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Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey

BACKGROUND: Active vector surveillance provides an efficient tool for monitoring the presence or spread of emerging or re-emerging vector-borne viruses. This study was undertaken to investigate the circulation of flaviviruses. Mosquitoes were collected from 58 locations in 10 provinces across the Ae...

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Autores principales: Ergünay, Koray, Litzba, Nadine, Brinkmann, Annika, Günay, Filiz, Sarıkaya, Yasemen, Kar, Sırrı, Örsten, Serra, Öter, Kerem, Domingo, Cristina, Erisoz Kasap, Özge, Özkul, Aykut, Mitchell, Luke, Nitsche, Andreas, Alten, Bülent, Linton, Yvonne-Marie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5360070/
https://www.ncbi.nlm.nih.gov/pubmed/28320443
http://dx.doi.org/10.1186/s13071-017-2087-7
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author Ergünay, Koray
Litzba, Nadine
Brinkmann, Annika
Günay, Filiz
Sarıkaya, Yasemen
Kar, Sırrı
Örsten, Serra
Öter, Kerem
Domingo, Cristina
Erisoz Kasap, Özge
Özkul, Aykut
Mitchell, Luke
Nitsche, Andreas
Alten, Bülent
Linton, Yvonne-Marie
author_facet Ergünay, Koray
Litzba, Nadine
Brinkmann, Annika
Günay, Filiz
Sarıkaya, Yasemen
Kar, Sırrı
Örsten, Serra
Öter, Kerem
Domingo, Cristina
Erisoz Kasap, Özge
Özkul, Aykut
Mitchell, Luke
Nitsche, Andreas
Alten, Bülent
Linton, Yvonne-Marie
author_sort Ergünay, Koray
collection PubMed
description BACKGROUND: Active vector surveillance provides an efficient tool for monitoring the presence or spread of emerging or re-emerging vector-borne viruses. This study was undertaken to investigate the circulation of flaviviruses. Mosquitoes were collected from 58 locations in 10 provinces across the Aegean, Thrace and Mediterranean Anatolian regions of Turkey in 2014 and 2015. Following morphological identification, mosquitoes were pooled and screened by nested and real-time PCR assays. Detected viruses were further characterised by sequencing. Positive pools were inoculated onto cell lines for virus isolation. Next generation sequencing was employed for genomic characterisation of the isolates. RESULTS: A total of 12,711 mosquito specimens representing 15 species were screened in 594 pools. Eleven pools (2%) were reactive in the virus screening assays. Sequencing revealed West Nile virus (WNV) in one Culex pipiens (s.l.) pool from Thrace. WNV sequence corresponded to lineage one clade 1a but clustered distinctly from the Turkish prototype isolate. In 10 pools, insect-specific flaviviruses were characterised as Culex theileri flavivirus in 5 pools of Culex theileri and one pool of Cx. pipiens (s.l.), Ochlerotatus caspius flavivirus in two pools of Aedes (Ochlerotatus) caspius, Flavivirus AV-2011 in one pool of Culiseta annulata, and an undetermined flavivirus in one pool of Uranotaenia unguiculata from the Aegean and Thrace regions. DNA forms or integration of the detected insect-specific flaviviruses were not observed. A virus strain, tentatively named as “Ochlerotatus caspius flavivirus Turkey”, was isolated from an Ae. caspius pool in C6/36 cells. The viral genome comprised 10,370 nucleotides with a putative polyprotein of 3,385 amino acids that follows the canonical flavivirus polyprotein organisation. Sequence comparisons and phylogenetic analyses revealed the close relationship of this strain with Ochlerotatus caspius flavivirus from Portugal and Hanko virus from Finland. Several conserved structural and amino acid motifs were identified. CONCLUSIONS: We identified WNV and several distinct insect-specific flaviviruses during an extensive biosurveillance study of mosquitoes in various regions of Turkey in 2014 and 2015. Ongoing circulation of WNV is revealed, with an unprecedented genetic diversity. A probable replicating form of an insect flavivirus identified only in DNA form was detected. