CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



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(2) Make up volume. 



(3) Strain thru cotton flannel. 



(4) Add 10.0 g. Witte's peptone. 



(5) Dissolve by heating on water 

 bath. 



(6) Heat over steam bath 30 minutes. 



(7) Restore volume. 



(8) Adjust reaction to +1 with phenol- 

 phthalein. 



(9) Boil 2 minutes over free flame. 



(10) Restore volume. 



(11) Filter thru absorbent cotton and 

 cotton flannel until clear. 



(12) Add 1.0% of any sugar to (11). 

 Jja. case of adding glucose the 

 muscle sugar must be removed by 

 fermentation with B. coli. 



(13) Sterilize in streaming steam. 



(c) Percival prepared the medium as 

 follows: 



(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 1 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) Add 2.0% of lactose, 2.0% glucose 

 or0.3%NaNO3. 



(d) Besson prepared the medium as 

 follows: 



(1) Remove all fat and tendons from 

 beef and chop into small pieces. 



(2) Allow 500.0 g. of (1) to macerate 

 with 1000.0 cc. of cold water for 

 6 hours, or if one wishes to remove 

 the sugar 12 hours at 37°C. 



(3) Place in an enamelled pot and 

 bring slowly to a boil. 



(4) Boil for ten minutes. 



(5) Throw on a thick cloth and press 

 the meat free from juice. 



(6) Filter the juice thru moistened 

 paper. 



(7) Add 10.0 g. Chapoteaut or De- 

 fresne peptone, 5.0 g. NaCl and 

 about 1.0 g. of sodium phosphate. 



(8) Boil stirring constantly until solu- 

 tion is complete. 



(9) Neutralize or make slightly alka- 

 line to litmus by the addition of 

 soda solution. 



(10) Heat at 115 to 117°C. for 5 minutes. 



(11) Filter until clear. 



(12) Make up to 1000.0 cc. by the addi- 

 tion of distilled water. 



(13) Add 2.0 to 4.0% of one of the 

 following: glucose, raffinose, lac- 

 tose, galactose, mannitol, dulcitol, 

 maltose, levulose or glycerol. 



(14) Distribute as desired. 



(15) Sterilize at 110 to 115°C. for 

 20 minutes. 



(e) Harvey prepared the meat infusion 

 peptone solution (see Dunham's 

 Meat Infusion Peptone Solution, 

 variant (bb) 779) and added 1.0% 

 glucose, 1.0% lactose, 2.0% starch, 

 1.0% mannitol or 5.0% glycerol. 

 Harvey also added 5.0 cc. of a 1.0% 

 neutral red solution to meat infusion 

 peptone solution and added 0.5% of 

 any desired sugar. 



(f) Stitt prepared the infusion broth as 

 in variant (hh) 779, or extract broth 

 variant (o) 689, and added 1.0 to 

 2.0% of any desired carbohydrate, 

 alcohol, etc. 



(g) Park, Williams and Krumwiede pre- 

 pared infusion broth as in variant 

 (11) 779, and proceeded as follows: 



(1) Adjust the reaction of meat ex- 

 tract of the infusion to pH = 7.0 

 (slightly alkaline to litmus). 



(2) To each liter add a broth culture 

 of B. coli or one of its allies. 



(3) Incubate for 48 hours. 



(4) Sterilize in the autoclave. 



(5) Test again for the production of 

 acid or gas before adding carbo- 

 hydrates. 



(6) Sterilize (5). (Method not given.) 



(7) Prepare 10.0 or 20.0% solutions 

 of the carbohydrates, alcohols, etc. 



