CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



361 



(5) Tube. 



(6) Autoclave for one hour. 



(7) Seed by means of a pipette or 

 needle, and mix the inoculum 

 thoroly with the medium. 



(b) Holman prepared a similar medium as 

 follows: 



(1) Free meat from fat and gross fibers, 

 mince finely. Grind in mortar and 

 mix with equal part of water. 



(2) Heat slowly to boiling, stir 

 constantly. 



(3) Neutralize or make slightly alkaline 

 while hot to phenolphthalein. 



(4) Tube at least 2 inches high and 

 autoclave at 115°C. for 30 minutes. 



(5) Just before use, place the tubes in 

 flowing steam for 30 minutes and 

 cool rapidly. 



(6) Seed with a pipette, swab or needle 

 and mix material with meat 

 particles. 



(c) Bachmann studied the effect of spices 

 on the growth of Clostridium botu- 

 linurn, using the following medium: 



(1) Grind hamburger and add enough 

 water to make 100.0 cc. 



(2) Add one of the following, cloves, 

 cinnamon, all-spice, ginger, nut- 

 meg, cayenne pepper, white mus- 

 tard, black mustard in 1.0, 2.0 or 

 2.5 g. quantity. 



(3) Mix thoroly. 



(4) Distribute into 6 inch test tubes. 



(5) Sterilize in the autoclave. 



(6) Inoculate when cool. 



He reported that the organisms were 

 not affected by any of the spices used 

 in amounts up to 2.0%. 2.5% 

 retarded the growth of some 

 organisms. 



(d) Burke isolated B. botulinus in a 

 medium prepared as follows: 



(1) Chop 500.0 g. of beef heart or other 

 lean beef with 1000.0 cc. of water. 



(2) Bring slowly to a boil, stirring 

 constantly. 



(3) Neutralize. 



(4) Tube. 



(5) Sterilize at 15 pounds pressure for 

 30 minutes. 



(e) Besson cultivated Vibrion septique 

 on a medium prepared as follows : 



(1) Add 500.00 to 600.00 g. of finely 

 chopped beef to a liter flask. 



(2) Add soda solution to neutralize or 

 make slightly alkaline. 



(3) Sterilize at 115°C. 



(f) Cunningham prepared a similar me- 

 dium as follows: 



(1) Cut up 500.0 g. of fat free bullock's 

 heart into pieces about the size of a 

 pea. 



(2) Add 500.0 cc. of water. 



(3) Steam until cooked. 



(4) Neutralize to turmeric paper (dis- 

 tinctly brown). 



(5) Stir well. 



(6) Steam for 30 minutes. 



(7) Pour off the fluid from the pieces of 

 tissue. 



(8) Tube the pieces of heart and just 

 cover them with the extract, taking 

 care to wash down any pieces of 

 tissue which adhere to the sides of 

 the tube. 



(9) Sterilize in the autoclave. 

 References: Migula (1901 p. 24), Holman 



(1918 p. 125), Bachmann (1918 p. 237), 

 Burke (1919 p. 557), Besson (1920 p. 55), 

 Cunningham (1924 p. 166). 



1240. Nevin's Glucose Veal Medium 



Constituents: 



1. Water 1000.0 cc. 



2. Chopped veal 1000.0 g. 



3. NaCl 10.0 g. 



4. Glucose 40.0 g. 



Preparation : 



(1) Mix equal parts chopped veal and 

 water. 



(2) Add 0.5% NaCl and 2.0% glucose. 



(3) Adjust reaction to 0.3% to 0.5% 

 alkaline to phenolphthalein. 



Sterilization: Method not given. 

 Use: Cultivation of B. botulinus. 

 Reference: Nevin (1924 p. 228). 



1241. Hueppe's Egg Meditim. 



Constituents: 



1. Eggs. 

 Preparation : 



(1) Wash the eggs thoroly. 



(2) Sterilize the shell by dipping in sub- 

 limate solution. 



(3) Wash in sterile water and dry with 

 sterile cotton. 



(4) Make a fine opening in the top of the 

 egg with an instrument heated to 

 glowing. 



