580 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(4) Pipette (3) into sterile tall test 

 tubes containing pieces of sterile 

 rabbit kidney and the inoculum. 



(5) Seal with sterile melted vaseline, 

 (b) Harvey cultivated anaerobic spiro- 



chaetes on the following medium: 



(1) Transfer small portions of organs 

 aseptically removed from a rabbit, 

 specially killed for the purpose 

 (2514 variant (a), to each sterile 

 test tube 20 x 2 cm. 



(2) Mix one part infusion agar, 

 1.0% acid to phenolphthalein at 

 45°C. (see medium 1661 variant (v) 

 for preparation) with one part 

 sterile ascitic fluid heated to 45°C. 



(3) Fill, with sterile precautions, about 

 20.0 cc. of (2) into each test tube 

 (1). 



(4) Cover the medium to a depth of 

 3 cm. with sterile liquid paraffin. 



(5) Test sterility by incubation 48 

 hours. 



References: Smillie (1918 p. 324), Olitsky ' 

 and Gates (1921 p. 715), Harvey (1921-22 

 p. 97). 



1854. van Riemsdyk's Liver Infusion Agar 



Constituents : 



1. Water 500.0 cc. 



2. Beef liver 0.5 lb. 



3. Peptone (Witte) 5.0 g. 



4. NaCl 2.5 g. 



5. Agar 10.0 g. 



6. 2.0% glucose agar 500.0 cc. 



7. Liver tissue 



8. Glucose 10.0 g. 



9. Chalk 

 Preparation : 



(1) Pass raw beef liver thru a fine meat 

 grinding machine. 



(2) Add 500.0 cc. of water to § pound of 

 (1). 



(3) Boil for 30 minutes with occasional 

 stirring. Allow to settle. 



(4) Pour off the liquid and force the liver 

 thru a very fine metallic sieve. 



(5) Add the pulverized liver and 3, 4 and 

 5 to the liquid. 



(6) Heat for one hour at 110°C. and make 

 alkaline to phenolphthalein using 

 N/10 NaOH. 



(7) Add 10,0 g. of glucose and mix with 

 an equal volume of 2.0% glucose agar. 



(8) Add sterile chalk in excess (about 5 

 spoonfulls) and distribute into sterile 

 tubes. 



(9) Add a sterile piece of liver to each 

 tube. 



Sterilization: Sterilize at 100°C. for one 



hour and slant. 

 Use: Cultivation of Bad. butyricus and 



other anaerobes. 

 Reference: van Riemsdyk (1922 p. 248). 



1855. Williams' Tissue Infusion Agar 

 Constituents : 



1. Water, tap 90.0 cc. 



2. Agar (1.0%) 1.0 g. 



3. Infusion broth (10.0%) 10.0 cc. 



4. Brain 

 Preparation : 



(1) Mix 90.0 cc. tap water, 10.0% ordinary 

 infusion broth and 1.0% agar. 



(2) Dissolve. 



(3) Reaction neutral to phenolphthalein. 



(4) Cut fresh sterile brain in small pieces 

 and place on the surface of sterile (3). 



Sterilization: Sterilize as usual (method 



not given). 

 Use: Cultivation of amoeba. 

 Variants: Park, Williams and Krumwiede 



substituted liver or kidney for brain. 

 Reference: Park, Williams and Krumwiede 



(1924 p. 134). 



1856. Olitsky and Gates' Ascitic Fluid 

 Kidney Agar 



Constituents : 



1. Beef infusion broth. 



2. Beef infusion (2.0%) agar. 



3. Human ascitic fiuid. 



4. Rabbit kidney. 

 Preparation : 



(1) Prepare beef infusion broth and 2.0% 

 beef infusion agar. 



(2) Adjust both the broth and agar to 

 pH = 7.4. 



(3) Place in each 50.0 cc. Florence flask 

 i of a moderate size rabbit kidney. 

 Sections are cut across the entire 

 kidney and placed with the cut sur- 

 face parallel to the base of the flask. 



(4) Inoculate the kidney. 



(5) Mix one part (2.0%) infusion agar, 

 (2), with two parts sterile human 

 ascitic fluid of pH = 7.8 to 8.0, in a 

 flask at 40°C. 



