The genus Alphavirus harbors arboviruses of great concern, such as the Chikungunya virus and the equine encephalitis viruses. Transmission of pathogenic alphaviruses by mosquitoes could be influenced by insect-specific alphaviruses such as Eilat virus (EILV). However, insect-specific alphaviruses are rarely found in wild mosquitoes and only a few have been described in the literature. Here, we report the detection of EILV in a Culex univittatus mosquito from the north-eastern Namibian Zambezi region. Full genome analysis of MP458-NA-2018 showed 94.5% nucleotide identity to an EILV isolate from Israel. MP458-NA-2018 grouped with EILV in phylogenetic analysis and was placed within the clade of insect-specific alphaviruses. The virus was isolated in mosquito cells and shown to be restricted to insects as hosts by the inability to infect different vertebrate cell lines and a complete block of virus replication at 34°C. We further showed that infection of cells with EILV MP458-NA-2018 reduced production of infectious particles of Sindbis virus by 2000-fold over the entire course of infection, whereas reduction rates of Bagaza and Middleburg virus were approximately 3-10-fold and dependent on time after infection. While production of infectious particles of cells superinfected with the Chikungunya virus were approximately 30-fold reduced and more pronounced at a lower multiplicity of infection of 0.01, EILV seemed to enhance production of West Nile virus infectious particles by >5-fold when superinfected at a multiplicitiy of infection of 0.1. In summary, EILV from the Namibian Zambezi region influences in vitro replication of endemic flavi- and alphaviruses.
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