270 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(6) Distribute in 100.0 cc. lots in 

 500.0 cc. Erlenmeyer flasks. 



(7) Seed half the sterilized flasks with 

 the grass bacillus of Karlensky and 

 the remainder with the glass bacillus 

 of Moeller. 



(8) Incubate at 37.5°C. for two weeks. 



(9) Then heat to 100°C. 



(10) Cultures are then filtered and fil- 

 trate made up to 1000.0 cc. 



(11) Add 5.0 g. peptone, 2.5 g. K2HPO4, 

 2.0 g. Liebig's beef extract, 50.0 cc. 

 glycerol and 5.0 g. of aminoid pep- 

 tone from beef (Arlco). 



(12) The reaction is left unchanged. 



(13) Filter and distribute 50.0 cc. lots 

 in 500.0 cc. Erlenmeyer flasks. 



(14) Add 5.0 cc. of sterile horse blood 

 serum (serum must be raw) just 

 before inoculation. 



Sterilization: Sterilize (6) and (13) sepa- 

 rately in the autoclave, time not given. 



Use: Preparation of johnin. Johnin is a 

 material used for detection of Johnin's 

 disease, much as tuberculin is used for 

 tuberculosis. The medium was inocu- 

 lated heavily with the organism causing 

 Johnin's disease. Cover the flasks with 

 tin foil to prevent evaporation and incu- 

 bate 10 to 16 weeks. 



Reference: Beach and Hastings (1922 

 p. 72). 



871. Harvey's Blood Infusion Broth 



Constituents : 



1. Water 1000.0 cc. 



2. Beef 500.0 g. 



3. NaCl 14.0 g. 



4. CaCla 0.25 g. 



5. KCl 0.42 g. 



6. Glucose 25.0 g. 



7. Blood, citrated plasma. . . . 200.0 cc. 

 Preparation : 



(1) Prepare meat infusion solution as 

 indicated in medium 729. 



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



(3) Tube in 5.0 cc. quantities. 



(4) Add 1.0 cc. of citrated humas plasma 

 to each tube of (3). 



Sterilization: Method not given. 

 Use: Cultivation of gonococci. 

 Reference:' Harvey (1921-22 p. 82). 



872. Park, Williams and Krumwiede's 



Potato Infusion Broth 



Constituents: 



1. Water 1000.0 cc. 



2. Meat infusion 1000.0 cc. 



3. Potato 1.0 lb. 



4. Peptone 20.0 g. 



5. NaCl 10.0 g. 



Preparation : 



(1) Grate 1.0 pound of white potatoes 

 or run thru a chopping machine. 



(2) Soak in a liter of water over night. 



(3) Heat to boiling. 



(4) Press thru cheese cloth. 



(5) Add one egg per liter. 



(6) Autoclave one hour to clarify. 



(7) Filter thru cotton (very tedious). 



(8) Flask. 



(9) Soak 1 pound of finely chopped lean 

 beef in a liter of tap water over night 

 in the ice box or at room tem- 

 perature. 



(10) Weigh (9) and heat at 45°C. for 1 

 hour and then boil for 30 minutes. 



(11) Make up the loss in weight by the 

 addition of hot water. 



(12) Strain thru cheese cloth and squeeze 

 by twisting the cloth or use a meat 

 press. 



(13) Mix equal parts of (8) and the juice 

 from (12). 



(14) Dissolve 20.0 g. peptone and 10.0 g. 

 NaCl in (13). 



(15) Distribute as desired. 

 Sterilization: Autoclave for 30 minutes at 



15 pounds pressure. 

 Use: General culture medium. 

 Reference: Park, Williams and Krum- 



wiede (1924 p. 122). 



873. Lubinski's Potato Infusion Broth 



Constituents : 



1. Water 1500.0 cc. 



2. Potato 1000.0 g. 



3. Meat 500.0 g. 



4. Peptone 10.0 g. 



5. NaCl 5.0 g. 



6. Glycerol 40.0 g. 



Preparation : 



(1) Wash potatoes clean and cut them 

 in small pieces. 



(2) Boil for 3 or 4 hours under a free 

 flame or in a steamer. 



