{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:36:32Z","timestamp":1785411392285,"version":"3.56.0"},"reference-count":65,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,4,25]],"date-time":"2026-04-25T00:00:00Z","timestamp":1777075200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003593","name":"National Council for Scientific and Technological Development","doi-asserted-by":"publisher","award":["141723\/2018-1"],"award-info":[{"award-number":["141723\/2018-1"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"award":["141723\/2018-1"],"award-info":[{"award-number":["141723\/2018-1"]}],"id":[{"id":"https:\/\/ror.org\/03swz6y49","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Evandro Chagas Institute\/SVSA\/MS"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Mosquitoes are recognized as the arthropod group with the greatest vectorial capacity, and the viruses they transmit constitute a significant concern in the context of global One Health. In addition, these insects act as hosts for a wide diversity of insect-specific viruses (ISVs), which exclusively infect arthropods. Expanding knowledge of ISVs is particularly relevant, given their potential influence on arbovirus replication and their role in elucidating the evolutionary processes that shape virus\u2013vector interactions. In this study, we report the isolation and molecular analysis of three negeviruses associated with different mosquito species of the genera Culex, Coquillettidia, Mansonia, and Ochlerotatus, collected in Bel\u00e9m, Par\u00e1 State, in the Brazilian Amazon: Loreto virus, Wallerfield virus, and a putative new species, designated Terra firme virus. Eleven pools exhibited cellular alterations consistent with cytopathic effects in invertebrate C6\/36 cells but showed no evidence of replication in vertebrate Vero cells. Notably, simultaneous infections by two or three negeviruses were detected in some mosquito pools, indicating the occurrence of multiple viral infections within individual samples. Genomic analyses revealed that the isolated strains share conserved domains with previously described isolates from other countries. Phylogenetic inferences demonstrated that the investigated strains are classified within the clades Nelorpivirus and Sandewavirus. Taken together, these findings expand the currently known diversity of the negevirus group and contribute to a more comprehensive understanding of its host range and geographic distribution.<\/jats:p>","DOI":"10.3390\/v18050501","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T07:43:51Z","timestamp":1777448631000},"page":"501","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Isolation and Molecular Analysis of Negeviruses in Mosquitoes (Diptera: Culicidae) from an Environmental Protection Area in the Brazilian Amazon"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4803-7825","authenticated-orcid":false,"given":"Bruna Alves","family":"Ramos","sequence":"first","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Virology, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel Damous","family":"Dias","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Parasitic Biology in the Amazon, State University of Par\u00e1, Perebebu\u00ed Bystreet, C Block, 2nd Floor, 2623, Bel\u00e9m 66095-662, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5981-0065","authenticated-orcid":false,"given":"Joaquim Pinto","family":"Nunes-Neto","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Parasitic Biology in the Amazon, State University of Par\u00e1, Perebebu\u00ed Bystreet, C Block, 2nd Floor, 2623, Bel\u00e9m 66095-662, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Wilson Rosa","family":"Junior","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Durval Bertram Rodrigues","family":"Vieira","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8385-8253","authenticated-orcid":false,"given":"Val\u00e9ria Lima","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Virology, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3473-1296","authenticated-orcid":false,"given":"Ana L\u00facia Monteiro","family":"Wanzeller","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Virology, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3319-1854","authenticated-orcid":false,"given":"Eliana Vieira Pinto da","family":"Silva","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Virology, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5834-9649","authenticated-orcid":false,"given":"Maria Nazar\u00e9 Oliveira","family":"Freitas","sequence":"additional","affiliation":[{"name":"Graduate Program in Virology, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Landeson Junior Leopoldino","family":"Barros","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Parasitic Biology in the Amazon, State University of Par\u00e1, Perebebu\u00ed Bystreet, C Block, 2nd Floor, 2623, Bel\u00e9m 66095-662, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maissa Maia","family":"Santos","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jamilla Augusta de Souza","family":"Pantoja","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erc\u00edlia de Jesus","family":"Gon\u00e7alves","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Claudia da Silva","family":"Ribeiro","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4607-9346","authenticated-orcid":false,"given":"Ana Cec\u00edlia Ribeiro","family":"Cruz","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Virology, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Parasitic Biology in the Amazon, State University of Par\u00e1, Perebebu\u00ed Bystreet, C Block, 2nd Floor, 2623, Bel\u00e9m 66095-662, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandro Patroca","family":"Silva","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carine Fortes","family":"Arag\u00e3o","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre do Ros\u00e1rio","family":"Casseb","sequence":"additional","affiliation":[{"name":"Institute of Animal Health and Production, Federal Rural University of Amaz\u00f4nia, President Tancredo Neves Boulevard, 2501, Bel\u00e9m 66077-830, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9400-2036","authenticated-orcid":false,"given":"Livia Caricio","family":"Martins","sequence":"additional","affiliation":[{"name":"Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Virology, Evandro Chagas Institute, BR 316, Km 07, s\/n, Ananindeua 67030-000, Par\u00e1, Brazil"},{"name":"Graduate Program in Parasitic Biology in the Amazon, State University of Par\u00e1, Perebebu\u00ed Bystreet, C Block, 2nd Floor, 2623, Bel\u00e9m 66095-662, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4911","DOI":"10.3390\/v7092851","article-title":"Insect-specific virus discovery: Significance for the arbovirus 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