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-017-2087-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-53600702017-03-24 Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey Ergünay, Koray Litzba, Nadine Brinkmann, Annika Günay, Filiz Sarıkaya, Yasemen Kar, Sırrı Örsten, Serra Öter, Kerem Domingo, Cristina Erisoz Kasap, Özge Özkul, Aykut Mitchell, Luke Nitsche, Andreas Alten, Bülent Linton, Yvonne-Marie Parasit Vectors Research BACKGROUND: Active vector surveillance provides an efficient tool for monitoring the presence or spread of emerging or re-emerging vector-borne viruses. This study was undertaken to investigate the circulation of flaviviruses. Mosquitoes were collected from 58 locations in 10 provinces across the Aegean, Thrace and Mediterranean Anatolian regions of Turkey in 2014 and 2015. Following morphological identification, mosquitoes were pooled and screened by nested and real-time PCR assays. Detected viruses were further characterised by sequencing. Positive pools were inoculated onto cell lines for virus isolation. Next generation sequencing was employed for genomic characterisation of the isolates. RESULTS: A total of 12,711 mosquito specimens representing 15 species were screened in 594 pools. Eleven pools (2%) were reactive in the virus screening assays. Sequencing revealed West Nile virus (WNV) in one Culex pipiens (s.l.) pool from Thrace. WNV sequence corresponded to lineage one clade 1a but clustered distinctly from the Turkish prototype isolate. In 10 pools, insect-specific flaviviruses were characterised as Culex theileri flavivirus in 5 pools of Culex theileri and one pool of Cx. pipiens (s.l.), Ochlerotatus caspius flavivirus in two pools of Aedes (Ochlerotatus) caspius, Flavivirus AV-2011 in one pool of Culiseta annulata, and an undetermined flavivirus in one pool of Uranotaenia unguiculata from the Aegean and Thrace regions. DNA forms or integration of the detected insect-specific flaviviruses were not observed. A virus strain, tentatively named as “Ochlerotatus caspius flavivirus Turkey”, was isolated from an Ae. caspius pool in C6/36 cells. The viral genome comprised 10,370 nucleotides with a putative polyprotein of 3,385 amino acids that follows the canonical flavivirus polyprotein organisation. Sequence comparisons and phylogenetic analyses revealed the close relationship of this strain with Ochlerotatus caspius flavivirus from Portugal and Hanko virus from Finland. Several conserved structural and amino acid motifs were identified. CONCLUSIONS: We identified WNV and several distinct insect-specific flaviviruses during an extensive biosurveillance study of mosquitoes in various regions of Turkey in 2014 and 2015. Ongoing circulation of WNV is revealed, with an unprecedented genetic diversity. A probable replicating form of an insect flavivirus identified only in DNA form was detected. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-017-2087-7) contains supplementary material, which is available to authorized users. BioMed Central 2017-03-20 /pmc/articles/PMC5360070/ /pubmed/28320443 http://dx.doi.org/10.1186/s13071-017-2087-7 Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Ergünay, Koray
Litzba, Nadine
Brinkmann, Annika
Günay, Filiz
Sarıkaya, Yasemen
Kar, Sırrı
Örsten, Serra
Öter, Kerem
Domingo, Cristina
Erisoz Kasap, Özge
Özkul, Aykut
Mitchell, Luke
Nitsche, Andreas
Alten, Bülent
Linton, Yvonne-Marie
Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey
title Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey
title_full Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey
title_fullStr Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey
title_full_unstemmed Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey
title_short Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey
title_sort co-circulation of west nile virus and distinct insect-specific flaviviruses in turkey
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5360070/
https://www.ncbi.nlm.nih.gov/pubmed/28320443
http://dx.doi.org/10.1186/s13071-017-2087-7
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