548 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



fuchsin in 95% alcohol and allow 

 to stand 24 hours. 



(7) Decant and filter the supernatant 

 liquid. 



(8) Add to each 100.0 cc, of sterile 

 melted (5), 1.0% of chemically 

 pure lactose in sterile solution 0.5 

 cc. of (7), and 0.125 g. anhydrous 

 Na2S03, dissolved in a small 

 amount of hot distilled water. 

 Mix thoroly. 



(9) Pour in plates. 



References: Heinemann (1905 p. 130), 

 Klinger (1906 p. 52), Kastle and Elvove 

 (1909 p. 622), Kendall and Day (1911-12 

 p. 96), Kinyoun and Deiter (1912 p. 979), 

 Committee A.P.H.A. (1913 p. 133), (1917 

 p. 97), (1920 p. 97), (1923 p. 96), (1925 p. 

 98), Robinson and Rettger (1916 p. 367), 

 Roddy (1917 p. 43), Krumwiede, Kohn, 

 Kuttner and Schumm (1918 p. 286), 

 Tanner (1919 p. 53), Ball (1919 p. 82), 

 Bezangon (1920 p. 342), Giltner (1921 pp. 

 384, 385, 387), Levine (1921 pp. 112, 113, 

 115), Abbott (1921 p. 524), Pitfield (1922 p. 

 120), Klimmer (1923 p. 209), Stitt (1923 p. 

 48), Park, Williams and Krumwiede 

 (1924 pp. 127, 131). 



1760. Hirschbruck and Schwer's Crystal 

 Violet Litmus Lactose Agar 



Constituents : 



1. Water 1000.0 cc. 



2. Agar 20.0 g. 



3. Meat extract, Liebig's 10.0 g. 



4. Peptone 10.0 g. 



5. NaCl 5.0 g. 



6. Litmus solution, Kubel and 

 Tiemann 130.0 cc. 



7. Crystal violet B (Hochst) 



0.1% 10.0 cc. 



Preparation : 



(1) Boil 2, 3, 4 and 5 in 1 for 1.5 hour. 



(2) Filter and boil again for 30 minutes. 



(3) Add 15.0 g. lactose to (2) and boil 

 for 15 minutes 



(4) Add a sterile soda solution (any 

 desired strength, but not too strong) 

 until red litmus paper is turned blue. 



(5) Add 130-0 cc. of Kubel-Tiemann's 

 litmus solution, and 10.0 cc. of a 

 crystal violet solution that has been 

 prepared by dissolving 0.1 g. Hochst 

 crystal violet in hot sterile distilled 

 water to (4) . 



(6) Mix (5) thoroly. 



(7) Pour into sterile plates, about 8.0 cc. 

 per plate, and allow to solidify with 

 cover removed. 



Sterilization : Not specified. 



Use: Diagnosis of cholera. Authors re- 

 ported that by transmitted and reflected 

 light after 10 hours the cholera colonies 

 were blue, and about 1.5 mm. in diameter. 

 The blue color of cholera colonies was 

 much darker than that of typhoid colo- 

 nies. After 20 hours the colonies were 

 about 2 mm. in diameter. By reflected 

 light they appeared milk blue, and by 

 transmitted light a deep i'ky blue. The 

 entire plate remained intensively blue 

 with a pure culture. Bad. colt colonies, 

 both by transmitted and reflected light, 

 were red. 



Reference: Hirschbruch and Schwer (1903 

 p. 587). 



1761. Chesney's Indicator Lactose Agar 



Constituents : 



1. Beef extract agar 



(3.0%) 1000.0 cc. 



2. Lactose 10.0 g. 



3. Brom-cresol purple 



(0.04%) 50.0 to 80.0 cc. 



Preparation : 



(1) E.xact method of preparation or 

 composition of 3.0% beef extract agar 

 not given. 



(2) Adjust (1) to pH of 7.2 to 7.4. 



(3) Add 1.0% lactose from a sterile 20.0% 

 solution to sterile (2). 



(4) Add from 5.0 to 8.0 cc. of a sterile 

 0.05% aqueous solution of brom cresol 

 purple, to every 100.0 cc. of sterile 

 (3). 



(5) Pour into sterile Petri dishes, about 

 20.0 cc. to each plate. 



Sterilization: Method of sterilization of 

 agar or lactose solution not given. 

 Sterilize the 0.04% brom cresol purple 

 in the autoclave. 



Use: Isolation of typhoid and dysentery 

 bacilli from stools. Author reported that 

 medium was clear and deep blue in color. 

 Lactose fermenters produced greenish 

 yellow colonies with a yellow zone. Non- 

 lactose fermenters produced bluish colo- 

 nies. Typhoid colonies have typical 

 woolly appearance. In thickly seeded 

 plates, typhoid colonies were bluish 



