256 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Variants : 



(a) Harvey prepared a similar medium 

 as follows: 



(1) Mince finely ox liver. 



(2) Add 500.0 g. to 1000.0 cc. distilled 

 water. 



(3) Heat the mixture 20 minutes at a 

 temperature not exceeding 50°C. 



(4) Skim off fat floating on surface. 



(5) Raise the temperature to boiling 

 point. 



(6) Boil 10 minutes. 



(7) Pour the mixture on to a wet, 

 thick, clean cloth. 



(8) Collect the fluid which drains thru 

 the cloth together with that ob- 

 tained by squeezing the meat in 

 the cloth. 



(9) Filter the fluid collected thru well- 

 wetted, thick filter paper. 



(10) Add to the filtrate, peptone 10.0 g. 



(11) Bring the volume up to 1000.0 cc. 

 by the addition of water. 



(12) Adjust the reaction. 



(13) Steam 30 minutes. 



(14) Filter, while hot, thru well-wetted, 

 thick filter paper. 



(15) Dissolve 10.0 g. glucose and 1.0 g. 

 K.HPO^ in the filtrate. 



(16) Harvey further modified the me- 

 dium in that he added 1.0 cc. of 

 defibrinated rabbit blood to each 

 10.0 cc. of the medium. 



(17) Sterilization not specified. 



(b) Klimmer prepared a medium as 

 follows: 



(1) Boil finely chopped liver with a 

 double amount of water. 



(2) Add 1.0% peptone and 0.5% NaCl 

 to the juice of (1). 



(3) Neutralize to phenolphthalein by 

 the addition of NaOH. 



(4) Boil. 



(5) Filter. 



(6) Add 2.0% glucose. 



(7) Cut the boiled liver in pieces 

 1-2 ccm. 



(8) Sterilize (7) (Method not given). 



(9) When ready for use, add about 4 

 pieces of (8) to a series of tubes and 

 add (6) until the pieces of liver are 

 covered to a depth of 3 cm. 



(10) Sterilize (method not given). 



(11) Add a 2 cm. layer of sterile paraffin 

 to each tube after inoculation. 



References: Jackson and Muer (1911 

 p. 290), (1911 p. 727), Levine (1921 p. 110), 

 Harvey (1921-22 p. 110), Klimmer (1923 

 p. 201). 



834. Haslam's Brain Liver Infusion Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Beef liver 500.0 g. 



3. Brain 500.0 g. 



4. Peptone (1.0%) 10.0 g. 



5. NaCl (0.5%) 5.0 g. 



Preparation : 



(1) Grind liver and to each liter of water 

 add 500.0 g. liver. 



(2) Bring to boil slowly and cook for 

 about 30 minutes or until the liquid 

 is clear. 



(3) Pour off the clear broth, and add 1.0% 

 peptone and 0.5% NaCl. 



(4) Adjust to pH = 8.0. 



(5) Grind brain, using the coarse grinder 

 and cook at 3 pounds pressure for 

 about one and one half hours. 



(6) Fill flasks f full with mixture of brain 

 tissue and liver broth (amounts of 

 each not given). 



Sterilization: Autoclave at 6 pounds pres- 

 sure for three hours. 



Use: Cultivation of B. chauvoei and 

 streptococci. 



Variants : 



(a) Gross, Barabin and Haines prepared 

 the medium as follows: 



(1) Grind beef liver and brain sepa- 

 rately. 



(2) To ground liver add 1000.0 cc. 

 water. 



(3) Cook ground brain and (2) in flow- 

 ing steam for 1 hour. 



(4) Strain liver broth thru cheese cloth 

 and cotton. 



(5) Add peptone (1.0%) and NaCl 

 (0.5%). 



(6) Titrate to pH = 8.2. 



(7) Tube, two parts (6) to one part 



brain. 



(8) Autoclave for one hour at 15 pounds 

 pressure. 



(b) Long and Cornwell attempted to cul- 

 tivate streptococci from a measles 

 patient on a medium prepared as 

 follows: 



(1) Heat 500.0 g. of ground fat free beef 



