CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



505 



Preparation : 



(1) Thoroly wash 15.0 g. of agar agar 

 shreds in running water. 



(2) Dissolve (1) in a liter of distilled 

 water and cool to between 50 and 

 60°C. 



(3) Add 500.0 g. of lean beef or beef 

 heart, chopped to moderate fine- 

 ness. 



(4) Bring to boil and cook slowly for 15 

 to 20 minutes. 



(5) Filter thru a 16 mesh sieve (cullander 

 type) until clear. 



(6) Add 10.0 g. peptone and 5.0 g. NaCl. 



(7) Boil for 5 minutes. 



(8) Adjust to desired reaction (pH 7.5). 



(9) Allow to stand several minutes and 

 decant the supernatant fluid. 



(10) Tube. 



Sterilization: Sterilize by the fractional 

 method or by autoclaving for about 

 20 minutes at 5 pounds pressure. 



Use: Hormone medium. 



Reference: Bailey (1925 p. 341). 



1674. Fasiani and Zironi's Veal Autolysate 



Peptone Agar 



Constituents : 



1. Autolysate of veal liver 1000.0 cc. 



2. Peptone (1.0%) 10.0 g. 



3. NaCl (0.5%) 5.0 g. 



4. Agar (2.0%) 20.0 g. 



Preparation : 



(1) Dissolve 1.0% peptone 0.5% NaCl and 

 2.0% agar in an autolysate of veal 

 liver. 



(2) Tube in 15 to 20.0 cc. lots. 

 Sterilization: Autoclave at 110°C. for 30 



minutes. 



Use: Isolation and cultivation of anae- 

 robes. 



Reference : Fasiani and Zironi (1918 p. 439). 

 Taken from (1919 p. 147). 



1675. Harvey's Basal Indicator Infusion 



Agar 



See medium 1661 variant (v) for prepara- 

 tion of Infusion Agar. Add 1.0% of any 

 desired carbohydrate, alcohol, etc., to the 

 agar, adjust the reaction with brom thymol 

 blue indicator to a pH value between 6.8 

 and 7.2. Add one of the following indi- 

 cators: 



(a) Brom cresol purple 0.001% 



(b) Brom cresol purple 0.0005% 



Cresol red 0.0005% 



(c) China blue 0.0025% 



(d) China blue 0.005% 



Sodium rosolate 0.005% 



(e) China green 0.003% 



(f ) China blue 0.0025 g. 



Phenol sulphonphthalein. . . 0.001 g. 



Keep the indicator in concentrated alco- 

 hol stock solution of such a strength that a 

 definite amount can be measured out per 

 liter of medium, e.g., 1.0 cc. of 1.6 per cent 

 brom cresol purple per liter of medium. 



1676. Jordan and Victorson's Lead Acetate 

 Infusion Agar 



Constituents : 



1. Water 1000.0 cc. 



2. Peptone Witte's 30.0 g. 



3. Beef, lean 1.0 lb. 



4. Agar 15.0 g. 



5. Lead acetate (10.0% soln.). 

 Preparation : 



(1) Dissolve 2 in fresh meat broth 

 (1 pound meat to 1 liter water, but 

 exact procedure not given) by boiling. 



(2) Filter. 



(3) Dissolve agar in (2). 



(4) Adjust to 1.0% acid to phenol- 

 phthalein. 



(5) Tube. 



(6) Cool sterile (5) to 43°C., add 2 drops 

 (0.1 cc.) of a 10.0% lead acetate solu- 

 tion prepared from recently sterilized 

 water to each tube and mix well. 



Sterilization: Sterilize (5) method not 

 given. 



Use : Differentiation of paratyphoid enteri- 

 tidis group. Inoculate by sliding the 

 needle in between the agar and wall of 

 the tube. Authors reported that B. para- 

 typhosus B and B. enteritidis produced 

 H2S, blackening the medium, while 

 B. paratyphosus A, B. suipestiver , did not 

 blacken the medium. Emile-Weil used 

 a similar medium to determine H2S pro- 

 duction by Bacillus leprae. 



Variants : 



(a) Emile-Weil prepared the medium as 

 follows: 



(1) Macerate 500.0 g. of finely divided 

 beef with one liter distilled water 

 for 12 hours. 



(2) Boil slowly for 15 minutes. 



