Fig 1.
Growth of Shuni virus (SHUV) in mammalian (Vero E6), mosquito (C6/36), and Culicoides biting midge (KC) cells.
Three different cell lines (Vero E6, C6/36, and KC cells) were inoculated with SHUV at an MOI of 0.01, and kept at 28°C (C6/36 and KC) or 37°C (Vero E6). Virus titers were determined at time points 0, 24, 48, and 72 h post infection. Mean virus titers ± SEM for three replicates are shown.
Fig 2.
Infection rates and virus titers of biting midges and of mosquitoes exposed to Shuni virus (SHUV) via an infectious blood meal.
(A-B) Mean infection rates of Culicoides nubeculosus (N = 243, 25°C) and C. sonorensis (N = 48, 28°C; panel A), and Culex pipiens pipiens (N = 211, 25°C) and Aedes aegypti (N = 149, 28°C; panel B) orally exposed to SHUV after 10 days of incubation at the respective temperatures. Infection rates are presented as the percentage of virus-positive females out of the total number of blood-fed females that remained alive at the end of the incubation period. Error bars indicate the SEM. (C-D) SHUV titers of virus-positive bodies of C. nubeculosus (N = 34, 25°C) and C. sonorensis (N = 29, 28°C; panel C), and Ae. aegypti (N = 3, 28°C; panel D) after 10 days incubation at the respective temperatures. Each dot represents one individual female, and the black bar indicates the mean. The detection limit of the endpoint dilution assay is indicated with the dashed line.
Fig 3.
Infection rates, transmission rates, and virus titers of mosquitoes intrathoracically injected with Shuni virus (SHUV).
Mean infection rates (A) and transmission rates (B) of Culex pipiens pipiens (N = 50) and Aedes aegypti (N = 50) intrathoracically injected with SHUV after 10 days incubation at 25°C and 28°C, respectively. Infection rates are presented as the percentage of virus-positive body or saliva samples, respectively, out of the total number of injected mosquitoes that remained alive at the end of the incubation period. Error bars indicate the SEM. SHUV titers of virus-positive body samples (C) and saliva samples (D) of Cx. p. pipiens (body samples: N = 26, saliva samples: N = 16) and Ae. aegypti (body samples: N = 14, saliva samples: N = 4) intrathoracically injected with SHUV after 10 days incubation at 25°C and 28°C, respectively. Each dot represents an individual sample, and the black bar indicates the mean. The detection limit of the endpoint dilution assay is indicated with the dashed line.