Novel Viruses Isolated from Mosquitoes in Pantanal, Brazil

Genomic sequences are described from five novel viruses and divergent strains of Brejeira and Guaico Culex viruses from mosquitoes collected in Pantanal, Brazil, in 2010.

Four BRJV isolates from Culex (Culex) spp. yielded 99% identity among themselves, but 88% to 90% identity with a BRJV isolated in northern Brazil, indicating the detection of a divergent strain of this recently described Negevirus-like virus (4).
We report here the first Brazilian detections of GCXV, a recently described multicomponent Jingmenvirus (5). The five genomic segments from Culex (Culex) spp. pool number 407 aligned with coverages from 78% to 87% and identities from 97% to 100% with published sequences. A second isolate from pool number 313, also Culex (Culex) spp., was the same strain, based on partial sequences (data not shown). In Brazil, the first detection of a Jingmenvirus was from tick collections in southeast Brazil, and called Mogiana tick virus (6).
Five novel viruses were detected and named to honor indigenous tribes of Mato Grosso do Sul state (Terena, Ofaié, Kadiwéu, Kaiowá, and Guató viruses). Translated nucleotide identities ranged between 32% and 73% to known viruses. The following abbreviations are suggested for the novel viruses: TERV, OFAV, KADV, KAIV, and GUTV.
TERV isolated from a Mansonia spp. pool aligns with viruses of a newly described genus within Bunyaviridae called Phasmavirus (7). It contains the standard three-segmented genome with the small (S), medium (M), and large (L) segments of other bunyaviruses.
OFAV derived from the same mosquito pool as TERV. OFAV has a large genome of 19.8 kb and most closely aligns (Ͻ66% predicted amino acid sequence identity) with Méno virus, recently described within the family Mesoniviridae (8). KADV is also related to Méno virus (Ͻ71% identical) and is similarly large (20.4 kb). OFAV and KADV align with each other with 70% nucleotide identity over a query coverage of 68%. GUTV and KAIV isolated from Culex (Culex) spp. most closely align with a recently described virus from China called Wuchang Cockroach virus 3. The three isolates of GUTV are 99% identical to each other, whereas KAIV and GUTV are 30% divergent at the nucleotide level. These viruses consistently accompanied isolates of BRJV. Whether the accompaniment of these viruses is coincidence, or they derived from coinfected mosquitoes within each pool, is unknown. However, KAIV and GUTV genomes could in fact be independent genomic segments within a multisegmented genome that includes BRJV.
To evaluate the host range of these virus isolates, attempts to generate cytopathic effect (CPE) were made with human, monkey, frog, bat, mosquito, and tick cell cultures. A clear CPE was observed only in mosquito cells. However, the capacity of these novel viruses to cause CPE in vertebrate and invertebrate cells was not fully evaluated.
Accession number(s). The sequences have been deposited in GenBank under the accession numbers listed in Table 1.

ACKNOWLEDGMENTS
Special thanks to Zilca Campos from EMBRAPA Pantanal for facilitation of the field collection effort and also Jason Velez from CDC for cell culture support.

FUNDING INFORMATION
This work was funded through institutional support of the authors, Centers for Disease Control and Prevention and Sandia National Laboratories' Laboratory-Directed Research and Development program. Sandia is a multiprogram laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000. The efforts of Alex Pauvolid-Corrêa were funded by Science without Borders Program, Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Government of Brazil.