CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



637 



1989. Fleming's Brilliant Green Blood Agar 



Constituents : 



1. Water 9.0 cc. 



2. Blood 1.0 cc. 



3. Nutrient agar 



4. Brilliant Green 1 to 500,000 



Preparation : 



(1) Boil 1.0 cc. of blood with 9.0 cc. water. 

 (Time not specified.) 



(2) Add some of the clear supernatant 

 liquid of (1) to nutrient agar. 

 (Amount of liquid not specified). 



(3) Add brilliant green in the ratio of 1 

 to 500,000. 



Sterilization: Not specified. 



Use: Isolation of B. influenzae. Author 

 reported that pneumococci, staphylococci 

 and streptococci were inhibited. B. influ- 

 enzae apparently was not inhibited. 



Reference: Fleming (1919 p. 139). 



1990. Hachla and Holobut's Alkaline Blood 



Agar 

 Constituents : 



1. Nutrient agar 700.0 cc. 



2. Blood, beef 200.0 cc. 



3. KOH N/1 100.0 to 150.0 cc. 



Preparation : 



(1) Prepare neutral nutrient agar. 



(2) Mix 200.0 cc. of beef blood with 

 100.0 cc. or 150.0 cc. of normal KOH. 



(3) Steam (2) for 30 minutes. 



(4) Mix 7 parts liquid (1) with 3 parts (3). 



(5) Pour into sterile plates. 



(6) Dry the plates for 24 hours at 37°C. 

 and then for 24 more hours at room 

 temperature. 



Sterilization : Method not given. 



Use: Enrichment medium for cholera vi- 

 brio. Authors reported that better re- 

 sults were obtained using 1 part blood to 

 0.75 normal KOH solution. Hog and 

 horse blood gave as good results as did 

 beef blood. 



Reference: Hachla and Holobut (1909 

 p. 299). 



1991. Matthews' Trypsinized Blood Agar 

 Constituents: 



1. Bouillon 4.75 cc. 



2. Trypsin (Allen and Han- 

 bury's) 0.25 cc. 



3. Blood 1.0 cc. 



4. Nutrient agar 30.0 cc. 



Preparation : 



(1) Add 0.25 cc. of Allen and Hanbury's 

 trypsin compound to a series of tubes 

 containing 4.75 cc. of sterile bouillon. 



(2) Incubate for 24 hours. 



(3) Discard tubes showing contami- 

 nation. 



(4) Add 1.0 cc. of blood from a vein 

 puncture to each tube. 



(5) Incubate the mixture for 3 or 4 days. 



(6) Mix 5.0 cc. of the finished product 

 with about 30.0 cc. of nutrient agar, 

 as in the preparation of ordinary 

 blood agar. 



Sterilization: Not specified. 



Use: Isolation of influenza bacillus. 

 Author reported that medium may be 

 prepared in larger quantities. This 

 amount is sufficient for a diagnosis, how- 

 ever. Do not allow the trypsin to act 

 longer than one week. Citrated blood 

 may be used. Pneumococci, streptococci 

 and other gram positive organisms were 

 inhibited. Staphylococci, however, were 

 favored. Gram negative cocci of the 

 catarrhalis group and bacilli of the coli 

 form group were not inhibited, but were 

 not favored as were the influenza. 



Reference: Matthews (1918 p. 104). 



1992. Bieling's Glucose Blood Agar 



Constituents: 



1. Nutrient agar 



2. Agar 150.0 cc. 



3. Blood, horse 50.0 cc. 



4. Glucose 2.0% 3.0 g. 



Preparation : 



(1) Mix one part horse blood with two 

 parts water. 



(2) Prepare nutrient agar. 



(3) Add 2.0% glucose to (2). 



(4) Melt the glucose agar and cool to 

 60°C. 



(5) Mix equal parts (1) and (4). 

 Sterilization: Not specified. 



Use: Differentiation of streptococci and 

 pneumococci. Author reported that 

 hemolytic streptococci did not change the 

 color of the medium. Colonies flat, about 

 2 mm. in diameter, with a central point 

 and a border. Colonies could easily be 

 picked from the agar with a platinum 

 loop aft?r 24 to 48 hours. Streptococcus 

 mitior colonies were about the same size, 



