CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



703 



2177. Wroblewski's Suprarenal Capsule 

 Agar 

 Constituents : 



1. Water 500.0 cc. 



2. Suprarenal capsule (ox) 100.0 g. 



3. Agar 7.5 g. 



4. Glycerol 18.0 g. 



5. NaCl 3.0 g. 



Preparation : 



(1) Remove the suprarenal capsule from 

 an ox immediately after death. 



(2) Chop to small bits and rub to a pulp. 



(3) To 100.0 g. of (2) add 200.0 cc. of 

 water. 



(4) Make (3) alkaline with soda and boil 

 for 2 hours. (Filtration not specified) . 



(5) Dissolve 7.5 g. agar, 18.0 g. glycerol 

 and 3.0 g. of NaCl in 300.0 cc. of water. 



(6) Mix equal quantities of (5) and (4). 



(7) Sterilize (method not given). 



(8) Filter two or three times to clarify. 

 Sterilization : Sterilize the filtrate (method 



not given) and store at 36°C. for several 

 days to test sterility. 



Use: General culture medium. Author re- 

 ported that the medium suported the 

 growth of some organisms better than 

 others. 



Reference: Wr6blewski (1896 p. 528). 



2178. Picker's Brain Agar 

 Constituents : 



1. Distilled water 1000.0 cc. 



2. Brain, beef 500.0 g. 



3. Agar (2.5%) 12-5 g. 



4. Glycerol 30.0 g. 



Preparation : 



(1) Pass 500.0 g. fresh brain thru a 

 meat chopping machine. 



(2) Add an equal weight (500.0 cc.) of 

 distilled water. 



(3) Stir and heat slowly to a boil. 



(4) Boil slowly for 15 minutes. 



(5) Filter thru a filtering cloth until the 

 filtrate assumes a pulpy character. 

 Press the coagulated mass free 

 from any liquid. 



(6) Prepare a 2.5% agar solution in 

 distilled water and filter. 



(7) Mix equal parts of (6) and sterile (5). 



(8) Add 3.0% glycerol. 



(9) Tube sterilized (8). 



(10) Cool to 45°C. Mix well and solidify 

 quickly in a slanted position. 



Sterilization: Sterilize the filtrate from (5) 

 by steaming for two hours. Sterilization 

 of the agar not specified. Sterilize (8) 

 by heating in steam for 30 minutes. 



Use: Cultivation of tubercle bacilli. 



Variants : 



(a) The author reported that human, 

 sheep, beef, calf and horse brain were 

 used, but all gave the same results. 



(b) Abel boiled the brain infusion for 45 

 minutes instead of 15 minutes as in 

 step (4) above. 



References: Ficker (1900 p. 593), Abel 

 (1912 p. 90), Kolle and Wasserman (1912 

 p. 410), Klimmer (1923 p. 224). 



2179. Picker's Spleen Agar 



Same as medium 2178 but substituting 

 beef spleen for brain. 



2180. Duval's Turtle Agar 



Constituents : 



1. Distilled water 500.0 cc. 



2. Agar 20.0 g. 



3. NaCl 3.0 g. 



4. Glycerol 30.0 cc. 



5. Muscle, turtle 500.0 g. 



Preparation : 



(1) Dissolve 2, 3 and 4 in 1. 



(2) Clarify and tube in 10.0 cc. lots. 



(3) Cut 500.0 g. turtle muscle into 500.0 

 cc. distilled water. 



(4) Keep in the ice chest 48 hours and 

 filter thru gauze to remove the tissue. 



(5) Add 5.0 cc. of sterile (4) to each tube 

 of sterile (2) that has been melted 

 and cooled to 42°C. 



(6) Agitate thoroly and allow to solidify 

 in the slanted position. 



Sterilization: Sterilize (2) in the usual 

 manner, method not given. Sterilize (4) 

 by filterting thru a Berkefeld filter. 



Use: Cultivation of B. leprae. Author 

 reported that growth was moist and 

 orange yellow in color. Growth reached 

 a maximum in 48 to 64 hours. 



Reference: Duval (1911 p. 370). 



2181. Loeffler's Malachite Green Nutrose 

 Agar 



Constituents : 



1. Water 2000.0 cc. 



2. Beef 1 lb. 



3. Agar 30.0 g. 



