264 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(4) Skim off fat floating on the surface, 



(5) Raise the temperature to boiling 

 point. 



(6) Boil 10 minutes. 



(7) Pour the mixture on to a wet, thick, 

 clean cloth. 



(8) Add sodium chloride 5.0 g. to the 

 filtrate. 



(9) Steam 45 minutes. 



(10) Bring the volume up to 1000.0 cc. 

 by the addition of water. 

 . (11) Estimate and adjust the reaction, 



(12) Steam for 30 minutes. 



(13) Filter while hot thru well-wetted, 

 thick filter paper. 



(14) Make a paste of 10.0 g. of corn flour 

 starch and 200.0 cc. of (13). 



(15) Heat 20 minutes at 20 pounds pres- 

 sure. 



(16) Add 10.0 g. peptone, 5.0 g. NaCl 

 and 800.0 cc. of (13) to (15). 



(17) Steam 45 minutes. 



(18) Make the reaction 0.2% acid to 

 phenolphthalein. 



(19) Steam 30 minutes. 



(20) Filter while hot thru well-wetted, 

 thick filter paper. 



(21) Add saturated litmus solution to 

 make a deep blue color. 



(22) Distribute into test tubes. 

 Sterilization: Sterilize at 100°C, for 20 



minutes on 3 days. 

 Use: Cultivation of meningococci. 

 Reference: Harvey (1921-22 p. 112), 



851. Loeffler's Malachite Green Infusion 

 Broth 



Constituents : 



1. Beef infusion 1000.0 cc. 



2. Peptone (2.0%) 20.0 g, 



3. Lactose (5.0%) 50.0 g. 



4. Dextrose (1.0%) 10.0 g. 



5. Sodium sulfate (0.5%) 5.0 g. 



6. KNO3 (2.0%) 20.0 g. 



7. KNO2 (1.0%) 10.0 g. 



8. Malachite green 2% solution 30.0 cc. 

 Preparation : 



(1) Prepare a beef infusion from one 

 pound of beef. 



(2) Dissolve 2, 3,t, 5, 6 and 7 in (1). 



(3) Add 3.0% of 2.0% malachite green 

 solution to (2), 



Sterilization: Sterilize on each of 3 suc- 

 cessive days for 10 minutes in streaming 

 steam. 



Use: Enrichment of the colon-typhoid 



group. 

 Variants: Klimmer prepared the medium 



as follows: 



(1) Dissolve 2, 3, 4, 5, 6 and 7 in meat 

 infusion broth. 



(2) Flask in 100.0 cc. quantities. 



(3) Sterilize on each of 3 successive days 

 for 10 minutes in streaming steam. 



(4) Add 3.0 cc. of a 2.0% solution of 120 

 Hochst malachite green to each 

 100.0 cc. lot of sterile (2). 



(5) Tube in 3.0 or 4.0 cc. lots in sterile 

 tubes. 



References: Loeffler (1906 pp. 289, 295), 

 Klimmer (1923 p. 213). 



852. Duval and Lewis' Inulin Bouillon 



Constituents : 



1. Meat infusion broth 1000.0 cc. 



2. Inulin 10.0 g. 



Preparation : 



(1) Prepare boiullon according to stand- 

 ard methods from beef. (Method or 

 reference not given.) 



(2) Adjust from 0.2 to 0.4% normal to 

 phenolphthalein. After sterilization 

 reaction from 0.5 to 0.8%, 



(3) Distribute into clean tubes in 9.0 cc. 

 lots. 



(4) Prepare a 10.0% inulin solution in 

 distilled water. 



(5) Add 1.0 cc. of sterile (4) to each 9.0 cc. 

 of sterile (3). 



Sterilization: Sterilize (3) method not 

 given. Sterilize (4) in the autoclave at 

 15 pounds pressure for 15 minutes. 



Use: Cultivation of pneumococci. 



Reference: Duval and Lewis (1905 p. 484). 



853. Bulir's Mannitol Infusion Broth 



Constituents : 



1. Water 2000.0 cc. 



2. Beef 1000.0 g. 



3. Peptone (Witte) 25.0 g. 



4. NaCl 15.0 g. 



5. Mannite 30.0 g. 



6. Neutral red. 

 Preparation : 



(1) Macerate 1000.0 g. finely chopped 

 lean beef with 2000.0 cc. water for 

 24 hours. 



(2) Filter thru linen, and press the meat 

 free from water. 



(3) To the liter of meat infusion thus 



