244 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Preparation : 



(1) Mince finely the flesh of fish. 



(2) Add 500.0 g. to the following solu- 

 tion: sodium chloride 26.5 g.; po- 

 tassium chloride 0.75 g.; magnesium 

 chloride 3.25 g. ; water 500.0 cc. 



(3) Heat the mixture 20 minutes over a 

 free flame at a temperature not ex- 

 ceeding 50°C. 



(4) Raise the temperature to boiling 

 point. 



(5) Boil 10 minutes. 



(6) Pour the mixture on to a thick, clean 

 cloth. 



(7) Collect the fluid which drains the 

 cloth together with that obtained by 

 squeezing the cloth. 



(8) Filter the fluid collected thru well 

 wetted, thick filter paper. 



(9) Add peptone, 5.0 g. 



(10) Steam 45 minutes. 



(11) Bring the volume up to 1000.0 cc. 

 by the addition of water. 



(12) Steam 30 minutes. 



(13) Filter while hot, thru well-wetted, 

 thick filter paper. 



(14) Distribute into flasks or test tubes. 

 Sterilization: Sterilize in streaming steam. 

 Use: General culture medium. 

 Reference: Harvey (1921-22 p. 69). 



800. Richardson's Mucosa Infusion Broth 



Constituents : 



1. Distilled water 500.0 cc. 



2. Mucosa, hog intestine 250.0 g. 



3. Peptone 5.0 g. 



4. NaCl 2.5 g. 



Preparation : 



(1) Take the small intestine of a hog in 

 fresh condition, without cutting, 

 wash thoroly in running water, until 

 the water runs perfectly clear. 



(2) Lay the intestine open and scrape 

 off the mucosa with a glass slide 

 (100.0 g. of mucosa is easily obtained 

 from one hog). 



(3) To 250.0 cc. of the mucosa add 

 500.0 cc. of distilled water. 



(4) Boil for 30 minutes and cool. 



(5) Boil again, neutralize and boil once 

 more. 



(6) Filter. 



(7) Boil and 5.0 g. peptone and 2.5 g. 

 NaCl. 



(8) Neutralize again and boil for 20 

 minutes. 



(9) Filter. 

 (10) Tube. 



Sterilization: Sterilize using the fractional 



method. 

 Use : Attempt to cultivate bodies found in 



carcinomatous tissue. Author reported 



no growth. 

 Reference: Richardson (1900-01 p. 73). 



801. Boyer's Bone Infusion Broth 



Constituents : 



1. Water 900.0 cc. 



2. Bones beef ribs 500.0 g. 



3. HCl 100.0 cc. 



4. Peptone 15.0 g. 



Preparation : 



(1) Grind beef bones to small pieces. 



(2) Mix 500.0 g. of (1) with 900.0 g. water 

 and 100.0 cc. normal HCl. 



(3) Allow to stand for 12 to 24 hours. 



(4) Heat in the autoclave at 125 to 

 130°C. for 30 minutes. 



(5) Cool and then filter thru linen. 



(6) Add 15 parts of peptone to 1000 

 parts of the liquid. 



(7) Neutralize by the addition of a 

 dilute soda solution. (Indicator not 

 specified.) 



(8) Heat in the autoclave at 125-130° 

 for 30 minutes. 



(9) Filter while hot. 



(10) Allow to stand for one day in the 

 cold. 



(11) Filter again. 



Sterilization: Sterilize at 120° (time not 



specified). 

 Use: General culture medium. Author 



cultivated streptococci, Streptococcus 



pyogenes and other organisms. 

 Reference: Boyer (1918 p. 230). 



802. Ball's Animal Infusion Broth 



Use the flesh of guinea pigs as well as 

 other experimental animals instead of beef 

 in the preparation of Dunham's Infusion 

 Broth (779). 

 Reference: Ball (1919 p. 84). 



803. Stutzer's Nitrate Extract Broth 



Constituents: 



1. Water. 



2. Peptone. 



