CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



599 



(2) Strain. 



(3) Add 20.0 g. of peptone, 5.0 g. NaCl, 

 10.0 cc. normal NasCOs solution 

 and 20.0 to 25.0 g. agar. 



(4) Prepare as for nutrient agar and 

 sterilize (method not given). 



(5) To one part melted and cooled to 

 60°C. (4) add two volumes of 

 defibrinated rabbit blood under 

 aseptic conditions. 



(6) Slant. 



(7) Cover the cotton plugs with 

 melted paraffin or use rubber 

 stoppers to seal the tubes. 



(u) Brown (Stitt) differentiated strepto- 

 cocci on the following medium: 



(1) Prepare an agar base using 500.0 g. 

 veal, 5.0 g. NaCl, 15.0 g. agar and 

 10.0 g. peptone in the same manner 

 as for the preparation of ordinary 

 nutrient agar. 



(2) Reaction of (1) to be between 

 +0.8 and +1.2. 



(3) Tube in 12.0 cc. amounts. 



(4) To prepare the blood agar melt the 

 tubes of agar and place in a water 

 bath at 45 °C. for 15 minutes. 



(5) Add 0.6 cc. defibrinated blood to 

 each tube and mix thoroly. 



(6) Inoculate. 



(7) Pour into plates. 



(v) Beilin (Klimmer) cultivated influenza 

 bacilli on a medium prepared as 

 follows: 



(1) Heat3.0%infusionagar with 1.0% 

 peptone to boiling. 



(2) Add 15.0% of sterile fresh defibri- 

 nated horse blood to boiling (1). 



(3) Remove from the flame. 



(4) Cool to 50 to 60°C. and mi.x 

 thoroughly. 



(5) Pour in plates. 



(w) Schottmuller (Klimmer) cultivated 

 gonococci, meningococci and strepto- 

 cocci on a medium prepared by mix- 

 ing five parts liquified agar at 45°C. 

 with two parts sterile defibrinated 

 human blood. 



(.\) Novy and MacNeal (Park, Williams 

 and Krumwiede) cultivated flagellate, 

 trypanosomes and leishmania on a 

 medium prepared as follows: 

 (1) Mix equal parts of nutrient agar 



and fresh defibrinated blood (rab- 

 bit or rat). 



(2) Allow the medium to stiffen 

 slanted so more of the water of 

 condensation will settle at the 

 bottom. 



(3) Sterilization not specified. 



(4) Inoculate with infected blood. 



(y) Soule cultivated Trypanosoma Lewisi 

 and Leishmania tropica on a medium 

 prepared as follows: 



(1) Infuse one part finely chopped 

 choice lean beef in two parts 

 distilled water in the ice box for 

 24 hours. 



(2) Strain. 



(3) Heat the juice to coagulate the 

 proteins. Filter. 



(4) Dissolve 1.0% Witte's peptone and 

 0.5% Kahlbaum's NaCl in the 

 filtered infusion. 



(5) Adjust to pH = 7.4. 



(6) Boil and filter. 



(7) Dissolve 2.0%, agar in (6). 



(8) Tube or distribute in flasks. 



(9) Cool sterile melted tubes of (8) 

 to 50°C. and add an equal volume 

 of sterile defibrinated rabbit 

 blood. 



(10) Mix well and slant, 

 ai) Autoclave (8) at 110°C. for 20 

 minutes. Blood collected and de- 

 fibrinated under aseptic con- 

 ditions. 

 References: Smedley (1905 p. 28), Gurd 

 (1908 p. 303), Duval (1910 p. 653), Hage- 

 meister (1914 p. 228), Warden (1915 p. 

 428), Holman (1916 p. 381), Kohlman 

 (1919 p. 574), Bernstein and Lowe (1919 

 p. 78), Wahl, White and Lyall (1919 p. 

 420), Yoshida (1920 p. 361), Bell (1920 p. 

 464), Anderson and Schultz (1921 p. 

 656), Harvey (1921-22 pp. 73, 74, 76), 

 Jones (1922 p. 363), Pitfield (1922 p. 119), 

 Stitt (1923 pp. 43, 44, 52), Klimmer (1923 

 p. 226), Park, Williams and Krumwiede 

 (1924 p. 133), Soule (1925 p. 248). 



1894. Harvey's Peptic Blood Digest Agar 



Constituents: 



1. Infusion agar 



2. NaCl (0.85% solution) 150.0 cc. 



3. HCl 6.0 cc. 



