Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)

Introduction: In Italy, despite the existence of an epidemiological surveillance system for Arboviruses, managed in a One Health perspective, the implementation of genomic characterisation remains fragmented and no systematic genomic surveillance initiative integrating human and animal data has been...

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Main Authors: Miss Martina Brandolini, Dr Alessandra Mistral De Pascali, Mr Giorgio Dirani, Ms Silvia Zannoli, Ms Irene Zaghi, Ms Ludovica Ingletto, Dr Antonio Lavazza, Dr Davide Lelli, Dr Michele Dottori, Dr Mattia Calzolari, Mr Massimiliano Guerra, Dr Carlo Biagetti, Prof Francesco Cristini, Dr Paolo Bassi, Dr Rino Biguzzi, Dr Andrea Porcellini, Dr Roberta Chicchi, Prof Monica Cricca, Prof Alessandra Scagliarini, Prof Vittorio Sambri
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:International Journal of Infectious Diseases
Online Access:http://www.sciencedirect.com/science/article/pii/S1201971224005174
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author Miss Martina Brandolini
Dr Alessandra Mistral De Pascali
Mr Giorgio Dirani
Ms Silvia Zannoli
Ms Irene Zaghi
Ms Ludovica Ingletto
Dr Antonio Lavazza
Dr Davide Lelli
Dr Michele Dottori
Dr Mattia Calzolari
Mr Massimiliano Guerra
Dr Carlo Biagetti
Prof Francesco Cristini
Dr Paolo Bassi
Dr Rino Biguzzi
Dr Andrea Porcellini
Dr Roberta Chicchi
Prof Monica Cricca
Prof Alessandra Scagliarini
Prof Vittorio Sambri
author_facet Miss Martina Brandolini
Dr Alessandra Mistral De Pascali
Mr Giorgio Dirani
Ms Silvia Zannoli
Ms Irene Zaghi
Ms Ludovica Ingletto
Dr Antonio Lavazza
Dr Davide Lelli
Dr Michele Dottori
Dr Mattia Calzolari
Mr Massimiliano Guerra
Dr Carlo Biagetti
Prof Francesco Cristini
Dr Paolo Bassi
Dr Rino Biguzzi
Dr Andrea Porcellini
Dr Roberta Chicchi
Prof Monica Cricca
Prof Alessandra Scagliarini
Prof Vittorio Sambri
author_sort Miss Martina Brandolini
collection DOAJ
description Introduction: In Italy, despite the existence of an epidemiological surveillance system for Arboviruses, managed in a One Health perspective, the implementation of genomic characterisation remains fragmented and no systematic genomic surveillance initiative integrating human and animal data has been established. The purpose of this study is to investigate the contribution of Whole Genome Sequencing (WGS) in the integrated genomic characterization of West Nile virus in target avian reservoirs, mosquito vectors and infected humans in Romagna (northeastern Italy). Methods: The study was conducted in Romagna, northeastern Italy, between May and October 2023 involving patients with encephalitis/meningitis and healthy blood donors who tested positive for WNV during screening programmes. Mosquitoes were sampled with attractive traps, and resident and migratory birds were collected as of active or passive surveillance. Samples were screened with WNV-specific PCR (Polymerase Chain Reaction) assays, and positive samples were subjected to WGS by tagmentation and hybrid-capture-mediated target enrichment (Viral Surveillance Panel, Illumina). Assemblies and consensus sequences were obtained with Bowtie2 v.2.4.1 and BCFtools. Maximum likelihood trees for viral phylogenetic inference were constructed with IQ-TREE 2.0.3 with automatic selection of the best nucleotide substitution model, 1000 ultrafast bootstrap replicates and midpoint rooted. Results: Eleven WNV lineage 2 sequences were obtained (4 meningitis cases, 3 blood donors, 3 Culex pipiens and 1 Pica pica). Genome sequencing yielded 98.8-100% coverage. Coding sequences maintained high nucleotide and deduced aminoacid conservation, with an average identity of 99.82% and 99.92%. Mutations of virulent clade 2d were identified, affecting infection rates in avian hosts and mosquito vectors and neuroinvasiveness in humans. Obtained sequences clustered within the Central-Southern European clade with robust bootstrap support, forming a distinct cluster with other viruses isolated in Italy from 2021 to 2023. Discussion: