602 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(6) Method of sterilization not given. 



(7) Medium is a faint yellow in color 

 and transparent. 



(b) Harvey prepared a medium as 

 follows: 



(1) Add 10.0 cc. of blood to 1.0 cc. 

 sterile 10.0% sodium citrate. 



(2) Add 1.0 cc. of (1) to a test tube of 

 melted infusion agar at 45°C. 

 (see variant (v) medium 1661 for 

 preparation) . 



(3) Rotate the test tubes between the 

 hands to distribute the blood thru 

 the agar. 



(c) Harvey gave the following method of 



preparation: 



(1) Add 10.0 cc. of pigeon's blood to 

 90.0 cc. of a 1.0% citrated 0.85% 

 sterile NaCl solution. 



(2) Heat 30 minutes at 65 to 70°C. 



(3) Filter thru thick filter paper. 



(4) Mix 1.0 cc. of the filtrate from (3) 

 with 10.0 cc. of melted infusion, 

 cooled to 45°C. with a reaction 

 slightly alkaline to litmus. (See 

 variant (v) medium 1661 for prep- 

 aration of agar.) 



(d) Soparkar (Harvey) prepared a similar 

 medium as follows: 



(1) Mix 10.0 cc. of fresh human blood 

 with 40.0 cc. of a 0.5% citrated 

 0.85% NaCl solution. 



(2) Heat in a water bath 30 minutes at 

 64 to 68°C. 



(3) Filter thru sterilized paper. 



(4) Mix 1.0 cc. of the filtrate from (3) 

 with 10.0 cc. of melted infusion, 

 cooled to 45°C. with a reaction 

 slightly alkaline to litmus. (See 

 variant (v) medium 1661 for prep- 

 aration of agar.) 



References: Liston (1918-19 p. 418), 

 Malone (1919-20 p. 504), Harvey (1921-22 

 p. 77). 



1901. Hirsch and McKinney's Chocolate 

 Agar 



Constituents : 



1. Infusion agar 1000.0 cc. 



2. Blood(citrated horse 



1.0 to 2.0%) 10.0 to 20.0 cc. 



Preparation : 



(1) Prepare beef infusion agar. 



(2) Adjust to a reaction of 0.2%. 



(3) Draw horse blood into flasks con- 

 taining sterile sodium citrate solution 

 to make the final concentration 1.5%. 



(4) After corpuscles have settled, the 

 plasma is removed and sterile distilled 

 water added to make up the original 

 volume of the blood. 



(5) Heat sterile (2) to 90°C. and add 

 sterile laked blood up to 1.0 to 2.0%. 



Sterilization: Method not specified. 

 Use: Isolate influenza bacilli. 

 Variants: Harvey prepared a similar 

 medium as follows: 



(1) Mix 5 parts infusion agar at 45°C. (see 

 variant (v) medium 1661 for prep- 

 aration) with 2/lOths part of citrated 

 blood at 45°C. (one part 10.0% 

 citrate in 0.95% sterile salt solution 

 to ten parts blood). 



(2) Place in a water bath at 80°C. until 

 the agar has become chocolate in 

 color. 



(3) Slope. 



References: Hirsch and McKinney (1919 

 p. 605), Harvey (1921-22 p. 77). 



1902. Harvey's Glucose Blood Agar 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Ox heart 1000.0 g. 



3. Peptone 10.0 g. 



4. NaoHP04 10.0 g. 



5. Agar 30.0 g. 



6. Glucose 25.0 g. 



7. Citrated blood 

 Preparation : 



(1) Mix 1000.0 cc. distilled water with 

 1000.0 g. ox heart. 



(2) Heat the mixture 20 minutes at a 

 temperature not exceeding 50°C., 

 with constant stirring. 



(3) Raise the temperature to boiling 

 point. 



(4) Boil 10 minutes. 



(5) Pour the mixture on to a wet, thick, 

 clean cloth. 



(6) Collect the fluid which drains thru 

 the cloth together with that obtained 

 by squeezing the meat in the cloth. 



(7) Add peptone 1.0%; di-sodium phos- 

 phate 1.0%, prepared agar (see 

 medium 1401) 3.0%. 



(8) Steam gently 2^ hours to dissolve the 

 agar. 



