Tracking arboviruses, their transmission vectors and potential hosts by nanopore sequencing of mosquitoes

Author(s):Show less -

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.

• Document type: Journal article
• Source: MICROBIAL GENOMICS; v. 10, n. 1, p. 14-pg., 2024-01-01.

Abstract

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)

• FAPESP's process: 18/14389-0 - Brazil-UK Centre for Arbovirus Discovery, Diagnosis, Genomics and Epidemiology (CADDE)
• Grantee: Ester Cerdeira Sabino
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 18/16232-1 - Haemosporidian parasites and emerging arbovirus in Mosquitoes (Diptera: Culicidae) and Culicoides biting midges (Diptera: Ceratopogonidae) from the Zoological Park of São Paulo
• Grantee: Lilian de Oliveira Guimarães
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 17/50345-5 - Institutional development plan in research and technology for vector surveillance and control of the Superintendência do Controle de Endemias - SUCEN (PDIp)
• Grantee: Vera Lucia Fonseca de Camargo Neves
• Support Opportunities: Research Grants - State Research Institutes Modernization Program