Table 1.
ZIKV isolates used in this study.
Fig 1.
ZIKV isolates grown on mammalian and mosquito cells show phenotypic differences.
A) ZIKV-PRV, ZIKV-PAN, ZIKV-FLR, and ZIKV-MR-766 stocks grown on either Vero mammalian or C6/36 mosquito cells were plaqued on Vero cells and visualized with neutral red staining 4–5 days after inoculation. B) The area of 30–50 individual plaques was measured for each isolate of ZIKV using ImageJ.
Table 2.
ZIKV titers by assay tested in this study.
Fig 2.
Replication kinetics of single-round infection on Vero or C6/36 cells is similar between ZIKVmam and ZIKVmos.
Vero or C6/36 cells were inoculated at an MOI 1–3 with ZIKV-PRV, ZIKV-PAN, ZIKV-FLR, and ZIKV-MR-766 grown on either Vero mammalian cells (ZIKVmam) or C6/36 mosquito cells (ZIKVmos). ZIKV-MR-766mos was omitted due to insufficient titer. Samples were collected at the indicated times and titered by plaque assay on Vero cells. Symbols represent the average, and error bars represent the standard deviation. Some error bars are smaller than the symbol and are not visible.
Fig 3.
ZIKVmam spread is reduced in C6/36 mosquito cells.
Vero or C6/36 cells were inoculated at an MOI 0.1 with Vero mammalian cell-derived (ZIKVmam) or C6/36 mosquito cell-derived (ZIKVmos) ZIKV-PRV, ZIKV-PAN, ZIKV-FLR, and ZIKV-MR-766. Samples were collected at the indicated times and titered by plaque assay on Vero cells. Symbols represent the average, and error bars represent the standard deviation. Some error bars are smaller than the symbol and are not visible.
Fig 4.
Plaque size is maintained in C6/36 cells, but not in Vero cells.
ZIKV-FLRmos was plaqued on Vero cells and representative tiny (L1), small (L2), or medium (L3) plaques were plaque purified over three rounds of passage through C6/36 cells. Similarly, ZIKV-FLRmam was plaqued on Vero cells and representative tiny (L1), small (L2), medium (L3), large normal (L4) or large with a fried-egg phenotype (L5) plaques were plaque purified over three rounds of passage through Vero cells. Each passage of virus was titered on Vero cells. A representative of the final passage of each lineage is shown (A). The area of 30–50 plaques from each passage of each lineage derived from ZIKV-FLRmos (B) and ZIKV-FLRmam (C) was measured using ImageJ.
Table 3.
Consensus mutations identified in plaque-purified ZIKV clones.