CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



615 



(8) Mix (7) with an equal volume of 

 ascitic fluid. 



(9) Distribute in tubes or plates. 

 Sterilization: Not specified. 



Use: Cultivation of gonococci. Author 

 reported that if the ascitic fluid be 

 strongly alkaline mix with unneutralized 

 or strongly acid agar. The final medium 

 should be slightly alkaline. 



Variants: Harvey prepared a similar me- 

 dium as follows: 



(1) Mix equal parts pure neutral glycerol 

 and ascitic fluid, (pleuritic fluid, 

 hydrocele fluid, ovarian fluid, milk, 

 urine, etc., may be used instead). 



(2) Leave until sterile on culture test. 



(3) Add (2) to sterile infusion agar (see 

 variant (v) medium 1661 for prepa- 

 ration). 



References: Abel (1912 p. 162), Harvey 

 (1921-22 p. 83). 



1929. Torrey and Buckell's Urine Ascitic 

 Fluid Agar 



Constituents : 



1. Distilled water 2000.0 cc. 



2. Veal, fat-free 1250.0 g. 



3. Peptone (Difco) 20.0 g. 



4. Urine, fresh 40.0 cc. 



5. NaCl 100 g- 



6. Glvcerol 40.0 cc. 



7. Flaked agar 36.0 g. 



8. Ascitic fluid 1000.0 cc. 



9. lodin-green 

 Preparation: 



(1) Place finely chopped lean veal in 

 water and bring slowly to a boil. 



(2) Allow to simmer 20 minutes with 

 occasional stirring. 



(3) Strain thru cotton flannel. 



(4) Cool and remove the fat. 



(5) Place in double boiler over a satu- 

 rated brine bath and raise the 

 temperature to about 60°C. 



(6) Add 3, 4, 5, 6 and 7 and boil 45 min- 

 utes to dissolve materials. 



(7) Adjust reaction to pH = 6.9, using 

 10.0% Na2C03. 



(8) Boil 30 minutes longer. 



(9) Remove from brine and add distilled 

 water to 2 liters. 



(10) Filter thru canton flannel and tube 

 in 10.0 cc. amounts. 



(11) In preparing plates 5.0 cc. of ascitic 



fluid free from bile and 0.5 cc. of a 

 1:3000 dilution of iodin-green (Gr ab- 

 ler) are added to each tube of sterile 

 melted (11) just before pouring. 

 (12) Final reaction pH = 7.2. 

 Sterilization: Autoclave (10) at about 



12 pounds pressure for 10 minutes. 

 Use: Isolation of gonococci. Authors re- 

 ported that after 48 hours gonococci 

 colonies were semi-translucent and had 

 raised or even indented edges. They 

 stood out from the medium. Centers 

 somewhat thickened. If used in slants 

 use 40.0 g. agar. 

 Variants: The authors omitted the iodin- 

 green. 

 Reference: Torry and Buckell (1922 p. 125). 



1930. Watabiki's Whey Ascitic Fluid Agar 



Constituents : 



1. Whey 200.0 cc. 



2. Ascitic fluid (5.0%) agar 



3. Nutrose (0.3%) agar 400.0 cc. 



4. Urea 2.0 g. 



Preparation : 



(1) Warm 200.0 cc. of cow's milk to 60°C. 



(2) Add 5.0% ascitic agar drop by drop. 

 (Composition or method of prepara- 

 tion not given) the milk being shaken 

 to cause precipitation of the casein. 



(3) Filter thru filter paper. 



(4) To filtrate add 10.0% caustic soda to 

 slightly alkaline. 



(5) Add urea (2.0 g.). 



(6) Mix sterile (5) with 0.3% nutrose agar. 

 (Composition or method of prepara- 

 tion not given.) One part fluid (5) 

 to 2 parts agar at 45°C. 



(7) Slant or plate as desired. 



(8) Incubate to insure sterility. 

 Sterilization: Sterilize (5) by heating at 



60°C. for 30 minutes on each of three 

 successive days. 



Use: Cultivation of gonococci. Author 

 reported that the medium was not as 

 satisfactory as blood agar or blood broth. 

 Ordinary broth or peptone water may be 

 mixed with the fluid or liquid medium. 



Reference: Watabiki (1916 p. 734). 



1931. Esch's Maltose Ascitic Fluid Agar 



Constituents : 



1. Infusion agar 



2. Peptone. Witte (1.0%) 



750.0 cc. 

 7.5 g. 



