Barcoded WNV efficiently infects, and replicates in cells and mosquitoes
(A) bcWNV (red) replication was evaluated relative to rWT-WNV (black) at MOIs of 0.1 (dashed line) and 1.0 (solid line) in mammalian (Vero) and insect cells (CT, Culex tarsalis-derived embryonic cell line) (n = 3, error bars denote mean ± SD). vRNA copies = WNV envelope copies per reaction. Cx. tarsalis (orange) and quinquefasciatus (blue), and Ae. aegypti (green) mosquitoes were infected with bcWNV. At the indicated days, saliva, salivary glands and midguts were collected from each species.
(B) Infection rates of midguts, salivary glands, and saliva were determined for all species at all timepoints.
(C) Levels of viral RNA were measured via qRT-PCR (n = 35–40), solid lines denote mean. vRNA copies = WNV envelope copies per sample. Within each time point, statistical significance between mosquito species was determined by two-way ANOVA with Tukey’s multiple comparisons test, ∗ = p < 0.05, ∗∗ = p < 0.005, ∗∗∗ = p < 0.0005, ∗∗∗∗ = p < 0.0001. Only significant relationships are shown.