258 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



an enzyme digest of milk to the amino 

 acid stage. Author reported no growth 

 in medium (15b). 

 Reference: Mueller (1922 p. 331). 



836. Mueller's Heart Infusion Peptone 

 Solutions 



Constituents : 



500.0 cc 

 1.0 lb 



10.0 



10.0 

 0.4 

 0.2 



Water (tap) 



Heart, beef 



Peptone (Difco) 



NaCl 



MgS04 



6. CaCl, 



7. K0HPO4 2.0 g. 



8. Glucose 2.0 g. 



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

 Preparation : 



(1) Prepare beef heart infusion by heat- 

 ing 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) Make up one of the following media: 

 Decolorized infu- 

 sion 25.0 cc. 



Glucose salt solu- 

 tion 25.0 cc. 



Peptone (Difco) .... 0.0 or 0.5 g. 

 Infusion (not de- 

 colorized) 25.0 cc. 



Glucose salt solu- 

 tion 25.0 cc. 



Peptone 0.0 or 0.5 g. 



'Water 25.0 cc. 



Glucose salt solu- 



jtion 25.0 cc. 



[Peptone 0.5 g. 



(5) Bring each lot to pH = 7.4 to 7.8. 



(6) Filter if necessary. 



(7) Tube. 



Sterilization: Sterilize at 10 pounds pres- 

 sure for ten minutes. 



Use: To study food requirements of strep- 

 tococci and pneumococci. Author re- 

 ported that no growth in (3) (a) occurred 

 without peptone, no growth in (3) (c). 

 All others gave growth. 



Reference: Mueller (1922 p. 325). 



(a) 



(b) 



(c) 



837. Huntoon's Hormone Heart Infusion 

 Broth 



Constituents : 



1. Water 1000.0 cc. 



2. Heart, beef 500.0 g. 



3. Peptone (Bacto) 10.0 g. 



4. Gelatin 10.0 g. 



5. Salt 5.0 g. 



6. Whole egg 1 



7. Glucose 1.5 g. 



8. Laked blood 

 Preparation : 



(1) Mix 10.0 g. Bacto peptone, 10.0 g. 

 gelatin, 5.0 g. NaCl, one whole egg 

 and 500.0 g. of finely chopped beef 

 heart in a liter of water. Place in an 

 enamel ware vessel or a large coffee 

 pot. 



(2) Heat over a free flame with constant 

 stirring until the red color of the 

 meat infusion changes to brown at a 

 temperature of about 68°C. Do not 

 go beyond this temperature. 



(3) Adjust to slightly alkaline to litmus 

 with N/1 NaOH and then add 1.0 cc. 

 per liter of medium. 



(4) Cover the vessel and place in an 

 Arnold sterilizer or in a water bath 

 at 100° for one hour. 



(5) Remove the vessel from the sterilizer 

 and separate with a glass rod the firm 

 clot which has formed from the side 

 of the vessel. 



(6) Return to the Arnold sterilizer at 

 100° for U hours. 



(7) Remove the vessel and allow to stand 

 at room temperature for about 10 

 minutes in a slightly inclined posi- 

 tion. 



(8) Pipette off the fluid portion or 

 decant. If it is poured thru a fine 

 wire sieve, many of the fine pieces 

 of meat clot may be caught. (Avoid 

 filtering thru cheese cloth, cotton 

 or other adsorptive materials.) 



(9) Allow (8) to stand in tall cylinders 

 for 15 to 20 minutes until the fat 

 present has risen to the surface and 

 removed. 



(10) The medium may be further cleared 

 by filtering thru glass wool, asbestos 

 wool, sedimentation or centrifu- 

 gation. 



