CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



241 



30 minutes on each of three successive 

 days. 



Use: Cultivation of streptococci. 



Reference: Krasnow, Rivkin and Rosen- 

 berg (1926 p. 389). 



790. Cesaris-Demel's Liver Infusion 

 Broth 



Constituents : 



1. Water 1000.0 cc. 



2. Liver (calf) 250.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



5. Litmus 

 Preparation: 



(1) Chop fresh calf liver into small pieces. 



(2) Add to 1000.0 cc. water, 250.0 g. of (1) 

 and infuse for 24 hours. 



(3) Press out the infusion and filter. 



(4) Boil the liquid for 1 hour at 100°C. 



(5) Filter and add 3 and 4. 



(6) Filter and neutralize with normal 

 soda solution (usually requires 3.0 cc. 

 solution to obtain the proper 

 reaction). 



(7) Place in the autoclave for ^ hour at 

 115°C. 



(8) Filter and add 20.0 cc. of neutral 

 litmus solution. 



(9) Tube in 10.0 cc. lots. 

 Sterilization: Sterilize in the autoclave. 

 Use: Cultivation of the colon-typhoid 



group and other organisms. 

 Variants: This medium has been prepared 

 in a variety of manners. The following 

 authors gave the various listed methods 

 of preparation: 



(a) Pfuhl cultivated anaerobes in a me- 

 dium prepared as follows: 



(1) Pass 500.0 g. of fresh beef liver thru 

 a meat grinding machine and add 

 to 1000.0 cc. of water. 



(2) Allow (1) to stand in the ice box for 

 1 to 2 hours. 



(3) Boil the mixture for 1 hour. 



(4) Strain thru a straining cloth. 



(5) Add peptone, NaCl and soda solu- 

 tion in the usual manner (amount 

 not given). 



(6) Boil and filter. 



(7) Distribute into test tubes in 

 10.0 cc. lots and sterilize in the 

 autoclave (time not given). 



(8) To each tube add 1.0 g. of plati- 



num sponge. Glucose (1.0 or 

 2.0%) may be added if desired. 

 (9) Boil the tubes for 10 minutes in the 

 water bath or steamer. 



(10) Cool quickly and inoculate. 



(11) "Hepin" a material from Much 

 and Romer, may replace the 

 platinum sponge. Boil the bouil- 

 lon and add 1 drop of the sterile 

 Hepin by means of a small pipette. 



(b) Kessler prepared the medium as fol- 

 lows and used it as an enrichment 

 medium for the typhoid bacilli from 

 the blood. 



(1) Chop 500.0 g. of beef liver into 

 small pieces and add to 500.0 g. 

 water. 



(2) Extract (1) at about 50°C. in a 

 steamer for 30 minutes. 



(3) Boil for 30 to 45 minutes. 



(4) Filter. 



(5) Make up the filtrate to 500.0 cc. 



(6) Dissolve 5.0 g. peptone (1%) and 

 2.5 g. NaCl (0.5%) in (5). 



(7) Neutralize with addition of NaOH 

 to litmus. 



(8) Boil 30 minutes in the steamer and 

 readjust the reaction. 



(9) Distribute in 9.0 cc. lots in tubes. 

 (10) Sterilize on 3 successive days for 



30 minutes each. 



(c) Harvey prepared the medium as 

 follows. He stated that other organs 

 may be similarly treated. 



(1) Make a sterilized liver extract in 

 the same way as a meat extract 

 with 1000.0 g. finely minced fresh 

 ox liver and 1000.0 cc. water. 



(2) Prepare a solution of peptone 2.0% 

 and NaCl 1.0%. 



(3) Sterilize the peptone solution. 



(4) Prepare while the solutions are 

 hot, and with sterile precautions: 

 sterilized liver extract 1; sterilized 

 peptone solution 1. 



(5) Distribute with sterile precautions 

 into test tubes. 



(d) Goss et al. cultivated B. chauvoei, on 



a medium prepared as follows: 



(1) Grind 500.0 g. beef liver and add 

 1000.0 cc. water. 



(2) Cook (1) in flowing steam 1 hour. 



(3) Strain thru cheese cloth and 

 cotton. 



