Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System

The Oropouche virus (OROV) is a segmented negative-sense RNA arbovirus member of the <i>Peribunyaviridae</i> family, associated with recurring epidemics of Oropouche fever in Central and South America. Since its identification in 1955, OROV has been responsible for outbreaks in both rura...

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Main Authors: Ana Luisa Teixeira de Almeida, Igor Pinto Silva da Costa, Maycon Douglas do Nascimento Garcia, Marcos Alexandre Nunes da Silva, Yasmim Gonçalves Lazzaro, Ana Maria Bispo de Filippis, Fernanda de Bruycker Nogueira, Debora Ferreira Barreto-Vieira
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Language:English
Published: MDPI AG 2025-03-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/17/3/373
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author Ana Luisa Teixeira de Almeida
Igor Pinto Silva da Costa
Maycon Douglas do Nascimento Garcia
Marcos Alexandre Nunes da Silva
Yasmim Gonçalves Lazzaro
Ana Maria Bispo de Filippis
Fernanda de Bruycker Nogueira
Debora Ferreira Barreto-Vieira
author_facet Ana Luisa Teixeira de Almeida
Igor Pinto Silva da Costa
Maycon Douglas do Nascimento Garcia
Marcos Alexandre Nunes da Silva
Yasmim Gonçalves Lazzaro
Ana Maria Bispo de Filippis
Fernanda de Bruycker Nogueira
Debora Ferreira Barreto-Vieira
author_sort Ana Luisa Teixeira de Almeida
collection DOAJ
description The Oropouche virus (OROV) is a segmented negative-sense RNA arbovirus member of the <i>Peribunyaviridae</i> family, associated with recurring epidemics of Oropouche fever in Central and South America. Since its identification in 1955, OROV has been responsible for outbreaks in both rural and urban areas, with transmission involving sylvatic and urban cycles. This study focuses on the characterization of an OROV isolate from a human clinical sample collected in the state of Rio de Janeiro, a non-endemic region in Brazil, highlighting ultrastructural and morphological aspects of the viral replicative cycle in Vero cells. OROV was isolated in Vero cell monolayers which, following viral inoculation, exhibited marked cytopathic effects (CPEs), mainly represented by changes in cell morphology, including membrane protrusions and vacuolization, as well as cell death. Studies by transmission electron microscopy (TEM) revealed significant ultrastructural changes, such as apoptosis, intense remodeling of membrane-bound organelles and signs of rough endoplasmic reticulum and mitochondrial stress. Additionally, the formation of specialized cytoplasmic vacuoles and intra- and extracellular vesicles emphasized trafficking and intercellular communication as essential mechanisms in OROV infection. RT-qPCR studies confirmed the production of viral progeny in high titers, corroborating the efficiency of this experimental model. These findings contribute to a better understanding of the cytopathogenic mechanisms of OROV infection and the contribution of cellular alterations in OROV morphogenesis.
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spelling doaj-art-e6c45e87af51437491c711847cd0df5e2025-08-20T02:43:05ZengMDPI AGViruses1999-49152025-03-0117337310.3390/v17030373Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro SystemAna Luisa Teixeira de Almeida0Igor Pinto Silva da Costa1Maycon Douglas do Nascimento Garcia2Marcos Alexandre Nunes da Silva3Yasmim Gonçalves Lazzaro4Ana Maria Bispo de Filippis5Fernanda de Bruycker Nogueira6Debora Ferreira Barreto-Vieira7Laboratory of Viral Morphology and Morphogenesis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilLaboratory of Viral Morphology and Morphogenesis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilLaboratory of Viral Morphology and Morphogenesis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilLaboratory of Viral Morphology and Morphogenesis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilLaboratory of Viral Morphology and Morphogenesis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilLaboratory of Arboviruses and Hemorrhagic Viruses, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilLaboratory of Arboviruses and Hemorrhagic Viruses, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilLaboratory of Viral Morphology and Morphogenesis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation—Fiocruz, Rio de Janeiro 21040-900, RJ, BrazilThe Oropouche virus (OROV) is a segmented negative-sense RNA arbovirus member of the <i>Peribunyaviridae</i> family, associated with recurring epidemics of Oropouche fever in Central and South America. Since its identification in 1955, OROV has been responsible for outbreaks in both rural and urban areas, with transmission involving sylvatic and urban cycles. This study focuses on the characterization of an OROV isolate from a human clinical sample collected in the state of Rio de Janeiro, a non-endemic region in Brazil, highlighting ultrastructural and morphological aspects of the viral replicative cycle in Vero cells. OROV was isolated in Vero cell monolayers which, following viral inoculation, exhibited marked cytopathic effects (CPEs), mainly represented by changes in cell morphology, including membrane protrusions and vacuolization, as well as cell death. Studies by transmission electron microscopy (TEM) revealed significant ultrastructural changes, such as apoptosis, intense remodeling of membrane-bound organelles and signs of rough endoplasmic reticulum and mitochondrial stress. Additionally, the formation of specialized cytoplasmic vacuoles and intra- and extracellular vesicles emphasized trafficking and intercellular communication as essential mechanisms in OROV infection. RT-qPCR studies confirmed the production of viral progeny in high titers, corroborating the efficiency of this experimental model. These findings contribute to a better understanding of the cytopathogenic mechanisms of OROV infection and the contribution of cellular alterations in OROV morphogenesis.https://www.mdpi.com/1999-4915/17/3/373Oropouche virusultrastructural studiestransmission electron microscopy
spellingShingle Ana Luisa Teixeira de Almeida
Igor Pinto Silva da Costa
Maycon Douglas do Nascimento Garcia
Marcos Alexandre Nunes da Silva
Yasmim Gonçalves Lazzaro
Ana Maria Bispo de Filippis
Fernanda de Bruycker Nogueira
Debora Ferreira Barreto-Vieira
Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System
Viruses
Oropouche virus
ultrastructural studies
transmission electron microscopy
title Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System
title_full Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System
title_fullStr Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System
title_full_unstemmed Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System
title_short Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System
title_sort oropouche virus isolation and ultrastructural characterization from a human case sample from rio de janeiro brazil using an in vitro system
topic Oropouche virus
ultrastructural studies
transmission electron microscopy
url https://www.mdpi.com/1999-4915/17/3/373
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