610 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(2) Filter. 



(3) Dissolve 2.5 g. agar, 0.5 g. NaCl, 

 1.0 g. glucose and 2.0 g. peptone in 

 each 100.0 cc. of the filtrate. 



(4) Heat beef serum at 60°C. for one hour 

 on each of 4 successive days. 



(5) To three parts (3) add one part (4). 



(6) Mix well. 



(7) Distribute in sterile containers. 

 Sterilization: Sterilization of (3) not spec- 

 ified. 



Use: General culture medium. 

 Reference: Bezangon (1920 p. 119). 



1919. Shmamine's Nucleic Acid Serum Agar 



Constituents: 



1. 3.0% agar with 0.5% 

 glucose 100.0 cc. 



2. Serum 100.0 cc. 



3. Physiological salt solu- 

 tion 10.0 cc. 



4. Sodium salt of nucleic 



acid 0.5 to 1.0 g. 



Preparation : 



(1) Dissolve 0.5 to 1.0 g. of the sodium 

 salt of nucleic acid in 10.0 cc. physio- 

 logical salt solution. 



(2) Add sterile (1) to 100.0 cc. sterile 

 clear transparent serum (or just 

 10.0 cc. of sterile physiological salt 

 solution may be added). 



(3) Distribute into sterile test tubes. 



(4) Prepare a 3.0% infusion agar contain- 

 ing 0.5% glucose. 



(5) Melt (4) and cool to 50°C. 



(6) Mix equal parts of sterile (5) and 

 sterile (3). 



Sterilization: Sterilize (1) for 15 minutes in 

 boiling water bath. Sterilize (3) by 

 heating for one hour on each of 3 suc- 

 cessive days at 60°C. Method of steri- 

 lization of (5) not given. 



Use : Isolation of spirochetes. 



Reference: Shmamine (1912 p. 317). 



1920. Robey, et al., Glucose Serum Agar 



Constituents : 



1. Veal infusion agar 1000.0 cc. 



2. Glucose 5.0 g. 



3. Serum, horse 50.0 g. 



Preparation : 



(1) Prepare veal infusion agar. 



(2) Adjust reaction to 0.3 to 0.5% acid to 

 phenolphthalein. 



(3) Add 2 and 3 to (2). 



Sterilization: Not specified. 



Use: Isolation of meningococci. Authors 

 reported that after 24 hours meningo- 

 cocci produced a colony 0.5-1.5 mm. in 

 diameter, easily recognizable from influ- 

 enza bacilli, streptococci and others. 



Reference: Robey, Saylor, Meleney, Ray 

 and Landmann (1918 p. 324). 



1921. Noguchi's Ringer Solution Serum 

 Agar 



Constituents : 



1. Ringer solution 45.0 cc. 



2. Rabbit serum 15.0 cc. 



3. Agar 2.0% lO.O cc. 



Preparation : 



(1) Prepare 2.0% agar. 



(2) Mix 4.5 parts Ringer's (see me- 

 dium 180) solution and 1.5 parts rab- 

 bit serum with 1.0 part 2.0% agar. 



Sterilization: Not specified. 



Use: Cultivation of Leptospira ictero- 

 haemorrhagiae. Medium was covered 

 with a sterile layer of paraffin oil after 

 inoculation. 



Variants : 



(a) Noguchi prepared a similar medium 

 as follows: 



(1) Prepare a 2.0% agar. 



(2) Mix 1.5 parts rabbit serum, 4.5 parts 

 Ringer's solution and 1.0 part 2.0% 

 agar. 



(3) Allow (2) to solidify. 



(4) Add to solidified (3) a mixture of 

 1.5 parts rabbit serum and 4.5 parts 

 Ringer's solution. 



(5) Cover with a layer of sterile par- 

 affin oil. 



(b) Harvey cultivated S. icterohaemor- 

 rhagiae on a medium prepared as 

 follows: 



(1) Dissolve 0.88 g. NaCl, 0.025 g. 

 KCl, 0.02 g. CaCl2 and 0.015 g. 

 NasCOa in 100.0 cc. distilled water. 



(2) Mix 1.5 parts of sterile rabbit serum 

 at 45°C., 4.5 parts (1) at 45°C. and 

 1 part of melted infusion agar at 

 45°C. (See variant (v) medium 

 1661.) 



(3) Cover with a thin layer of paraffin 

 oil. 



References: Noguchi (1918 p. 606), Harvey 

 (1921-22 p. 80). 



