CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



775 



(4) Place in a cool place for 24 hours. 



(5) Transfer the separated serum with a 

 sterile pipette in quantities of 5.0 

 cc. to sterile test tubes. 



(6) Sterilize 30 minutes at 58°C. on 

 each of 8 successive days or 20 

 minutes at 100°C. three days. 



(7) Test sterility by incubation for 48 

 hours. 



(n) Harvey. 



(1) Prepare: chloroform 2; blood serum 

 100. 



(2) Keep in a well stoppered bottle in 

 the dark. 



(3) Distribute in sterile test tubes with 

 sterile precautions. 



(4) Coagulate in a slanting position in 

 an inspissator or over steam at 

 temperatures varying from 65 to 

 90°C. according to the degree of 

 transparency required. 



(o) Harvey. 



(1) Coagulate serum in a slanting 

 position in an inspissator or over 

 steam at temperatures varying 

 from 65 to 90°C. according to the 

 degree of transparency required. 



(2) Add 0.85 sterile salt solution to each 

 test tube, slope to cover the 

 medium. 



(3) Sterilize 60 minutes at 115°C. 



(4) Pour off the salt solution at the time 

 of use. 



Note: The addition of salt solu- 

 tion allows for satisfactory sterili- 

 zation and keeps the medium moist 

 till required, 

 (p) Harvey cultivated spirochaetes on 

 the following medium: 



(1) Fill the serum into tall test tubes. 



(2) Heat in the upright position at 

 65°C. 



(3) Remove as soon as the serum begins 

 to set. 



Note: The heat retained in the 



tube will complete the coagulation. 



A soft, almost transparent coagu- 



lum is formed. 



(q) Harvey prepared a medium as follows 



for the cultivation of spirochaetes: 



(1) Prepare: horse serum 3; distilled 

 water 1. 



(2) Distribute in quantity to nearly 

 fill test tubes. 



(3) Close the test tube with a rubber 

 cork. 



(4) Heat one hour at 60°C. in a water 

 bath. 



(5) Heat, 24 hours later one hour at 

 70°C. in a water bath. 



(6) Heat, 24 hours later at 70°C. until 

 the medium becomes syrupy. 



(7) Keep in the ice chest till required 

 for use. 



(r) Pitfield. 



(1) Obtain dog, sheep or cow blood 

 under aseptic conditions. 



(2) Pipette off the serum with a sterile 

 pipette. 



(3) Distribute into sterile test tubes. 



(4) Coagulate by heat. 



(s) Jones cultivated an organism re- 

 sembling Bacillus actinoides from 

 pneumonic rat lungs on a medium 

 prepared as follows: 



(1) Coagulate horse serum in tubes. 



(2) Remove a small portion of the lung 

 of an infected rat under sterile 

 precautions. 



(3) Add 0.5 cc. of sterile calf serum 

 water to the water of condensation. 



(4) Push the piece of tissue down the 

 tube and into the liquid at the 

 bottom. 



(5) Seal the tubes with sealing wax. 



(t) Vardon prepared a desiccated 

 medium as follows: He reported that 

 the dried serum when dissolved in 

 water and brought to its original 

 volume can be used for all purposes 

 where serum is used in media. 



(1) Measure out a quantity of serum. 



(2) Pour into shallow trays. 



(3) Dry at a temperature not exceeding 

 50°C. 



Note: Coagulation of the serum 

 should not occur in the process 

 of drying. The drying process may 

 be continued overnight by placing 

 the trays in a large incubator. 

 To use powder. 



(4) Dissolve by shaking the requisite 

 amount, calculated from amount 

 of dry material obtained from the 

 liquid, of (3) in cold water. 



(5) Distribute when dissolved in 

 amounts of about 7.0 cc. into test 

 tubes. 



