CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



565 



1806. Scales' Salt Cellulose Agar 

 Constituents : 



1. Water. 



2. Filter paper 



3. Nutrient agar. 



4. Salts. 

 Preparation : 



(1) Dilute 100.0 cc. of concentrated 

 H2SO4 with 60.0 cc. distilled water in 

 a two liter Erlenmeyer flask. 



(2) Cool to about 60 or 65°C. 



(3) Moisten 5.0 g. of filter paper with 

 water. 



(4) Add (3) to (2) and shake until the 

 paper is dissolved. 



(5) When solution is complete fill the 

 flask as quickly as possible with cold 

 tap water. The task of dissolving 

 the filter paper and filling the flask 

 requires about one minute. 



(6) The rapid addition of cold water 

 precipitates the cellulose in small 

 flocks. 



(7) Throw the precipitate on a filter 

 paper and wash with distilled water 

 until free from H2SO4. This re- 

 quires about 3 hours time and only 5 

 liters of distilled water. A 12 inch 

 funnel with a folded filter is the best 

 apparatus to filter with. 



(8) During the washing do not allow the 

 volume of water in the funnel to 

 get below 100.0 cc. 



(9) When the wash water is free from 

 H2SO4 as shown by the addition of 

 BaCl2 solution, allow the volume in 

 the funnel to drain to about 200.0 cc. 



(10) Brush any cellulose particles cling- 

 ing on the dry filter paper into the 

 suspension. 



(11) Punch a hole in the bottom of the 

 filter, collecting the cellulose sus- 

 pension. 



(12) Wash the filter with a stream of 

 water from a wash bottle. 



(13) Make the suspension to 500.0 cc. 



(14) Prepare 500.0 cc. of nutrient agar 

 with salts (composition not spe- 

 cified). 



(15) Mix (13) and (14). 

 Sterilization: Not specified. 



Use: Cultivation of organisms capable of 



utilizing cellulose. 

 Reference: Scales (1916 p. 662). 



1807. Cantani's Basal Glycerol Agar 

 Constituents : 



1. Nutrient agar. 



2. Glycerol. 

 Preparation : 



(1) Mi.x equal amounts of glycerol and 

 one of the added nutrients. Place in 

 an Erlenmeyer flask. 



(2) Melt tubes of sterile nutrient agar and 

 add 0.5 to 0.75 cc. of sterile (1) to 

 each tube. 



(3) Incubate 24 hours to test sterility. 

 Sterilization: Storing the fluid in glycerol 



tends to sterilize it. After a time test the 

 sterility of the mixture. Method of 

 sterilization of agar not given. 



Use: Cultivation of parasitic and patho- 

 genic forms. 



Added nutrients: The author added one 

 of the following: Urine, pus, sperm, milk, 

 egg white and other albuminous materials. 



Reference: Cantani (1910 p. 471). 



1808. Wurtz's Glycerol Agar 



Constituents : 



1. Nutrient agar 1000.0 cc. 



2. Glycerol (6.0%) 60.0 g. 



Preparation : 



(1) Add 6.0% sterile glycerol to sterile 

 nutrient agar under aseptic con- 

 ditions. 

 Sterilization: Method not specified. 

 Use: Cultivation of tubercle bacilli. Simi- 

 lar media have been used to cultivate a 

 large variety of other pathogenic forms. 

 Variants : 



(a) Gllicksmann (1897) added 7.7% gly- 

 cerol to 1.5% nutrient agar and 

 cultivated diphtheria bacilli. 



(b) Thoinot and Masselin added 4.0 to 

 6.0% glycerol to nutrient agar, 



(c) Dalton and Eyre added 5.0% glycerol 

 to nutrient agar and adjusted the 

 reaction to -}-10 on Eyre's scale. 

 They cultivated Micrococcus meli- 

 tensis. 



(d) Smith cultivated plant parasites in a 

 medium prepared by the addition of 

 50.0 cc. of Schering's c.p. twice dis- 

 tilled glycerol to nutrient agar. 



(e) Roux and Rochaix added 1.0 to 5.0% 

 glycerol to agar. They suggested 

 the addition of several drops of a 

 saturated gum arable solution. This 



