CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



319 



(1) Free 750.0 g. meat from fascia and 

 cut in finger-thick pieces. 



(2) Drop (1), piece by piece, into 

 1500.0 g. of boiling water, stirring 

 constantly. 



(3) Boil up strongly and take from 

 fire. 



(4) Take out the meat and put thru 

 a chopping machine. 



(5) Cool the water to 37°C. and add 

 1.5 g. NaoCOs per liter. 



(6) Put the chopped meat in flasks 

 (2 liter Erlenmeyer) 550.0 g. in 

 each flask. 



(7) Add (5) (the water) at 37°C. to the 

 flask, filling them up to the narrow 

 neck. 



(8) Add 3.0 g. of pancreatin, 10.0 cc. 

 chloroform and 10.0 cc. toluol to 

 each flask. 



(9) Cork tightly. 



(10) Shake thoroly. 



(11) Incubate at 37° over night. 



(12) Shake the next morning and add 

 more pancreatin unless the fluid 

 shows a yellow color, and particles 

 of meat look smaller, indicating 

 digestion. 



(13) Digest for 4 or 5 days at room 

 temperature or for 2 or 3 days in 

 the incubator. Shake the flasks 

 each day. The meat should be in 

 a finely divided mass, giving off a 

 very offensive odor, when digestion 

 is complete. 



(14) The medium may be stored in the 

 ice box, after acidifying to litmus 

 by the addition of HCl, if neces- 

 sary. (Hottinger's statement.) 



(15) As soon as the digestion is com- 

 plete decant the liquid thru cheese 

 cloth. (This process gives better 

 results according to the author.) 



(16) Add an equal amount of water 

 to the residue. 



(17) Shake (16) thoroly. 



(18) Allow to settle and again decant. 



(19) Place the meat on the cheese cloth 

 and allow to drain. 



(20) Boil the filtrate a few minutes. 



(21) Filter thru absorbent cotton and 

 paper until clear. 



(22) Autoclave at 15 pounds pressure 

 for 30 minutes. 



(23) Store as stock broth. 



(24) Dilute the stock broth as desired 

 for use. According to Hottinger 

 it may be diluted 10, 20 or more 

 times. Excellent results were ob- 

 tained, however, by diluting 1 part 

 stock broth with 1 part water, 

 (b) Park, Williams and Krumwiede pre- 

 pared a similar medium as follows: 



(1) Add 300 to 500.0 g. of fat-free 

 chopped meat to 1000.0 g. of water 

 to which 0.4% Na-iCOj has been 

 added. 



(2) Soak over night. 



(3) Heat at 80 °C. 



(4) Cool to 38°C. and add 15.0 cc. of 

 liquid trypsin. 



(5) Keep at 38°C. for 5 hours stirring 

 frequently. If kept over night at 

 this temperature add 10.0 cc. of 

 toluol (or thymol crystals). 



(6) Add normal HCl to neutralize 

 (indicator not specified). 



(7) Boil 7 minutes. 



(8) Strain. 



(9) Adjust the reaction. 



(10) Boil 30 minutes. 



(11) Filter. 



(12) Sterilize (method not given). 

 References: Klimmer (1923 p. 175), Park, 



Williams and Krumwiede (1924 pp. 118, 

 119). 



1120. Peckham's Trypsinized Beef Solution 



Constituents : 



1. Water 1000.0 cc. 



2. Beef 500.0 g. 



3. NaCl 5.0 g. 



4. Trypsin 4.0 g. 



Preparation : 



(1) Place 500.0 g. of finely chopped beef 

 which is as old as can be obtained 

 from the shops, in order that it be 

 free from muscle sugar, in 500.0 cc. 

 of water. 



(2) Make slightly alkaline with sodium 

 carbonate. 



(3) Place in a water bath and raise tem- 

 perature to 40°C. and add trypsin. 



(4) After an hour the mixture must be 

 again made alkaline with sodium 

 carbonate. 



(5) Allow to digest only from one to Ih 

 hours, or traces of indol may be 

 detected. 



(6) Boil and strain thru gauze. 



