400 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



than in the kidney medium. In the buff 

 medium the spirochaetes kept their 

 regular forms of life for a longer period 

 of time and kept their virulence for at 

 least two months. 



Variants : In place of the kidney 2 or 3 small 

 pieces of buff coagulum cut in 1 cc. 

 dimensions may be added. Buff co- 

 agulum is a part of the blood clot. The 

 clot separates into two layers, the cruor, 

 consisting of red corpuscles and a buff 

 coagulum consisting of white corpuscles 

 and blood platelets. These pieces of 

 buff coagulum must be pushed to the 

 bottom of the medium with a sterile 

 glass rod. 



Reference: Hata (1913 p. 109). 



1367. Noguchi's Ascitic Fluid Tissue 

 Medium 



Constituents : 



1. Ascitic fluid. 



2. Tissue (rabbit). 

 Preparation : 



(1) Place sterile pieces of tissue, usually 

 rabbit kidney, in test tubes 2 x 20 cm. 



(2) After inoculation add quickly about 

 15.0 cc. of sterile unheated unfiltered 

 ascitic fluid. 



Sterilization : Materials are obtained under 

 aseptic conditions. 



Use: Cultivation of spirochaetae, Spiro- 

 chaeta dutioni, Spirochaeta kochi, Spiro- 

 chaeta ohermeieri, Spirochaeta novyi, 

 treponemata and others. A few centi- 

 meters of citrated blood from the heart 

 of an infected rat or mouse was added to 

 the rabbit tissue before the addition of 

 ascitic fluid. Incubate under anaerobic 

 as well as aerobic conditions. Flexner 

 and Noguchi inoculated the medium 

 with a piece of cerebrum or other part of 

 the brain or spinal cord of an infected 

 animal to cultivate the organism causing 

 epidemic poliomyelitis. Noguchi iso- 

 lated and cultivated Spirochaeta galli- 

 narum, inoculating the medium with a 

 blood emulsion containing the spiro- 

 chaeta. Loewe and Strauss cultivated 

 the organism causing epidemic en- 

 cephalitis. Olitsky and Gates isolated 

 and cultivated anaerobic organisms found 

 in influenza, in the medium. 



Variants : 



(a) Noguchi substituted hydrocele fluid 

 for ascitic. 



(b) Stitt gave the following method of 

 preparation, of a medium for the 

 cultivation of treponemata. 



(1) Fit a test tube with a perforated 

 rubber stopper which can be 

 pushed down into the tube. 



(2) Pass a piece of glass tubing thru 

 the stopper to project slightly into 

 the test tube. 



(3) Draw out the other end of the glass 

 tube into a capillary tube and 

 bend at an acute angle. 



(4) Boil the apparatus. 



(5) When cool a piece of sterile tissue is 

 dropped into the tube. 



(6) Draw a strip of gauze thru a glass 

 bead and soak the gauze in the 

 material to be cultured. 



(7) Drop (6) into the bottom of the 

 tube. 



(8) Run in ascitic fluid to the point 

 where it would be reached by the 

 bottom of the rubber stopper. 



(9) Push in the stopper as quickly as 

 possible and when fluid appears in 

 the capillary tube seal off quickly 

 with a small flame. 



References: Noguchi (1912 p. 201), Flexner 

 and Noguchi (1913 p. 463), Noguchi 

 (1916 p. 622), Rosenow and Towne (1917 

 p. 177), Loewe and Strauss (1920 p. 252), 

 Olitsky and Gates (1921 p. 715), Abbott 

 (1921 p. 635), Stitt (1923 p. 53). 



1368. Kligler and Robertson's Ascitic Fluid 

 Egg Medium. (Stitt) 



Constituents : 



1. Egg albumin 300.0 cc. 



2. Ascitic fluid 100.0 cc. 



Preparation : 



(1) ]\Iix three parts egg albumin with one 

 part ascitic fluid (horse or rabbit 

 serum may be used instead). 



(2) Tube in tall tubes. 

 Sterilization: Not specified. 



Use: Cultivation of Borrelia recurrentis. 

 The medium was covered with a layer 

 of oil not greater than 1.5 cm. high. 



Reference: Stitt (1923 p. 54). 



