Microbial exposure across life reduces susceptibility ofAedes aegyptito Zika virus by enhancing blood digestion and limiting midgut cell infection

Abstract

The worldwide expansion of mosquito-borne pathogens necessitates improved control measures, including approaches to restrict infection and transmission by mosquito vectors. Reducing transmission is challenging because determinants of vector competence for viruses like Zika (ZIKV) are poorly understood. Our previous work established thatAedes (Ae.) aegyptilarvae reared in microbe-rich environmental water are less susceptible to ZIKV as adults compared to cohorts reared in microbe-deficient laboratory tap water. Here, we explain the association by identifying a mechanism by which environment-derived microbes reduce susceptibility ofAe. aegyptifor ZIKV. Provided that the midgut represents the first barrier to mosquito infection, we hypothesized that microbial exposure modulates midgut infection by ZIKV. Since mosquitoes live in water as larvae and pupae and then transition to air as adults, we also define the stage in the life of a mosquito when microbial exposure reduces ZIKV susceptibility.Ae. aegyptilarvae were reared in microbe-rich water and then treated with antibiotics during the pupal and adult stages, adult stage only, or provided no antibiotics at any stage. Vector competence was next evaluated in mosquitoes that ingested ZIKV-spiked bloodmeals. Antibiotic treatment enhanced ZIKV infection and dissemination rates, especially inAe. aegyptitreated as both pupae and adults. Antibiotic treated adult mosquitoes also had increased midgut epithelium permeability, higher numbers of ZIKV-infected midgut cells, and impaired bloodmeal digestion. Consistent with these changes,Ae. aegyptitreated with antibiotics as pupae and adults that ingested ZIKV in bloodmeals showed reduced expression of genes associated with bloodmeal digestion and metabolism relative to mosquitoes that were not antibiotic treated. Together, these data show that exposure to microbes throughout the life ofAe. aegyptirestricts ZIKV dissemination by facilitating blood digestion and limiting midgut cell infection. Understanding the roles mosquito microbiota play in determining midgut physiology and arbovirus susceptibility can lead to novel approaches to decrease mosquito transmission and will improve understanding of vector competence in microbe-rich environmental habitats.Author SummaryMosquito-transmitted viruses like Zika continue to threaten human health. Absent vaccines or treatments, controlling mosquitoes or limiting their ability to transmit viruses represents a primary way to prevent mosquito-borne viral diseases. The role mosquito microbiota play in shaping transmission of Zika virus has been limited to association-based studies. Our prior work showed thatAedes aegyptimosquito larvae that develop in bacteria-rich water are less susceptible to Zika virus compared to larvae reared in microbe-poor laboratory tap water. Here we identify a mechanism that explains this association. Since mosquitoes are aquatic as larvae and pupae and terrestrial as adults, we also define the life stage when microbes need be present to reduce Zika virus susceptibility. We used antibiotics to reduce environmental water-derived microbes at pupal and adult or only adult stages and observed that, compared to mosquitoes with microbes, antibiotic treatment increases Zika virus dissemination, increases permeability and infection of the midgut, and impairs bloodmeal digestion. These data show that microbial exposure throughout the life of a mosquito restricts Zika virus dissemination by facilitating blood digestion and limiting midgut cell infection. These findings advance understanding of microbiota-mosquito-virus interactions by defining how microbes reduce susceptibility ofAedes aegyptito Zika virus.