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Mirza, Jeremy D. ; Guimaraes, Lilian de Oliveira ; Wilkinson, Sam ; Rocha, Esmenia C. ; Bertanhe, Mayara ; Helfstein, Vanessa Christe ; de-Deus, Juliana Telles ; Claro, Ingra M. ; Cumley, Nicola ; Quick, Joshua ; Faria, Nuno R. ; Sabino, Ester C. ; Kirchgatter, Karin ; Loman, Nicholas J.

• Tipo de documento: Artigo Científico
• Fonte: MICROBIAL GENOMICS; v. 10, n. 1, p. 14-pg., 2024-01-01.

Resumo

The risk to human health from mosquito - borne viruses such as dengue, chikungunya and yellow fever is increasing due to increased human expansion, deforestation and climate change. To anticipate and predict the spread and transmission of mosquito - borne viruses, a better understanding of the transmission cycle in mosquito populations is needed. We present a pathogen- agnostic combined sequencing protocol for identifying vectors, viral pathogens and their hosts or reservoirs using portable Oxford Nanopore sequencing. Using mosquitoes collected in Sao Paulo, Brazil, we extracted RNA for virus identification and DNA for blood meal and mosquito identification. Mosquitoes and blood meals were identified by comparing cytochrome c oxidase I (COI) sequences against a curated Barcode of Life Data System (BOLD). Viruses were identified using the SMART - 9N protocol, which allows amplified DNA to be prepared with native barcoding for nanopore sequencing. Kraken 2 was employed to detect viral pathogens and Minimap2 and BOLD identified the contents of the blood meal. Due to the high similarity of some species, mosquito identification was conducted using BLAST after generation of consensus COI sequences using RACON polishing. This protocol can simultaneously uncover viral diversity, mosquito species and mosquito feeding habits. It also has the potential to increase understanding of mosquito genetic diversity and transmission dynamics of zoonotic mosquito - borne viruses. (AU)

• Processo FAPESP: 18/14389-0 - Centro Conjunto Brasil-Reino Unido para Descoberta, Diagnóstico, Genômica e Epidemiologia de Arbovírus (CADDE)
• Beneficiário: Ester Cerdeira Sabino
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/16232-1 - Hemosporídeos e arbovírus emergentes em culicídeos e culicoides do Parque Zoológico de São Paulo
• Beneficiário: Lilian de Oliveira Guimarães
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/50345-5 - Plano de desenvolvimento institucional em pesquisa e tecnologia para a vigilância e controle de vetores da Superintendência de Controle de Endemias - SUCEN (PDIp)
• Beneficiário: Vera Lucia Fonseca de Camargo Neves
• Modalidade de apoio: Auxílio à Pesquisa - Programa Modernização de Institutos Estaduais de Pesquisa