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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



923. Rivas' Glucose Bouillon 

 Constituents: 



1. Bouillon 958.0 cc. 



2. Glucose 10.0 g. 



3. Peptone 15.0 g. 



4. Ammono-sulpho-hydrate 



water 40.0 cc. 



5. Sodium indigo sulphuric 



acid (10.0% soln.) 2.0 cc. 



Preparation : 



(1) Prepare bouillon. 



(2) Add 1.0% dextrose and 1.5% pep- 

 tone. 



(3) Prepare a 10.0% sodium indigo sul- 

 phuric acid solution in distilled 

 water. 



(4) Heat (3) at 100°C. for one hour. 



(5) Prepare a 1.0% ammonia solution in 

 sterile distilled water. 



(6) Place 150-200.0 g. of distilled water 

 in an Erlenmeyer flask and stopper 

 with cotton. Place a bent rod thru 

 the cotton reaching the bottom of the 

 flask. Plug the other end of the 

 rod with cotton. 



(7) When cool pass a stream of washed 

 hydrogen sulphide from a Kipp 

 generator thru the water for 5 

 minutes. 



(8) Prepare 10 test tubes, containing 

 10.0 cc. of (7) each. 



(9) To a series of tubes add 1 drop, 

 2 drops, 3 drops, etc., of (5) to each 

 tube. Shake thoroughly. 



(10) To each tube of (9) add 3 drops of a 

 10.0% methylene blue solution in 

 50.0% alcohol. 



(11) Observe in which tube a complete 

 decolorization takes place in about 

 1 minute (usually required 3 to 6 

 drops) . 



(12) When the proper amount of am- 

 monia has been determined, pour 

 20.0 cc. of (7) into a sterile cylinder. 



(13) Add the proper amount of am- 

 monia (usually 10 to 20 drops) to the 

 20.0 cc. of hydrogen sulphite water. 

 This solution is to be kept cool or a 

 turbidity will develop. 



(14) To 958.0 cc. of sterile (2) add 21.0 

 cc. of (13) (20.0 cc. of H2S water 

 and 1.0 cc. of indigo sulphuric acid 

 solution). The medium should be- 

 come clear after 1 minute. 



(15) Distribute in tubes, and cover with 

 a layer of sterile olive oil. Plug 

 with cotton and seal with gum. 

 This process should be done quickly. 



(16) Incubate at 37°C. for 48 hours dis- 

 carding any tubes showing turbidity 

 or any sign of color. 



Sterilization: Sterilize (6) at 120°C. for 

 15 minutes. 



Use: Cultivation of anaerobes. 



Variants : The author omitted the ammono- 

 sulpho-hydrate (steps (5) thru (14)), but 

 prepared a 1.0% NaiSOs solution and 

 added 50.0 cc. to 948.0 cc. of glucose 

 bouillon containing 2.0 cc. of a 1.0% 

 solution of sodium indigo sulphuric. 

 The medium was covered with olive oil 

 as specified above. This medium is in- 

 ferior to the one described above. 



Reference: Rivas (1902 p. 836). 



924. MacConkey's Iodide Glucose Bouillon 



Constituents: 



1. Bouillon 1000.0 cc. 



2. Glucose 20.0 g. 



3. KI 5.0 g. 



Preparation : 



(1) Prepare bouillon. 



(2) Dissolve 2 and 3 in (1). 



(3) Tube in fermentation tubes. 

 Sterilization: Sterilize in a steamer for 10 



minutes on each of two successive days, 



taking care not to over heat. 

 Use: Enrichment of colon-typhoid group. 

 Variants: Author added 10.0 or 20.0 g. KI 



instead of 5.0 g. 

 Reference: MacConkey (1905 p. 337). 



925. Kitasato's Formate Glucose Bouillon 



(Tanner) 

 Constituents : 



1. Bouillon 1000.0 cc. 



2. Glucose 20 g. 



3. Sodium formate 4.0 g. 



Preparation : 



(1) Dissolve 2 and 3 in bouillon. 



(2) Tube. 



Sterilization: Method not given. 

 Use: General culture medium. 

 Reference: Tanner (1919 p. 47). 



926, Heymann's Acetic Acid Glucose 

 Bouillon (Finkelstein) 

 Constituents : 

 1. Bouillon 1000.0 cc. 



