CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



399 



Preparation : 



(1) Break up the white and yolk of one 

 egg in a flask containing beads and 

 shake thoroly. 



(2) Add 300.0 cc. of water and again 

 shake. 



(3) Place the flask in a water bath and 

 bring to boiling point. 



(4) Maintain this temperature for 30 

 minutes, shaking from time to time. 



(5) Tube in 5.0 cc. amounts. 



(6) Add a few drops of human blood. 



Sterilization: Sterilize in the autoclave. 



Use: Cultivation of Entamoeba his- 

 tolytica and other intestinal protozoa as 

 amoeba, etc. 



References: Cutler (1918 p. 22), Stitt (1923 

 p. 51). 



1364. Bruschettini's Egg Yolk Blood 



Infusion Solution (Kolle and 



Wassermann) 



Constituents : 



1. Veal infusion solution 100.0 cc. 



2. Blood, defibrinated, rabbit 



or dog 10.0 cc. 



3. Egg yolk 5.0 cc. 



Preparation : 



(1) Mix 1, 2 and 3. 

 Sterilization: Not specified. 

 Use: Cultivation of tubercle bacilli. 

 Reference: Kolle and Wassermann (1912 p. 



413). 



1365. Noguchi's Serum Tissue Medium 



Constituents: 



1. Distilled water 300.0 cc. 



2. Serum 100.0 cc. 



3. Tissue, (rabbit) 

 Preparation : 



(1) Tube a mixture of one part serum 

 (sheep, horse or rabbit) and three 

 parts distilled water in test tubes 20 

 cm. high and 1.5 cm. wide to a depth 

 of about 16 cm. 



(2) Add to each sterile tube a small piece 

 of freshly removed sterile rabbit 

 tissue (testicles, kidney or heart 

 muscle) (liver being unsuitable). 



(3) Incubate at 37° for 2 days to test 

 sterility. 



Sterilization: Sterilize (1) at 100°C. for 15 



minutes on 3 successive days. 

 Use: Cultivation of Treponema pallidum 



and spirochaetes. The author added a 

 layer of sterile paraffin oil to each tube 

 and incubated under strict anaerobic 

 conditions at 35° to 37°C. 

 Variants: Pitfield prepared a similar me- 

 dium as follows: 



(1) Obtain dog, sheep or cow blood under 

 aseptic conditions. 



(2) Pipette off the serum with a sterile 

 pipette. 



(3) Mix 3 to 4 parts of distilled water with 

 1 part (2). 



(4) Tube. 



(5) Add a bit of sterile animal tissue to 

 each tube. 



References: Noguchi (1911 p. 101), (1911 

 pp. 1550-1551), Pitfield (1922 p. 119). 



1366. Hata's Serum Tissue Medium 



Constituents: 



1. Physiological salt solution 80.0 cc. 



2. Horse serum 40.0 cc. 



3. Kidney 

 Preparation : 



(1) Draw blood from the vein of a normal 

 horse into tall glass cjdinders and 

 allow the serum to separate. 



(2) Pipette 4.0 cc. of the serum into tubes 

 having a diameter of 1.5 to 1.7 cm. 



(3) To each tube add 8.0 cc. of physiologi- 

 cal salt solution and mix well. The 

 contents of the tube are about 6.5 to 

 7.0 cm. high. 



(4) Place in a water bath at 58°C. raising 

 the temperature gradually until it 

 reaches 70° or 71 °C. in three hours. 



(5) Heat for 30 minutes at 7rC. The 

 contents present a translucent milky 

 appearance with a semi-solid surface. 



(6) Immerse one or two small pieces of 

 rabbit kidney in each tube. It may 

 be necessary to push the kidney to 

 the bottom by means of a sterile glass 

 rod. 



Sterilization: Not specified. 



Use: Cultivation of Spirochaeta recurrens. 

 Inoculate the medium with infected 

 blood by means of a sterile capillary tube. 

 Author reported that in the kidney 

 medium growth appeared after 24 hours 

 and reached a maximum on the 4th day. 

 In the buff coagulum medium growth was 

 at a maximum at the end of the 5th to 

 7th days. The number seemed to be less 



