CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



363 



(4) Allow to settle for one day. 



(5) Filter. 



(6) Distribute into test tubes. 

 Sterilization: Sterilize in the usual manner 



(method not given). 

 Use: Cultivation of diphtheria, cholera, 



influenza and other pathogenic forms. 

 Variants: Besredka cultivated tubercle 



bacilli on a medium prepared as follows: 



(1) Remove the yolks from 20 eggs (350.0 



CO.). 



(2) Add one liter of neutral distilled 

 water (neutralize acid if necessary. 

 Indicator not specified). 



(3) Prepare a 1.0% soda solution. 



(4) Add 175.0 cc. of (3) to (2). 



(5) Take a small portion of (4) in a 

 pipette and determine its trans- 

 parency. 



(6) Add (3) to (4) in 1.0 cc. lots until the 

 greatest amount of transparency is 

 obtained, testing as above. (The 

 quantity of soda necessary to be 

 added varies with the yolk.) 



(7) Dilute (6) until distilled water has 

 been added to the egg yolk in the 

 ration of 1:20. In this case dilute to 

 7.0 liters. 



(8) Distribute in Roux flasks in 50 to 

 150.0 cc. lots. 



(9) Sterilize at 110°C. for 20 minutes. 

 References: Nastiukoff (1893 *33 and 34), 



Besredka (1921 p. 291). 



1245. Harvey's Egg Yolk Solution. 



Constituents: 

 1. Egg yolk. 

 Preparation : 



(1) Extract 20.0 g. of egg yolk with 100.0 

 cc. of 30.0% alcohol for 4 days at 

 37°C. 



(2) Allow to deposit. 



(3) Decant the supernatant fluid. 

 Sterilization: Not specified. 



Use: Add 0.25% supernatant fluid to any 

 desired medium. The author treated 

 ascitic fluid in exactly the same manner. 



Reference: Harvey (1921-22 p. 121). 



1246. Hollande and Fumey's Basal Albumin 

 Solution. 



Constituents: 



1. Water 



2. NaCl 



1000.0 cc. 

 9.0 g. 



3. Egg white 



4. Litmus 

 Preparation: 



(1) Mix the whites of 2 eggs and measure 

 30.0 cc. into a flask. 



(2) Add 70.0 cc. of a physiological salt 

 solution prepared by dissolving 9.0 

 in a liter of distilled water. 



(3) Shake from time to time. 



(4) After 15 minutes filter on wet ab- 

 sorbent cotton. 



(5) Add 8.0 cc. of a solution containing 

 4.0 g. soda to 1000.0 cc. distilled 

 water to the filtrate. 



(6) Make up the volume of 200.0 cc. by 

 the addition of physiological salt 

 solution (see (2) for preparation). 



(7) Heat in the autoclave at 120° for 20 

 minutes. 



(8) Decant. 



(9) Distribute in test tubes. 



(10) Boil 10.0 g. of litmus cubes in 30.0 

 cc. ethyl alcohol at 80°C. 



(11) Decant, filter and save the filtrate. 



(12) Treat the residue on the water bath 

 for 45 minutes with 40.0 cc. distilled 

 water. 



(13) Filter and save the filtrate. 



(14) Treat the residue with 10.0 or 20.0 

 cc. of ethyl alcohol at 80°C. 



(15) Mix the filtrates and washings. 



(16) Neutralize the litmus solution by the 

 addition of 5.0% H2SO4 solution. 



(17) Dissolve 2.5 g. of one of the added 

 nutrients in 25.0 cc. of (16) by 

 heating. 



(18) Allow to cool and immerse strips of 

 bibulous paper in the solution. 



(19) Suspend the paper until dryness is 

 reached. 



(20) Mix equal parts of ether and alcohol. 



(21) Add one part collodion to 9 parts 

 (20). 



(22) Dip the paper (19) into (21). 



(23) Allow the paper to dry a second time. 



(24) Cut the paper into small rectangles 

 1 by 3 or 4 centimeters. 



(25) Add one or two of the rectangles to 

 each tube of (9). 



Sterilization: Sterilize for 20 minutes at 



118°C. 

 Use: To study fermentation by dysentery 



bacillus. The paper is turned red if the 



sugar is fermented. 



