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. 2014 Feb;95(Pt 2):481-485.
doi: 10.1099/vir.0.058412-0. Epub 2013 Nov 21.

Characterization of a novel Negevirus and a novel Bunyavirus isolated from Culex (Culex) declarator mosquitoes in Trinidad

Albert J Auguste  1 , Christine V F Carrington  2 , Naomi L Forrester  1 , Vsevolod L Popov  1 , Hilda Guzman  1 , Steven G Widen  3 , Thomas G Wood  3 , Scott C Weaver  1 , Robert B Tesh  1
Affiliations

Characterization of a novel Negevirus and a novel Bunyavirus isolated from Culex (Culex) declarator mosquitoes in Trinidad

Albert J Auguste et al. J Gen Virol. 2014 Feb.

Abstract

Pools of mosquitoes were tested for insect-specific viruses using cytopathic effect (CPE) assays on Aedes albopictus (C6/36) cells. Illumina sequencing of RNA from pool TR7094, which produced extensive CPE 2 days post-infection, yielded the complete genome sequences of a previously unknown Bunyavirus, designated Cumuto virus (CUMV), and a second virus designated Wallerfield virus (WALV). WALV shared highest amino acid identity (60.1 %) with Dezidougou virus from Côte d'Ivoire, a positive-sense, single-strand RNA, insect-specific virus belonging to the newly proposed genus Negevirus associated with mosquitoes and phlebotomine sandflies. The S, M and L segments of CUMV were most closely related to those of Gouleako virus, also from Côte d'Ivoire (amino acid identities of 36 %, 38% and 54 % respectively). Neither virus produced CPE on vertebrate cells, or illness in newborn mice. Isolation and characterization of these viruses increase our knowledge of the geographical distribution, diversity and host range of mosquito-specific bunyaviruses and negeviruses.

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Figures

Fig. 1.
Fig. 1.
Ultrastructure of C6/36 cells infected with Wallerfield and Cumuto viruses. (a) Expansions of perinuclear space and granular endoplasmic reticulum filled with tubular and vesicular structures. Arrows show points of perinuclear membrane expansion. Bar, 0.5 µm. (b) Large vacuoles with spherules of Wallerfield virus at their periphery and expansions of granular endoplasmic reticulum filled with vesicles 20–40 nm in diameter. Bar, 100 nm. (c) Two vacuoles with spherical enveloped particles ~75 nm in diameter possibly representing Cumuto virus. Bar, 100 nm.
Fig. 2.
Fig. 2.
(a) Midpoint rooted ML phylogeny estimated from amino acid sequences for phlebovirus L segment; (b) midpoint rooted ML phylogeny estimated from nucleotide sequences of negevirus RdRp genes. Bootstrap support values >50 % indicated at relevant nodes. Taxon names include virus name, strain and corresponding accession numbers used for analysis. Scale bars represent percentage divergence.
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