582 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Use: To determine food requirements for 

 Bacillus pyogenes. Authors reported 

 that this medium supported the growth 

 of Bacillus pyogenes nearly as well as did 

 blood agar. 



Variants: The authors prepared a similar 

 medium as follows: 



(1) Prepare veal infusion agar. 



(2) Defibrinate blood and wash repeat- 

 edly with sterile physiological salt 

 solution. 



(3) Lake the washed corpuscles with 

 distilled water and remove the cor- 

 puscle stroma by centrifugation. 

 (Centrifuge the laked blood cor- 

 puscles until the supernatant hemo- 

 globin solution no longer gives a 

 clouding reaction with salt.) 



(4) Wash the corpuscle stroma repeatedly 

 in sterile distilled water until no 

 visible trace of hemoglobin remains. 



(5) Add the stroma suspension to melted 

 agar (amount not given) and pour into 

 plates. 



This medium was inferior to blood cell 

 medium above. 

 Reference: Brown and Orcutt (1920 pp. 

 222-223). 



1860. Behren's Blood Cell Agar 



Constituents : 



1. Water 1000.0 cc. 



2. Chopped beef 125.0 g. 



3. Peptone 20.0 g. 



4. NaCl 5.0 g. 



5. CaCl, 0.1 g. 



6. N/1 NaaCOs solution 10.0 cc. 



7. Agar 20.0 g. 



8. Red blood cells from 2000.0 

 cc. Rabbit blood 



Preparation : 



(1) Digest chopped beef in 250.0 cc. 

 water over night in the cold, or for 

 one hour at 55°C. 



(2) Strain (1), boil the infusion and 

 filter 



(3) Dialyze the filtrate in a large col- 

 lodium sac against running distilled 

 water for 24 to 48 hours. 



(4) Dilute (3) to 1000.0 cc. 



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



(6) Distribute in test tubes in 1.0 cc. 

 lots. 



(7) Centrifuge defibrinated rabbit blood 

 and draw off serum. 



(8) Dilute the red blood cells, freed 

 from serum, with 0.5% NaCl solution 

 to the original blood volume. 



(9) Shortly before use melt the desired 

 tubes of sterile agar, cool to 60°C. 

 and add two volumes of (8) to each 

 tube. 



(10) Mix well and solidify in slanting 

 position. 



Sterilization: Sterilize (6) in autoclave by 

 heating to 105-108° for 15 minutes. 



Use: Special culture medium for Trypano- 

 soma Brucei. 



Reference: Behrens (1914 p. 29). 



1861. Sacquepee and Delater's Egg Albumin 

 Infusion Agar 



Constituents : 



1. Distilled water 



2. Egg white 



3. Meat 500.0 g. 



4. Peptone 10.0 g. 



5. NaCl 5.0 g. 



6. Agar 30.0 g. 



Preparation : 



(1) Place the whites of two eggs in a 

 graduated glass cylinder. 



(2) Shake constantly and add little by 

 little three times the volume of 

 distilled water. 



(3) Make alkaline by the addition of 

 about 0.5 cc. of a 10.0% soda solution 

 for each 100.0 cc. of (2). 



(4) Shake well. 



(5) Heat in the autoclave at 115° for 15 

 minutes. 



(6) Macerate 500.0 g. of meat in a liter of 

 water. 



(7) Allow to stand in the cold for 24 

 hours and then bring slowly to boil. 



(8) Filter. 



(9) Dissolve 10.0 g. peptone, 5.0 g. 

 NaCl in (8). 



(10) Dissolve 30.0 g. agar in (9). 



(11) Exactly neutralize to litmus and 

 then add 9.0 cc. of a 10.0% soda 

 solution. 



(12) Clarify by the addition of an egg 

 white to the agar at 60°C. Heat in 

 the autoclave and filter thru paper. 



(13) Mix (5) and (12). Approximately 

 one part (5) to five parts (12). 



(14) Distribute as desired. 

 Sterilization: Sterilize for 15 minutes at 



112°C. 



