224 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(1) Prepare an extract broth using 

 Liebig's extract and peptone. 



(2) Titrate and adjust reaction of (1) 

 to 0.6-0.8 acid to phenolphthalein. 



(3) Add 2.0% glucose, if desired, and 

 10.0% ascitic fluid. 



(4) Distribute in six ounce nursing 

 bottles containing 150.0 cc. of fluid 

 in a column 4.5 cc. wide and 8.75 cc. 

 high. 



(5) Fresh sterile pieces of rabbit kidney 

 may be added, if desired. 



(6) Method of sterilization not given. 

 References: Symmers and Wilson (1909 



p. 9), Rosenow and Towne (1917 p. 176). 



768. Holman's Basal Serum Extract Broth 



Constituents : 



1. Distilled water 500.0 cc. 



2. Beef serum 100.0 cc. 



3. Peptone (Witte) 4.0 g. 



4. Meat extract (Liebig's) 1.2 g. 



5. NaCl 2.0 g. 



6. Andrade's Indicator 4.0 cc. 



Preparation : 



(1) Collect beef blood from abattoir in 

 sterile quart jars. 



(2) Allow blood to clot in a cool room of 

 the abattoir for about 15 minutes. 



(3) Take to laboratory and allow to 

 stand over night in a cool place. 



(4) Obtain clear serum by centrifuga- 

 tion. 



(5) Add 100.0 cc. of serum to 300.0 cc. 

 distilled water. 



(6) Dissolve 3, 4 and 5 in 200.0 cc. of dis- 

 tilled water. 



(7) Adjust to +1.2% acid. 



(8) Add 4.0 cc. of Andrade's indicator 

 and 4.0 g. of one of the nutrients 

 listed below. 



(9) To cool sterile (8) add 500.0 cc. of 

 sterile (5). 



(10) Tube by means of a sterile tubing 

 funnel or by means of a sterile 

 syphon. 



(11) Incubate several days to insure 

 sterility. 



Sterilization : Sterilize (5) by filtering thru 

 a Berkefeld filter. Sterilize (8) in flowing 

 steam 15 to 20 minutes on each of 3 suc- 

 cessive days. 



Use: To study fermentation of carbohy- 

 drates, alcohols, etc. by bacteria. The 

 author used streptococci. 



Added nutrients and variants: 



(a) The author added 4.0 g. of any de- 



sired carbohydrate, alcohol, etc. 



(b) The author prepared a similar me- 

 dium in the following manner: 



(1) Collect beef blood from abattoir 

 in sterile quart jars. 



(2) Allow blood to clot in the cool 

 room of the abattoir for about 15 

 minutes. 



(3) Take to laboratory and allow to 

 stand over night in a cool place. 



(4) Obtain clear serum by centrifu- 

 gation. 



(5) Add 100.0 cc. of serum to 300.0 cc. 

 distilled water. 



(6) Sterilize for 15 to 20 minutes in 

 flowing steam on 3 successive days. 



(7) Dissolve 40.0 g. Witte's peptone, 

 12.0 g. Liebig's meat extract and 

 20.0 g. NaCl in 1000.0 cc. distilled 

 water. 



(8) Titrate while hot to neutral to 

 phenolphthalein. 



(9) Add 4.0 g. of any desired carbo- 

 hydrate and 4.0 cc. of Andrade's 

 indicator per 100.0 cc. 



(10) Sterilize in flowing steam for 15 to 

 20 minutes on 3 successive days. 



(11) The remaining method of prepara- 

 tion is somewhat ambiguous. 



"The sterile serum water was then 

 mi.xed with this quadruple strength 

 carbohydrate broth, and the medium, 

 consisting of 1 part serum to 4 parts 

 1% carbohydrate broth, was tubed 

 into sterile tubes by means of a 

 sterile tubing-funnel, or by use of a 

 sterile syphon." 

 Reference: Holman (1914 pp. 210, 211). 



769. MacConkey's Basal Bile Salt Iodide 

 Bouillon 



Constituents: 



1. Nutrient bouillon 100.0 g. 



2. Sodium taurocholate (Com- 

 mercial) 0.5 g. 



3. KI 0.5 g. 



Preparation : 



(1) Preparation of nutrient bouillon not 

 given. 



(2) Dissolve 2. 3 and one of the added 

 nutrients in (1). 



(3) Tube in fermentation tubes. 

 Sterilization: Sterilize in a steamer on 2 



