664 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



agar to give the desired consistency 

 and 0.5% galactose. 



Sterilization: Sterilize (9) in tlie autoclave. 

 Final sterilization not given. 



Use: Cultivation of Lactobacillus acid- 

 ophilus and Lactobacillus bulgaricus. 

 The authors reported that any other 

 carbohydrate might be used instead of 

 galactose, but galactose favored the 

 growth of the organisms most. This 

 medium is slightly superior to preceding 

 casein digest medium. 



Variants: Any other carbohydrate may be 

 used instead of galactose. The carbo- 

 hydrate may be used in 1.0% instead of 

 0.5% concentration if desired. 



Reference: Kulp and Rettger (1924 p. 363). 



2061. Harvey's Trypsinized Ox Heart Agar 



Constituents : 



1. Water 400.0 cc. 



2. Heart, ox 



3. NaCl 1.0 g. 



4. CaCl2 0.5 g. 



5. Blood (citrated laked) 10.0 cc. 



Preparation : 



(1) Mince finely an averaged sized ox 

 heart. 



(2) Add to 400.0 cc. tap water. 



(3) Heat slowly to 75°C. 



(4) Allow to cool to 45°C. 



(5) Add trypsin solution to 1.0%. 



Note: e.g. Liq. trypsin Co. (A 

 and H). 



(6) Place in incubator 150 minutes. 



(7) Test for peptone by the Biuret test. 



Note: Add 1.0 cc. 5.0% copper 

 sulphate to 5.0 cc. trj^psin digest, 

 followed by 5.0 cc. N/1 potassium 

 hydroxide. Note the color change. 

 If the color is pink, peptonization is 

 complete, if bluish purple incom- 

 plete. 



(8) Make faintly acid to litmus with 

 4.0% acetic acid. 



(9) Boil 15 minutes. 



(10) Allow the solid matter to settle. 



Note: Or simply strain thru cloth. 



(11) Pour off the supernatant fluid. 



(12) Add: sodium chloride 1.0 g., calcium 

 chloride 0.5 g. 



(13) Make faintly alkaline to litmus. 



(14) Steam 45 minutes. 



(15) Bring up to original volume. 



(16) Adjust the reaction. 



(17) Steam 30 minutes. 



(18) Clarify and filter. 



(19) Bleed a rabbit directly from the 

 carotid into 1.5% sodium citrate 

 solution. 



(20) Dilute with 0.85% sterile salt solu- 

 tion to give a 5.0% suspension of 

 blood. 



(21) Add 10.0% ether. 



(22) Shake to mix. 



(23) Leave the sediment 24 hours. 



(24) Draw off the laked blood into a 

 sterile bottle. 



(25) Add an excess of ether. 

 Sterilization: Final sterilization not spec- 

 ified. 



Use: Cultivation of highly pathogenic 

 organisms. 



Variants: The author cultivated meningo- 

 cocci in a medium prepared as follows : 



(1) Steam 50.0 g. pea flour and 100.0 g. 

 NaCl in 1000.0 cc. water for 30 min- 

 utes, with occasional stirring. 



(2) Filter thru thick filter paper. 



(3) Sterilize (method not given). 



(4) Mix 5.0% of (3) with the agar ob- 

 tained at step (18) in the medium 

 given above. 25.0% rabbit or horse 

 serum may be added if desired. 



Reference: Harvey (1921-22 pp. 75, 77, 

 114, 119). 



2062. Norris' Trypsinized Mutton Agar 



Constituents : 



1. Water 2000.0 cc. 



2. Mutton, fat-free 2.0 lbs. 



3. NaCl 5.0 g. 



4. CaCls 0.25 g. 



5. Agar 80.0 g. 



Preparation : 



(1) Mince two pounds fat free mutton. 



(2) Place 2 liters of water and (1) in a 

 3 liter pot. 



(3) Autoclave at 130°C. for one hour. 



(4) Keep over night, but if urgently 

 wanted, cool to 45°C. and continue 

 as below. 



(5) Test amino acidity by Sorensen's 

 method. 



(6) Add 40.0 cc. pancreatic extract per 

 liter. 



(7) Digest four hours at 37°C. 



(8) Retest amino acid acidity. 



