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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(5) Solidify in a Koch inspissator at 68 

 to 70°C. 

 (g) Heinemann. 

 (1) Coagulate dog's blood serum in 

 slanted tubes in the Koch inspis- 

 sator for three hours at Ib^C. 

 (h) Schereschewsky cultivated Syphilis 

 spirochaete on the following medium : 



(1) Draw off normal horse serum, by 

 means of a syphon under aseptic 

 conditions into thin walled sterile 

 tubes about 10 to 15 cm. long. 

 Allow the serum to run down the 

 wall of the tube so as not to mix the 

 serum with air. 



(2) Seal the tubes with cotton or cork 

 and gum. 



(3) Heat in a water bath at 57-58 °C. for 

 one hour. 



(4) Raise the temperature slowly to 

 70 °C. 



(5) When the serum has solidified, 

 incubate at 37°C. over night to 

 test sterility. 



(i) Sowade used the following medium to 

 cultivate syphilis spirochaete: 



(1) Collect horse blood in long measur- 

 ing cylinders. 



(2) Place in the ice box until the serum 

 separates. 



(3) Distribute into test tubes to a 

 depth of 12.0 cm. 



(4) Heat in a water bath on 3 successive 

 days for two hours at 58°C. 



(5) On the third day slowly raise the 

 temperature until the serum 

 reaches a gelatinous consistency. 

 Raise the temperature slowly or an 

 intensive turbidity will take place. 



(6) Medium is honey yellow, trans- 

 parent and reacts slightly alkaline. 



(j) Abbott. 



(1) Obtain blood serum from the 

 slaughter house or anti-toxin 

 manufacturers. 



(2) Tube in sterile tubes and plug. 



(3) Slant. 



(4) Place in a dry air sterilizer and 

 slowly raise the heat to 80-90 °C. 

 Keep at this temperature until the 

 serum has solidified. 



(5) Steam on three successive days for 

 20 minutes at 100 °C. 



(k) Ball. 



(1) Collect blood under aseptic con- 

 ditions at the slaughter house, if 

 possible, in large tall sterile flasks. 



(2) Place on ice for 48 hours. 



(3) Draw out the serum by means of 

 sterile pipettes into test tubes. 

 Do not shake the jar. 



(4) Place the tubes in an inspissator 

 in a slanted position. 



(5) Heat to 65° to 68° until coagulation 

 occurs. 



(6) Remove the tubes and sterilize by 

 the fractional method. 



(7) Keep for 3 or 4 days in the incubator 

 at 58°C. and discard those tubes 

 showing growth. These tubes are 

 transparent and straw colored. 



(8) The serum may be prepared by 

 coagulating at a temperature short 

 of the boiling point— temperature 

 not specified. Sterilize by exposing 

 the tubes to a temperature of about 

 90°C. on each of three successive 

 days for 5 minutes. These tubes 

 are opaque and white. 



(1) Harvey. 



(1) Collect ox or sheep blood at the 

 slaughter house in a sterile blood 

 jar. 



(2) Allow the blood to coagulate. 



(3) Detach the clot. 



(4) Place in the ice chest. 



(5) Pipette off the serum with a sterile 

 pipette. 



(6) Keep the serum in the ice chest till 

 required. 



(7) Distribute a portion of the serum 

 in quantities of 5.0 cc. in test tubes. 



(8) Sterilize 2 days 30 minutes at 60 °C. 



(9) 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. 



(m) Harvey. 



(1) Collect ox or sheep blood at the 

 slaughter house in a sterile blood 

 jar or in a bucket. 



(2) Allow the blood to coagulate before 

 removing it. 



(3) Separate the clot from the sides of 

 the containing vessel with a sterile 

 glass rod. 



