Fig. 3. In vitro and in vivo infection of TMUV SD in C. pipiens mosquitoes. (a) Replication kinetics: growth curves of cluster 3 TMUV (SD strain) in C6/36 mosquito cells infected at an MOI of 0.0001. Supernatant samples were collected at various time points and titrated on BHK cells. (b) Viral load in mosquitoes: Chickens were intramuscularly inoculated with 103.5TCID50 of the SD strain. The chickens were bitten by 4-day-old female C. pipiens mosquitoes at 3 dpi. Blood-engorged mosquitoes were selected and reared as the infected group. The viral E gene copy number in five mosquitoes was quantified by qPCR at 0 and 10 dpi. (c) Dissemination of TMUV SD in mosquito tissues: at 0 and 10 dpi, viral E gene copies were quantified in the head, thorax and abdomen of five infected mosquitoes using qPCR.
