CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(2) Leave overnight to allow for sep- 

 aration of clot. 



(3) Add 50.0 cc. N/1 sodium hydroxide 

 per liter. 



(4) Steam 20 minutes. 



(5) Add 1.0 g. agar per liter after 

 making it into a paste or suspen- 

 sion with a little of the alkaline 

 ascitic fluid. 



(6) Steam to dissolve the agar. 



(7) Filter, while hot, thru thick, filter 

 paper by placing filter funnel, 

 stand and receptacle for filtrate in 

 the sterilizer. 



(8) Dissolve 10.0 g. glucose per liter 

 in the hot, filtered, nutrient agar 

 (7). (Add 50.0 cc. glycerol per 

 liter also if desired.) 



(9) Distribute into test tubes. 

 (10) Sterilize. 



(c) Harvey prepared a similar medium 

 from 20.0 g. glycerol, 20.0 cc. of 10.0% 

 NaOH, 20.0 g. agar and 1000.0 cc. of a 

 body fluid (ascitic fluid, pleuritic 

 fluid, hydrocele fluid, ovarian fluid, 

 milk, urine, etc.). 

 References: Kanthack and Stephens (1896 



p. 609), (1896 p. 835), Harvey (1921-22 p. 



83). 



2156. Beck's Glycerol Serum Agar 

 (Klimmer) 



Constituents : 



1. Water 1800.0 cc. 



2. Serum (beef) 200.0 cc. 



3. KHSO4 10.0 g. 



4. MgS04 5.0 g. 



5. Asparagin 4.0 g. 



6. Glycerol 40.0 g. 



7. Agar (3.0%) 60.0 g. 



Preparation : 



(1) Steam 200.0 cc. beef serum (con- 

 taining no chloroform) with 1800.0 cc. 

 of water for 1.0 to 1.5 hours. 



(2) Filter. 



(3) Add 3, 4, 5 and 6 to the filtrate and 

 steam for 2 to 3 hours. 



(4) Filter while hot. 



(5) Add 3.0% fiber agar, and heat until 

 dissolved. 



(6) Tube. 



(7) Reaction should be slightly acid. 

 Sterilization: Not specified. 



Use: Cultivation of tubercle bacilli. 

 Reference: Klimmer (1923 p. 224). 



2157. Ecker's Bile Agar 

 Constituents: 



1. Bile (ox) 1000.0 cc. 



2. Agar 15.0 g. 



Preparation : 



(1) Dissolve 2 in 1 as rapidly as possible 

 by gently boiling. 



(2) Filter and allow to cool and again boil 

 and refilter without titration. 



(3) Tube. 



Sterilization: Sterilize in autoclave for 3 

 minutes at 15 pounds pressure. 



Use: To study effect of bile on B. typhosus. 

 The author reported that some strains 

 were completely inhibited while others 

 were only partially inhibited. Approxi- 

 mately the same results were obtained 

 using 50.0% or 10.0% bile. (Bile diluted 

 with distilled water.) 



Reference: Ecker (1918 p. 97). 



2158. Raskin's Milk Egg Albumin Agar 



Constituents : 



1. Milk 1000.0 cc. 



2. Egg albumin 



3. NaCl 5.0 g. 



4. Agar 5.0 to 7.0 g. 



Preparation : 



(1) Stir thoroughly with a glass rod 

 100.0 g. albumin from fresh eggs in a 

 flat bottomed dish and add drop by 

 drop concentrated NaOH until a 

 solid opaque gelatinous material is 

 obtained. 



(2) Cut this solid egg albumin in small 

 pieces and place in distilled water. 

 Do not allow the alkaline albuminate 

 to stand or the albuminate will 

 become liquid in several hours. 



(3) Shake thoroughly and pour off the 

 water, continue this washing until 

 the wash water is only slightly 

 alkaline. 



(4) Place the coagulated sodium al- 

 buminate in a flask of distilled water, 

 plug with cotton and heat in the 

 steamer for 15 to 30 minutes. The 

 albumin dissolves. 



(5) Filter. 



(6) Add 50.0 cc. glycerol and 5.0 to 7.0 g 

 of finely divided dried agar to 1000.0 

 cc. of fresh milk. 



(7) Allow to stand for 12 to 14 hours (at 

 room temperature in the winter) 

 and then boil for 75 to 90 minutes 



