542 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



pounds for 15 minutes, or in the 

 Arnold, 

 (d) Conradi-Drigalski (Stitt). 



(I) Add 1.0 cc. of a 1 to 100 solution of 

 crystal violet in distilled water to 

 100.0 cc. of the medium as prepared 

 by Stitt above (variant (c)). 



References: Committee A.P.H.A. (1917 

 p. 97), Meyer (1917 p. 238), Tanner (1919 

 p. 48), Committee A.P.H.A. (1920 p. 97), 

 Stitt (1923 p. 49). 



1759. Endo's Fuchsin Sulphite Agar 

 (Heinemann) 



Constituents : 



1. Extract agar (3.0%) 1000.0 cc. 



2. Lactose 10.0 g. 



3. Fuchsin (ale. soln.) 5.0 cc. 



4. NaaSOa (10.0% soln.) 25.0 cc. 



5. NaOH (10.0% soln.) 10.0 cc. 



Preparation : 



(1) Prepare a medium from 2, 3 and 4 

 using 3.0% extract agar as a base. 



(2) Add 10.0 cc. of a 10.0% NaOH 

 solution. 



Sterilization: Not specified. 



Use: Isolation of typhoid bacilli. Coli 

 colonies large and red, typhoid colonies 

 small, colorless and bluish. 



Variants: The following authors have pre- 

 pared similar media as indicated: 

 (a) Klinger. 



(1) Add 20.0 g. Liebig's meat extract, 

 20.0 g. Witte's peptone, 10.0 g. 

 NaCl and 80.0 g. agar to 1000.0 cc. 

 of water. 



(2) Heat for either 3 hours in stream- 

 ing steam, or 2 hours at 110°, or 

 one hour at 120°C. 



(3) Filter thru a thick layer of cbtton. 



(4) Neutralize to litmus. 



(5) Add 20.0 cc. of a sterile 10.0% soda 

 solution to (4). 



(6) Add 20.0 g. of chemical pure lac- 

 tose to (5). 



(7) Add 10.0 g. of crystalline fuchsin to 

 100.0 cc. of 96% alcohol and allow 

 to stand for 20 hours. 



(8) Pour off the saturated solution. 



(9) Add 10.0 cc. of (8) to (6). 



(10) Add 50.0 cc. of a freshly prepared 

 10.0% NasSOs solution to (9). 



(II) INIix well. 



(12) Distribute in 200-400 cc. lots and 

 store in the dark. 



(13) Sterilization not specified. 



(b) KastleandElvove. 



(1) Dissolve 10.0 g. Liebig's extract, 

 10.0 g. peptone and 5.0 g. NaCl in 

 1000.0 cc. distilled water by heat. 



(2) Cool and add 40.0 g. powdered 

 agar. Allow agar to settle. 



(3) Place in Arnold and cook 3 hours. 



(4) Neutralize to litmus with NaoCOs. 



(5) Filter thru cotton on a Buchner 

 filter or allow to settle, rejecting 

 turbid bottom part. 



(6) Add 10.0 cc. of a sterile 10.0% 

 Na2C03 solution to^(5) after 

 filtering. 



(7) This medium may be storedTin 

 100-200 or 400.0 cc. flasks until 

 desired for use. 



(8) When desired for use melt (7) and 

 to each liter add c.p. 10 g. lactose 

 and 5.0 cc. of a 10.0% alcoholic 

 fuchsin solution (freshly | pre- 

 pared). 



(9) Prepare a 10.0% alcoholic fuchsin 

 solution by shaking 10.0 g fuchsin 

 (not acid fuchsin) with 100.0 cc. 

 of 96% alcohol, allow to stand 24 

 hours. Decant supernatant fluid 

 and filter this fluid each time 

 immediately before use. 



(10) After fuchsin and sulphite have 

 been added to (7), shake vigor- 

 ously and place unstoppered in 

 sterilizer for 5 to 10 minutes to 

 allow foam to settle. 



(11) Add 25.0 cc. of a freshly prepared 

 sterile 5.0% anhydrous Na.SO, 

 solution to (10) and mix by rotat- 

 ing flask. 



(12) Sterilize in Arnold for a few minutes 

 and pour in Petri dishes while 

 steaming hot. Solidified medium 

 nearly colorless to transmitted 

 light— and rose or flesh to reflected 

 light. 



(c) Kendall and Day. 



(1) "Dust" 15.0 g. powdered agar 

 upon the surface of cold tap water. 

 Allow to settle into the medium 

 before heat is applied or other 

 ingredients added. 



