216 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(8) Heat (7) in small containers in the 

 Arnold sterilizer for 30 minutes on 

 3 successive days. (Sterilize the 

 inulin solution in the autoclave.) 



(9) Add sufficient of (8) to sterile (6) 

 to give a 1.0% concentration of 

 sugar (under aseptic conditions), 

 generally 5.0% glycerin is used 

 instead of 1.0%. 



(10) In routine work with glucose, lac- 

 tose, sucrose, mannitol and dul- 

 citol it is generally sufficient to 

 add the sugar to the medium and 

 sterilize in the Arnold for 30 

 minutes on each of 3 successive 

 days. 

 They also used an extract broth, see 

 variant (Q) 779. 

 References: Committee A. P. H. A. (1899 

 p. 77), (1905 p. 107), Percival (1920 p. 56), 

 Besson (1920 p. 31), Harvey (1921-22 

 pp. 87, 112), Stitt (1923 p. 34), Park, 

 Williams and Krumwiede (1924 p. 123). 



749. Heinemann's Basal Sugar Free 

 Infusion Broth 



Constituents : 



1. Water 1000.0 cc. 



2. Chopped beef 1.0 lb. 



3. Peptone (Witte) 10.0 g. 



4. NaCl 5.0 g. 



5. CaCl2 6.0 g. 



Preparation : 



(1) Chop beef fine and soak with water 

 over night. 



(2) Add a culture of B. coli and incubate 

 the mixture at 37° for 24 hours. 



(3) Then heat to 60 °C. for two hours. 



(4) Raise to boiling point and boil 

 30 minutes. 



(5) Press out meat, and the remaining 

 meat mixed again with water and 

 pressed a second time. 



(6) Mix both fluids and bring to 

 1000.0 cc. volume. 



(7) Dissolve 3 and 4 in boiling (6) . 



(8) Add 5 and boil until the reaction is 

 about 0.3 acid to phenolphthalein. 

 (Requires from 30 to 60 minutes.) 



(9) Filter sterile (8) while boiling hot, 

 under aseptic conditions. 



(10) Dissolve one of the added nutrients 

 in (9). 



(11) Distribute in small sterile Erlen- 

 meyer flasks or sterile Nessler tubes. 



Sterilization: Method of sterilization not 

 given. Sterilize in large Erlenmeyer 

 flasks before filtering. 



Use: To study the fermentation of sugars, 

 alcohols, etc. 



Added nutrients and variants: 



(a) The author added 1.0% of one of the 

 following to the basic solution. 



glucose inulin 



lactose mannitol 



sucrose salicin 



raffinose 



(b) Besemer did not specify the prepara- 

 tion or composition of meat infusion 

 bouillon, and adjusted the reaction 

 to -hO.3 Fuller's scale. He added 

 1.0% of one of the following: 

 fructose amylose 

 glucose dextrin 

 galactose inulin 

 mannose glycerol 

 sorbose erythritol 

 arabinose adonitol 

 xylose dulcitol 

 rhamnose mannitol 

 lactose sorbitol 

 sucrose amygdalin 

 raffinose maltose 



(c) Kan Ichiro Morishima, prepared 

 media containing indicators as fol- 

 lows: 



(1) Prepare meat infusion broth 

 (method not given). 



(2) Inoculate with B. coli, incubate 

 for 24 hours, autoclave and filter. 



(3) Add 1.0% peptone and 0.5% NaCl. 



(4) Sterilize at 15 pounds for 15 

 minutes. 



(5) To broth add sterile sugar solu- 

 tion in concentration of 1.0% sugar 

 solution prepared by dissolving 

 sugar in sterile water and heating 

 in autoclave for 10 minutes at 

 10 pounds. 



(6) Add 5.0 cc. sterile litmus or 5.0 cc. 

 of 2.0% phenol red and 1.2 cc. of 

 decolorized 1.0% aqueous solution 

 of china blue per 100.0 cc. of broth. 



(7) Adjust to pH = 7.0-7.1 by means 

 of phenol red. 



(d) Giltner prepared a neutral red me- 

 dium as follows: 



(1) Soak one pound of lean finely 

 chopped beef in 1000.0 cc. of dis- 

 tilled water over night (24 hours). 



