CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



227 



The medium was also employed with- 

 out the addition of carbohydrates, 

 alcohols, etc. 

 (c) Klimmer added 0.5 to 1.0% of any 

 desired carbohydrate, alcohol, etc., 

 to bouillon, and mixed one part of 

 serum with one to twenty parts of 

 the sugar bouillon. 

 References: Hiss (1901-05 p. 323), Koch 

 and Pokschischewsky (1913 p. 10), Klim- 

 mer (1923 p. 200). 



775. Holman's Basal Serum Bouillon 



Constituents: 



1. Water 400.0 cc. 



2. Bouillon, double strength. . . 400.0 cc. 



3. Andrade's Indicator 80.0 cc. 



4. Beef serum 200.0 cc. 



Preparation : 



(1) Preparation of double strength bouil- 

 lon not given. 



(2) Adjust (1) to +1.2. 



(3) Add 200.0 cc. of water to 400.0 cc. of 

 (1). 



(4) Dissolve 8.0 g. of one of the added 

 nutrients in 80.0 cc. of Andrade's 

 indicator and add to (3). 



(5) Add 200.0 cc. of water to 200.0 cc. of 

 beef serum. 



(6) Mix 400.0 cc. of sterile (5) and 600.0 cc. 

 sterile (3). 



(7) Tube into sterile test tubes. 



(8) Incubate 2 days to determine sterility. 

 Sterilization: Sterilize (4) by heating in 



flowing steam on each of 3 successive days. 

 Sterilize (5) by filtering thru a Berke- 

 feld filter. 



Use: To study fermentation of carbo- 

 hydrates, alcohols, etc., by bacteria. 

 The author used streptococci. The pro- 

 duction of acid coagulates the serum. 



Added nutrients and variants: The author 

 added 8.0 g. of any desired carbohydrate, 

 alcohol, etc. to the basic solution. 



References: Holman (1916 p. 385), Stitt 

 (1923 p. 36). 



776. Ragit's Dehydrated Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Ragit Bouillon (Marck) .... 22.0 g. 

 Preparation: (1) Dissolve 2 in 1. 

 Sterilization: Not specified. 



Use: This solution is equivalent to a 1.0% 



peptone broth. 

 Variants: Lohnis prepared the medium as 



follows : 



(1) Heat 22.0 g. of Ragit bouillon in a 

 liter of tap water in the steam steri- 

 lizer for an hour. 



(2) Filter thru filter paper. 



(3) Add concentrated soda solution until 

 red litmus paper is turned slightly 

 blue. 



(4) Heat for 30 to 45 minutes in the steam 

 sterilizer. 



(5) Filter. 



(6) Sterilization not specified. 

 References: Marx (1910 p. 361), Lohnis 



(1913 p. 14). 



777. Bacto Dehydrated Broth 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Bacto bouillon (0.8%) 8.0 g. 



Preparation : 



(1) Dissolve 2 in 1. (Prepared accord- 

 ing to directions given on the bottle). 



(2) Initial pH = 7.1. (If sterilized at 

 15 pounds pressure for 20 minutes 

 pH = 6.5+.) 



Sterilization: Sterilize in the usual manner. 

 Use: General culture medium. 

 References: Brown (1921 p. 562), Digestive 

 Ferments Co. (1925 p. 10). 



778. Harvey and Iyengar's Dehydrated 

 Broth 



Constituents: 1. Trypsinized mutton in- 

 fusion broth. 

 Preparation : 



(1) Preparation of tryptic digest of 

 mutton infusion broth not given. 



(2) Adjust (1) topH = 8.0. 



(3) Cut up agar fibre into small pieces. 



(4) Add (3) to (2) (6.0% by weight). 



(5) Autoclave at 120° for an hour to 

 melt the agar. 



(6) Filter thru cotton, wool and muslin 

 into a tin receptacle. 



(7) Cut the agar out of the receptacle 

 and into slices. 



(8) Pass slices thru meat mincing ma- 

 chine with a finely perforated outlet 

 disc. 



(9) Spread minced nutrient agar on 

 metal or other type of trays. 



