238 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



5. Sodium or potassium 



phosphate 2.0 g. 



Preparation : 



(1) Chop 500.0 g. of fat and tendon free 

 beef into very small pieces. 



(2) Add 1 liter of water and allow to 

 stand over night in the cold. 



(3) Press the juice thru a linen cloth by 

 means of a meat press. 



(4) Make up the volume to 1000.0 cc. 

 by the addition of water. 



(5) Add 5.0 g. NaCl, 2.0 g. sodium or 

 potassium phosphate and 20 to 25 

 grams of peptone. 



(6) Boil for an hour. 



(7) Filter thru a linen cloth and then 

 thru paper. 



(8) Adjust the reaction slightly alkaline 

 to litmus by the addition of KHCO3 

 solution, drop by drop. 



(9) Boil for 15 minutes. 



(10) Allow to settle. Skim off the fat if 

 any be present. 



(11) Distribute as desired. 

 Sterilization: Sterilize at 115 to 120°C. for 



30 minutes or by the fractional method. 



Use: General culture medium. 



Variants: Besson prepared a similar me- 

 dium 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, 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 10 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. of Chapoteaut's or De- 

 fresne's 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 alkaline 

 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) Distribute as desired. 



(14) Sterilize at 100 to 115"C. for 20 

 minutes. 



Reference : Roux and Rochaix (1911 p. 106), 

 Besson (1920 p. 26). 



783. Zielleczky's Phenolphthalein Infusion 

 Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Beef... 500.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



5. Phenolphthalein solution 

 Preparation : 



(1) Prepare bouillon from 1, 2, 3 and 4. 

 (Exact method not given.) 



(2) Make alkaline with NasCO,. 



(3) Prepare a mixture of 50.0 cc. of water 

 and 50.0 cc. of absolute alcohol. 



(4) Dissolve 1.0 g. of phenolphthalein 

 in (3). 



(5) Just before use add 1.0 cc. of (4) to 

 19.0 cc. of water. 



(6) Add 0.7 to 0.8 cc. of (5) to 5.0 cc. of 

 bouillon. 



Sterilization: Not specified. 



Use: Differentiation of coli and typhoid 

 organisms. Coli decolorized the red 

 medium completely and more thoroly 

 than did typhoid. Coli caused decolori- 

 zation after 5 to 7 hours. 



Reference: Zielleczky (1902 p. 153). 



784. Muller's Indicator Infusion Broth 



Constituents : 



1. Meat Infusion 1000.0 cc. 



2. Peptone 10.0 g. 



3. NaCl 5.0 g. 



4. Indicator 

 Preparation : 



(1) Exact method of preparation of meat 

 infusion from fresh meat not given. 



(2) Dissolve 2 and 3 in 1. 



(3) Make slightly alkaline. 



(4) Add 10 drops (12 drops = 1.0 cc.) of 

 a 1:100 solution of methylene blue, 

 10 drops of 1 : 100 solution of rosanalin, 

 12.0 cc. of a concentrated watery solu- 

 tion of litmus or 10.0 cc. of a 2:100 

 solution of indigo carmine. 



Sterilization: Not specified. 

 Use: To study the reduction ability of the 

 colon-typhoid group and other bacteria. 



