232 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(16) Adjust the reaction. 



(17) Sterilize in the autoclave, 

 (s) Mcintosh and Smart (1918). 



(1) Mince 1.0 pound of fat free meat 

 and macerate in 1000.0 cc. cold 

 water over night. Or the mix- 

 ture may be macerated at 40°C. 

 for 30 minutes instead. 



(2) Boil for 15 minutes. 



(3) Filter thru muslin and then 

 paper. 



(4) Add 10.0 g. peptone and 5.0 g. 

 NaCl. 



(5) Make up to 1000.0 cc, dissolve 

 and steam for 45 minutes. 



(6) Measure 10.0 cc. of the broth into 

 a porcelain evaporating dish, and 

 add 25.0 cc. of distilled water. 



(7) Prepare another dish in the same 

 way as a control. 



(8) Add 5 drops of a 0.5% alcoholic 

 solution of thymolphthalein to 

 one dish. 



(9) Add N/10 NaOH to (8) until the 

 color changes from the yellow to 

 a bluish tint. 



(10) Calculate and add the amount of 

 normal NaOH that would be re- 

 quired to give the same reaction 

 to the remainder of the broth. 



(11) Boil and filter. 



(12) Add 10.0 cc. of N/1 HCl per liter 

 of filtrate. This gives a pH 

 = 7.6. 



(13) Tube or flask and sterilize in the 

 autoclave for 20 minutes at 115°C. 



(t) BezauQon (1920). 



(1) Soak 500.0 g. of finely chopped 

 lean beef (or other meat) in 

 1000.0 cc. of water for 12 hours. 



(2) Collect the liquid from the meat 

 and add 10.0 g. peptone and 5.0 g. 

 NaCl. 



(3) Autoclave for 10 minutes at 

 115°C. 



(4) Filter thru a wetted filter. 



(5) Make slightly alkaline to litmus 

 by the addition of 10.0% soda 

 tiolution. 



(6) Autoclave for 15 minutes at 

 120°C. 



(7) Filter. 



(8) Distribute as desired 



(9) Sterilize in the autoclave at 115° 

 for 15 minutes. 



(u) Percival (1920). 



(1) Chop one pound of lean, fat free 

 beef with a knife or mincing 

 machine. 



(2) Place (1) in a porcelain dish or 

 glass beaker and add 1 liter of 

 water. 



(3) Allow to soak over night in a cool 

 place. 



(4) Strain thru muslin, and boil for 

 one hour. 



(5) Filter thru filter paper into a 

 large flask. 



(6) Make the volume up to one liter. 



(7) Add 10.0 g. Witte's peptone and 

 5.0 g. NaCl to (6). 



(8) Neutralize to phenolphthalein. 



(9) Steam for 15 minutes. 



(10) Neutralize again and adjust the 

 reaction to +10. 



(11) Filter into sterile flasks or test 

 tubes. 



(12) Sterilize on 3 successive days for 

 20 minutes each time. 



(v) Giltner (1921). 



(1) Add 500.0 cc. of tap water to 

 500.0 g. of finely chopped fresh 

 lean beef in a 3.5 liter agate-ware 

 pail. 



(2) Mix thoroly and allow to stand 

 in a cool place (refrigerator) for 

 not more than 16 to 24 hours. 



(3) Strain the infusion thru a cheese 

 cloth, thoroly pressing out all 

 the juice. 



(4) Make up to 500.0 cc. by the addi- 

 tion of tap water if necessary. 



(5) Place (4) in a sterile liter Erlen- 

 meyer flask. 



(6) Heat in the autoclave at 120°C. 

 for 30 minutes. 



(7) Pour the contents of the flask into 

 an agate pail and add 500.0 oc. 

 of tap water. 



(8) Add 1.0% Witte's peptone and 

 0.5% NaCl. 



(9) Thoroly mix 10.0 g. of egg albu- 

 min with 100.0 cc. of tap water 

 and add to (8) or one well beaten 

 egg white may be used. 



(10) Heat in the autoclave at 120°C. 

 for one hour. 



