522 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(6) Test the reaction and adjust if 

 necessary. 



(7) If more than 0.2% normal soda is 

 required per liter, heat again for 

 10 minutes. 



(8) Filter thru cotton. 



(9) Distribute in tubes or flasks. 

 (10) Sterilize at 15 pounds pressure for 



30 minutes. 



(k) Park, Williams and Krumwiede (1924) 

 reported a very satisfactory medium 

 can be made by simply using 0.5% or 

 less of agar instead of the usual 1.5% 

 employed. This can be diluted by 

 the addition of i of its bulk of an 

 enrichment fluid. The finished 

 medium should just "set" suffi- 

 ciently for stab culture purposes. 

 Nutrose (1.0%) may also be added. 



(1) Park, Williams and Krumwiede (1924) 

 gave the following medium as Mus- 

 grave and Clegg's Agar for the culti- 

 vation of amoeba: 



(1) Mix 90.0% tap water, 10.0% ordi- 

 nary nutrient broth (preparation 

 not given) and 1.0% agar. 



(2) Dissolve. 



(3) Reaction neutral to phenol- 



phthalein. 



(4) Sterilize as usual (method not 

 given). 



(m) Cunningham (1924). 



(1) Add 1.5% agar to bouillon. 



(2) Steam for 30 minutes to dissolve 

 the agar. 



(3) Boil over an open flame for 

 15 minutes, stirring constantly. 



(4) Adjust the reaction to a slight 

 alkalinity using turmeric paper 

 as an indicator (distinctly brown). 



(5) Filter while hot thru a plug of 

 cotton-wool in the bottom of an 

 enamelled funnel. 



(6) Tube. 



(7) Sterilize intermittently in steam. 

 References: Wurtz (1897 p. 29), Smith 



(1902 p. 92), Roux and Rochaix (1911 

 pp. 115, 116), Hoffmann (1912 p. 387), 

 Lohnis (1913 p. 16), Roddy (1917 p. 43), 

 Bezan^on (1920 p. 112), Hilgermann and 

 Weissenberg (1917-18 p. 470), Klimmer 

 (1923 p. 229), Stitt (1923 p. 36), Park, 

 Williams and Krumwiede (1924 pp. 117, 

 118, 134), Cunningham (1924 p. 15). 



1711. Warden's Salt Agar 

 Constituents : 



1. Distilled water 1000.0 



2. Bouillon 200.0 



2.5 g 

 0.2 g 

 0.25 g 

 0.45 g 

 10.8 g, 



3. Agar 



4. Sodium bicarbonate 



5. CaCla 



6. KCl 



7. NaCl 



Preparation : 



(1) Dissolve 3, 4, 5, 6 and 7 in 1 by aid of 

 heat until fluid shows a translucent 

 ground glass appearance. 



(2) Add bouillon (composition or method 

 of preparation not specified). 



(3) Filter while hot, through gauze or 

 cotton into test tubes or flasks. 



(4) Adjustment of reaction not specified. 

 Sterilization: Sterilize once in autoclave 



(time not specified). 



Use: Culture medium for gonococci. 

 Author reported that when cool medium 

 was semi-solid, and had a translucent 

 silvery appearance. Nearly all strains 

 of gonococci did not grow on this me- 

 dium. If a few loopfuls of sterile human 

 blood be placed on this surface, all 

 strains grew. 



Reference: Warden (1913 p. 94). 



1712. Stroszner's Regenerated Agar 



Constituents : 



1. Bouillon. 



2. Agar (used). 

 Preparation: 



(1) Place the used agar in an enamel con- 

 tainer and melt in streaming steam 

 (add no water). 



(2) Measure the agar in a graduated 

 cylinder. 



(3) To each liter of agar add 40.0 g. of 

 powdered charcoal and boil for 30 to 

 40 minutes in the steamer. 



(4) Cool to 50°C. and add 40.0 cc. defi- 

 brinated blood per liter of agar. 



(5) Boil in streaming steam once more 

 for 40 minutes. 



(6) Filter (method not given). 



(7) Add 300.0 cc. meat infusion or 1 to 

 1.5% Liebig's bouillon to each liter 

 of agar. 



Sterilization: Method not given. 

 Use: Regenerated agar. 



