CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



793 



(3) Thoroly mix the yolk and albuminous 

 portion by gently stirring. 



(4) Strain the mixture thru sterile gauze. 



(5) Add 1 part sterile physiological salt 

 solution (0.85%) NaCl containing 

 6.0% glycerol to 3 parts (4). 



(6) Tube. 



(7) Coagulate in 3 to 5 minutes by heat- 

 ing in the steamer. 



Sterilization: Methods not given. 

 Use: Cultivation of tubercle bacilli. 

 Variants : The author added sterile glycerol 



bouillon to prevent drying. 

 References: Abel (1912 p. 97), Harvey 



(1921-22 p. 85), Cunningham (1924 p. 165). 



2440. Twort and Ingram's Basal Glycerol 

 Egg Medium 



Constituents : 



1. Physiological salt solution 30.0 cc. 



2. Glycerol 4.0 cc. 



3. Eggs (fresh) 66.0 cc. 



Preparation : 



(1) Add 4.0 cc. of glycerol to 30.0 cc. of 

 physiological salt solution. 



(2) Add one of the added nutrients 

 (amount not given) to (1). 



(3) Steam for one-half hour. 



(4) Add 66.0 cc. of whole freshly laid 

 eggs. Mix well. 



(5) Distribute into sterile tubes. 



(6) Plug with cotton wool and sterilize 

 at 61°C. for one hour on 3 successive 

 days. Incubate the tubes over night 

 between the second and third heating. 



(7) Immediately after the third heating 

 slope the tubes and inspissate at 

 about 85°C. 



(8) Inoculate and incubate for 4 months 

 at 39 °C. 



Sterilization: See step (6) above. 



Use: Cultivation of Johnes bacilli. 

 Author reported that media prepared 

 from vegetable extracts were not so good 

 as media prepared from acid fast bacilli 

 or their extracts. Growth was sometimes 

 obtained with the extracts listed. Many 

 others were tried with negative results. 



Added nutrients: Alcoholic extracts of 

 currant grapes, figs, oats, linseed, acid 

 fast bacilli, and Contharelhis aurantiacus 

 (a fungus) were prepared using a Soxhlet 

 extractor. The materials were extracted 



for 3 hours, filtered and the extract evap- 

 orated to dryness. One of these ex- 

 tracts was added to the basic medium. 

 Reference: Twort and Ingram (1914 p. 



278). 



2441. Roddy's Glucose Bouillon Egg Medium 



Constituents : 



1. Bouillon 1000.0 cc. 



2. Glucose 1.0% 10.0 g. 



3. Egg 

 Preparation : 



(1) Prepare a 1.0% glucose bouillon. 



(2) Add 10.0 to 15.0 cc. of (1) to the 

 white and yolk of one egg. 



(3) Make a smooth mixture in a mortar. 



(4) Tube. 



(5) Inspissate. 



Sterilization: Sterilize as for ordinary 



serum slants. (Method not given). 

 Use: General culture medium. 

 Variants : 



(a) Roddy added 1.0 cc. of glycerol 

 bouillon to each tube before final 

 sterilization in the autoclave when 

 used to cultivate tubercle bacilli. 

 The cotton plugs were paraffined. 



(b) Stitt cultivated anaerobes and pre- 

 pared the medium as follows: 



(1) Preparation of 1.0% glucose bouil- 

 lon not given. 



(2) Flame a mortar wet with alcohol. 



(3) Crack a clean egg with a sterile 

 knife and allow the contents to 

 drop into the sterile mortar. 



(4) Add from 10.0 to 15.0 cc. of sterile 

 (1) to (3). 



(5) Add 5 drops of a 1.0% neutral red 

 solution for each egg added to (4). 



(6) Mix into a smooth paste. 



(7) Inspissate for 2 hours in the rice 

 cooker. If the mortar and knife 

 are not sterilized it is necessary to 

 sterilize after inspissation. 



(c) Harvey cultivated anaerobes and 

 isolated B. diphtheriae on the follow- 

 ing medium. 



(1) Add 1.0% glucose to infusion broth 

 (See variant (bb) medium 779). 



(2) Make a suspension of 12.0 cc. of 

 (1) and the contents of one egg. 



(3) Place the suspension in test tubes. 



(4) Coagulate. 



