics as the cholera colonies 

 described by Koch, shiny with irregular 

 edge. Liquefaction took place after 24 

 to 30 hours at 20-22°C. 



Reference: Deycke (1895 p. 244). 



1631a. Deycke's Albuminate Glycerol Agar 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Alkali albuminate (from 



veal) 10.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



5. Agar 20.0 g. 



6. Glycerol 10.0 g. 



Preparation : 



(1) Dissolve 2, 3, 4, 5 and 6 in 1. 



(2) Make alkaline by adding 33.0% soda. 



(3) Filter. 



(4) Tube. 



Sterilization: Method not specified. 

 Use: Isolation of cholera vibrios. Author 

 reported that only cholera, anthrax, 

 diphtheria and tubercle bacilli grew. 

 Variants : 



(a) The author used the following me- 

 dium for the cultivation of diphtheria 

 bacilli. 



(1) Dissolve 10.0 g. alkaline albumi- 

 nate, 10.0 g. peptone, 5.0 g. NaCl, 

 20.0 g. agar-agar and 5.0 g. glycerol 

 in 1000.0 cc. of distilled water. 



(2) Neutralize the alkaline mixture by 

 the careful addition of HCl, drop 



by drop, using litmus paper as an 

 indicator. 



(3) Make alkaline by adding 1.0% of a 

 soda solution which contains two 

 parts water and one part soda. 



(4) Soak at room temperature for one 

 or more hours. 



(5) Boil in the steamer for I to one 

 hour. 



(6) Filter thru a layer of sterile cotton 

 (may be filtered thru paper in a 

 Unna steam funnel). 



(7) Distribute into sterile test tubes. 



(8) Sterilize once more in streaming 

 steam for one-half hour. 



(9) Slant. 



(b) Deycke and Voigtlander prepared a 

 similar medium as follows: 



(1) Pour 300.0 cc. Ba(OH)ii solution on 

 finely chopped horse heart (or other 

 fat free horse meat) and place in 

 the incubator at 37°C. for 48 hours. 

 This solution is usually complete 

 at the end of this time. 



(2) Dilute (1) with 600.0 cc. of water. 



(3) Filter. 



(4) Heat to avoid the splitting of 

 ammonia. 



(5) Neutralize the filtrate with HCl 

 using litmus as an indicator. 



(6) To each 100.0 cc. of (5) add 400.0 

 cc. of water, 1.5 g. NaCl, 5.0 g. 

 peptone, 25.0 g. glycerol and 10.0 

 g. agar. 1.5 to 2.0% glucose may be 

 added. 



(7) Sterilization not specified. 

 References: Deycke (1894 p. 528), (1896 



p. 242), Deycke and Voigtlander (1901 

 p. 621). 



1632. Krumwiede and Pratt's Gelatin 

 Serum Agar 



Constituents: 



1. Water 1000.0 cc. 



2. Peptone 20.0 g. 



3. Agar 10.0 g. 



4. Gelatin SO.O g. 



5. Serum, horse 500.0 cc. 



Preparation : 



(1) Dissolve 2, 3 and 4 in 1. '^'"^ 



(2) Mix one part of 5 with two parts (1). 



(3) Adjust to slightly alkaline to litmus. 

 Sterilization: Method not specified. 



