﻿ix.B, 1 Boynton: Cultivation of Virus of Rinderpest 41 



and Wijnikewitch(2) and with the medium used by Bass and 

 Johns. (4) This report is based upon work with these modified 

 media. 



TECHNIQUE 



The first medium tested was a salt-peptone preparation advo- 

 cated by Nencki, Sieber, and Wijnikewitch. It is composed of 

 900 cubic centimeters of water to which are added 100 grams of 

 peptone Witte and 20 grams of sodium chloride. The mixture 

 is filtered, placed in test tubes, and sterilized. I have been 

 unable to obtain successful results in the cultivation of rinderpest 

 virus in the medium described. This medium was modified by 

 adding 0.1 cubic centimeter of a 33i per cent solution of glucose 

 to each 10 cubic centimeters of the salt-peptone mixture de- 

 scribed by Nencki, Sieber, and Wijnikewitch. Test tubes 1.5 

 centimeters in diameter and 15 centimeters long were employed. 

 The glucose solution to the amount of 0.1 centimeter was added 

 to each tube, and to this were added 10 cubic centimeters of 

 the salt-peptone solution. It was sterilized for one hour in an 

 autoclave at 135° C. on the day previous and one-half hour just 

 prior to inoculation. Normal blood was drawn under aseptic 

 conditions from a nonimmune animal and defibrinated by shaking 

 with glass beads. One cubic centimeter of this blood was added 

 to each tube immediately before it was inoculated. Blood from 

 an animal suffering with rinderpest was then drawn under 

 aseptic conditions and defibrinated. Each tube was inoculated 

 with either 0.5 or 1 cubic centimeter of the infective blood. 

 Anaerobic conditions were produced by covering the surface of 

 the inoculated media with from 1.5 to 2 cubic centimeters of 

 sterile paraffin oil. The tubes were then placed in the incubator 

 at 40° C. 



In making transfers, a sterile pipette of 1 cubic centimeter 

 capacity was inserted into the culture tube from which the 

 transfer was to be made and the medium was thoroughly agitated 

 by filling the pipette with the medium and emptying. This was 

 repeated four or five times. Then either 0.3 or 0.5 cubic centi- 

 meter of the culture was transferred to the prepared culture 

 medium, after which the tubes were sealed with paraffin oil 

 and placed in the incubator. It was found best to make these 

 transfers every three or four days. 



The medium which has given the best results is a modifica- 

 tion of the one used by Bass and Johns for cultivating the 

 Plasmodium of malaria. To each of a series of test tubes was 



