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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(2) Add 1.0% sugar and some litmus 

 solution to (1). 



(3) Steam for 10 minutes on 3 suc- 

 cessive days to sterilize. 



(4) Add one part of sterile ascitic fluid 

 to two parts (3). 



(5) Tube and slant. 



(6) Incubate 2 days to test sterility. 



(b) Gordon (Abel) used a medium pre- 

 pared as follows to study fermenta- 

 tion of meningococci and gonococci. 



(1) Prepare 10.0% solution of any de- 

 sired carbohydrate, alcohol, etc., in 

 Kahlbaum's litmus solution and 

 boil two minutes. 



(2) When cool add 0.5 cc. of normal soda 

 solution to each 10.0 cc. of (1). 



(3) Mix 3 parts 3.0% nutrient agar with 

 • one part ascitic fluid. 



(4) Add 1.5 cc. of (2) to each 13.5 cc. 

 of (3)_. 



(5) Pour into plates. 



(c) V. Przewoski used the following me- 

 dium to determine fermentation 

 ability of diphtheria and pseudo 

 diphtheria: 



(1) Prepare nutrient agar. 



(2) To 100.0 cc. of liquid (1) add 5.0 cc. 

 litmus solution (Kahlbaum), 2.0 cc. 

 ascitic fluid and 1.0 g. of one of 

 the following: 



fructose mannitol 



glucose lactose 



mannose inulin 



dulcitol 



(d) Hancken studied the fermentation 

 ability of meningococci on a medium 

 prepared as follows: 



(1) Prepare nutrient agar. 



(2) Dissolve 2.5 g. of any desired carbo- 

 hydrate sugar or alcohol in 50.0 cc. 

 of Kahlbaum's litmus solution. 

 Boil for a few minutes. 



(3) Melt the sterile agar and cool to 

 55°C. and add 3.3 cc. ascitic fluid 

 and 4.0 cc. of (2) to 9 or 10.0 cc. of 

 the agar under aseptic conditions. 



(4) Pour into sterile plates. 



(e) Eastwood studied the fermentation 

 of carbohydrates, alcohols, etc., on 

 Lingelsheim's medium prepared as 

 follows: 



(1) Prepare a 10.0% solution of one 

 of the following: maltose, glucose, 



levulose, galactose, mannitol, dul- 

 citol, sucrose, lactose, inulin with 

 Kubel-Tiemann's litmus and steri- 

 lize. (Method not given.) 



(2) Add (1) to ascitic agar so that the 

 agar content is 1.0%. 



(3) Final sterilization not given. 

 Eastwood reported that meningococci 

 gave acid (red colonies) with glucose 

 and maltose only. 



References: Salomon (1908 p. 2), Symmers 

 and Wilson (1909 p. 10), Abel (1912 

 p. 159), V. Przewoski (1912 p. 15), Hancken 

 (1916 p. 368), Eastwood (1916 p. 408). 



2016, Rosenow's Glucose Ascitic Fluid Agar 

 Constituents : 



1. Nutrient agar (2.0%). 1000.0 cc. 



2. Glucose 2.0 to 10.0 g. 



3. Ascitic fluid 300.0 cc. 



Preparation : 



(1) Prepare 2.0% nutrient agar with a 

 reaction of 0.4 to 0.6% acid to phenol- 

 phthalein. 



(2) Dissolve 2 in (1). 



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



(4) Boil sterile (3) to drive off o.xygen 

 and cool to 50°C. 



(5) Add 2 to 3.0 cc. heated ascitic fluid 

 (60°C. for 24 hours) to each tube. 



(6) Cool to 40°C. 



Sterilization: Method of sterilization of 

 (3) not given. See step (5) above for 

 sterilization of ascitic fluid. 



Use: To isolate organism causing rheuma- 

 tism, author added some of rheumatic 

 material to medium and mixed well; 

 plunged tube into cold water to "set" 

 and incubated at 37°C. Author reported 

 that largest number of colonies developed 

 between 1.5 cm. from the top and 3.5 cm. 

 from the bottom. A similar medium was 

 used for the cultivation of meningococci 

 and gonococci by Tilmant and Carrien. 



Variant: Tilmant and Carrien cultivated 

 meningococci and gonococci on a medium 

 prepared as follows: 



(1) Collect about 100.0 cc. of ascitic fluid 

 as carefully as possible. 



(2) Estimate the albumin content and 

 dilute with physiological saline so 

 that there will be 3.0 g. of albumin 

 per liter. 



(3) Add 1.0 cc. of a 10.0% soda solution 



