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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



2010. Meyer's Tuberculin Agar 



Constituents : 



1. 5.0% glycerol bouillon with 

 heavy growth of tubercle 



bacilli 200.0 cc. 



2. Glycerol bouillon (4.0 or 



5.0%) 200.0 cc. 



3. Agar (2.0%) 8.0 g. 



4. Serum (10.0%) 40.0 cc. 



Preparation : 



(1) Tuberculin flasks containing about 

 250.0 cc. of a 5.0% glycerol peptone 

 bouillon, with a heavy growth of 

 human or bovine tubercle bacilli, 

 about 6 to 8 weeks old are steamed in 

 a Board of Health sterilizer for two 

 hours. 



(2) Cool and filter thru fine filter paper 

 and centrifuge to eliminate every 

 tubercle bacillus. 



(3) Mix the clear liquid with an equal 

 volume of the same 4.0 or 5.0% 

 glycerol peptone bouillon and 2.0% 

 agar is added and dissolved. 



(4) Filter and correct the reaction to 1.5. 



(5) Distribute in 9.0 cc. quantities in 

 test tubes. 



(6) Before being slanted 1.0 cc. (or 10.0%) 

 of fresh sterile horse or cattle serum 

 is added to each tube of sterile agar. 



Sterilization: Sterilize (5) in the autoclave. 

 Use: Cultivation of Bacillus para tuber- 

 culosis. 

 Reference: Meyer (1913 p. 175). 



2011. Douglas' Tellurite Trypsinized Serum 

 Agar 



Constituents : 



1. Nutrient agar (2.0%) 1000.0 cc. 



2. Potassium tellurite 



3. Serum 



4. Trypsin 

 Preparation: 



(1) Preparenutrient (2.0%) agar. (Pref- 

 erably a trypsin digest agar, but 

 ordinary peptone agar gave almost 

 identical results). 



(2) Add 1.0% potassium tellurite to 

 100.0 cc. of water and dissolve as 

 much as possible. 



(3) Allow the insoluble portion of (2) 

 to settle and pour off the clear 

 supernatant layer. 



(4) Rub up the deposit with 10.0% KOH 

 solution added drop by drop to pre- 

 vent e.xcess of alkali. 



(5) Add HCl to (4) until a precipitate 

 begins to appear. Add this fraction 

 to the clear supernatant fluid 

 from (3). 



(6) Add sufficient glycerinated com- 

 mercial trypsin to serum (horse 

 generally) to partially neutralize 

 the antitryptic power. (Requires 

 from 2.0 to 8.0 cc. of trypsin per 

 100.0 cc. depending on the strength 

 of the trypsin, 2 to 4.0 cc. of Fair- 

 child & Co. and Digestive Ferments 

 Co. trypsin being sufficient and 5.0 

 to 8.0 cc. of Allen and Hanbury's 

 trypsin). 



(7) Filter (6) thru a porcelain candle of 

 medium porosity and distribute in 

 sterile flasks. 



(8) Melt sterile (1) and add 4.0 cc. of (5) 

 for each 100.0 cc. of agar. 



(9) Cool to between 65° and 60°C. and 

 add 15.0 cc. of trypsinized serum per 

 100.0 cc. agar. 



(10) Mix thoroughly. 



(11) Tube in sterile tubes. 



(12) Slant. 



(13) Incubate to test sterility. 

 Sterilization: Method of sterilization of (1) 



and (5) not given. 

 Use: Cultivation of B. diphtheriae. 

 Reference: Douglas (1922 p. 263). 



2012. Czaplewski's Alkaline Serum Glucose 

 Agar 



Constituents : 



1. Glucose agar 



2. Serum 300.0 cc. 



3. NaOH (normal) 30.0 cc. 



Preparation : 



(1) Add 30.0 cc. normal NaOH to 

 300.0 cc. sheep serum. 



(2) Sterilize (method not given). 



(3) Allow to stand and pour off the clear 

 supernatant liquid. 



(4) Mix sterile (3) with sterile glucose 

 agar (1.0, 2.0 or 3.0 cc. serum to 

 10.0 cc. agar). 



Sterilization: Sterilize (3) in the autoclave. 

 Method of sterilization of glucose agar 

 not given. 



