568 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(3) Adjust the reaction by adding sodium 

 hydrate until the mixture is neutral to 

 litmus. 



(4) Add 1.0% lactose and 0.1% glucose 

 (dissolved in a small amount of hot 

 water) to (3). 



(5) Slant the tubes following sterilization 

 and store in small quantities in a 

 dark place. 



Sterilization: Sterilize in the Arnold. 

 Pack the tubes loosely in the sterilizer 

 basket to allow good circulation of the 

 steam. Under these conditions 10 

 minutes on the first day and 15 minutes 

 the second are sufficient. 



Use: Isolation of typhoid, paratyphoid 

 and dysentery from feces and urine. 



Variants : 



(a) Kligler gave the following variant: 



(1) Prepare the nutrient agar from 

 meat infusion (or beef extract). 

 (Preferably clear sugar free beef 

 infusion.) Exact composition not 

 given. 



(2) Adjust to pH = 7.4 or neutral to 

 Andrade's indicator. 



(3) Add 1.0% by volume of Andrade's 

 indicator. 



(4) Tube in 5.0 cc. lots and sterilize. 



(5) Prepare a solution containing 20.0% 

 lactose and 20.0% glucose and 

 sterilize. 



(6) Add 0.25 cc. of sterile (5) to each 

 tube of melted sterile (4), cooled to 

 60°C. 



(7) Prepare and sterilize a 0,25% basic 

 lead acetate solution and add 1.0 

 cc. to each tube of (4) cooled to 

 60°C. 



(8) Slant so as to have a butt of at 

 least § to f inches and a slant of 

 about IJ inches. 



Kligler gave the following special 

 reactions : B. coli reddened whole tube 

 and gas was produced. Bacillus 

 typhosus reddened butt, colorless 

 slant and produced browning particu- 

 larly near the surface of the stab. 

 The dysentery bacillus reddened the 

 butt but did not produce browning. 

 Paratyphosus bacilli produced gas 

 while Bacillus hjphosus and Bacillus 

 dysenteriae did not. Bacillus para- 

 typhosus B. and allied bacilli, Bacilli 



enteritidis and Bacillus murium pro- 

 duced browning while A type did not. 

 (b) Giltner prepared a similar medium 

 as follows: 



(1) Prepare a 1.5% nutrient agar. 



(2) Add 1.0% glucose and 1.0%o lactose 

 to (1). 



(3) Make up a 0.5% solution of basic 

 lead acetate. 



(4) Sterilize (3). (Method not given.) 



(5) Tube. 



(6) Add the necessary amount of (4) 

 to each tube of agar to give 0.05% 

 basic lead acetate. 



(7) Sterilize (method not given). 



(c) Pitfield substituted Andrade's in- 

 dicator for litmus as an indicator. 

 References: Russell (1911 p. 226), Roddy 

 (1917 p. 43), Kligler (1918 p. 321), Tanner 

 (1919 p. 52), Ball (1919 p. 84), Giltner 

 (1921 pp. 385, 386), Harvey (1921-22 p. 

 109), Pitfield (1922 p. 120), Stitt (1923 

 pp. 50, 51). 



1819. Thoinot and Masselin's Glucose 

 Glycerol Agar 



Constituents : 



1. Agar 1000.0 cc. 



2. Glucose (5.0 to 



10.0%) 50.0 to 100.0 g. 



3. Glycerol (4.0 to 



6.0%) 40.0 to 60.0 g. 



Preparation : 



(1) Add 5.0 to 10.0% glucose and 4.0 to 

 6.0% glycerol to agar. 

 Sterilization: Not specified. 

 Use: General culture medium. 

 Variants : 



(a) Besson dissolved 20.0 g. agar in a 

 liter of bouillon, and added 2.0 to 

 4.0% glucose and 5.0% glycerol. 

 The medium was sterilized at 115°C. 

 for 20 minutes. 



(b) Dopter and Sacquep^e solidified 

 bouillon with agar, and added 5.0% 

 glycerol and 2.0% glucose. 



References: Thoinot and Masselin (1902 p. 

 35), Besson (1920 p. 43), Dopter and 

 Sacquepee (1921 p. 128). 



1820. Thoinot and Masselin's Sucrose 

 Glycerol Agar 



Same as medium 1819 but substituting 

 5.0 to 10.0 g. sucrose for glucose. 



