670 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Variants: If desired, glycerol, peptone, 

 salt, etc., may be added in the preparation 

 of this agar. 



Reference: Duval and Harris (1911 p. 169). 



2071. Distaso's Trypsinized Serum Agar 



Constituents : 



1. Water 1500.0 cc. 



2. Serum (beef or sheep). 500.0 cc. 



3. Agar 30.0 to 40.0 g. 



Preparation : 



(1) Mix equal parts (500.0 cc.) of water 

 and sheep or beef serum. 



(2) Sterilize at 120°C. for 15 minutes. 



(3) Digest for 24 hours at 60°C. with a 

 pancreatic extract from a hog in the 

 presence of chloroform. Activate the 

 extract with an extract of the upper 

 portions of the small intestine. 



(4) Filter on paper. 



(5) Add 30.0 or 40.0 g. of agar to 1000.0 cc. 

 of water. 



(6) Sterilize (5). (Method not given.) 



(7) Mix equal parts (4) and (6). 



(8) Tube. 



Sterilization: Method of final sterilization 



not given. 

 Use: Culture medium for tubercle bacilli, 



strict anaerobes, B. proteus and others. 

 Reference: Distaso (1916 p. 600). 



2072. Stickel and Meyers' Trypsinized 



Blood Clot Agar 



Constituents: 



1. Tap water 1000.0 cc. 



2. Blood clots 500.0 g. 



3. K2HPO4 2.0 g. 



4. Pancreatic extract 10.0 g. 



5. Agar 20.0 g. 



Preparation : 



(1) Obtain 10 liters fresh beef blood from 

 the abbatoir. 



(2) Decant and store the serum (which 

 has separated on standing) in a 

 refrigerator. 



(3) Weigh the blood clots and mix 

 500.0 g. with 1 liter tap water. 



(4) Place the mixture in an enameled 

 pot, bring slowly to a boil and boil 

 slowly for 5 minutes stirring con- 

 stantly. 



(5) Strain fluid thru cheese cloth and 

 pass the residue thru a fruit press, 

 cool to 37°C. 



(6) Make the thick brownish fluid 

 slightly alkaline to litmus. 



(7) Add 1.0% pancreatic extract and 

 incubate at 37°C. for 5-24 or 48 hours. 



(8) When the process is sufficiently ad- 

 vanced, render slightly acid with 

 glacial acetic acid and boil slowly 

 15 minutes. 



(9) Either filter or decant the clear 

 fluid which results on placing the 

 digest over night in a cool place. 



(10) Adjust the reaction as desired. 



(11) Dissolve 3 and 5 in (10). 



(12) Adjust to desired reaction using lit- 

 mus or preferably to a definite H-ion 

 concentration (pH = 7.0 to 7.5). 



(13) Clear (12) by adding 5-10.0% of the 

 decanted beef serum. Steam 45 to 

 60 minutes. 



(14) Remove (13) from steamer and 

 allow the clot to form as a compact 

 mass. Decant or better centrifuge 

 the medium to remove it. 



Sterilization: Sterilize at 100° for 30 min- 

 utes on two successive days. 



Use: General inexpensive culture medium. 

 Authors reported that this medium was 

 excellent for primary isolation of highly 

 parasitic organisms. 



Reference: Stickel and Meyer (1918 p. 81). 



2073. Stickel and Meyer's Tryptic and 

 Peptic Digest Agar 



Constituents : 



1. Tap water 4000.0 cc. 



2. Pig's stomach (minced) .... 400.0 g. 



3. Beef, liver, placenta, blood 



clots (minced) 400.0 g. 



4. HCl (Baker Chemical Co.) . . 40.0 g. 



5. Pancreatic extract or 

 "Bacto" trypsin 40.0 g. 



6. K2HPO4 8.0 g. 



7. Agar 80.0 g. 



Preparation : 



(1) Wash clean and mince fine 5 or more 

 large pigs stomachs. Mince an 

 equal amount of clean pig or beef 

 liver, cheap fat-free beef, placenta 

 or blood clots. 



(2) Mix 2, one of 3, and 4 at 50°C. and 

 keep at 50°C. for 18 to 24 hours. 



(3) Make a biuret and tryptophane test. 

 When both are + the digest is yel- 



