600 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



4. Blood, defibrinated sheep . . . 50.0 cc. 



5. Pepsin 1.0 g. 



6. NaOH (20.0%) 12.0 cc. 



Preparation : 



(1) Prepare by addition in the order 

 given: 0.85% sterile salt solution 

 150.0 cc; hydrochloric acid 6.0 cc; 

 defibrinated sheep blood 50.0 cc, 

 granulated pepsin B.P. 1.0 g. 



(2) Shake to dissolve. 



(3) Heat in the water bath at 55°C. for 

 2 to 24 hours with occasional shak- 

 ing. (Note: The exact time is im- 

 material.) 



(4) Add 12.0 cc. of 20.0% sodium 

 hydroxide. 



(5) Adjust the reaction by addition of 

 20.0% sodium hydroxide until a 

 sample gives with cresol red indi- 

 cator solution (0.02%) the color of 

 permanganate (corresponding to 

 pH = 7.6). 



(6) Add pure Hydrochloric acid drop by 

 drop until cresol red indicator solu- 

 tion gives practically no change of 

 color but phenol red gives red. 

 Note: Corresponding to pH = 7.0 

 to 7.2. 



(7) Add chloroform to 0.25%. 



(8) Shake to mix. 



(9) Keep in a tightly stoppered bottle 

 till required for use. 



(10) When ready for use, mix at 45°C. 

 100 parts melted infusion agar (see 

 variant (v) medium 1661) with 3.5 

 parts 9. 



Sterilization: Not specified. 



Use: Cultivation of B. tn^uenzae. 



Reference: Harvey (1921-22 p. 100). 



1895. Bailey's Hormone Blood Agar 

 Add 5 parts per 100 of fresh defibrinated 

 human blood to Bailey Hormone Agar 

 (1673). This medium is recommended to 

 preserve stock cultures of pneumococci and 

 streptococci. 



1896. Harvey's Ascitic Fluid Blood Agar 



Constituents: 



1. Infusion broth 30.0 cc. 



2. Infusion agar 600.0 cc. 



3. Blood, defibrinated, sheep... 200.0 cc. 



4. Ascitic fluid 100.0 cc 



5. Maltose 10.0 g. 



Preparation : 



(1) See variant (bb) medium 779 for 

 preparation of infusion broth. 



(2) See variant (v) medium 1661 for 

 preparation of infusion agar. 



(3) Mix 100 parts ascitic fluid, 200.0 cc. of 

 defibrinated sheep blood, 10.0 g. 

 maltose, dissolved in 30.0 cc. (1) with 

 600.0 cc. of (2). 



Sterilization: Not specified. 

 Use: General culture medium. 

 Reference: Harvey (1921-22 p. 84). 



1897. Harvey's Oxalated Blood Agar 



Constituents : 



1. Infusion agar 



2. Blood, ox or sheep 400.0 cc. 



3. NaCl (0.85% solution) 



4. Ammonium oxalate (1.0% 

 solution) 30.0 cc. 



Preparation : 



(1) Collect 400.0 cc. ox or sheep blood 

 at the slaughter house in a sterile 

 flask containing 30.0 cc. of a 1.0% 

 solution of ammonium oxalate and 

 0.5 cc. formalin. 



(2) Mix well. 



(3) Allow to stand 30 minutes. 



(4) Dilute i with 0.85% sterile salt 

 solution. 



(5) Leave for 48 hours. 



(6) Mix one part of oxlated blood with 

 fifteen parts of infusion agar at 45°C. 

 (see variant (v) medium 1661 for 

 preparation). 



Sterilization: Not specified. 

 Use: Cultivation of gonococci, meningo- 

 cocci and pneumococci. 

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



1898. Wolbach, Chapman and Steven's 

 Citrated Blood Agar 



Constituents: 



1. Distilled water 1000.0 cc 



2. Veal 500.0 g. 



3. Glucose 1.5 g. 



4. NaCl 6.0 g. 



5. Agar agar 14.0 g. 



6. Blood, citrated rabbit 

 Preparation : 



(1) Prepare a veal infusion from 500.0 cc 

 of distilled water and 500.0 g. of lean 

 veal. Exact method not given. 



(2) Evaporate (1) to 100.0 cc. 



