776 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(6) Inspissate the medium in a sloping 

 position at a temperature of 75°C. 

 for 60 minutes on three successive 

 days. 



(7) Incubate tubes 24 hours and reject 

 any which are not sterile. 



(u) Park, Williams and Krumwiede cul- 

 tivated spirochaetes on a medium 

 prepared as follows: 



(1) Tube serum in tall tubes. 



(2) Warm the tubes to 65°C. 



(3) Place a few of the tubes at a time 

 in water at 75°C. 



(4) Tilt these tubes every few seconds 

 and remove as soon as they start to 

 set. The heat in the tube com- 

 pletes the coagulation. 



They reported that the serum may be 



diluted with saline or broth and 



treated in the same way. 



References : Frothingham (1895 pp. 55, 57), 



Marmier (1895 p. 569), Michel (1896 



p. 261), Wurtz (1897 p. 33), Glucksmann 



(1897 p. 436), Smith (1898 p. 456), Thoinot 



and Masselin (1902 p. 42), Heinemann 



(1905 p. 131), Schereschewsky (1912 p. 



1335), Sowade (1912 p. 797), Abbott 



(1921 p. 135), Ball (1919 p. 78), Harvey 



(1921-22 pp. 78-81), Pitfield (1922 p. 119), 



Jones (1922 p. 363), Vardon (1923-24 p! 



431), Park, Williams and Krumwiede 



(1924 p. 125). 



2399. Proco, et al., Gentian Violet Serum 



Constituents: 



1. Water. 



2. Serum. 



3. Gentian violet. 



4. NaCl. 

 Preparation : 



(1) Dissolve 0.1 centigrams of gentian 

 violet in 300.0 cc. of physiological salt 

 solution. 



(2) Mix equal parts of tyndallized serum 

 and (1). 



(3) Coagulate at 80 °C. 

 Sterilization: Sterilization given in the 



preparation. 

 Use: Cultivation of Syphilus spirochaete 

 References: Proco, Danila and Stroe (1912 



p. 896), Harvey (1921-22 p. 81). 



2400. Lorrain Smith's Alkali Serum 



(Harvey) 

 Constituents : 



1. 10.0%NaOH 1.5,, 



2. Ox serum 100.0 cc. 



Preparation : 



(1) Add 15.0 cc. of 10.0% NaOH to 1000.0 

 cc. of ox serum. 



(2) Coagulate in an inspissator or over 

 steam at 65 to 90°C. according to the 

 degree of transparency required. 



Use: General culture medium. 

 Reference: Harvey (1921-22 p. 78). 



2401. Duval's Trypsinized Serum 



Constituents : 



1. Serum (human). 

 Preparation : 



(1) Pour human blood serum into sterile 

 petri dishes and inspissate for three 

 hours at 70°C. (Egg albumin or 

 nutrient agar with or without 1.0% 

 tryptophane may be treated in the 

 same manner.) 



(2) Cut an excised leprous nodule into 

 thin slices, two to four millimeters in 

 breadth and 0.5 to 1.0 millimeters in 

 thickness and distribute over the 

 surface of the coagulated serum 

 (albumin or agar). 



(3) The medium is now seeded. Bathe 

 the surface with a 1.0% sterile trypsin 

 solution, taking care not to submerge 

 the piece of leprous tissue. Add 

 sufficient trypsin solution to com- 

 pletely cover the medium. 



(4) Incubate in a moist chamber at 37°C. 

 for a week or ten days. Remove the 

 plates from time to time and add more 

 trypsin as is necessary. 



Sterilization : Method not given. 



Use: Isolation and cultivation of B. leprae. 

 The author reported that the colonies at 

 first were grayish white, but after several 

 days they assumed a distinct orange 

 yellow tint. Transfers may be made 

 from this medium to slants or other 

 plates. 



Reference: Duval (1911 p. 369). 



2402. Carrel and Burrows' Plasma Medium 

 Constituents: 1. Blood plasma 



