552 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(6) Neutralize with 10.0% soda solution. 



(7) Boil one hour. 



(8) Filter and sterilize. (Method not 

 given). 



(9) Add 10.0 cc. of 10.0% soda solution 

 and 1.0 g. of sodium taurocholate 

 to one liter of (8). 



(10) Heat at 100°C. in the steamer for 

 20 minutes. 



(11) Add 10.0 g. lactose, 5.0 cc. of a 

 saturated alcoholic solution of fuch- 

 sin, and 25.0 cc. of a 10.0% sodium 

 sulphite solution to each liter of 

 (10). 



(12) Distribute in small flasks. 

 Sterilization: Method of sterilization not 



specified. 

 Use: Water analysis. Enrichment medium 



for Bacterium coli. 

 Reference : Olszewski and Kohler (1923 p. 3) . 



1772. Bacto Saccharose-Mannitol Agar 



(Dehydrated) 



Constituents : 



1. Distilled water 



2. Beef extract (Bacto) 3.0 g. 



3. Peptone (Bacto 5.0 g. 



4. Agar (Bacto) 25.0 g. 



5. Sucrose (Bacto) 10.0 g. 



6. Mannitol (Bacto) 1.0 g. 



7. Andrade Indicator (Difco) . 0.0275 g. 

 Preparation : 



(1) Dissolve 44.0 g. of Bacto Saccharose- 

 Mannitol Agar (Dehydrated) in 

 1000.0 cc. of distilled water by boiling 

 or autoclaving, preferably the latter. 



(2) Tube. 



(3) Final pH = 7.5±. 



Sterilization : Sterilize in the usual manner, 



avoiding excessive heat. 

 Use: Differential tube medium. 

 Reference: Digestive Ferments Co. (1925 



p. 12). 



1773. Hesse's Lactose Glycerol A^ar 



(Stokes and Hachtel) 

 Constituents : 



1. Distilled water 1000.0 cc. 



2. Agar 6.0 g. 



3. Beef extract (Liebig's) 5.0 g. 



4. Peptone (Witte's) 10.0 g. 



5. NaCl 8.5 g. 



6. Lactose (1.0%) 10.0 g. 



7. Glycerol (5.0%) 50.0 g. 



8. Azolitmin 



Preparation : 



(1) Dry agar at 150°C. for 30 minutes. 



(2) Dissolve 6.0 g. of (1) in 500.0 cc. 

 distilled water by boiling and make 

 up the loss in weight by adding dis- 

 tilled water. 



(3) Dissolve 3, 4, 5, 6 and 7 in 500.0 cc. 

 of distilled water. 



(4) Mix (3) and (2) and boil 30 minutes. 



(5) Make up the loss in weight by the 

 addition of distilled water. 



(6) Filter. 



(7) Adjust the reaction to neutrality 

 (indicator not specified). 



(8) Add sufficient azolitmin to give the 

 desired color. 



(9) Tube in 9.0 cc. lots. 

 Sterilization: Autoclave at 15 pounds pres- 

 sure for 20 minutes. 



Use: Isolation of typhoid bacillus and 

 cholera spirillum. The author reported 

 that typhoid colonies were pink. They 

 had an opaque nucleus surrounded by a 

 translucent area. Bacillus alcaligenes 

 formed a blue concentric colony. Other 

 motile organisms produced a blue colony. 



Variants: The authors prepared a similar 

 medium as follows: 



(1) Dry agar at 150°C. for 30 minutes. 



(2) Dissolve 5.5 g. of (1) in 500.0 cc. of 

 distilled water. 



(3) Add 5.0 g. beef extract to 500.0 cc. 

 of distilled water. 



(4) Filter (3) into a sterile flask and 

 inoculate with the colon bacillus and 

 incubate at 37°C. for 24 hours. 



(5) Filter the sugar-free bouillon and 

 dissolve 10.0 g. Witte's peptone, 

 10.0 g. lactose and 8.5 g. NaCl in it. 



(6) Make up the loss due to evaporation 

 by addition of distilled water. 



(7) Mix (6) and (2) and boil for 30 

 minutes, making up the loss in 

 weight by the addition of distilled 

 water. 



(8) Filter. 



(9) Adjust the reaction to neutral. 



(10) Add 50.0 cc. of glycerol. 



(11) Color with Kahlbaum's azolitmin 

 solution. 



(12) Distribute in 10.0 cc. lots. 



(13) Autoclave at 16 lbs. pressure for 20 

 minutes. 



(14) Store in the ice box until ready for 

 use. 



