CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(4) Dilute (3) to 1000.0 cc. 



(5) Dissolve 20.0 g. peptone, 5.0 g. 

 NaCl, 0.1 g. CaCls, 10.0 cc. N/1 

 Na2C03 solution, and 20.0 g. agar 

 in (4). 



(6) Distribute in test tubes in 1.0 cc. 

 lots. 



(7) Sterilize in autoclave by heating 

 to 105-108° for 15 minutes. 



(8) Centrifuge defibrinated rabbit 

 blood and draw off serum. 



(9) Dilute serum with 0.5% NaCl 

 solution to original blood volume. 



(10) Shortly before use melt the desired 

 tubes of agar, cool to 60°C. and 

 add two volumes of (9) to each 

 tube. 



(11) Mix well and solidify in slanting 

 positions. 



(c) Ferry and Noble cultivated Diplococ- 

 cus pneumoniae, Streptococcus viri- 

 dans and Streptococcus hemolyticus 

 on a medium prepared as 



(1) Add 500.0 g. of finely chopped lean 

 veal to 1000.0 cc. of water. 



(2) Macerate and allow to stand over 

 night. 



(3) Strain thru cheese cloth and bring 

 to a boil. 



(4) Filter. 



(5) Add 20.0 g. peptone (this may be 

 omitted) 5.0 g. NaCl and 30.0 g. 

 of finely chopped agar. 



(6) Boil and adjust the reaction to the 

 neutral point, using phenolphthal- 

 ein as an indicator. 



(7) Filter. 



(8) Tube in 3.0 cc. quantities and 

 sterilize the fractional method in 

 the steamer. 



(9) When ready for use, melt the agar, 

 cool to 45°C. and add 2.0 cc. of 

 sterile normal horse serum to each 

 tube. 



(d) Ball. 



(1) Add 500.0 g. lean chopped veal to 

 1000.0 cc. water, 20.0 g. peptone, 

 5.0 g. NaCl and 30.0 g. finely 

 chopped agar. 



(2) Adjust reaction. 



(3) Sterilize by fractional method. 



(4) Cool to 45°C. 



(5) Add 2.0 cc. of sterile normal horse 

 serum. 



(e) Harvey. 



(1) Mix two parts melted infusion 

 agar at 45 °C. (see variant (v) 

 medium 1661) with one part sterile 

 human serum at 45 °C. 



(f) Harvey cultivated meningococci on a 

 medium prepared as follows: 



(1) Collect ox or sheep blood at the 

 slaughter house in a sterile blood 

 jar. 



(2) Leave the jar at the slaughter 

 house to avoid the shaking up con- 

 sequent on transportation. 



(3) Transfer the separated serum 

 24 hours later with a sterile pipette 

 to a sterile flask. 



(4) Transport to laboratory. 



(5) Add ether to 0.5%. 



(6) Cork tightly. 



(7) Leave 24 hours. 



(8) Replace the cork with a sterile 

 wool plug. 



(9) Keep the flask and contents in a 

 water bath for 3 hours at 50 °C. to 

 drive off the ether. 



(10) Mix 25 parts (9) at 45°C. with 100 

 parts melted infusion agar (see 

 variant (v) medium 1661). 



(g) Jones cultivated an organism re- 

 sembling Bacillus actinoides from 

 pneumonic rat lungs on a medium 

 prepared as follows : 



(1) Prepare veal infusion agar, 



(2) Slant. 



(3) Add 0.5 cc. of calf serum water 

 (or defibrinated horse blood) to the 

 water of condensation. 



(4) Remove small pieces of lung tissue 

 from an infected rat under aseptic 

 conditions. 



(5) Push the pieces of tissue down in 

 the tube into the liquid. 



(6) Seal the tubes with sealing wax. 

 (h) Fitch cultivated Bact. abortus Bang 



on a medium prepared as follows: 



(1) Prepare beef infusion broth. 



(2) Dissolve 20.0 g. agar in (1). 



(3) Adjust to pH from 6.8 to 7.2. 



(4) Tube. 



(5) Method of sterilization not given. 



(6) At time of use melt agar and add 

 (10.0%) naturally sterile horse 

 serum. 



