Despite the fact learn more that the authors used another mosquito strain in their studies, they also used a non-EGFP expressing virus and higher virus concentrations in their bloodmeals, ranging from 108-109 pfu/ml. In our study the virus concentrations in bloodmeals ranged from 1.7-2.7 × 107 pfu/ml. In the presence

of a functional RNAi mechanism as in HWE mosquitoes, the lower virus concentration in the bloodmeal was probably approaching the threshold for midgut infection. In the RNAi pathway impaired Carb/dcr16 mosquitoes however, this virus concentration was sufficient to cause productive midgut infections. Between 7 and 14 days pbm a strong reduction of virus infection intensity was observed in midguts of Carb/dcr16 mosquitoes, causing

a decrease in average SINV titers from 14,000 to 2400 pfu/ml. Such strong reduction of virus infection intensity was not observed in the RNAi pathway competent HWE control. After 7 days pbm Autophagy signaling pathway inhibitors the RNAi pathway in Carb/dcr16 mosquitoes was no longer compromised as it was during virus acquisition. It appears that the RNAi mechanism, when functional, down-regulated the unusually high SINV concentration in midguts of the transgenic mosquitoes to levels similar to those of the HWE control. This strongly suggests that the task of the RNAi pathway in the mosquito midgut is to keep arbovirus replication at a level that can be tolerated by the mosquito. Modulation of arbovirus infections in mosquitoes has been reported for several virus-vector combinations and research of the last

few years eventually confirmed that the RNAi pathway of the mosquito is a major driving force behind this modulation [2, 3, 6, 14, 16, 32]. Nevertheless, recent studies indicate that other innate immune pathways, such as JAK-STAT and/or Toll also contribute to the modulation of arbovirus infections in insects [33–37]. Since a proposed role for the RNAi pathway in mosquitoes is to protect the insect selleckchem from pathogenic effects of replicating arboviruses [4–6], we investigated whether SINV-TR339EGFP causes such effects in HWE or Carb/dcr16 mosquitoes. Our survival curve data indicate that the initial increase in virus titer in Carb/dcr16 females did not cause obvious pathogenic effects. It needs to be pointed out that after 7 days pbm the RNAi pathway was no longer impaired in midguts of Carb/dcr16 mosquitoes and the intensity of infection was strongly modulated. Thus, the RNAi pathway activation in the transgenic mosquito line could have been similar to that in the control for the latter 21 days of the survival study. Our observations confirm those by Campbell and co-workers [3] that transient silencing of the RNAi pathway in Ae. aegypti did not affect longevity of the mosquitoes for seven days after infection with SINV. However, several authors have described pathological effects caused by alphaviruses in mosquito midguts and salivary glands, claiming that these effects could be virus dose-dependent [38–41].