Genetic Characterization of Goutanap Virus, a Novel Virus Related to Negeviruses, Cileviruses and Higreviruses

Abstract

Pools of mosquitoes collected in Côte d’Ivoire and Mexico were tested for cytopathic effects on the mosquito cell line C6/36. Seven pools induced strong cytopathic effects after one to five days post infection and were further investigated by deep sequencing. The genomes of six virus isolates from Côte d’Ivoire showed pairwise nucleotide identities of ~99% among each other and of 56%–60% to Dezidougou virus and Wallerfield virus, two insect-specific viruses belonging to the proposed new taxon Negevirus. The novel virus was tentatively named Goutanap virus. The isolate derived from the Mexican mosquitoes showed 95% pairwise identity to Piura virus and was suggested to be a strain of Piura virus, named C6.7-MX-2008. Phylogenetic inferences based on a concatenated alignment of the methyltransferase, helicase, and RNA-dependent RNA polymerase domains showed that the new taxon Negevirus formed two monophyletic clades, named Nelorpivirus and Sandewavirus after the viruses grouping in these clades. Branch lengths separating these clades were equivalent to those of the related genera Cilevirus, Higrevirus and Blunervirus, as well as to those within the family Virgaviridae. Genetic distances and phylogenetic analyses suggest that Nelorpivirus and Sandewavirus might form taxonomic groups on genus level that may define alone or together with Cilevirus, Higrevirus and Blunervirus a viral family.

Negeviruses found in multiple species of mosquitoes from southern Portugal: Isolation, genetic diversity, and replication in insect cell culture

“…Ever since they were first described in 2013 (Vasilakis et al, 2013), several studies have systematically highlighted not only the genetic diversity, but also the wide geographic dispersal, and unusual wide host range of negeviruses (Ferreira et al, 2013;Vasilakis et al, 2013;Auguste et al, 2014;Kallies et al, 2014;Nabeshima et al, 2014). The experimental approach used in this study allowed the confirmation of these observations, by supporting the detection of Negev-like viruses in four different species of mosquitoes (Oc.…”

“…In both phylogenetic trees, the common ancestry shared by Piura, Ngewotan, Negev, and Loreto viruses, including our OCNV and CUNV, was evident, backing up the tentative designation of this clade as Nelorpivirus (Kallies et al, 2014). Despite the genetic distances involved, this clade was found to be more closely related to plant cileviruses ( Fig.…”

“…This is certainly due to the costs associated with the production of a sufficient number of reads required for de novo assembly of unknown viral sequences in complex mixtures. Although the potential of NGS has been clearly demonstrated for the rapid characterization of new mosquito viruses once isolated in cell culture (Nasar et al, 2012;Vasilakis et al, 2013;Kallies et al, 2014), virus detection based on conventional RT-PCR is still used on a regular basis in most laboratory settings, and such an approach has allowed us to recently identify ISF from different species of mosquitoes collected in southern Portugal (Parreira et al, 2012;Ferreira et al, 2013). Unexpectedly, the exuberant CPE observed when C6/36 cells were exposed to a macerate of Oc.…”

“…6A and B) and higreviruses (Fig. 6B) than to the Santana/Wallerfield/Dezidougou group, the latter forming a second cluster of Negev-like viruses, tentatively designated Sandewavirus (Kallies et al, 2014). When the sequences of genetically distant RdRp domains of plant viruses (including tobamoviruses, such as the cactus mild mottle and the rattail cactus necrosis-associated viruses), and endogenous viral elements (EVE) found in the genomes of Aedes aegypti, Drosophila rhopaloa, and Bombus terrestris, were included in the analysis (Fig.…”

“…Although they do not share overall sequence homology with other known viruses, at least two of the conserved domains in ORF1 (helicase and RdRp) allow for stable phylogenies to be reconstructed (Vasilakis et al, 2013;Kallies et al, 2014), and indicate a shared evolutionary history with plant cile-, higre-, and blunerviruses. Given the high nt.…”