CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



393 



tion of toxin by diphtheria bacilli, 

 and cultivated them on a medium 

 prepared as follows: 



(1) Mix 4 liters of medium fine placenta 

 with 4 liters of water. 



(2) Boil for about 3 hours with the 

 addition of water. 



(3) Filter. 



(4) Concentrate the filtrate to 3 liters. 



(5) Add N/1 10.0% NaOH until phenol- 

 phthalein is turned weakly red. 

 (Hot titration). 



(6) Distribute into flasks and sterilize. 

 (Method not given). 



(b) Park, Williams and Krumwiede pre- 

 pared a similar medium as follows: 



(1) Add 500.0 g. of ground up tissue 

 (placenta) to a liter of water. 



(2) Soak for 90 minutes. 



(3) Strain thru cheese cloth and 

 squeeze by twisting the cloth or 

 use a meat press. 



(4) Filter thru paper or sand. 



(5) Sterilize by filtration thru a filter 

 candle. 



References: Wellman (1912 p. 143), Groer 

 and Srnka (1918-19 p. 334), Park, Williams 

 and Krumwiede (1924 p. 125). 



1338. Moon's Brain Infusion 



Constituents: 



1. Physiological salt solution.. . 900.0 cc. 



2. Brain (dog) 100.0 g. 



Preparation : 



(1) Remove brain matter from normal 

 dog under the most careful aseptic 

 precautions. 



(2) Add 900.0 cc. of physiological salt 

 solution to 100.0 g. of (1). 



(3) Emulsify by shaking with glass beads. 

 Sterilization: Filter thru a coarse Berke- 



feld filter. 

 Use: Cultivation of Negri bodies. Author 



reported that growth was poor, both 



anaerobically and aerobically. 

 Reference: Moon (1913 p. 233). 



1339. Wolbach and Saiki's Liver Infusion 

 Constituents: 



1. Distilled water 1000.0 cc. 



2. Liver (dog) 1000.0 g. 



Preparation : 



(1) Pass fresh dog's liver thru the meat 

 grinder. 



(2) Mix equal parts of (1) and distilled 

 water. 



(3) Shake vigorously. 



(4) Filter thru Buchner filters and then 

 thru Berkefeld filters (finest grade). 



(5) Distribute to fermentation tubes. 



(6) Incubate for 24 hours to determine 

 sterility. 



Sterilization: Sterilization given under 

 preparation. 



Use: Cultivation of a spore bearing an- 

 aerobe from a dog's liver. The medium 

 obtained at first is rather viscous, red and 

 slightly acid in reaction. After 48 hours 

 incubation proteins are precipitated. 



Reference: Wolbach and Saiki (1909 p. 

 270). 



1340. Brieger's Thymus Gland Infusion 



(Besson) 



Constituents: 



1. Distiled water 1000.0 cc. 



2. Thymus gland (beef) 1000.0 g. 



Preparation : 



(1) Remove the thymus glands from dead 

 beef. 



(2) Chop into a pulp and add an equal 

 weight of distilled water. 



(3) Allow to soak for 12 hours. 



(4) Filter thru a gauze, pressing out the 

 liquid. 



(5) Add an equal weight of water to the 

 turbid viscous liquid. 



(6) Make slightly alkaline by the addition 

 of a 1.0% solution of sodium bicar- 

 bonate. 



(7) Heat at 100°C. for 15 minutes in the 

 autoclave or salt bath. 



(8) Filter thru a fine linen cloth. 



(9) Distribute in sterile tubes. 

 Sterilization: Sterilize at 100° for 15 



minutes on two days. 



Use: General culture medium. Besson re- 

 ported that the cholera vibrio did not 

 develop in this medium unless it was 

 diluted with 5 or 6 volumes of sterile 

 water before use. 



Reference: Besson (1920 p. 30). 



1341. Nencki, Sieber and Wyznikiewicz's 



Salivary Gland Infusion 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Salivary glands 200.0 g. 



