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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



821. Dalimier and Lancereau's Opsine 

 Bouillon 



Constituents : 



1. Bouillon. 



2. Opsine. 

 Preparation : 



(1) Method of preparation of bouillon 

 not specified. Reaction alkaline. 



(2) Add opsine. (Opsine is a commercial 

 mixture of amino acids.) 



Sterilization: Method not given. 



Use: General culture medium. Authors 

 reported the growth of staphylococci, 

 typhoid and paratyphoid bacilli, Micro- 

 coccus melitensis, B. prodigiosus, Vibrio 

 cholerae, pneumococci and others. 



Variants : The authors suggested the addi- 

 dion of glycerol. 



Reference: Dalimier and Lancereau (1913 

 p. 419). 



822. 



Frieber's Trypsinized Bouillon 

 (Committee S. A. B.) 



Constituents : 



1. Bouillon 1000.0 cc. 



2. Physiological salt Solution. . 3000.0 cc. 



3. Trypsin 0.2 g. 



Preparation: 



(1) Add 0.2 g. trypsin to aliter of peptone 

 bouillon. 



(2) Add chloroform and toluol. 



(3) Incubate for 24 to 48 hours at 37°C. 



(4) Filter. 



(5) Dilute with 3 parts physiological salt 

 solution. 



Sterilization: Method not given. 

 Use: To determine the production of indol. 

 Reference: Committee Society American 

 Bacteriologists (1923 p. 31). 



823. Wilcox's Glucose Veal Infusion Broth 



Constituents : 



1. Water 1000.0 cc. 



2. Veal (Market) 1.0 lb. 



3. Peptone (Witte) 10.0 g. 



4. Glucose (anhydrous) 10.0 g. 



5. NaCl 5.0 g. 



Preparation : 



(1) Add water to chopped veal and let 

 stand in ice chest over night. 



(2) Place infusion over free flame and 

 heat to 45°C. for 1 hour, 



(3) Boil briskly for ^ hour. 



(4) Strain broth thru cheese cloth. 



(5) Measure broth and add 3, 4 and 5. 



(6) Boil until 3, 4 and 5 are in solution. 



(7) Correct to +1, phenolphthalein as 

 indicator, with N/10 NaOH. 



(8) Filter thru 2 layers absorbent cotton 

 directly into 2 liter Erlenmeyer flask, 

 leaving only enough space to allow 

 for expansion of broth in Arnold. 



Sterilizatiori : Sterilize in Arnold 90 minutes 

 on first day and 1 hour on second day. 



Use: Study toxin production by B. tetani 

 and preliminary cultures for inoculation 

 in toxin broth. 



Variants : 



(a) Walbum studied diphtheria toxin pro- 

 duction on a veal infusion medium 

 containing 1.5% Witte peptone, 0.5% 

 NaCl and 0.2% glucose. The me- 

 dium was sterilized unadjusted, and 

 the sterile medium adjusted to pH 

 = 7.0 by adding sterile calcinated 

 soda. He reported that the addition 

 of small amounts of manganese chlo- 

 ride (final concentration 0.01 molar) 

 in many cases markedly increased the 

 toxin production. 



(b) Park, Williams and Krumwiede pre- 

 pared the medium as above, but did 

 not give the adjustment of reaction 

 and heated for 90 minutes at 90°C. 

 instead of 90 minutes in Arnold. 



(c) Park, Williams and Krumwiede used 

 a similar medium for the production 

 of diphtheria toxin. This medium 

 was prepared as follows: 



(1) Mix 20 pounds of lean chopped veal 

 with 20 liters of tap water. 



(2) Place in the ice box over night. 



(3) Strain the next morning. 



(4) When chilled add 10.0 cc. of a 

 24 hour culture of B. coli for each 

 liter and incubate at 37°C. over 

 night. 



(5) In the morning add 1 egg for each 

 2 liters. 



(6) Boil 20 minutes and strain. 



(7) Adjust the reaction to 0.5 acid to 

 phenolphthalein. 



(8) Add 1.0% Park Davis & Co. pep- 

 tone, and 0.5% NaCl and heat to 

 boiling to dissolve. 



(9) Readjust the reaction. 



(10) Filter thru paper and cotton. 



