CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



673 



small pieces, and store in the ice 

 box for 24 hours. 



(8) Add 40.0 cc. of glycerol and 100.0 

 cc. water to (7) and place in the 

 ice box for several days. 



(9) Filter the juice from (8). 



(10) Add 15.0 cc. of (9) to (6) and 

 incubate at 37°C. for 6 hours. 



(11) Sterilize (method not given). 



(12) Neutralize with 5.0% HCl. 



(13) Add 1950.0 cc. water, 6.0 g. NaCl, 

 and 39.0 g. agar, and prepare as 

 ordinary nutrient agar. 



References: Deycke and Voigtlander (1901 

 p. 624), Bosse (1903 p. 472), Kolle and 

 Wass&rmann (1912 p. 413), Harvey (1921- 

 22, p. 100), Klimmer (1923 p. 221). 



2075. Kligler's Yeast Autolysate Agar 



Constituents : 



1. Yeast autolysate 1000.0 cc. 



2. Agar 15.0 g. 



Preparation : 



(1) Prepare yeast autolysate as in 

 medium 982, but do not filter. 



(2) Add agar to (1) and stir thoroly to 

 soften agar thread or powder. 



(3) Heat over a free flame to dissolve 

 agar. 



(4) Readjust the reaction to pH = 7.4. 



(5) Heat on a water bath or in an Arnold 

 for ^ hour and decant the partially 

 clear fluid to an Erlenmeyer flask or 

 other vessel. 



(6) Cool the agar (exact temperature not 

 given), and add one whole egg to 

 clear. 



(7) Steam in the Arnold for 30 to 45 

 minutes. 



(8) Filter and tube. 

 Sterilization: Method not given. 

 Use: General culture medium. 

 Variants : 



(a) Abt and Blanc solidified medium 984 

 with the addition of agar. 



(b) Harvey prepared a similar medium 

 as follows: 



(1) Keep 100.0 g. yeast at 50°C. for 24 

 hours or until liquefaction is com- 

 plete by autolysis. 



(2) Dilute with water to 400.0 cc. 



(3) Steam 30 minutes. 



(4) Adjust reaction. 



(5) Filter. 



(6) Make up the volume to 1500.0 cc. 



(7) Solidify with agar. 

 References: Kligler (1919 p. 186), Abt and 



Blanc (1921 p. 452), Harvey (1921-22 p. 

 120). 



2076. van Steenberge's Yeast Extract Agar 



Constituents : 



1. Yeast water agar 1000.0 cc. 



2. Esculin 1.0 g. 



3. Iron citrate 1-0 g. 



Preparation : 



(1) Dissolve 2 and 3 in yeast water agar. 

 Sterilization: Not specified. 

 Use: Cultivation of lactic acid bacteria. 

 Variants : The author added 50.0 g. glucose. 

 Reference: van Steenberge (1920 p. 841). 



2077. Jotten's Yeast Autolysate Agar 



Same as medium 983, and variant (b) 

 medium 983, but solidified by the addition 

 of 2.2% agar. 



2078. Couret and Walker's Autolyzed 



Tissue Agar 



Constituents : 



1. Agar solution. 



2. Tissue. 

 Preparation : 



(1) Collect bits of sterile liver, kidney 

 and brain of healthy rabbits, guinea 

 pigs, kittens and human placenta, 

 in sterile flasks. 



(2) Seal and place in the thermostat at 

 40 °C. for 10 to 20 days. 



(3) Titrate the fluid tissue to a neutral or 

 slightly acid reaction (not exceeding 

 1.5% acid) by the addition of N/1 

 NaOH. 



(4) The agar is to be prepared in the 

 usual manner (exact method not 

 given) but is to contain no peptone, 

 beef extract, etc. 



(5) Adjust the reaction to neutral or to 

 an alkalinity not to exceed 0.5%. 

 Tube and slant. 



(6) Distribute 0.1 to 0.5 cc. of (3) (auto- 

 lyzed tissue) over the surface of each 

 agar slant. 



(7) Keep several hours in the horizontal 

 position to allow the agar to absorb 

 the tissue juice. 



(8) Incubate to determine sterility. 



