592 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



zone will appear about the colony on the 

 plate flooded with acid bichloride of 

 mercury. The plate flooded with tannic 

 acid presents a white to clear zone about 

 the colony depending on the vigor with 

 which the organism attacked the gelatin. 

 Reference: Frazier (1926 p. 302). 



1881. Duval and Lewis' Glucose Blood Agar 



Constituents : 



1. Infusion agar 



2. Glucose (0.5 to 1.0%)... 5.0 to 10.0 g. 



3. Blood, defibrinated 



(2.0 to 5.0%) 20.0 to 50.0 cc. 



Preparation : 



(1) Prepare beef infusion agar. 



(2) Adjust reaction of (1) but with one 

 change if possible between 0.4 and 

 0.8% acid to phenolphthalein (cold 

 titration). 



(3) Add glucose from 0.5 to 1.0%, also 

 from 2.0 to 5.0% "fresh" defibrinated 

 blood. 



Sterilization: Not specified. 



Use: Cultivation of pneumococci. Similar 



media were employed to cultivate a large 



variety of organisms. 

 Variants : 



(a) Novy (Hagemeister) cultivated 

 pathogenic trypanosomes on a 

 medium prepared as follows: 



(1) Prepare an extract of 125.0 g. beef 

 (or horse meat) in one liter of water. 



(2) Dissolve 20.0 g. agar, 20.0 g. peptone 

 and 5.0 g. NaCl in (1). 



(3) Neutralize to litmus by the addition 

 of N/1 NaOH. 



(4) Add 5.0 cc. of N/1 NaOH per liter. 



(5) Dissolve 2.0% glucose in (4). 



(6) Distribute in 3.0 or 4.0 cc. lots in 

 test tubes. 



(7) Add a double or triple amount of 

 sterile defibrinated blood to each 

 tube of sterile agar, melted and 

 cooled to 60°C. Mix well. 



(8) Solidify in a slanted position. 



(9) After inoculation seal the tubes 

 with paraffin. 



(b) Bushnell used the following medium 

 for the isolation of meningococci. 

 Author reported that by reflected 

 light, meningococci colonies appeared 

 raised, somewhat convex, glistening. 



moist, translucent and somewhat 

 brownish, with uniform slightly 

 granular structure. By transmitted 

 light they were translucent, homo- 

 geneous and showed no zone of 

 hemolysis. Best results were ob- 

 tained by using blood as fresh as 

 possible. Laking was not necessary. 

 If plates are warmed before use, more 

 rapid and abundant growth of menin- 

 gococci was obtained. 



(1) Exact method of preparation or 

 exact composition of 2.0% meat 

 infusion agar not given. 



(2) Dissolve 1.0 peptone in (1). 



(3) Adjust to exactly neutral to 

 phenolphthalein. 



(4) Distribute in 300.0 cc. lots in 500.0 

 cc. flasks. 



(5) Autoclave at 15 pounds for 30 

 minutes. 



(6) Sterilize 5.0 g. glucose in 50.0% 

 solution. 



(7) Cool the melted agar to 45°C. 

 and add the slightly warmed glucose 

 solution and 50.0 cc. sterile defibri- 

 nated sheep's (or whole) blood. 



(8) Mix by a whirling motion avoiding 

 air bubbles. 



(9) Pour in sterile plates. 



(c) Hertig and Wolbach cultivated Rick- 

 ettsia melpohagi on a medium pre- 

 pared as follows : 



(1) Prepare beef bouillon in the usual 

 manner (exact procedure not 

 given). 



(2) Dissolve 20.0 g. glucose and 25.0 g. 

 agar in (1). 



(3) Method of sterilization not given. 



(4) To melted (3) at 100°C. add sterile 

 defibrinated rabbit blood in equal 

 volume to double volume. 



(5) Slant the tubes and seal with rubber 

 caps to prevent evaporation. 



References: Duval and Lewis (1905 p. 

 474), Hagemeister (1914 p. 228), Bushnell 

 (1918 p. 2), Hertig and Wolbach (1924 p. 

 337). 



1S82. Torrey's Glucose Blood Agar 

 Constituents: 



1. Distilled water 1000.0 cc, 



2. Liver, beef 500.0 g. 



