556 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(8) Prepare an alcoholic saturated solu- 

 tion of fuchsin by pulverizing com- 

 mercial fuchsin in a mortar and 

 adding absolute alcohol and placing 

 in the incubator for a day shaking 

 occasionally, 



(9) Prepare a 10.0% solution of sodium 

 sulphite. Boil several times to 

 sterilize. 



(10) To 100.0 cc. of liquid and sterile (4) 

 add 6.0 cc. of sterile (5). 



(11) Heat in streaming steam for 15 min- 

 utes. The agar becomes dark brown 

 and turbid. 



(12) Add 5.0 cc. sterile (6), 5.0 cc. sterile 

 (7), 0.4 cc. (8) and 2.0 cc. (9) to each 

 100.0 cc. lot immediately after it is 

 removed from the steamer. 



(13) Slant the flasks so that the precipi- 

 tate settles quickly to the bottom. 



(14) Pour into plates, but leaving the 

 settled precipitate in the flasks. 



(15) Dry in the incubator or at 50°C. for 

 30 minutes. 



(16) The plates are transparent and 

 yellowish brown in color. 



Sterilization: Sterilize (5), (6) and (7) by 

 heating in steam for 30 minutes. 



Use: Diagnosis of cholera. Author re- 

 ported that cholera organisms gave large 

 red colonies. Other organisms were 

 inhibited. The more alkali added, the 

 more coli colonies were inhibited. 



Variants : Klimmer prepared the medium as 

 follows : 



(1) Soak 35.0 g. agar in a liter of water 

 over night. 



(2) Add 10.0 g. peptone, 5.0 g. NaCl and 

 10.0 g. meat extract to (1). 



(3) Boil for 4 or 5 hours in the steamer. 



(4) Distribute in 100.0 cc. lots in 200.0 

 cc. flasks. 



(5) Add 6.0 cc. of a 10.0% solution of 

 water free soda, 5.0 cc. of a 20.0% 

 sucrose solution, 5.0 cc. of a 20.0% 

 dextrin solution, 0.25 cc. of a satu- 

 rated alcoholic fuchsin solution and 

 2.5 cc. of a 10.0% NasSOa solution to 

 each 100.0 cc. of agar. 



(6) Sterilization not specified. 

 References: Aronson (1915 p. 1028), Klim- 

 mer (1923 p. 218). 



1780, Hesse's Malachite Green Agar 

 (Klimmer) 

 Constituents : 



1. Water 1000.0 cc. 



2. Peptone 10.0 g. 



3. NaCl 5.0 g. 



4. Meat extract 10.0 g. 



5. Agar 35.0 g. 



6. NazCOa (10.0%) 



soln.) 30.0 to 40.0 cc. 



7. Sucrose (20.0%) 



soln.) 50.0 cc, 



8. Dextrin (20.0% 



soln.) 50.0 cc. 



9. Malachite green 

 (chlorzinc salt c.p. 



sat. ale. soln.) 4 cc. 



10. Na-^SOs (10.0% 



soln.) 25.0 cc. 



Preparation : 



(1) Soak 35.0 g. of agar in a liter of water 

 over night. 



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



(3) Boil for 4 or 5 hours in the steamer. 



(4) Distribute in 100.0 cc. lots in 200.0 

 cc. flasks. 



(5) Add 3.0 to 4.0 cc. of a 10.0% water 

 free soda solution, 5.0 cc. of a 20.0% 

 sucrose solution, 5.0 cc. of a 20.0% 

 dextrin solution, 0.4 cc. of a con- 

 centrated alcoholic solution of mala- 

 chite green (chlorzinc, double salt, 

 crystalline, c.p.) and 2.5 cc. of a 

 10.0% NajSOg solution to each 100.0 

 cc. of agar. 



Sterilization: Not specified. 



Use: Cholera diagnosis. Author reported 



that cholera colonies were green after 24 



to 48 hours. 

 Reference: Klimmer (1923 p. 219). 



1781, Bacto Krumwiede Triple Sugar Agar 

 (Dehydrated) 



Constituents: 



1. Distilled water 



2. Peptone (Bacto) 10.0 g. 



3. Agar (Bacto) 15.0 g. 



4. Lactose (Bacto) 10.0 g, 



5. Sucrose (Bacto) 10.0 g. 



6. Glucose (Bacto) 1.0 g. 



7. NaCl 5.0 g. 



8. Andrade indicator 0.025 g. 