Our study, albeit with preliminary and limited results, provides an insight into integrated genomic characterisation of WNV strains circulating in the study area, where WNV is endemic and which is considered a high-transmission-risk hotspot due to its geoclimatic characteristics and its proximity with one of the main stop-overs and breeding ground for migratory birds in Central and Southern Europe (Delta of Po river). Continued genomic surveillance, throughout future seasons, will support the tracking of arboviruses genetic diversity, evolution, emergence of new variants or introduction of genetically diverse strains. Conclusion: Our approach suggests the importance of adopting an interdisciplinary multistakeholder strategy to integrate the steps from clinical diagnosis to entomological and ornithological monitoring, up to epidemiological and genomic surveillance to create an operational process involving human and animal health suitable for the genomic characterisation of both autochthonous and imported Arboviruses. Engaging both human and animal health sectors, will facilitate a comprehensive characterisation of endemic and emerging Arboviruses to mitigate their impact on public health.
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spelling doaj-art-7c337f2a70d14e89938b288ef69b0bb82025-08-20T02:17:08ZengElsevierInternational Journal of Infectious Diseases1201-97122025-03-0115210744210.1016/j.ijid.2024.107442Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)Miss Martina Brandolini0Dr Alessandra Mistral De Pascali1Mr Giorgio Dirani2Ms Silvia Zannoli3Ms Irene Zaghi4Ms Ludovica Ingletto5Dr Antonio Lavazza6Dr Davide Lelli7Dr Michele Dottori8Dr Mattia Calzolari9Mr Massimiliano Guerra10Dr Carlo Biagetti11Prof Francesco Cristini12Dr Paolo Bassi13Dr Rino Biguzzi14Dr Andrea Porcellini15Dr Roberta Chicchi16Prof Monica Cricca17Prof Alessandra Scagliarini18Prof Vittorio Sambri19Department of Medical and Surgical Sciences, University of Bologna, Bologna, ItalyDepartment of Medical and Surgical Sciences, University of Bologna, Bologna, ItalyUnit of Microbiology, The Greater Romagna Area Hub Laboratory, Cesena, ItalyUnit of Microbiology, The Greater Romagna Area Hub Laboratory, Cesena, ItalyUnit of Microbiology, The Greater Romagna Area Hub Laboratory, Cesena, ItalyDepartment of Medical and Surgical Sciences, University of Bologna, Bologna, ItalyIstituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna “Bruno Ubertini”, Brescia, ItalyIstituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna “Bruno Ubertini”, Brescia, ItalyIstituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna “Bruno Ubertini”, Brescia, ItalyIstituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna “Bruno Ubertini”, Brescia, ItalyUnit of Microbiology, The Greater Romagna Area Hub Laboratory, Cesena, ItalyUnit of Infectious Diseases, Infermi Hospital, Rimini, ItalyUnit of Infectious Diseases, Morgagni-Pierantoni Hospital, Forlì, ItalyUnit of Infectious Diseases, Santa Maria delle Croci Hospital, Ravenna, ItalyUnit of Transfusion Medicine, The Greater Romagna Area Hub Laboratory, AUSL Romagna, Cesena, ItalyUnit of Transfusion Medicine, The Greater Romagna Area Hub Laboratory, AUSL Romagna, Cesena, ItalyUnit of Transfusion Medicine, The Greater Romagna Area Hub Laboratory, AUSL Romagna, Cesena, ItalyDepartment of Medical and Surgical Sciences, University of Bologna, Bologna, ItalyDepartment of Medical and Surgical Sciences, University of Bologna, Bologna, ItalyDepartment of Medical and Surgical Sciences, University of Bologna, Bologna, ItalyIntroduction: In Italy, despite the existence of an epidemiological surveillance system for Arboviruses, managed in a One Health perspective, the implementation of genomic characterisation remains fragmented and no systematic genomic surveillance initiative integrating human and animal data has been established. The purpose of this study is to investigate the contribution of Whole Genome Sequencing (WGS) in the integrated genomic characterization of West Nile virus in