CULTURE MEDIA FOR CULTIVATION' OF MICROORGANISMS 



605 



6. Agar (2.5%) 25.0 g. 



7. Blood, human (0.5 to 



2.5%) 5.0 to 25.0 cc. 



Preparation : 



(1) Remove all connective tissue from 

 beef testicles and put thru a meat 

 grinder. 



(2) Cover (1) with twice its weight of 

 distilled water. 



(3) Infuse on ice over night. 



(4) Heat to 50°C. in a double boiler on 

 the following morning. 



(5) Allow to stand for an hour and then 

 raise to the boiling point. 



(6) Allow to stand for another hour to 

 permit the settling of the solid 

 particles. 



(7) Decant the liquor. 



(8) Add 2.0% peptone, 0.5% glucose, 

 0.2 to 0.3% NaHaPOi and 2.5% 

 granular agar. 



(9) Heat over a free flame until solution 

 is complete, stirring constantly. 



(10) Adjust the reaction to pH = 7.4 to 

 7.8, using phenol red as an indicator. 



(11) Tube. 



(12) Check the reaction after steriliza- 

 tion. 



(13) Add human blood in the proportion 

 of 0.5 to 2.5%, to the tubes while 

 they are still liquid (just before the 

 agar solidifies). 



Sterilization: Sterilize (1) by heating at 15 

 pounds pressure for 15 minutes. 



Use: Cultivation of gonococci. 



Variants: The author gave the following 

 variants: 



(a) Defibrinated rabbits blood (1.0 to 

 5.0%) may be added instead of 

 human blood. 



(b) The blood may be added before 

 sterilization. This gives a "choc- 

 olate" colored medium. 



Reference: Stitt (1923 p. 46). 



1908. Cutler's Blood Clot Infusion Agar 

 (Klinuner) 



Constituents : 



1. Water 1000.0 cc. 



2. Blood clot 500.0 g. 



3. Peptone (1.0%) 10.0 g. 



4. NaCl (0.5%) 5.0 g. 



5. Blood 



Preparation : 



(1) Boil 500.0 g. human blood clot with 

 a liter of water. 



(2) Filter. 



(3) Add 0.5%, NaCl and 1.0% peptone to 

 the filtrate. 



(4) Distribute as desired (tube). 



(5) Add several drops of blood to each 

 tube of sterile (4) when ready for use- 

 Sterilization : Method of sterilization of (4) 



not given. 



Use: Cultivation of amoeba. 



Variants: Klimmer infused placenta in- 

 stead of blood clot. 



Reference: Klimmer (1923 p. 229). 



1909. Harvey's Trypsinized Blood Agar 



Constituents : 



1. Infusion agar. 



2. Trypsin. 



3. Blood, human. 

 Preparation : 



(1) Add 0.25 g. of anhydrous Na2C03 to 

 100.0 cc. of a 5.0%o trypsinized 0.85% 

 sterile salt solution under aseptic 

 conditions. 



(2) Distribute in quantities of 10.0 cc. 

 in sterile test tubes. 



(3) Add to each test tube 2.0 cc. sterile 

 human blood. 



(4) Incubate 8 days or longer with occa- 

 sional shaking. 



(5) Add one part of (4) at 45''C. to nine 

 parts of melted infusion agar at 45 ^C. 

 (see variant (v) medium 1661 for 

 preparation). 



(6) Distribute into test tubes or plates. 

 Sterilization: Method not given. 



Use: Cultivation of B. influenzae. 

 Reference: Harvey (1921-22 p. 77). 



1910. Harvey's Hydrolyzed Blood Agar 

 Constituents : 



1. Infusion agar 350.0 cc. 



2. Water 50.0 cc. 



3. Blood 10.0 cc. 



4. H2SO4 (normal) 10.0 cc. 



Preparation : 



(1) Mix one part of blood, one part nor- 

 mal H2SO4 with 5 parts water. 



(2) Boil. 



(3) Store for use. 



(4) Make faintly alkaline to litmus at the 

 time of use. 



