CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



269 



of a mixture of immune and plain 

 serum. 



(b) Lucke and Rea cultivated strepto- 

 cocci on a similar medium. They 

 reported that typical streptococci 

 gave a heavy white sediment consist- 

 ing of albumin. Their medium was 

 prepared as follows: 



(1) Prepare a meat infusion broth (2.0% 

 peptone). 



(2) Boil 1000.0 CO. of (1) to 80.0% of its 

 original volume and adjust to 0.4+. 



(3) Tube and sterilize for 3 days in 

 steam. 



(4) Obtain horse serum aseptically and 

 heat to 60°C. for 1 hour in a water 

 bath to destroy antihemolysins. 



(5) Add 2.0 cc. of (4) to each 8.0 cc. of 

 (3). 



(c) Stevens, Brady and West studied the 

 relation between virulence and hemol- 

 ysin. They reported that a strain 

 of streptococcus whose virulence had 

 been increased did not produce 

 greater concentrations of hemolysin. 

 Their medium was prepared as 

 follows : 



(1) Prepare a meat infusion broth with 

 2.0% peptone. 



(2) Titrate (1) so that it is pH = 7.6 

 after sterilization (method not 

 mentioned). 



(3) Distribute into 250.0 cc. Pyrex 

 flasks in 80.0 cc. lots. 



(4) Add 20.0 cc. of fresh horse serum to 

 each flask. 



(5) Inactivate the contents at 56°C. on 

 3 successive days, and store on ice 

 until ready for use. 



(d) Harvey mixed 2 parts serum with 

 1 part infusion broth, (see variant 

 (bb) 665). The mixture was steri- 

 lized for 60 minutes at 57°C. on each 

 of 2 successive days. 



References: Stryker (1916 p. 50), Lucke 

 and Rea (1919 p. 535), Stevens (1921 

 p. 224), Harvey (1921-22 p. 82). 



869. Foster's Serum Infusion Broth 



Constituents: 



1. Beef infusion broth 1000.0 cc. 



2. Glucose 10.0 g. 



3. Horse serum broth 5.0 g. 



Preparation : 



(1) Prepare beef infusion broth. 



(2) Adjust (1) to pH = 6.3. 



(3) Prepare (2) 10.0% glucose solution. 



(4) Mix (1) and (2). 



(5) Add sterile 3 to sterile (4) in suflB- 

 cient quantity to give a 1.0% con- 

 centration of glucose. 



Sterilization: Sterilize (3) method not 

 given. Sterilize (4) at 15 pounds pressure 

 for 15 minutes. 



Use: To study protein and carbohydrate 

 metabolism of Streptococcus hemolyticus . 

 General culture medium. 



Variants: Harvey cultivated meningococci 

 on a medium containing 1 part unheated 

 clear sterile horse serum to 20 parts in- 

 fusion broth (see variant (bb) 770) to 

 which was added 1.0% glucose. 



References: Foster (1921 p. 222), Harvey 

 (1921-22 p. 80). 



870. Beach and Easting's Serum Infusion 

 and Extract Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Lean beef 450.0 g. 



3. Peptone (Difco) 15.0 g. 



4. K2HPO4 7.5 g. 



5. Liebig's beef extract 4.0 g. 



6. Glycerol 55.0 cc. 



7. Aminoid peptone from beef 

 (Arlco) 5.0 g. 



8. Serum (sterile blood) 100.0 cc. 



Preparation : 



(1) Extract 450.0 g. lean beef with 

 1000.0 cc. water for 3 hours. Bring 

 the water to 45°C. After an hour 

 raise the temperature to 50°C. and 

 during the third hour to 55°C. Stir 

 frequently. 



(2) Remove the major portion of the 

 liquid from the meat. 



(3) Heat meat and remaining liquid to 

 100°C. This causes the meat par- 

 ticles to shrink giving the maximum 

 amount of liquid. 



(4) To the liquid (2) plus that of (3) 

 add 10.0 g. peptone, 5.0 g. K2HPO4, 

 2.0 g. Liebig's beef extract and 

 50.0 cc. of glycerol. 



(5) Adjust the reaction to pH of 7.5 

 to 7.8. 



