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Home / Equine Research Bank / Virol J / Phylogenetic and molecular characterization of equine H3N8 i...
Virology journal2011; 8; 350; doi: 10.1186/1743-422X-8-350

Phylogenetic and molecular characterization of equine H3N8 influenza viruses from Greece (2003 and 2007): evidence for reassortment between evolutionary lineages.

Abstract: For first time in Greece equine influenza virus infection was confirmed, by isolation and molecular analysis, as the cause of clinical respiratory disease among unvaccinated horses during 2003 and 2007 outbreaks. Methods: Equine influenza virus (EIV) H3N8 was isolated in MDCK cells from 30 nasal swabs from horses with acute respiratory disease, which were tested positive by Directigen Flu A. Isolation was confirmed by haemagglutination assay and RT-PCR assay of the M, HA and NA gene. Results: HA sequences of the Greek isolates appeared to be more closely related to viruses isolated in early 1990s in Europe. These results suggested that viruses with fewer changes than those on the main evolutionary lineage may continue to circulate. On the other hand, analysis of deduced NA amino acid sequences were more closely related to viruses isolated in outbreaks in Europe and Asia during 2003-2007. Phylogenetic analysis characterized the Greek isolates as a member of the Eurasian lineage by the haemagglutinin (HA) protein alignment, but appeared to be a member of the Florida sublineage clade 2 by the neuraminidase (NA) protein sequence suggesting that reassortment might be a possible explanation. Conclusions: Our findings suggest that the Greek strains represent an example of "frozen evolution" and probably reassortment between genetically distinct co-circulated strains. Therefore expanding current equine influenza surveillance efforts is a necessity.
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Publication Date: 2011-07-14 PubMed ID: 21756308PubMed Central: PMC3158125DOI: 10.1186/1743-422X-8-350Google Scholar: Lookup
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Summary

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This study details the first confirmation of equine influenza virus infection in Greece from 2003 and 2007 outbreaks in unvaccinated horses. The researchers discovered that the Greek strains could be an instance of “frozen evolution” or likely reassortment between genetically distinct co-circulating strains.

Methods

  • The researchers isolated the Equine Influenza Virus (EIV) H3N8 from 30 nasal swabs taken from horses exhibiting acute respiratory disease.
  • The subjects were tested for influenza using Directigen Flu A, confirming the presence of the virus.
  • Isolation was then substantiated via a haemagglutination assay, a method used to measure the presence and amount of viruses, and a reverse transcription polymerase chain reaction (RT-PCR) assay of the M (matrix), HA (haemagglutinin), and NA (neuraminidase) genes. These are key genetic elements fundamental to a viruses’ functionality and propagation.

Results

  • The HA sequences of the Greek isolates were found to have more in common with viruses isolated in early 1990s in Europe.
  • This suggests that viruses with fewer evolutionary changes than those on the main evolutionary lineage could still be in circulation.
  • However, the NA sequences were more closely related to viruses isolated during 2003-2007 outbreaks in Europe and Asia.
  • From a phylogenetic perspective, which deals with the evolutionary history and relationships among individuals or groups of organisms, the Greek isolates have been classified as a part of the Eurasian lineage by their HA protein alignment but seem to be a part of the Florida sublineage clade 2 by their NA protein sequence.
  • This finding implies that reassortment, a process where different strains of a virus exchange genetic material, might be a probable explanation.

Conclusions

  • The researchers concluded that the Greek strains could represent what they call a “frozen evolution,” where the virus is not significantly changing or evolving over time.
  • It was also concluded that the Greek strains could be an example of reassortment between genetically different co-circulating strains, suggesting the possibility of two genetically different strains of the virus being present in Greece, one from the Eurasian lineage and the other from the Florida clade.
  • These findings emphasize the importance of expanding the current surveillance efforts for equine influenza, given the implications of the virus’s complex evolution and the potential presence of multiple strains.

Cite This Article

APA
Bountouri M, Fragkiadaki E, Ntafis V, Kanellos T, Xylouri E. (2011). Phylogenetic and molecular characterization of equine H3N8 influenza viruses from Greece (2003 and 2007): evidence for reassortment between evolutionary lineages. Virol J, 8, 350. https://doi.org/10.1186/1743-422X-8-350

Publication

Virology journal
ISSN: 1743-422X
NlmUniqueID: 101231645
Country: England
Language: English
Volume: 8
Pages: 350

Researcher Affiliations

Bountouri, Maria
  • Department of Anatomy and Physiology of Farm Animals, Faculty of Animal Science and Aquaculture, Agricultural University of Athens, Greece. eftychia.xylouri@yahoo.gr
Fragkiadaki, Eirini
    Ntafis, Vasileios
      Kanellos, Theo
        Xylouri, Eftychia

          MeSH Terms

          • Animals
          • Cell Line
          • Cluster Analysis
          • Dogs
          • Greece
          • Hemagglutination Inhibition Tests
          • Horse Diseases / virology
          • Horses / virology
          • Influenza A Virus, H3N8 Subtype / genetics
          • Influenza A Virus, H3N8 Subtype / isolation & purification
          • Molecular Sequence Data
          • Nasal Mucosa / virology
          • Orthomyxoviridae Infections / virology
          • Phylogeny
          • RNA, Viral / genetics
          • Reassortant Viruses / genetics
          • Reassortant Viruses / isolation & purification
          • Reverse Transcriptase Polymerase Chain Reaction
          • Sequence Analysis, DNA
          • Virus Cultivation

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          This article has been cited 6 times.
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