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Background   Wolbachia ’s ability to restrict arbovirus transmission makes it a promising tool to combat mosquito-transmitted diseases.  Wolbachia -infected  Aedes aegypti  are currently being released in locations such as Brazil, which regularly experience concurrent outbreaks of different arboviruses.  A .  aegypti  can become co-infected with, and transmit multiple arboviruses with one bite, which can complicate patient diagnosis and treatment.    Methodology/principle findings  Using experimental oral infection of  A .  aegypti  and then RT-qPCR, we examined ZIKV/DENV-1 and ZIKV/DENV-3 co-infection in  Wolbachia -infected  A .  aegypti  and observed that  Wolbachia -infected mosquitoes experienced lower prevalence of infection and viral load than wildtype mosquitoes, even with an extra infecting virus. Critically, ZIKV/DENV co-infection had no significant impact on  Wolbachia ’s ability to reduce viral transmission.  Wolbachia  infection also strongly altered expression levels of key immune genes  Defensin C  and  Transferrin 1 , in a virus-dependent manner.    Conclusions/significance  Our results suggest that pathogen interference in  Wolbachia -infected  A .  aegypti  is not adversely affected by ZIKV/DENV co-infection, which suggests that  Wolbachia -infected  A .  aegypti  will likely prove suitable for controlling mosquito-borne diseases in environments with complex patterns of arbovirus transmission.

Pathogen blocking in Wolbachia-infected Aedes aegypti is not affected by Zika and dengue virus co-infection