502 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



cultures. Bronfenbrenner and Schles- 

 inger reported that the medium gave a 

 large amount of water of condensation. 

 To keep a pure culture on this medium 

 for several weeks, inoculate the water of 

 condensation and tilt the tubes every day 

 or so. Other investigators cultivated a 

 variety of organisms as B. welchii, B, 

 chauvoei, the gonococcus, etc., on sim- 

 ilar media. 

 Variants : 



(a) Bronfenbrenner and Schlesinger pre- 

 pared the mediima as follows: 



(1) Prepare a liver infusion using 

 1000.0 g. liver to 1000.0 cc. water. 

 The extract is collected thru a 

 single layer of cheese cloth. Boil 

 and stir continually (Time not 

 given). 



(2) Strain thru a single layer of cheese 

 cloth and a thin layer of cotton. 



(3) Do not adjust the reaction, and 

 sterilize at 10 pounds pressure for 

 10 minutes. 



(4) Prepare a 3.0% agar solution in 

 water (30.0 g. agar to 1000.0 cc. 

 water. 



(5) Filter thru cotton. 



(6) Add 2.0% (20.0 g.) peptone and 

 1.0% (10.0 g.)NaCl. 



(7) Do not adjust the reaction. 



(8) Mix equal parts of sterile (3) and 

 sterile (7) while hot. 



(9) Tube in sterile tubes. 



(b) Heller isolated B. Welchii, B. oede- 

 matiens and other soil anaerobes on 

 the following medium: 



(1) Grind 250.0 g. beef liver and infuse 

 with 4 parts water over night. 

 Boil and strain. 



(2) Add 15.0 g. peptone, 5.0 g. NaCl 

 and 2.0 g. Agar. 



(3) pH 7.2 (faintly alkaline to litmus). 



(4) Tube in deep tubes. 



(5) Method of sterilization not given. 

 He reported that if large amount of 

 H2O be produced, add KNO3 1.0%. 

 Use 3.0% agar if rapid growers out- 

 grow others. 



(c) Goss et al. cultivated B. chauvoei and 

 other organisms on a medium pre- 

 pared as follows: 



(1) Grind 500.0 g. beef liver and add 

 1000.0 cc. water. 



(2) Cook (1) in flowing steam one 

 hour. 



(3) Strain thru cheese cloth and 

 cotton. 



(4) Add peptone (1.0%) and NaCl 

 (0.5%). 



(5) Add 20.0 g. agar and heat in flow- 

 ing steam until all is dissolved. 



(6) Adjust to pH = 8.2. 



(7) Clarify with egg albumin. 



(8) Filter and tube. 



(9) Autoclave at 15 pounds pressure 

 for 20 minutes. 



(d) Harvey prepared the medium as 

 follows: 



(1) Prepare an infusion from 1000.0 g. 

 liver (or other organ, placenta, 

 etc.) and 1000.0 cc. water in the 

 same manner as for ordinary infu- 

 sion broth. (See variant (bb) 

 medium 779.) 



(2) Dissolve 3.0% agar, 2.0% peptone 

 and 1.0% NaCl in water. 



(3) Sterilize (2). 



(4) Mix equal parts sterile (1) and (3) 

 while hot under aseptic conditions. 



(5) Tube in sterile tubes. 



(e) Heller cultivated pathogenic anae- 

 robes in the following medium: 



(1) Infuse one part beef liver with 

 4 parts distilled water in the 

 refrigerator over night. 



(2) Bring to boil and strain thru cheese 

 cloth. 



(3) Add 15.0 g. Peptone (Difco), 5.0 g. 

 salt and 20.0 g. agar and dissolve 

 by heating. 



(4) Titrate and adjust to pH = 7.2. 



(5) Cool to 60° and add white of egg 

 and serum. 



(6) Autoclave for one hour. 



(7) Titrate and adjust to pH = 7.2. 



(8) Filter through cotton. 



(9) Tube. 



(10) Sterilize in the autoclave (time not 

 given). 



(f) Park, Williams and Krumwiede culti- 

 vated gonococci on the following 

 medium: 



(1) Soak 5.0 pounds of finely chopped 

 liver in 5000.0 cc. of tap water 

 over night in an ice box or at room 

 temperature. 



