498 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(2) Heat then to a boil stirring con- 

 stantly. 



(3) Filter thru a filtering cloth, wash- 

 ing the residue with one liter of 

 water and squeeze out the residue. 



(4) Distribute into glass flasks and 

 sterilize twice before use (This 

 constitutes a stock meat infusion.) 



(5) For the preparation of one liter of 

 agar medium add 11.5 g. fibre agar 

 to 500.0 g. water. Allow the agar 

 to grow and swell in the water. 



(6) Add 8.0 g. ovi albumin (siccum) to 

 about 40.0 cc. of water. It dissolves 

 slowly. 



(7) Place 1000.0 g. of (4) (meat infusion) 

 in a weighed kettle. 



(8) Add 250-300.0 cc. water to (7) and 

 1.0 g. peptone. 



(9) Heat nearly to boiling and neutralize 

 with NaOH until litmus paper be- 

 comes strongly blue. 



(10) Add (5) (the soaked agar) to (9) and 

 boil over a free flame to dissolve the 

 agar. Do not over heat. 



(11) Add 1.0 g. (NH4)2S04 and 1.0 g. 

 KNO3 to (10). 



(12) Weigh and add distilled water until 

 the contents of the kettle weigh 

 1000.0 g. 



(13) Cool to 50-55°C. 



(14) Add (6) (albumin solution) or the 

 whites of two eggs and place in a 

 steamer for 50-60 minutes. 



(15) Filter thru a hot water funnel. 

 (May use a folded felty, so-called 

 coffee filter paper.) 



Sterilization: Method not given. 



Use : Cultivation of Spirillum undula majus 



and other spirilla. 

 Reference: Zettnow (1895 p. 394). 



1663. Besson's Phosphate Infusion Agar 



Constituents: 



1. Water 1000.0 cc. 



2. Beef 500.0 g. 



3. Peptone 10.0 g. 



4. Agar 20.0 g. 



5. Sodium phosphate 1.0 g. 



6. NaCl 5.0 g. 



Preparation : 



(1) Remove all fat and tendons from 

 beef and chop into small pieces. 



(2) Allow 500.0 g. of (1) to macerate with 

 1000. cc. cold water for 6 hours, or 



if one wishes to remove the sugar, 

 12 hours at 37°C. 



(3) Place in an enamelled pot and bring 

 slowly to a boil. 



(4) Boil for 10 minutes. 



(5) Throw on a thick cloth and press 

 the meat free from juice. 



(6) Filter the juice thru moistened 

 paper. 



(7) Add 10.0 g. of Chapoteaut's or 

 Defresne peptone, 5.0 g. XaCl and 

 about 1.0 g. of sodium phosphate. 



(8) Boil stirring constantly until solu- 

 tion is complete, 



(9) Neutralize or make slightly alkaline 

 to litmus by the addition of soda 

 solution. 



(10) Soak 20.0 g. of chopped thread agar 

 in cold water for several hours. 



(11) Heat (10) and (9) at 100°C. until 

 the agar is dissolved. 



(12) Readjust the reaction if necessary. 



(13) Allow to cool to 55 or 60°C. 



(14) Beat the white of an egg in 100.0 cc. 

 of water and add to (13). 



(15) Mix well. 



(16) Autoclave at 120°C, for one hour. 



(17) Filter thru a moistened Chardin 

 filter using a hot water funnel. 



(18) Tube. 



Sterilization: Sterilize at 115° for 20 



minutes. 

 Use: General culture medium. 

 Variants : 



(a) The agar may be treated with a 6 to 

 100 solution of HCl (500.0 cc. water 

 to 30.0 cc. HCl for 24 hours and then 

 with a 5.0% ammonia solution. Then 

 thoroly wash the agar with water. 



(b) Besson also used 20.0 g. peptone in- 

 stead of 10.0 g. 



Reference: Besson (1920 p. 41). 



1664. Smith's Infusion Agar 



Constituents: 



1. Distilled water 2000.0 cc. 



2. Beef 1000.0 g. 



3. Peptone (Merck, brown) . . . 20.0 g. 



4. Agar 20.0 g. 



Preparation : 



(1) Soak finely chopped fat free beef with 

 2000.0 cc. distilled water for some 

 hours and then slowly raise to 65°C. 

 on the water bath. 



(2) Finally steam and filter. 



