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Memorias do Instituto Oswaldo Cruz2019; 114; e180332; doi: 10.1590/0074-02760180332

First isolation of West Nile virus in Brazil.

Abstract: Serological evidence of West Nile virus (WNV) infection has been reported in different regions of Brazil from equine and human hosts but the virus had never been isolated in the country. Objective: We sought to identify the viral etiology of equine encephalitis in Espírito Santo state. Methods: We performed viral culture in C6/36 cells, molecular detection of WNV genome, histopathology and immunohistochemistry from horse cerebral tissue. We also carried out sequencing, phylogenetic analysis and molecular clock. Results: Histopathologic analysis from horse cerebral tissue showed injury related to encephalitis and WNV infection was confirmed by immunohistochemistry. The virus was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) from brain tissue and subsequently isolated in C6/36 cells. WNV full-length genome was sequenced showing the isolated strain belongs to lineage 1a. The molecular clock indicated that Brazilian WNV strain share the same common ancestor that were circulating in US during 2002-2005. Conclusions: Here we report the first isolation of WNV in Brazil from a horse with neurologic disease, which was clustered into lineage 1a with others US WNV strains isolated in beginning of 2000's decade.
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Publication Date: 2019-01-17 PubMed ID: 30672980PubMed Central: PMC6343470DOI: 10.1590/0074-02760180332Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses the first-ever isolation of the West Nile Virus (WNV) in Brazil, a virus typically reported through serological evidence from equine and human hosts. The study involved methods such as viral culture in C6/36 cells, molecular detection of WNV genome, histopathology, immunohistochemistry, sequencing, phylogenetic analysis, and molecular clock techniques.

Objective and Methods

  • The primary aim of the article was to identify the viral etiology of equine encephalitis in Espírito Santo state in Brazil. Equine Encephalitis is an inflammation of the brain and spinal cord in horses and can be caused by various viruses.
  • The researchers applied a range of methods in their study, including viral culture in C6/36 cells; a standard process that involves culturing of virus in cells to increase their number, and molecular detection of WNV genome to confirm the virus’s presence.
  • Histopathologic analysis was performed on the horse’s cerebral tissue to study the structural changes caused by the virus, and immunohistochemistry was done to visibly detect the presence of WNV in the tissue samples.
  • The researchers also employed sequencing, to determine the order of nucleotides in the virus’s RNA; phylogenetic analysis, to understand the evolutionary relationships and origin of the isolated WNV; and a molecular clock, to estimate the timeframe in which the virus might have evolved.

Results

  • The histopathologic analysis verified injury in the horse’s brain tissue, consistent with encephalitis, and the WNV infection was confirmed using immunohistochemistry.
  • WNV was detected in the brain tissue through the reverse transcription quantitative polymerase chain reaction (RT-qPCR) – a technique used to amplify and quantify targeted DNA molecules, allowing them to be detected more easily. The virus was subsequently isolated in C6/36 cells.
  • The sequencing of WNV full-length genome showed that the isolated strain belongs to lineage 1a, one of the genetic groupings of the virus.
  • The molecular clock indication presented that the Brazilian strain of WNV shares a common ancestor with the strains that were circulating in the United States from 2002-2005.

Conclusions

  • The research concludes by confirming the first successful isolation of WNV in Brazil from a horse with a neurological disease.
  • The isolated virus is shown to belong to lineage 1a and shares common ancestry with US strains from the early 2000s, providing insights into the virus’s evolution and dispersion.

Cite This Article

APA
Martins LC, Silva EVPD, Casseb LMN, Silva SPD, Cruz ACR, Pantoja JAS, Medeiros DBA, Martins Filho AJ, Cruz EDRMD, Araújo MTF, Cardoso JF, Cunha MACRD, Almada GL, Romano APM, Santos MGDP, Rodrigues GAP, Chiang JO, Quaresma JAS, Carvalho VL, Vasconcelos PFDC. (2019). First isolation of West Nile virus in Brazil. Mem Inst Oswaldo Cruz, 114, e180332. https://doi.org/10.1590/0074-02760180332

Publication

Memorias do Instituto Oswaldo Cruz
ISSN: 1678-8060
NlmUniqueID: 7502619
Country: Brazil
Language: English
Volume: 114
Pages: e180332
PII: e180332

Researcher Affiliations

Martins, Lívia Caricio
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Silva, Eliana Vieira Pinto da
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Casseb, Livia Medeiros Neves
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Silva, Sandro Patroca da
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Cruz, Ana Cecília Ribeiro
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Pantoja, Jamilla Augusta de Sousa
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Medeiros, Daniele Barbosa de Almeida
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Martins Filho, Arnaldo Jorge
  • Instituto Evandro Chagas, Seção de Patologia, Ananindeua, PA, Brasil.
Cruz, Ermelinda do Rosário Moutinho da
  • Instituto Evandro Chagas, Seção de Patologia, Ananindeua, PA, Brasil.
Araújo, Marialva Tereza Ferreira de
  • Instituto Evandro Chagas, Seção de Patologia, Ananindeua, PA, Brasil.
Cardoso, Jedson Ferreira
  • Instituto Evandro Chagas, Centro de Inovação tecnológica, Ananindeua, PA, Brasil.
Cunha, Marcos Antônio Correia Rodrigues da
  • Secretaria de Estado da Saúde do Espírito Santo, Vitória, ES, Brasil.
Almada, Gilton Luiz
  • Secretaria de Estado da Saúde do Espírito Santo, Vitória, ES, Brasil.
Romano, Alessandro Pecego Martins
  • Ministério da Saúde, Secretaria de Vigilância em Saúde, Brasília, DF, Brasil.
Santos, Maria Guadalupe Dias Pestana
  • Secretaria de Saúde do Município de Venécia, Venécia, ES, Brasil.
Rodrigues, Gilsa Aparecida Pimenta
  • Secretaria de Estado da Saúde do Espírito Santo, Vitória, ES, Brasil.
Chiang, Jannifer Oliveira
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Quaresma, Juarez Antonio Simões
  • Instituto Evandro Chagas, Seção de Patologia, Ananindeua, PA, Brasil.
Carvalho, Valéria Lima
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.
Vasconcelos, Pedro Fernando da Costa
  • Instituto Evandro Chagas, Seção de Arbovirologia e Febres Hemorrágicas, Ananindeua, PA, Brasil.

MeSH Terms

  • Animals
  • Brazil
  • Encephalomyelitis, Equine / veterinary
  • Encephalomyelitis, Equine / virology
  • Horse Diseases / diagnosis
  • Horse Diseases / virology
  • Horses
  • Immunohistochemistry
  • Male
  • Phylogeography
  • RNA, Viral / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • West Nile Fever / diagnosis
  • West Nile Fever / veterinary
  • West Nile virus / genetics
  • West Nile virus / isolation & purification

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