CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



309 



999. Martin's Liver Digest Solution 



Constituents : 



1. Water 2000.0 cc. 



2. Stomach (hog) 200.0 g. 



3. Liver, beef or hog 200.0 g. 



Preparation : 



(1) Mi.x 200.0 g. of chopped hog stomach 

 and 200.0 g. chopped hog or beef liver, 

 and 20.0 cc. of commercial HCl in 2 

 liters of water. 



(2) Digest (1) at 50°C. for 12 to 24 hours. 



(3) Boil the liquid, at the end of 24 hours. 



(4) Allow to cool and settle for 24 hours 

 or less. 



(5) Decant or siphon off the clear liquid. 



(6) Neutralize and make slightly alkaline 

 by the addition of soda. Indicator 

 not specified. 



(7) Heat at 120 degrees. 



(8) Filter and distribute as desired. 

 Sterilization: Method not given. 



Use: Cultivation of typhoid bacilli. 

 Author reported that typhoid and para 

 typhoid bacilli developed rapidly in this 

 medium, producing gas, and acid and 

 finally the reaction became alkaline. 



References: Martin (1915 p. 261), Harvey 

 (1921-22 p. 99). 



1000. Sellards and Bigelow's Liver Digest 

 Blood Solution 



Constituents: 



1. Water 1000.0 cc. 



2. Veal 100.0 g. 



3. Stomach (hog) 150.0 g. 



4. Liver 200.0 g. 



5. Blood, horse or rabbit 

 (10.0%) 100.0 cc. 



Preparation : 



(1) Prepare a peptone by digesting 

 100.0 g. veal, 150.0 g. hog stomach and 

 200.0 g. liver in a liter of water con- 

 taining 1.0% concentrated HCl, for 

 19 hours at 53°C. 



(2) Heat in an Arnold to destroy the pep- 

 sin. 



(3) Decant the supernatant fluid and 

 neutralize at room temperature first 

 to litmus and then to china blue and 

 rosalic acid. 



(4) Again decant and dilute with an equal 

 volume of water. 



(5) Add 10.0%, horse or rabbit blood to 

 sterile (4). 



Sterilization: Sterilize (4) in the Arnold. 

 Method of sterilization of blood not 

 given. Sterilize the mi.xture of blood and 

 digest for 15 to 20 minutes in the Arnold. 



Use: Cultivation of measles virus from 

 patients' blood. 



Reference: Sellards and Bigelow (1920-21 

 p. 242). 



1001. Dubovsky and Meyer's Liver Digest 

 Solution 



Constituents : 



1. Water, tap 5000.0 cc. 



2. Stomach, hog 400.0 g. 



3. Liver 400.0 g. 



4. K2HPO4 (0.2%). 



5. Heart 1000.0 g. 



Preparation : 



(1) Wash clean and mince finely 5 or 

 more large hogs' stomachs. 



(2) Mince an equal amount of clean 

 hog or beef liver. 



(3) Mi.x 400.0 g. (1), 400.0 g. (2), 40.0 g. 

 HCl and 4000.0 cc. tap water at 

 50°C. 



(4) Keep in glass or porcelain receptacles 

 for 18-24 hours. 



(5) Make Biuret and tryptophan test, 

 when both are + the digest is green- 

 yellowish and contains little un- 

 digested debris. 



(6) Transfer to large bottles and steam 

 10 minutes at 100.0°C. to stop 

 digestion. 



(7) Strain the digest thru cotton or 

 preferably store over night in the 

 ice chest and decant after 24 hours. 



(8) Warm the digest to 70°C. and neu- 

 tralize with 2 N Na2C03 using 

 litmus. 



(9) Filter the desired amount, add 0.2% 

 K2HPO4 and adjust to "pH = 7.4. 



(10) Slowly heat to boiling finely ground 

 1000.0 g. fat-free heart and tap water 

 1000.0 cc. 



(11) Adjust to pH = 8.0 or 8.2. 



(12) Cool and carefully skim off the layer 

 of fat which floats on medium. 



(13) To each liter of (12) add 2 liters 

 of (9). 



(14) Adjust to pH = 7.2 or 7.4. 

 Sterilization: Sterilize at 18 pounds pres- 

 sure for one hour. Incubate 5 days and 

 repeat the same sterilization for one hour 

 at 18 pounds pressure. 



