316 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(10) When ready for use, use in 40.0% 

 strength. 



(11) Sterilization of stock solution not 

 specified. 



References: Stickel and Meyer (1918 p. 81), 

 Knorr (1921 p. 598), Harvey (1921-22 

 p. 117), Klimmer (1923 p. 176). 



1114. Spray's Blood Clot Digest Solution 



Constituents: 



1. Water. 



2. Blood clot. 



3. NajCOa. 

 Preparation : 



(1) Remove the serum from the blood 

 clot and allow to drain. 



(2) Pass the clot thru a wire gauze. 



(3) Boil and divide finely again. 



(4) Place one liter of this semi fluid ma- 

 terial in a two liter flask. 



(5) Add 20.0 g. anhydrous Na2C03, 5.0 g. 

 trypsin, and 15.0 to 20.0 cc. chloro- 

 form. 



(6) Incubate for 15 days with a second 

 addition of 3.0 g. trypsin on the 

 tenth day. Chloroform may be 

 added on the fifth and tenth days. 



(7) Make strongly acid by the addition 

 of 50.0 to 75.0 cc. HCl. 



(8) Steam in the water bath to drive off 

 the chloroform. 



(9) Adjust to pH = 7.4 to 7.5. 

 (10) Tube. 



Sterilization: Sterilize in the autoclave. 



Use: The author reported that this blood 

 clot digest may be added in from 5.0 to 

 10.0% to melted North's or other agar for 

 growth of H. influenzae, pneumococcus 

 and streptococcus. When sodium oleate 

 is added H. influenzae grows very vigor- 

 ously. 



Reference: Spray (1927 p. 14). 



1115. Loffl's Trypsinized Blood Solution 



Constituents: 



1. Water. 



2. Blood. 

 Preparation : 



(1) Allow 9 liters of blood to flow into a 

 liter of 0.8% ammonium oxalate. 



(2) Mix well. 



(3) Centrifuge at 3000 revolutions per 

 minute. 



(4) This yields about 6 liters of plasma. 



(5) Precipitate out the oxalate by the 

 addition of calcium acetate. 



(6) Add 40.0 g. trypsin to the plasma and 

 allow it to act for 8 days. 



(7) At the end of this time, boil for a few 

 minutes, allow to stand and filter. 



Sterilization: Method not given. 



Use: Inexpensive medium. Author re- 

 ported that this medium contains about 

 8.0% nutrient materials and should be 

 diluted 1:3 before use. 



Reference: Loffl (1915-16 p. 109). 



1116. Gordon et al. Trypsinized Heart 

 Solution (Tanner) 



Constituents: 



1. Water 1000.0 cc. 



2. Heart, beef 500.0 g. 



Preparation : 



(1) Free fresh bullock's heart from fat 

 and blood vessels. 



(2) Mince (1) very finely and weigh. 



(3) To each 0.5 kilo add 1 liter of water, 

 and make faintly alkaline to litmus 

 by the addition of 20.0% KOH 

 solution. 



(4) Heat slowly to 75 to 80°C. for 

 5 minutes. 



(5) Cool to 37°C. and add 1.0% of liquid 

 trypsine comp., and keep at 37° for 

 2.5 to 3 hours. 



(6) When trypsinizing is finished test for 

 peptone with copper sulphate and 

 KOH (Method not given). 



(7) Render slightly acid by the addition 

 of glacial acetic acid. 



(8) Bring slowly to boil and boil for 

 25 minutes. 



(9) Leave over night in a cool place. 



(10) Siphon off the clear liquid in the 

 morning. 



(11) Make faintly alkaline to litmus. 

 Sterilization: Sterilize in an autoclave at 



118° for one hour on each of two days. 

 Use: General culture medium. 

 Reference: Tanner (1919 p. 45). 



1117. Harvey's Trypsinized Heart Solution 



Constituents: 



1. Water 400.0 cc. 



2. Heart, ox 1 



3. NaCl 1.0 g. 



4. CaCli 0.5 g. 



