796 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



minutes or by dipping in boiling 

 water for 5 seconds or so. 



(5) Mix the whites and yolks of these 

 eggs in a sterile mortar. 



(6) Add 1.0% alcoholic solution of a 

 1:100 gentian violet solution to 

 the filtrate of (3). 



(7) Mix equal parts (6) and (5). 



(8) Tube ip 3.0 to 4.0 cc. quantities 

 and slant. 



(9) Inspissate at 85° until the medium 

 has solidified. 



(10) Heat the slants at 75°C. for one 

 hour on the second and third days, 

 (g) Klimmer. 



(1) Soak 250.0 g. of finely chopped lean 

 veal in 212.0 cc. of water and 

 37.5 g. glycerol in the ice box over 

 night. 



(2) Filter 16 to 20 fresh thoroly mixed 

 eggs thru gauze. 



(3) Add 200.0 cc. of the juice from (1) 

 to 400.0 cc. of (2). 



(4) To each 100.0 cc. of (3) add 1.0 cc. 

 of a 1.0% gentian violet solution. 



(5) Distribute ih tubes. 



(6) Heat in the serum inspissator on 

 each of 3 successive days for 30 

 minutes at 85°, 75° and 75°C. 



(h) Park, Williams and Krumwiede. 



(1) Extract 500.0 g. of veal or beef in 

 500.0 cc. of 15.0% glycerol in water 

 for 24 hours. 



(2) Collect the fluid by means of a 

 press. 



(3) Sterilize by filtering. 



(4) Add 1.0 cc. of a 1.0% alcoholic 

 solution of gentian violet to each 

 lOO.Occ. of (3). 



(5) Thoroly cleanse egg with water. 



(6) Wash with 2.5% carbolic acid 

 solution. 



(7) Allow partially to dry. 



(8) Dry the ends in the flame, and 

 pierce with a small flamed sharp 

 forcep. The hole should be | of 

 an inch in diameter and the 

 membrane broken. 



(9) Make a smaller hole in the other 

 end and do not break the mem- 

 brane. This hole is to blow into. 



(10) Blow the contents of the eggs into 

 sterile Erlenmeyer flasks. Blow 

 from the cheeks. 



(11) Mix 1 part (4) to 2 parts (10). 



(12) Tube in sterile tubes. 



(13) Inspissate on 3 days for 45 minutes 

 at 80 to 85°C. 



References: Petroff (1915 p. 39), Roddy 

 (1917 p. 46), Waksman (1919 p. 307), 

 Giltner (1921 p. 368), Limousin (1921 p. 

 559), Harvey (1921-22 p. 85), Despeignes 

 (1922 p. 931), Stitt (1923 p. 47), Klimmer 

 (1923 p. 224), Park, Williams and Krum- 

 wiede (1924 p. 127). 



2444. Harvey's Sperm Egg Medium 

 Constituents: 



1. Physiological Salt Solution. 



2. Glycerol. 



3. Sperm. 



4. Egg. 

 Preparation : 



(1) Add 5.0% sperm oil to beaten and 

 filtered egg medium. 



(2) Add glycerol 0.85 sterile salt solution 

 to give 5.0% of glycerol. 



(3) Shake the mixture in a bowl of hot 

 water with a Bunsen burner under- J 

 neath, to melt the wax. I 



(4) Distribute into test tubes and keep * 

 in a bowl of hot water to prevent the 

 wax separating out. 



(5) Shake to mix. 



(6) Place low down in a steamer and 

 steam 4 minutes to coagulate the 1 

 egg. I 



Sterilization: Sterilize by placing high up 

 in the steamer for 1 hour on 3 successive 

 days. 



Use : Cultivation of tubercle bacilli. 



Reference: Harvey (1921-22 p. 87). 



2445. Harvey's Olive Oil Egg Medium 



Prepare in the same manner as Harvey's 

 Sperm Egg Medium, see medium 2444, but 

 use olive oil instead of sperm. 



2446. Lubenau's Glycerol Bouillon Egg 

 Medium (Park and Krumwiede) 



Constituents : 



1. Whole eggs 10 



2. 5.0% glycerol bouillon 250.0 cc. 



Preparation : 



(1) Sterilize the outside of 10 eggs by 

 washing with water and then 5.0% 

 phenol. 



