CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



589 



Sterilization: Sterilize (7) by heating in the 



autoclave at 120°C. for 20 minutes. 

 Use: Cultivation of gonococci. 

 Reference: Leboeuf (1924 p. 768). 



1873. Besson's Gelatin Infusion Agar 



Constituents : 



1. Infusion broth 1000.0 cc. 



2. Gelatin 80.0 g. 



3. Agar 5.0 g. 



Preparation : 



(1) Dissolve 80.0 g. of gelatin in 1000.0 

 cc. of infusion broth. 



(2) Neutralize. 



(3) Dissolve 5.0 g. of agar in (2). 



(4) Allow to cool to 55 or 50°C. 



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

 of water and add to (4) . 



(6) Mix well. 



(7) Autoclave at 115°C. for 20 minutes. 



(8) Filter thru a moistened Chardin filter 

 paper using a hot water funnel. 



(9) Tube. 



Sterilization: Sterilize at 115° for 20 

 minutes. 



Use: General culture medium. 



Variants: Author reported that 50.0 g. of 

 gelatin and 8.0 g. of agar may be used 

 instead of the amounts indicated. 



Reference: Besson (1920 p. 43). 



1874. Frothingham's Gelatin Infusion Agar 



Constituents: 



1. Infusion broth 1000.0 cc. 



2. Agar 7.5 g. 



3. Gelatin 50.0 g. 



Preparation: 



(1) Dissolve 7.5 g. agar in 1000.0 cc. of 

 infusion broth. (A 0.5% solution of 

 Liebig's meat extract solution may 

 be used.) 



(2) Dissolve 50.0 g. of gelatin in (1). 

 Sterilization: Method not given. 



Use : General culture medium and to main- 

 tain stock cultures of streptococci, 

 pneumococci and other pathogens. North 

 reported that the addition of 1.0% glucose 

 made the growth more favorable at times, 

 but reduced vitality. 



Variants : 



(a) North gave the following method of 

 preparation: 



(1) Extract one pound of lean chopped 

 beef or veal in 500.0 cc. of water 

 for 18 hours. 



(2) Add and dissolve in this infusion, 

 made up to 1 liter with water, 

 10.0 g. agar, 20.0 g. gelatin, 20.0 g. 

 Witte peptone and 5.0 g. salt. 



(3) Adjust to the neutral point using 

 phenolphthalein as an indicator. 



(4) Tube. 



(b) Dopter and Sacqu6p^e prepared the 

 medium as follows: 



(1) Add 1000.0 cc. of water to 500.0 

 g. of finely chopped fat and tendon 

 free beef. 



(2) Allow to stand in the ice box for 

 12 hours, or heat at 50 to 55° for 

 30 minutes. 



(3) Heat slowly to boiling. 



(4) Boil slowly for 10 minutes, stirring 

 constantly. 



(5) Strain thru a cloth. 



(6) Dissolve 80.0 g. gelatin, 5.0 g. 

 NaCl and 20.0 g. peptone in (5). 



(7) Neutralize. 



(8) Dissolve 5.0 g. agar in (7). 



(9) Clarify by the addition of egg 

 white. 



(10) Autoclave. 



(11) Filter. 



(12) Distribute 



(13) Sterilize (method not given). 



(c) Guarniari (Abbott) gave the follow- 

 ing method of preparation : 



(1) Dissolve 3.0 to 4.0 g. agar sep- 

 arately in about 100.0 cc. of water 

 while the other materials are being 

 prepared. 



(2) Prepare meat infusion. 



(3) Dissolve 5.0 g. NaCl, 25.0 to 30.0 

 g. peptone and 40.0 to 60.0 g. 

 gelatin in (2) . 



(4) Mix (3) and (1). 



(5) Sterilization not specified. 



(d) Harvey solidified infusion broth, see 

 variant (bb) medium 779 by the 

 addition of 10.0 or 12.0%, gelatin 

 and 0.5% agar. 



(e) Park, Williams and Krumwiede gave 

 the following method of preparation: 



(1) Prepare a meat infusion using 1.0 

 pound of meat to 500.0 cc. of water 

 (Method not given). 



