CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



257 



liver in 1000.0 cc. of water in flowing 

 steam for two hours. 



(2) Add 10.0 g. peptone and 5.0 g. NaCl. 



(3) Adjust to pH = 8.2. 



(4) Strain through gauze. 



(5) Free beef brain from blood and 

 membrane and make a mixture of 

 medium consistency of 500.0 g. 

 brain and 200 to 300.0 cc. water. 



(6) Heat (5) in flowing steam for two 

 hours. 



(7) Mix 120.0 cc. of (4) with 60.0 cc. 

 of (6). 



(8) Final reaction, following sterilizing 

 pH = 7.6. 



(9) Autoclave at 15 pounds pressure for 

 one hour. 



References: Haslam (1920 p. 540), Goss, 

 Barabin and Haines (1921 p. 615), Long 

 andCornwell (1927 p. 409). 



835. Mueller's Heart Infusion Aminoid 

 Solution 



Constituents : 



1. Water 600.0 cc. 



2. Heart, beef 1.0 lb. 



3. Aminoids 10.0 g. 



4. NaCl 10.0 g. 



5. MgS04 0.4 g. 



6. CaClz 0.2 g. 



7. K2HPO4 2.0 g. 



8. Glucose 2.0 g. 



9. Phenol red (0.02% soln.) .... 80.0 cc. 

 Preparation : 



(1) Prepare a beef heart infusion by 

 heating 1 pound of chopped beef 

 heart with 500.0 cc. of water to the 

 boiling point. Strain and filter. 



(2) Decolorize by boiling for 25 minutes 

 with 10.0% "Norit" a commercial 

 grade of wood charcoal. 



(3) Filter thru paper. 



(4) Dissolve 10.0 g. aminoids in 100.0 cc. 



of water. 



(5) Precipitate aminoids from (4) with 

 100.0 cc. of 10.0% solution of HgSOi 

 in 5.0% H2SO4. 



(6) Allow to stand over night and filter 

 on a Buchner funnel and wash with 

 water. 



(7) Suspend filtrate in water, make 

 slightly alkaline with Ba(0H)2 and 

 decomposed with H2S with warming. 



(8) Filter off the precipitated HgS and 

 free the heavily pigmented filtrate 

 from Ba with H2SO4. Dilute to 

 100.0 cc. 



(9) Dissolve 4, 5, 6, 7, 8 and 9 in 1000.0 cc. 

 of water. 



(10) Make the following mixture: 

 Decolorized infusion 



(2) 25.0 cc. 



Glucose salt solution 



(9) 25.0 cc. 



HgS04 precipitate frac- 

 tion (8) 0.25 or 2.5 cc. 



(11) To the remainder of the precipitate 

 fraction add 10.0 cc. of a 10.0% solu- 

 tion of HgS04 and allow to stand 

 over night. 



(12) Filter on Buchner funnel. 



(13) Free precipitate and filtrate from 

 H2SO4 and Hg by making slightly 

 alkaline with Ba(0H)2 and bubbling 

 in H2S. 



(14) Free from Ba with H2SO4 and dilute 

 to 100.0 cc. 



(15) Make the following mixtures: 

 Decolorized infu- 

 sion (2) 25.0 cc. 



Glucose salt solu- 



] tion (9) 25.0 cc. 



HgS04 filtrate frac- 



[tion (14) 0.5 or 1.0 cc. 



Decolorized infu- 

 sion (2) 25.0 cc. 



Glucose salt solu- 

 tion (9) 25.0 cc. 



HgS04 filtrate frac- 

 tion (14) 1.0 cc. 



HgS04 precipitate 



Infraction (8) 1.0 cc. 



Decolorized infu- 

 sion (2) 25.0 cc. 



Glucose salt solu- 

 tion (9) 25.0 cc. 



HgS04 filtrate frac- 

 tion (14) 0.5 or 1.0 cc, 



HgS04 precipitate 



fraction (8) 0.5 or 1.0 cc. 



(16) Bring (10) and (15) (a), (b), (c) to 

 pH of 7.4 to 7.8. Filter if necessary. 



Sterilization: Tube and sterilize at 10 

 pounds pressure for 10 minutes. 



Use: To study food requirements of strep- 

 tococci and pneumococci. Aminoids are 



(a) 



(b) 



(c) 



