496 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Broth (see variant (bb) medium 

 779) for 2 hours. 



(2) Bring the agar into solution by 

 heating. 



(3) Sterilization not specified, 

 (z) Harvey (1921-22). 



(1) Add 500.0 cc. of water to 500.0 cc. 

 of the variant given by Harvey 

 above, 

 (aa) Abbott (1921). 



(1) Add 500.0 g. chopped lean beef 

 to 1 liter of water and soak for 

 24 hours, kept at ice box tem- 

 perature. 



(2) Strain thru a coarse towel and 

 press until a liter of fluid is 

 obtained. 



(3) Dissolve 1.0% peptone, and 0.5% 

 NaCI in (2). 



(4) Dissolve 1.0 to 1.5% agar in (3) by 

 boiling in a porcelain lined iron 

 vessel. Add 250.0 to 300.0 cc. 

 of water and boil until this vol- 

 ume of water has evaporated, 

 leaving one liter volume. 



(5) Place the vessel in a large dish 

 of cold water. 



(6) Stir the agar constantly until 

 cooled to 68 to 70°C. 



(7) Add the white of one egg which 

 has been beaten up in about 

 50.0 cc. of water (a 10.0% dry 

 albumin solution may be used). 



(8) Mix (7) thoroly with (6). 



(9) Allow to boil slowly for 30 min- 

 utes. Do not allow the volume of 

 the liquid to fall below the liter 

 mark. 



(10) Filter thru a heavy folded filter 

 paper. 



(11) Sterilize by steam (method not 

 given). 



(bb) Dopter and Sacquepee (1921). 



(1) Add 1000.0 cc. of water to 500.0 g. 

 of finely chopped fat and tendon 

 free beef. 



(2) Allow to stand in ice box for 

 12 hours, or heat at 50 to 55° for 

 30 minutes. 



(3) Heat slowly to boiling. 



(4) Boil slowly for 10 minutes, stir- 

 ring constantly. 



(5) Press the liquid thru a clean 

 cloth. 



(6) Add 20.0 g. peptone and 5.0 g. 

 NaCl to 1000.0 cc. of the filtrate 

 and dissolve by shaking. 



(7) Filter thru a wetted filter paper. 



(8) Make slightly alkaline to litmus 

 by the addition of NaOH or 

 NaaCOs. 



(9) Soak 20.0 g. of agar in (8) for 

 several hours. 



(10) Boil until solution is complete, 

 stirring constantly. 



(11) Test, and readjust the reaction if 

 necessary. 



(12) Cool to 60°C. 



(13) Add the white of one egg beaten 

 up in 50 to 100.0 cc. of water. 



(14) Add (13) to (12). 



(15) Heat in the autoclave at 118" 

 for 15 minutes. 



(16) Filter while hot thru paper. 



(17) Tube. 



(18) Sterilize at 115° for 20 minutes, 

 (cc) Pitfield (1922). 



(1) Cover 500.0 g. of finely cut fat 

 free beef with 1000.0 cc. of water. 



(2) Shake well and place on ice over 

 night. 



(3) Squeeze out the fluid by means 

 of a cloth and make up the volume 

 to 1 liter. 



(4) Inoculate with a culture of the 

 colon bacillus. 



(5) Allow to stand at room tempera- 

 ture over night. 



(6) Boil and add 10.0 g. Witte's pep- 

 tone and 5.0 g. NaCl. 



(7) Weigh the saucepan and contents 

 and heat to 60°C. 



(8) Make up the loss in weight by 

 the addition of water. 



(9) Neutralize to litmus, 

 (dd) Klimmer (1923). 



(1) Preparation of meat water not 

 given. 



(2) Soak the agar in water from 1 to 

 12 hours. 



(3) Pour off the water and press 

 the agar free from liquid. 



(4) Add (3) to (1) and boil until solu- 

 tion is complete (steamer or free 

 flame). 



(5) ]\Iake up the loss in weight by the 

 addition of water. 



