CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



635 



(7) Incubate over night at 37°C. 



(8) To 100.0 cc. of sterile melted and 

 cooled to 50°C. agar containing 0.5 

 to 1.0% lactose, add 3.0 cc. of (7), 

 1.0 cc. of a 0.1% crystal violet solu- 

 tion and 12.0 cc. of Kubel and Tie- 

 mann's litmus solution. 



(9) Pour into plates. 



(10) Incubate over night at 37° to drive 

 off the ether. 

 Sterilization: Method of sterilization not 



given. 

 Use: Cultivation of colon-typhoid group. 

 Reference: Wilson and Darling (1918 



p. 105). 



1986. Stitt's Glycerol Blood Agar (Chocolate 

 Agar) 



Constituents: 



1. Glycerol agar 100.0 cc. 



2. Citrated blood 

 Preparation : 



(1) Prepare glycerol agar. 



(2) Melt (1) and while the tubes are at 

 a temperature of about 90 °C. add 

 about 4.0 or 5.0% citrated blood 

 (0.5 cc. to 10.0 cc. of agar in a tube). 



(3) Mix thoroly avoiding bubbles. 



(4) Pour in sterile plates. 

 Sterilization: Not specified. 



Use: Cultivation of Pfeiffer's bacilli. 

 Author reported that this medium was 

 less satisfactory for isolation of Pfeiffer's 

 bacillus than plain blood agar (due to its 

 brown color). 



Variants: Stitt employed nutrient agar 

 instead of glycerol agar. 



Reference: Stitt (1923 p. 43). 



1987. Besson's Citrated Blood Agar 



Constituents : 



1. Nutrient agar. 



2. Blood, citrated. 

 Preparation : 



(1) Collect 400.0 cc. beef blood in a 

 sterile flask containing citrate solu- 

 tion and 0.1 cc. of commercial formol. 



(2) Shake thoroly. 



(3) Store for several days at room tem- 

 perature. 



(4) Dilute (3) with three volumes dE 

 sterile physiological salt solution. 



(5) Add one part (4) to 15 parts sterile 

 agar. 



Sterilization: Method of sterilization of 

 agar not given. 



Use: Cultivation of pneumococci, menin- 

 gococci and gonococci. 



Variants : 



(a) Stitt prepared an opaque medium 

 as follows: 



(1) Prepare 2.0 to 3.0% nutrient agar. 



(2) The reaction of (1) should be -0.3 

 to phenolphthalein. 



(3) To 100.0 cc. of (2) add 20.0 cc. of a 

 mixture of equal parts blood and 

 sodium citrate solution (1.0% in 

 normal saline). 



(4) Pour into 10 plates. 



(b) Stitt prepared a transparent medium 

 as follows : 



(1) Prepare nutrient agar. 



(2) Mix equal parts 1.0% sodium ci- 

 trate solution and blood. 



(3) Add 20 to 30 drops of (2) to 100.0 cc. 

 of (1). 



(4) Pour into 10 plates. 

 References: Besson (1920 p. 31), Stitt 



(1923 p. 44). 



1988. Bezangon, Griffon and LeSourd's 

 Blood Agar 



Constituents: 



1. Nutrient agar. 



2. Blood, rabbit. 

 Preparation : 



(1) Prepare nutrient agar. 



(2) Cool sterile melted agar tubes to 50°C. 



(3) Add fresh rabbit blood, drawn under 

 aseptic conditions. (Amount not 

 given.) 



(4) Mix the blood and agar. 



(5) Slant and cool. 



Sterilization: Method of sterilization of (2) 

 not given. 



Use: Cultivation of the Ducrey bacillus, 

 A large variety of organisms were culti- 

 vated on the same or similar media by a 

 number of investigators. 



Variants : 



(a) Thoinot and Masselin streaked agar 

 slants or agar plates with whole or 

 defibrinated blood, using a sterile 

 pipette. 



(b) Davis specified that one volume of 

 freshly drawn rabbit blood be added 

 to two volumes of 2.0% nutrient agar. 



(c) Czaplewski specified the use of pigeon 