target avian reservoirs, mosquito vectors and infected humans in Romagna (northeastern Italy). Methods: The study was conducted in Romagna, northeastern Italy, between May and October 2023 involving patients with encephalitis/meningitis and healthy blood donors who tested positive for WNV during screening programmes. Mosquitoes were sampled with attractive traps, and resident and migratory birds were collected as of active or passive surveillance. Samples were screened with WNV-specific PCR (Polymerase Chain Reaction) assays, and positive samples were subjected to WGS by tagmentation and hybrid-capture-mediated target enrichment (Viral Surveillance Panel, Illumina). Assemblies and consensus sequences were obtained with Bowtie2 v.2.4.1 and BCFtools. Maximum likelihood trees for viral phylogenetic inference were constructed with IQ-TREE 2.0.3 with automatic selection of the best nucleotide substitution model, 1000 ultrafast bootstrap replicates and midpoint rooted. Results: Eleven WNV lineage 2 sequences were obtained (4 meningitis cases, 3 blood donors, 3 Culex pipiens and 1 Pica pica). Genome sequencing yielded 98.8-100% coverage. Coding sequences maintained high nucleotide and deduced aminoacid conservation, with an average identity of 99.82% and 99.92%. Mutations of virulent clade 2d were identified, affecting infection rates in avian hosts and mosquito vectors and neuroinvasiveness in humans. Obtained sequences clustered within the Central-Southern European clade with robust bootstrap support, forming a distinct cluster with other viruses isolated in Italy from 2021 to 2023. Discussion: Our study, albeit with preliminary and limited results, provides an insight into integrated genomic characterisation of WNV strains circulating in the study area, where WNV is endemic and which is considered a high-transmission-risk hotspot due to its geoclimatic characteristics and its proximity with one of the main stop-overs and breeding ground for migratory birds in Central and Southern Europe (Delta of Po river). Continued genomic surveillance, throughout future seasons, will support the tracking of arboviruses genetic diversity, evolution, emergence of new variants or introduction of genetically diverse strains. Conclusion: Our approach suggests the importance of adopting an interdisciplinary multistakeholder strategy to integrate the steps from clinical diagnosis to entomological and ornithological monitoring, up to epidemiological and genomic surveillance to create an operational process involving human and animal health suitable for the genomic characterisation of both autochthonous and imported Arboviruses. Engaging both human and animal health sectors, will facilitate a comprehensive characterisation of endemic and emerging Arboviruses to mitigate their impact on public health.http://www.sciencedirect.com/science/article/pii/S1201971224005174
spellingShingle Miss Martina Brandolini
Dr Alessandra Mistral De Pascali
Mr Giorgio Dirani
Ms Silvia Zannoli
Ms Irene Zaghi
Ms Ludovica Ingletto
Dr Antonio Lavazza
Dr Davide Lelli
Dr Michele Dottori
Dr Mattia Calzolari
Mr Massimiliano Guerra
Dr Carlo Biagetti
Prof Francesco Cristini
Dr Paolo Bassi
Dr Rino Biguzzi
Dr Andrea Porcellini
Dr Roberta Chicchi
Prof Monica Cricca
Prof Alessandra Scagliarini
Prof Vittorio Sambri
Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)
International Journal of Infectious Diseases
title Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)
title_full Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)
title_fullStr Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)
title_full_unstemmed Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)
title_short Pushing Forward West Nile Virus Understanding: An Integrated One Health Genomic Surveillance Journey in Romagna (Italy)
title_sort pushing forward west nile virus understanding an integrated one health genomic surveillance journey in romagna italy
url http://www.sciencedirect.com/science/article/pii/S1201971224005174
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