This study was performed in order to examine the method of cultivating vaccinia virus, and to obtain the egg-adapted strain of vaccinia virus, and practically to evaluate the possibility of vaccine lymph production using the embryonated hen's egg.
1) When 5, 000 units of streptomycin and 500 units of penicillin G crystal were added into lml of vaccine lymph Tyrode suspension and then the mixture was inoculated into the allantoic fluid of chick embryo. As a result, it was found that the vaccinia virus could be propagated and all inoculated eggs were free from bacilli.
2) The distribution of vaccinia virus at the each portion of the egg was examined. It was the infectious chorioallantoic membrane to possess the highest infective unit. The embryo was the same or a little lower compared with the chorioallantoic membrane. The allantoic fluid possessed the low unit and the yolk lower. This distribution of vccinia virus in the egg was principally the same when the egg addapted strain of vaccinsa virus (20 passages) was inoculated.
3) The testicular strain of vaccinia virus pssessing the high infective unit (10
-7) was inoculated into the allantoic fluid of 12 day's embry and incubated at 37°C for 3-4 days. The infectious chorioallantoic membrane which was obtained consequently, was used as the next inoculum. In the same way, the infectious chorioallantoic membrane of inoculated egg was used as an inoculum of the next generation. In this way, the author has succeeded the successine cultivation of vaccinia virus for 23 generations and obtained the egg adapted strain possessing high infective unit (10
-15).
4) The culture method of influenza virus in the empty egg which was devised by Bernkopf (1950) was used at the cultivation of vaccinia virus. As a result, the favorable multiplication of the virus was found in it.
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