Abstract
Insect specific viruses (ISVs) of the yellow fever mosquito Aedes aegypti have been demonstrated to modulate transmission of arboviruses such as dengue virus (DENV) and West Nile virus by the mosquito. The diversity and composition of the virome of Ae. aegypti, however, remains poorly understood. In this study, we characterised Aedes anphevirus (AeAV), a negative-sense RNA virus from the order Mononegavirales. AeAV identified from Aedes cell lines were infectious to both Ae. aegypti and Aedes albopictus cells, but not to three mammalian cell lines. To understand the incidence and genetic diversity of AeAV, we assembled 17 coding-complete and two partial genomes of AeAV from available RNA-Seq data. AeAV appears to transmit vertically and be present in laboratory colonies, wild-caught mosquitoes and cell lines worldwide. Phylogenetic analysis of AeAV strains indicates that as the Ae. aegypti mosquito has expanded into the Americas and Asia-Pacific, AeAV has evolved into monophyletic African, American and Asia-Pacific lineages. The endosymbiotic bacterium Wolbachia pipientis restricts positive-sense RNA viruses in Ae. aegypti. Re-analysis of a small RNA library of Ae. aegypti cells co-infected with AeAV and Wolbachia produces an abundant RNAi response consistent with persistent virus replication. We found Wolbachia enhances replication of AeAV when compared to a tetracycline cleared cell line, and AeAV modestly reduces DENV replication in vitro. The results from our study improve understanding of the diversity and evolution of the virome of Ae. aegypti and adds to previous evidence that shows Wolbachia does not restrict a range of negative strand RNA viruses.
Importance The mosquito Aedes aegypti transmits a number of arthropod-borne viruses (arboviruses) such as dengue virus and Zika virus. Mosquitoes also harbour insect-specific viruses that may affect replication of pathogenic arboviruses in their body. Currently, however, there are only a handful of insect-specific viruses described from Ae. aegypti in the literature. Here, we characterise a novel negative strand virus, Aedes anphevirus (AeAV). Meta-analysis of Ae. aegypti samples showed that it is present in Ae. aegypti mosquitoes worldwide and is vertically transmitted. Wolbachia transinfected mosquitoes are currently being used in biocontrol as they effectively block transmission of several positive sense RNA viruses in mosquitoes. Our results demonstrate that Wolbachia enhances the replication of AeAV and modestly reduces dengue virus replication in a cell line model. This study expands our understanding of the virome in Ae. aegypti as well as providing insight into the complexity of the Wolbachia virus restriction phenotype.