Detection and characterization of the Dezidougou virus (genus Negevirus) in mosquitoes (Ochlerotatus caspius) collected in the Republic of Sakha (Yakutia)

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Abstract

Introduction. Monitoring and research on arthropod-borne microorganisms is important. Recently, with the development of next-generation sequencing methods, many previously unknown viruses have been identified in insects.

Aim of the study. Isolation of viruses from mosquitoes sampled in the Republic of Sakha (Yakutia), followed by the study of a new for Russia negevirus isolated from mosquitoes of the species Ochlerotatus caspius, including determination of its complete nucleotide sequence, phylogenetic and virological characteristics.

Materials and methods. Dezidougou virus isolation was performed on C6/36 (Aedes albopictus) cell culture. Electron microscopy was performed using a JEM 1400 electron microscope. Nucleotide sequence screening was performed by NGS on a high-throughput sequencer MiSeq, Illumina (USA). Full genome nucleotide sequence was determined by Sanger sequencing. Phylogenetic analysis was performed using GenBank database, using Vector NTI Advance 11 and MEGA 11 programs.

Results. The virus isolated from mosquitoes replicated efficiently in C6/36 cells, causing their death. However, it did not replicate in the mammalian cell cultures used. The isolated virus did not cause pathologic manifestations in suckling mice when infected intracerebrally. Electron microscopic examination of the purified virus-containing suspension showed the presence of spherical viral particles with a diameter of 45‒55 nm. The results of full genome sequencing identified it as belonging to Dezidougou virus, first isolated in Côte d’Ivoire. The nucleotide sequence of the genome of Yakutsk 2023 strain of Dezidougou virus was deposited in GenBank (PP975071.1).

Conclusion. Dezidougou virus of genus Negevirus was isolated and characterized for the first time in the Russian Federation. Further studies on the prevalence of negeviruses, their virological features, potential importance for public health and their impact on vector competence of vectors are important and promising.

About the authors

Marina A. Stepanyuk

State Research Center of Virology and Biotechnology «Vector»

Email: stepanyuk_ma@vector.nsc.ru
ORCID iD: 0009-0002-2658-7746

junior researcher of the department of molecular virology of flaviviruses and viral hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk region

Stanislav S. Legostaev

State Research Center of Virology and Biotechnology «Vector»

Email: legostaev_ss@vector.nsc.ru
ORCID iD: 0000-0002-6202-445X

trainee researcher of the department of molecular virology of flaviviruses and viral hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk region

Kristina V. Karelina

State Research Center of Virology and Biotechnology «Vector»

Email: karelina_kv@vector.nsc.ru
ORCID iD: 0009-0003-1421-1765

trainee researcher of the department of molecular virology of flaviviruses and viral hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk region

Nina F. Timofeeva

M.K. Ammosov North-Eastern Federal University

Email: niakswan@mail.ru
ORCID iD: 0000-0001-9895-5873

Researcher

Russian Federation, 677000, Republic of Sakha (Yakutia), Yakutsk

Ksenia F. Emtsova

State Research Center of Virology and Biotechnology «Vector»

Email: k.emtsova@g.nsu.ru
ORCID iD: 0009-0003-5165-5357

trainee researcher of microscopic research department

Russian Federation, 630559, Koltsovo, Novosibirsk region

Olesia V. Ohlopkova

State Research Center of Virology and Biotechnology «Vector»

Author for correspondence.
Email: ohlopkova_ov@vector.nsc.ru
ORCID iD: 0000-0002-8214-7828

Candidate of Biological Sciences, Senior Researcher, Department of Biophysics and Environmental Research

Russian Federation, 630559, Koltsovo, Novosibirsk region

Oleg S. Taranov

State Research Center of Virology and Biotechnology «Vector»

Email: taranov@vector.nsc.ru
ORCID iD: 0000-0002-6746-8092

head of microscopic research department

Russian Federation, 630559, Koltsovo, Novosibirsk region

Vladimir A. Ternovoi

State Research Center of Virology and Biotechnology «Vector»

Email: tern@vector.nsc.ru
ORCID iD: 0000-0003-1275-171X

Candidate of Biological Sciences, Leading Researcher, Department of Molecular Virology of Flaviviruses and Viral Hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk region

Albert V. Protopopov

M.K. Ammosov North-Eastern Federal University

Email: a.protopopov@mail.ru
ORCID iD: 0000-0001-6543-4596

Doctor of Biological Sciences, Chief Researcher

Russian Federation, 677000, Republic of Sakha (Yakutia), Yakutsk

Valery B. Loktev

State Research Center of Virology and Biotechnology «Vector»

Email: loktev@vector.nsc.ru
ORCID iD: 0000-0002-0229-321X

MD, PhD, DSc, Prof., academician RANS, Chief Researcher, Department of Molecular Virology of Flaviviruses and Viral Hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk region

Victor A. Svyatchenko

State Research Center of Virology and Biotechnology «Vector»

Email: svyat@vector.nsc.ru
ORCID iD: 0000-0002-2729-0592

Candidate of Biological Sciences, Leading Researcher, Department of Molecular Virology of Flaviviruses and Viral Hepatitis

Russian Federation, 630559, Koltsovo, Novosibirsk region

Alexander P. Agafonov

State Research Center of Virology and Biotechnology «Vector»

Email: agafonov@vector.nsc.ru
ORCID iD: 0000-0003-2577-0434

Doctor of Biological Sciences, Director General, Federal Biotechnology and Biotechnology

Russian Federation, 630559, Koltsovo, Novosibirsk region

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2. Fig. 1. Light microscopy (×200) of C6/36 cell culture infected with DEZV Yakutsk 2023 strain 120 hours after infection. On the left is a control of C6/36 cell culture.

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3. Fig. 2. Transmission electron microscopy of a purified virus suspension. Rounded particles with a diameter of 45‒55 nm and an electron-dense region in the central part. Contrasted with 2% uranyl acetate. The scale is indicated on images.

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4. Fig. 3. Phylodendrogram showing the maximum likelihood analysis of full-length viral sequences of DEZV and viruses of the genus Negevirus. The sequence characterized in this study is highlighted with the symbol (●). A, B, and C are the main branches of negeviruses.

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5. Appendix 1
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Copyright (c) 2025 Stepanyuk M.A., Legostaev S.S., Karelina K.V., Timofeeva N.F., Emtsova K.F., Ohlopkova O.V., Taranov O.S., Ternovoi V.A., Protopopov A.V., Loktev V.B., Svyatchenko V.A., Agafonov A.P.

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