206 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(3) Boil for one hour. 



(4) Remove from the flame and add 10.0 g. 

 of powdered lactose. 



(5) Filter thru cotton flannel until clear. 



(6) To each liter of the filtrate add 10.0 cc. 

 of a 1.0% solution of brilliant green. 



(7) Tube. 



Sterilization: Sterilize in the autoclave for 

 15 minutes at 15 pounds pressure. 



Use: Presumptive test for B. coli in water 

 analysis. Author reported that B. coli 

 produced gas in this medium. Other 

 forms were inhibited, especially B. welchii 

 and other anaerobes. 



References: Muer and Harris (1920 p. 875), 

 Levine (1921 p. 112), Park, Williams and 

 Krumwiede (1924 p. 132). 



736. Conradi's Bile Peptone Solution 

 Constituents: 



1. Bile (fresh beef) 900.0 cc. 



2. Peptone (Witte) 100.0 g. 



3. Glycerol 100.0 g. 



Preparation : 



(1) Mix 1, 2 and 3. 



(2) Place the unfiltered sterile (1) in 2.0 

 or 3.0 cc. quantities in sterile glass 

 tubes 9.0 cm. high and 18 mm. in 

 diameter with rubber stoppers. 



Sterilization: Sterilize (1) in streaming 

 steam for 2 hours. Heat (2) at 100°C. 

 for 30 minutes. 



Use: Enrichment and isolation of typhoid 

 bacilli from patient's blood. About 0.5 

 to 2.0 cc. of the patient's blood is added 

 to each tube of medium. 



Variants: Roddy and Stitt used 2.0% 

 peptone. 



References: Conradi (1906 p. 58), (1906 

 p. 1655), Roddy (1917 p. 42), Harvey 

 (1921-22 p. 89), Stitt (1923 p. 47), Klim- 

 mer (1923 p. 214). 



737. Vierling's Whey Peptone Solution 

 Constituents: 



1. Distilled water. . . . 350.0 cc. 



2. Skim milk powder. 30.0 g. 



3. Physiological salt 

 solution 150.0 cc. 



4. Peptone 0.02 to 0.03 g. 



5. Kahlbaum's litmus 



solution 20.0 to 30.0 cc. 



Preparation : 



(1) Dissolve 30.0 g. of skim milk powder 



in 200.0 cc. distilled water by heat- 

 ing on a water bath. (Milk powder 

 is prepared by evaporating skim 

 milk to 5 its volume in a vacuum, 

 powdered, and pass the powder thru 

 a hot stream of air to dry.) 



(2) Add 4.0 cc. of an 18.0% CaClo solu- 

 tion to (1) and heat in the steamer 

 for 40 minutes to precipitate the 

 casein. 



(3) Add 2.0 cc. N/1 soda solution and 

 heat for 25 more minutes. 



(4) Filter thru a folded filter, pressing 

 out the whey by means of cloth. 



(5) If not clear filter thru moistened 

 filter paper shreds using suction. 

 The whey will not become bright 

 and clear unless it is allowed to 

 stand in the ice box for 3 to 5 days. 



(6) Add 150.0 g. distilled water and 

 150.0 cc. physiological salt solution 

 making the volume to 500.0 cc. 



(7) Dissolve 0.02 or 0.03 g. Witte pep- 

 tone in (6). 



(8) Sterilize for 30 minutes. A tur- 

 budity is formed which settles out 

 by standing in the ice box over 

 night. Filtering is unsatisfactory. 



(9) Add 20.0 to 30.0 cc. sterile Kahl- 

 baum's litmus tincture to (8) by 

 means of a sterile pipette. 



(10) Standardize the tint by the addition 

 of N/10 soda solution or lactic acid. 

 The medium will become somewhat 

 bluer by sterilization so a rather red 

 tinge is to be desired. 



(11) Prepare the tubes in which the whey 

 is to be distributed by adding about 

 10 mg. CaCOs to each tube and heat- 

 ing for 1 hour, dry heat at 120 to 

 140°C. The tubes should be of Jena 

 glass or tubes that have been used 

 a great deal. The CaCOa tends to 

 neutralize the acids formed from 

 galactose and glucose which have 

 come from hydrolysed lactose. It 

 does not injure the reddening or the 

 whey. The tubes may be used with- 

 out the CaCOa. 



(12) Distribute into 5.0 cc. lots into 

 sterile test tubes. 

 Sterilization: Sterilize for 5 minutes 



(method not given). 

 Use: General culture medium. 



