598 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(3) Mix thoroly and pour in plates. 



(i) Bell cultivated influenza bacilli in a 

 medium containing 4.0% defibrinated 

 rabbit blood to a 2.0% infusion agar. 



(j) Yoshida cultivated Entamoeba tetra- 

 gena and Entamoeba coli from their 

 cysts in the water of condensation of 

 an agar containing one part defibri- 

 nated horse blood to two parts agar. 

 The blood and agar were mixed at 

 60°C. 



(k) Anderson and Schultz added 2.0% 

 defibrinated blood to infusion agar 

 and heated to 80°C. until a brown 

 color is produced. They cultivated 

 Bacillus ■pjeifferi on this medium. 



(1) Harvey mixed one part of sterile 

 defibrinated blood at 45°C. with two 

 parts infusion agar (see variant (v) 

 medium 1661). 

 (m) Novy-MacNeal (Harvey) cultivated 

 leishmania on a medium prepared by 

 mixing one part melted infusion agar 

 at 45 °C. containing 2.0% peptone (see 

 variant (aa) medium 1375) with two 

 parts sterile defibrinated rabbit blood 

 at 45°C. 



(n) Harvey cultivated gonococci on a 

 medium prepared by adding 5 drops 

 of sterile defibrinated blood at 45°C. 

 to a tube of infusion agar (see variant 

 (v) medium 1661) with a reaction of 

 0.6% acid to phenolphthalein at 

 45°C. Preferably infusion agar in 

 which sodium chloride is replaced by 

 di-sodium phosphate. The substitu- 

 tion of phosphate is important. The 

 blood is sterilized by heating 8 days 

 at 57°C. Serum may be used instead 

 of blood. 



(o) Harvey cultivated B. influenza, etc., 

 on a medium prepared as follows: 



(1) Boil 1.0 cc. blood with 9.0 cc. 

 water. 



(2) Allow to deposit. 



(3) Add 0.5 cc. clear colorless super- 

 natant fluid to 5.0 cc. melted 

 infusion. (See variant (v) me- 

 dium 1661.) 



(p) Harvey used the following medium to 

 cultivate B. influenza etc.: 

 (1) Mix 1.0 cc. of defibrinated rabbit 

 or human blood with 20.0 cc. of 



infusion agar at 70°C. (See 

 variant (v) medium 1661.) 



(2) Raise the temperature of the 

 mixture to boiling point over a 

 free flame. 



(3) Shake to mix. 



(4) Raise to boiling point twice again. 



(5) Allow to deposit. 



(6) Decant the clear supernatant fluid 

 into test tubes while the agar is 

 still melted, or filter thru glass 

 wool. 



(7) Slope. 



(q) Harvey prepared a medium as 

 follows: 



(1) Mix one part defibrinated ox blood 

 at 70°C. with 20 parts of infusion 

 agar at 70°C. (See variant (v) 

 medium 1661.) 



(2) Steam 45 minutes. 



(3) Allow to solidify. 



(4) Steam to melt the medium. 



(5) Strain thru glass wool, filled to a 

 depth of l/8th inch into a Buchner 

 funnel, under slightly reduced 

 pressure. 



(6) Distribute into test tubes. 



(7) Sterilize at 100°C. on each of 3 

 successive days. 



(r) Jones cultivated an organism re- 

 sembling Bacillus actinoides from 

 pneumonic rat lung on a medium 

 prepared as follows: 



(1) Prepare veal infusion agar. 



(2) Slant. 



(3) Add 0.5 cc. of defibrinated horse 

 blood, or calf serum, to the water 

 of condensation. 



(4) Remove a small piece of lung 

 tissue from an infected rat under 

 aseptic conditions and push the 

 tissue down into the tubes. 



(5) Seal the tubes with sealing wax. 

 (s) Pitfield added 1 part sterile defibri- 

 nated blood to 5 parts sterile melted 

 agar. 



(t) Stitt prepared a N. N. N. medium 

 for the cultivation of trypanosomes, 

 leishmania and protozoa in the 

 following manner: 



(1) Cover 125.0 g. chopped beef with 

 1000.0 cc. of water and place over 

 night in the refrigerator. 



