CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



313 



(3) Heat to 100°C. and then filter on a 

 Chardin paper. 



(4) Distribute the opalescent fluid in 

 flasks or tubes in 20.0 cc. lots. 



(5) Dilute egg yolks with 10 volumes of 

 distilled water. 



(6) Clarify the egg yolk by the addition 

 of normal soda solution. Generally 

 1.0 cc. of normal soda solution to 

 100.0 cc. suspension of egg yolk will 

 give the proper solution. However, 

 different yolks may require only 

 half as much. The suspension 

 should be slightly yellow in a thin 

 layer. 



(7) Heat at 100°C. 



(8) Filter on Chardin paper. 



(9) Distribute in flasks or tubes in 

 20.0 cc. lots. 



(10) Prepare a bouillon from 500 to 

 750.0 g. chopped beef or veal with 

 1000.0 cc. water and Martin's pep- 

 tone, by heating to boiling under a 

 low flame and then boiling for 

 30 minutes. See med. (1006). 



(11) Filter and make slightly alkaline. 



(12) Heat at 120°C. to activate pre- 

 cipitation. 



(13) Filter thru double paper. 



(14) Distribute in flasks or Rou.x tubes. 



(15) Add to 500.0 cc. of (14), 400.0 cc. of 

 (4) (sterile egg white solution), and 

 100.0 cc. (9) (sterile egg yolk solu- 

 tion) under aseptic conditions. 



(16) Distribute in sterile tubes in 10.0 cc. 

 lots under aseptic conditions. 



Sterilization: Sterilize (4) and (9) at 115°C. 

 for 20 minutes. Sterilize (14) at 115°C. 

 for 25 minutes. 



Use: General culture medium, for highly 

 parasitic and saprophytic forms. To cul- 

 tivate tubercle bacilli, add cold, and 

 under aseptic conditions, 20.0 cc. of 

 (4) (egg white solution) and 5 to 20.0 cc. 

 of (9) (egg yolk solution) to 100.0 cc. of 

 the bouillon, prepared as before but con- 

 taining no peptone. 



Reference: Besredka and Jupille (1913 

 p. 1009). 



1008. Stickel and Meyer's Meat Digest 

 Solution 



Constituents: 



1. Water, tap 4000.0 cc. 



2. Stomachs (hog) 400.0 g. 



3. Liver, beef placenta or blood 



clots 400.0 g. 



4. K2HPO, 8.0 g. 



Preparation : 



(1) Wash clean and mince finely 5 or 

 more large pig's stomachs. Mince 

 an equal amount of clean pig's or 

 beef liver, cheap fat-free beef, pla- 

 centa or blood clots. 



(2) Mix 2, one of 3 and 40.0 g. of Baker's 

 Chemical Co. HCl in 1 at 50°C. and 

 keep at 50°C. for 18 to 24 hours. 



(3) Make a Biuret and tryptophane 

 test. When both are + the digest 

 is yellowish green and contains very 

 little undigested debris. 



(4) Transfer to large bottles and steam 

 for 10 minutes at 100°C. to stop 

 digestion. 



(5) Strain thru cotton or preferably 

 store over night in the ice chest and 

 decant after 24 hours. 



(6) Warm (5) to 70°C. and neutralize 

 with 2 N Na2C03, to litmus. 



(7) Filter into a flask. 



(8) AddS.Og. K2HPO4. 



(9) Adjust to desired reaction using lit- 

 mus or preferably to a definite H-ion 

 concentration (pH 7.0 to 7.5). 



(10) Heat in steamer at 100° for 15 

 minutes. 



(11) Correct reaction and filter thru 

 paper. 



(12) Distribute in receptacles used for 

 cultures. 



Sterilization: Sterilize at 100° for 30 min- 

 utes on 2 successive days or at 10 pounds 

 pressure for 15 minutes. 

 Use: General inexpensive culture medium. 

 Variants : 



(a) The authors prepared a medium as 

 indicated from step (1) thru (6). 

 The remainder of the preparation was 

 as follows: 



(7) Sterilize at 10 pounds pressure for 

 15 minutes in the autoclave or for 

 30 minutes at 100 °C. on 2 suc- 

 cessive days. 



(8) Inoculate (7) with 1.0% of a 

 24 hour old broth culture of B. 

 saccharolyte or B. coli and incu- 

 bate for 12-18 hours at 37°C. 



(9) Steam for 20 minutes. 



