Variable Inhibition of Zika Virus Replication by Different Wolbachia Strains in Mosquito Cell Cultures

Mosquito-borne arboviruses are a major source of human disease. One strategy to reduce arbovirus disease is to reduce the mosquito's ability to transmit virus. Mosquito infection with the bacterial endosymbiont Mel is a novel strategy to reduce mosquito competency for flavivirus infection. However, experiments investigating cyclic environmental temperatures have shown a reduction in maternal transmission of Mel, potentially weakening the integration of this strain into a mosquito population relative to that of other strains. Consequently, it is important to investigate additional strains. All Zika virus (ZIKV) suppression studies are limited to the Mel strain. Here we show ZIKV inhibition by two different strains: AlbB (isolated from mosquitoes) and Stri (isolated from the planthopper ) in mosquito cells. strain Stri inhibited ZIKV most effectively. Single-cycle infection experiments showed that ZIKV RNA replication and nonstructural protein 5 translation were reduced below the limits of detection in Stri-containing cells, demonstrating early inhibition of virus replication. ZIKV replication was rescued when was inhibited with a bacteriostatic antibiotic. We observed a partial rescue of ZIKV growth when -infected cells were supplemented with cholesterol-lipid concentrate, suggesting competition for nutrients as one of the possible mechanisms of inhibition of ZIKV. Our data show that AlbB and Stri infection causes inhibition of ZIKV, making them attractive candidates for further mechanistic and studies and future vector-centered approaches to limit ZIKV infection and spread. Zika virus (ZIKV) has swiftly spread throughout most of the Western Hemisphere. This is due in large part to its replication in and spread by a mosquito vector host. There is an urgent need for approaches that limit ZIKV replication in mosquitoes. One exciting approach for this is to use a bacterial endosymbiont called that can populate mosquito cells and inhibit ZIKV replication. Here we show that two different strains of , AlbB and Stri, are effective at repressing ZIKV in mosquito cell lines. Repression of virus growth is through the inhibition of an early stage of infection and requires actively replicating Our findings further the understanding of viral inhibition and provide novel tools that can be used in an effort to limit ZIKV replication in the mosquito vector, thereby interrupting the transmission and spread of the virus.