CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



647 



for each 200.0 cc. of fluid and sterilize 

 in the autoclave. 

 (4) Add sterile (3) to sterile melted glu- 

 cose agar (or gelatin) in proportion 

 of 3 to 7. 

 References: Rosenow (1914 p. 61), Tilmant 

 and Carrien (1917 p. 499) taken from 

 (1917 p. 222). 

 2017. Ruediger's Inulin Ascitic Fluid Agar 



Constituents: 



1. Distilled water 200.0 cc. 



2. Peptone (Witte) 10.0 g. 



3. Agar 15.0 g. 



4. Bouillon (sugar free) 800.0 cc. 



5. Inulin 15-0 g. 



6. Litmus (5.0% soln.) 20.0 cc. 



7. Ascitic fluid 145.0 cc. 



Preparation: 



(1) Dissolve 2 and 3 in sugar free bouil- 

 lon by boiling for one hour. Add 

 water to make the volume to 800.0 cc. 



(2) Heat in autoclave for 15 to 20 

 minutes. 



(3) Clarify with egg. 



(4) Filter thru cotton. 



(5) Make up volume to 800.0 cc. with 

 distilled water. 



(6) Dissolve 15.0 g. pure inulin in 

 200.0 cc. distilled water. 



(7) Mix (6) with (5). 



(8) Add 20.0 cc. of 5.0% solution of 

 litmus (Merck's highest purity). 



(9) Tube in 7.0 to 8.0 cc. lots. 



(10) Just before use, melt sterile agar, 

 cool to 45° and add 1.0 cc. of heated 

 (65°) fluid to each tube. 

 Sterilization: Sterilize (9) in autoclave 

 under 10 pounds pressure for 15 minutes. 

 Use: Isolation of pneumococci. Acid pro- 

 duction. Ruediger reported the pneu- 

 mococci colonies were red. 

 Reference: Ruediger (1906 p. 184). 



2018. Klimenko's Glycerol Ascitic Fluid 

 Agar 



Constituents : 



1. Nutrient agar (3.0 or 4.0%) . 100.0 cc. 



2. Glycerol (1.0%) 10.0 cc. 



3. Ascitic fluid (human) 100.0 cc. 



Preparation : 



(1) Prepare 3.0 or 4.0% nutrient agar con- 

 taining 1.0% glycerol. 



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

 ascitic fluid (or blood serum). 



Sterilization: Not specified. 



Use : Cultivation of whooping cough bacilli. 

 Author reported that whooping cough 

 bacilli grew as a glistening opalescent 

 covering of a grayish shade. 



Variants : 



(a) Cantani prepared a similar medium 

 as follows: 



(1) Mix equal amounts of glycerol and 

 the albuminous fluid (urine, pus, 

 milk, egg white, etc.). Place in an 

 Erlenmeyer flask. 



(2) Storing the fluid in glycerol tends 

 to sterilize the fluid. 



(3) After a time test the sterility of the 

 mixture. 



(4) Prepare nutrient agar. 



(5) Mix 6 parts ascitic fluid (with or 

 without glycerol) with one part 

 of (3). 



(6) Add to each tube of sterile (4) 0.5 

 to 0.75 cc. of (5). 



(7) Incubate 24 hours to test sterility. 



(b) Cantani (Besson) prepared a medium 

 as follows: 



(1) Mix equal parts of sterile ascitic 

 fluid and sterile glycerol. . 



(2) Allow to stand several hours. 



(3) Add 0.5 to 0.75 cc. of (2) to tubes of 

 liquified sterile agar. 



References: Klimenko (1909 p. 311), Can- 

 tani (1910 p. 472), Besson (1920 p. 54). 



2019. Vellion's Ascitic Fluid Agar 



Constituents: 



1. Nutrient agar (2.0%). 



2. Ascitic fluid. 

 Preparation: 



(1) Prepare 2.0% nutrient agar with a 

 slightly alkaline reaction. 



(2) Tube. 



(3) Cool melted sterile (2) to about 60°C. 



(4) Add an equal amount or even § 

 amount of ascitic fluid. 



(5) Slant. 



Sterilization: Method of sterilization of 



(2) not given. 

 Use: Cultivation of gonococci. 

 Variants : 



(a) Thoinot and Masselin prepared the 

 medium as follows: 



(1) Liquify sterile agar and cool to 

 about 50°C. 



(2) Add a known quantity of sterile 



