Mosquito Infection and Fitness Evaluation

Adult mosquitoes were kept in cages with free access to sugar solution (10%) until they were 5–6 days old. One day before virus experimental infection, female mosquitoes were deprived from sugar solution and transferred to infection cages (8 cm height, 6 cm diameter). The blood meal (1 mL of erythrocytes, 1 mL of virus suspended in L15 medium for infected groups; 1 mL of erythrocytes, 1 mL of L15 medium for uninfected groups) was offered using an artificial feeder (Hemotek, Great Hardwood, UK) at 37 °C for approximately 30 min. Only mosquitoes which were visually blood-engorged proceeded to further investigations. Aedes aegypti used in fitness experiments (longevity, fecundity and fertility evaluation) were individualized in plastic vials (6.5 cm height, 2.5 cm diameter) [33 (link),36 (link),37 (link)]. Females used for the egg quiescence assessment were transferred to cylindrical cages (25 cm height, 20 cm diameter) with oviposition cups. In both experiments, mosquitoes were kept inside an incubator (26 ± 1 °C, 75% ± 5 humidity) with free access to sugar solution (10%).