218 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



2. Peptone 100 g. 



3. Beef infusion 500.0 cc. 



Preparation : 



(1) Composition of beef infusion not 

 given. Author also used a 1.0% 

 solution of Liebig's beef extract 

 instead of infusion. 



(2) Prepare 500.0 cc. of a 2.0% peptone 

 solution. 



(3) Mix (1) and (2). 



(4) Dissolve one of the combinationg 

 given under added nutrients in (3). 



(5) Steam 15 minutes and check the 

 reaction. 



(6) Distribute as desired. 

 Sterilization: Sterilize at 115° for 20 



minutes. 



Use: Cultivation of Bad. diphtheriae and 

 toxin production. 



Added nutrients: The authors added one 

 of the following materials or combina- 

 tions of materials. 



(a) xanthine 0.05 g, 



hypoxanthine 0.05 g, 



(b) glucose amine hydrochloride. 2.0 g 



(c) sodium asparaginate 1.5 g 



(d) creatin 0.2 g 



creatinin 0.15 g 



(e) cystine 0.5 g 



(f) glutaminic acid hydro- 

 chloride 2.5 g 



(g) glycocoll 0.75 g 



(h) histidine dichloride 0.5 g 



(i) leucine 30.0 g, 



(j) tryptophane 

 (k) tyrosine 

 Reference: Davis and Ferry (1919 pp. 235, 

 236). 



753. Grimbert's Basal Carbonate Extract 

 Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Peptone 3.0 g. 



3. Meat extract (Liebig's) .... 2.0 g. 



4. CaC03 10.0 g. 



Preparation: 



(1) Dissolve 2 and 3 in 1. 



(2) Dissolve 3.0% of one of the added 

 nutrients in (1). 



(3) AddtheCaCOs to (2). 

 Sterilization: Method not given. 



Use: To study fermentation. Author used 

 Friedlander's pneumobacillus. 



Added nutrients: The author added 3.0% 



of any desired fermentable sugar. 

 Reference: Grimbert (1895 p. 840). 



754. Committee A. P. H. A. (1917) Basal 

 Extract Broth 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Beef extract 3.0 g. 



3. Peptone 5.0 g. 



Preparation: 



(1) Add 3.0 g. of beef extract and 5.0 g. 

 of peptone to a liter of distilled 

 water. 



(2) Heat slowly on a steam bath to at 

 least 65°C. 



(3) Neutralize to phenolphthalein. 



(4) Cool to 25°C. and filter thru paper 

 until clear. 



(5) Add 1.0% of one of the added nu- 

 trients. 



Sterilization: Sterilize in the autoclave at 

 15 pounds (120°C.) for 15 minutes after 

 the pressure reaches 15 pounds. 



Use: To study fermentation of carbohy- 

 drates, alcohols, etc., by bacteria. 



Added nutrients and variants: 



(a) The committee added 1.0% of any 

 desired carbohydrate. 



(b) Mudge specified the use of Liebig's 

 meat extract, and used 1.0% Witte's 

 peptone. He neutralized to phenol- 

 phthalein, added 1.0% of any desired 

 carbohydrate, tubed and sterilized 

 for 15, 30, 60 or 120 minutes in the au- 

 toclave, or 15 minutes in the Arnold 

 on each of 3 successive days. 



(c) Committee S. A. B. used the same 

 constituents as Committee A. P.H. A. 

 They specified that the medium might 

 be clarified by the addition of egg 

 and adjusted to pH = 6.6 to 7.4. 

 One per cent of any desired carbo- 

 hydrate was added. 



(d) Tanner and Ball added 1.0% of any 

 desired carbohydrate, alcohol, etc., 

 to variant (b) 695. 



(e) Committee A. P. H. A. (1920) ad- 

 justed to a faint pink using phenol 

 red as an indicator or neutralize to 

 phenolphthalein if the reaction does 

 not come between neutral and +1.0. 

 The committee used 0.5% carbo- 

 hydrate instead of 1.0% as in 1917. 



