CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



535 



egg albumin stirred up well in a little 

 water. 



(6) Add soda solution until the reaction 

 is distinctly alkaline and heat in the 

 autoclave at 120°C. for 20 minutes. 



(7) When taken from the autoclave pour 

 immediately upon a Chardin filter 

 paper. 



(8) Distribute in about 10-12 cm. layer in 

 a 20 to 22 mm. by 22 cm. test tubes. 



Sterilization: Sterilize at 112 to 115°C. for 



15 to 20 minutes. 

 Use: Cultivation of anaerobes. Meier 



made bacterial counts from milk and 



whey in a similar medium. 

 Variants : Meier prepared a similar medium 



as follows: 



(1) Boil 500.0 g. of fat and tendon-free 

 beef in one liter of water. 



(2) Filter. 



(3) Dissolve 2.5 g. lactose, 2.5 g. glucose, 

 5.0 g. Witte's peptone, 5.0 g. NaCl 

 and 5.0 g. agar in 1. 



(4) Neutralize by the addition of KOH. 

 Add KOH until turmeric paper is 

 turned quite weakly brownish red. 



(5) Sterilization not specified. 

 References: de Gasperi and Savini (1911 p. 



248), Meier (1918 p. 454). 



1743. Teague and Clurman's Eosin Brilliant 

 Green Agar 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Beef, chopped 500.0 g. 



3. Peptone (Witte) 10.0 g. 



4. XaCl(C. P.) 5.0 g. 



5. Agar 15.0 g. 



6. Lactose 10.0 g. 



7. Saccharose 10.0 g. 



8. 3.0% 3'ellowish eosin 

 solution 20.0 cc. 



9. 1.0% brilliant green solu- 

 tion 20.0 cc. 



Preparation : 



(1) Soak beef in water in the ice chest 

 over night. 



(2) Squeeze infusion thru cheese cloth, 

 heat in Arnold sterilizer and pass 

 thru filter paper. 



(3) Add 3, 4 and 5 to warm (2). Rub 

 peptone into paste before adding. 



(4) Heat in autoclave for 30 minutes at 

 120°C. 



(5) Adjust to +1 with 2X, NaOH. 



(6) Heat in Arnold sterilizer for ^ hour. 



(7) Adjust again to -|-1. 



(8) Cool to 55°C., clear with egg white 

 and filter thru cotton. 



(9) Flask in 100.0 cc. or 200.0 cc. lots. 



(10) When ready for use melt sterile (9), 

 add 1.0% lactose and 1.0% saccha- 

 rose. (1.0 g. to 100.0 cc. medium of 

 each sugar.) 



(11) Prepare a 3.0% stock solution of 

 yellowish eosin in distilled water. 



(12) Prepare a 1.0% stock solution of 

 brilliant green in 50.0% alcohol, a 



g% solution in distilled water. 



(13) Add 1.0 cc. of (11) and 1.0 cc. of (12) 

 to each 50.0 cc. of medium. 



(14) Shake well. 



(15) Pour into sterile Petri dishes. 

 Sterilization: Sterilize (9) by heating in the 



autoclave at 120°C. for 20 minutes. 



Use: Isolation of typhoid bacilli from 

 stools. Author reported that typhoid 

 colonies after 18 hours were grayish in 

 color by reflected light. B. coli colonies 

 were red. In transmitted light typhoid 

 were colorless and transparent. Liebig's 

 meat extract cannot be substituted for 

 beef infusion. 



References: Teague and Clurman (1916 p. 

 651), Tanner (1919 p. 54). 



1744. Kan-Ichiro Morishima's Lead 

 Acetate China Blue Agar 



Constituents : 



1. Meat infusion agar (compo- 

 sition not given) 2000.0 cc. 



2. Neutral 2.0% solution lead 



acetate in distilled water. . . 50.0 cc. 



3. 1.0% solution China blue 



4. N sodium hydrate solution 



5. Lactose 10.0 g. 



6. Glucose 1-0 g. 



Preparation : 



(1) Preparation or composition of meat 

 infusion agar not given except that 

 it is to be prepared in the usual way. 



(2) Clear with egg white. 



(3) Titrate while hot to -0.2 to -0.4 

 with phenolphthalein. 



(4) Prepare 2.0% neutral lead acetate 

 in sterile distilled water. Heat for 

 5 hour at 100°C. in water bath. 



(5) Melt (3), and add 5.0 cc. of (4) to 



