CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



557 



Preparation : 



(1) Dissolve 52.0 g. of Bacto Krumwiede 

 Triple Sugar Agar (Dehydrated) in 

 1000.0 cc. distilled water by boiling 

 or autoclaving. Avoid excess heat. 



(2) Tube. 



Sterilization: Sterilize in the usual manner. 



Use: To detect intermediates of the colon- 

 typhoid group. 



Reference: Digestive Ferments Co. (1925 

 p. 12). 



1782. Krumwiede and Kohn's Triple Sugar 

 Agar 



Constituents : 



1. Water 1000.0 cc. 



2. Agar 15.0 g. 



3. Meat extract 3.0 g. 



4. Peptone lO'O S- 



5. NaCl (salt) 5.0 g. 



6. Lactose 100 g- 



7. Sucrose 10-0 g- 



8. Glucose 1-0 g- 



9. Andrade indicator 10.0 cc. 



Preparation : 



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



(2) Adjust slightly alkaline to litmus 

 (or 0.6% acid to phenolphthalein hot 

 titration). 



(3) Distribute in 100.0 cc. lots. 



(4) At the time of use, add 1.0% lactose, 

 1.0% saccharose and 0.1% glucose to 

 melted sterile (3). (These amounts 

 may be conveniently added from a 

 stock solution containing 20.0% lac- 

 tose, 20.0% saccharose and 2.0% 

 glucose that has been sterilized inter- 

 mittently. Five cc.'s of this solution 

 gives the desired concentration. 



(5) Add 1.0% Andrade indicator and 

 adjust reaction. The medium is to 

 be red when hot and colorless when 

 cold. (If it is not red when warm the 

 medium is too alkaline, and if it fails 

 to decolorize when cool it is too acid.) 



(6) Tube from a sterile covered funnel, 

 preferably in narrow tubes. 



(7) Steam for 20 minutes. 



(8) Slant leaving a generous butt for 

 inoculation. 



(9) Incubate over night to test sterility. 

 Sterilization: Sterilize (3) in the autoclave. 

 Use: Isolation of para-typhoid bacilli. 



Used especially to isolate the inter- 

 mediates of the colon-typhoid group. 

 References: Krumwiede and Kohn (1917 p. 

 226), Park, Williams and Krumwiede 

 (1924 p. 128). 



1783. Amoss' Four Sugar Agar 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Beef extract 3.0 g. 



3. Peptone 100 g. 



4. NaCl 5.0 g. 



5. Agar 150 g. 



6. Lactose 2.5 g. 



7. Rafhnose 2.5 g. 



8. Sucrose 2.5 g. 



9. Salicin 2.5 g. 



10. Lead acetate (0.25% 



soln.) 250.0 cc. 



Preparation : 



(1) Dissolve 2, 3, 4, 5, 6, 7, 8, 9 and 10 in 1. 



(2) Adjust to pH = 7.2. 



(3) Distribute in 4.0 cc. lots. 



(4) Add to each tube 1.0 cc. of a 0.25% 

 solution of lead acetate. 



Sterilization: Not specified. 



Use: Cultivation of Bacillus typhi murium. ^ 

 Author reported that Bacillus typhi 

 7miritim produced browning of medium. 



Reference: Amoss (1922 p. 28). 



1784. Amoss' Sucrose Salicin Agar (Topley 

 and Ayrton) 



Constituents : 



1. Water 1000.0 cc. 



2. Beef extract (Lemco) 5.0 g. 



3. Peptone (Witte) 10 g. 



4. NaCl 5.0 g. 



5. Lactose 10 g. 



6. Sucrose 10-0 g- 



7. Salicin 100 g. 



8. Andrade indicator 10.0 cc. 



9. Agar 25.0 g. 



10. Lead acetate (1.0%) 50.0 cc. 



11. Na2HP04 (1.0%) 50.0 cc. 



Preparation : 



(1) Dissolve 2, 3, and 4 in 1 by heating 

 in the steamer. 



(2) Add 25.0 g. agar and steam until 

 dissolved. 



(3) Remove from the steamer and add 

 the white of an egg. 



(4) Coagulate in the steamer. 



