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Isolation of virus was attempted by suckling mouse brain passage and cell culture (neuroblastoma cells; Diagnostic Hybrids, Athens, OH, USA); both methods were successful for the cat sample.
The glycoslyation motif in WNV may be lost during extensive mouse brain passage; however, no direct evidence exists to support this hypothesis (21 ).
Peak titers in individual monkeys inoculated with the ninth infant mouse brain passage of the TP21 strain of Langat virus ranged from 10 0.5 10 1.5 and were observed on days 2 or 3 pi.
Most of the experiments were performed with strain P9605 derived from serum of a febrile individual in the endemic area and isolated and maintained by infant mouse brain passage in Swiss albino strain of mice.
This process may occur in ZIKV; the glycoslyation motif in MR 766 may have been present in earlier passages of prototype MR766 and lost during extensive mouse brain passage.
The TP 21 isolate of LGTV derived from a tick pool was used in its 8 th and 9 th infant mouse brain passage (LGTV, M8 or M9), or as the 14 th chick embryo tissue culture passage following nine infant mouse brain passages (LGTV, CE14).
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South African WNV isolates SPU116/89, SA93/01, SA381/00, and H442 were obtained from the Special Pathogens Unit, National Institute for Communicable Diseases, South Africa, as freeze-dried mouse brain passages 2 4.
The Nakayama strain of the JEV was isolated from the cerebral spinal fluid of a patient in 1935 and was maintained by continuous mouse-brain passage.
This virus was initially isolated in mouse brain, further passaged once in mouse brain and three times in BHK before used in our mouse experiments.
Isolates 97-103 (passage 5 in suckling mouse brain [SMB]) and 99-222 (passage 4 in SMB) were freeze-dried in October 2000 (14 ).
Passages 4 12 in infant mouse brain were employed with passage 9 used for a majority of the animals.
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