CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



583 



Use: Cultivation of Meningococci, Micro- 

 coccus catharralis and Diplococcus flavus. 



References : Sacquepee and Delater (1914 p. 

 224), Dopter and Sacquepee (1921 p. 

 139). 



1862. Emile-Weil Egg Yolk Infusion Agar 



Constituents : 



1. Distilled water 250.0 cc. 



2. Sea water 750.0 cc. 



3. Veal 500.0 g. 



4. Glycerol 40.0 g. 



5. Glucose 80 S- 



6. Peptone 100 S- 



7. Agar 20.0 g. 



8. Egg yolk 250.0 cc. 



Preparation : 



(1) Prepare infusion broth from 500.0 g. 

 veal, 250.0 cc. distilled water and 

 750.0 cc. sea water. Details of 

 method not given. 



(2) Make distinctly alkaline. 



(3) Add and dissolve 4, 5, 6 and 7 in (2). 



(4) Add to cool agar, one part egg yolk to 

 four parts agar. (Not clearly stated 

 if the agar is in a liquid or solid state.) 



Sterilization: Not specified. 

 Use: Cultivation of B. leprae. 

 Reference: Emile-Weil (1905 p. 799). 



1863. Huntoon's Hormone Agar 



Constituents : 



1. Water 1000.0 cc. 



2. Beef, fresh heart or steak. . 500.0 g. 



3. Peptone (Bacto) 10.0 g. 



4. Agar (Bacto) or soaked 



thread 16.0 g. 



5. Salt 5.0 g. 



6. Whole egg 1 



Preparation : 



(1) Chop 2 and mix 1, 2, 3, 4, 5 and 6. 

 Place in an enameled ware vessel or a 

 large coffee pot. 



(2) Heat over a free flame with constant 

 stirring until the red color of the 

 meat infusion changes to brown at a 

 temperature of about 68°C. Do not 

 go beyond this temperature. 



(3) Adjust to slightly alkaline to litmus 

 with N/1 NaOH and then add 1.0 

 cc. per liter of medium. 



(4) Cover the vessel and place in an 

 Arnold sterilizer or in a water bath 

 at 100°C. for one hour. 



(5) Remove the vessel from the sterilizer 

 and separate with a glass rod, the 

 firm clot which has formed from the 

 side of the vessel. 



(6) Return to the Arnold sterilizer at 

 100°C. for U hours. 



(7) Remove the vessel and allow to stand 

 at room temperature for about 10 

 minutes in a slightly inclined 

 position. 



(8) Pipette off the fluid portion or 

 decant. If it is poured thru a fine 

 wire sieve, many of the fine pieces of 

 meat clot may be caught. (Avoid 

 filtering thru cheese cloth, cotton or 

 other adsorptive materials.) 



(9) Allow (8) to stand in tall cylinders 

 for 15 to 20 minutes until the fat 

 present has risen to the surface and 

 removed. The medium may be 

 further cleared by filtering thru 

 glass wool, asbestos wool, sedimenta- 

 tion or centrifugation. 



(10) Tube in 10.0 cc. lots. 

 Sterilization : Sterilize intermittently in the 



steamer. 

 Use: Substitute for media containing ser- 

 ous fluids. Author reported that the 

 medium had a growth value 10 times as 

 great as standard agar and was at least 

 as good as the average grade serum agar. 

 Variants : 



(a) Huntoon prepared a semi-solid hor- 

 mone agar as indicated above but 

 using only 5.0 g. agar instead of 16.0 g. 

 Huntoon reported that this medium 

 was as good as average grade serum 

 agar, can be employed as stab cul- 

 tures, and very useful both for 

 anaerobic and aerobic cultures. It 

 is especially suitable for preservation 

 of stock cultures. 



(b) Huntoon added 5.0% glycerol to the 

 medium just before tubing and 

 reported that after ten days growth of 

 tubercle bacilli almost as heavy as in 

 Dorset's egg medium. 



(c) Harvey gave the following method 

 of preparation: 



(1) Prepare finely minced ox heart 

 500.0 g.; 10.0 g. peptone, sodium 

 chloride 5.0 g., contents of one 

 egg; prepared agar (see medium 

 1401) 16.0 g., water 1000.0 cc. 



