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A highly divergent Encephalomyocarditis virus isolated from nonhuman primates in Singapore.

Yeo DS, Lian JE, Fernandez CJ, Lin YN, Liaw JC, Soh ML, Lim EA, Chan KP, Ng ML, Tan HC, Oh S, Ooi EE, Tan BH - Virol. J. (2013)

Bottom Line: In 2001 and 2002, fatal myocarditis resulted in the sudden deaths of four, two adult and two juvenile, orang utans out of a cohort of 26 in the Singapore Zoological Gardens.When compared with existing EMCV sequences in the VP1 and 3Dpol gene regions, the nucleotide divergence were at a maximum of 38.8% and 23.6% respectively, while the amino acid divergence were at a maximum of 33.9% and 11.3% respectively.The etiological agent responsible for the fatal myocarditis cases among two of the four orang utans in the Singapore Zoological Gardens was a highly divergent variant of EMCV.

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ABSTRACT

Background: In 2001 and 2002, fatal myocarditis resulted in the sudden deaths of four, two adult and two juvenile, orang utans out of a cohort of 26 in the Singapore Zoological Gardens.

Methods: Of the four orang utans that underwent post-mortem examination, virus isolation was performed from the tissue homogenates of the heart and lung obtained from the two juvenile orang utans in Vero cell cultures. The tissue culture fluid was examined using electron microscopy. Reverse transcription and polymerase chain reaction with Encephalomyocarditis virus (EMCV)-specific primers targeting the gene regions of VP3/VP1 and 3D polymerase (3Dpol) confirmed the virus genus and species. The two EMCV isolates were sequenced and phylogenetic analyses of the virus genes performed. Serological testing on other animal species in the Singapore Zoological Gardens was also conducted.

Results: Electron microscopy of the two EMCV isolates, designated Sing-M100-02 and Sing-M105-02, revealed spherical viral particles of about 20 to 30 nm, consistent with the size and morphology of members belonging to the family Picornaviridae. In addition, infected-Vero cells showed positive immunoflorescence staining with antiserum to EMCV. Sequencing of the viral genome showed that the two EMCV isolates were 99.9% identical at the nucleotide level, indicating a similar source of origin. When compared with existing EMCV sequences in the VP1 and 3Dpol gene regions, the nucleotide divergence were at a maximum of 38.8% and 23.6% respectively, while the amino acid divergence were at a maximum of 33.9% and 11.3% respectively. Phylogenetic analyses of VP1 and 3Dpol genes further grouped the Sing-M100-02 and Sing-M105-02 isolates to themselves, away from existing EMCV lineages. This strongly suggested that Sing-M100-02 and Sing-M105-02 isolates are highly divergent variants of EMCV. Apart from the two deceased orang utans, a serological survey conducted among other zoo animals showed that a number of other animal species had neutralizing antibodies to Sing-M105-02 isolate, indicating that the EMCV variant has a relatively wide host range.

Conclusions: The etiological agent responsible for the fatal myocarditis cases among two of the four orang utans in the Singapore Zoological Gardens was a highly divergent variant of EMCV. This is the first report of an EMCV infection in Singapore and South East Asia.

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Electron micrographs showing viruses from the Sing-M105-02 and Sing-M100-02 isolates. Viruses were concentrated from the tissue culture fluid of virus-infected Vero cells and negatively-stained. Magnification at x50,000. Scale bars represent 100 nm.
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Figure 1: Electron micrographs showing viruses from the Sing-M105-02 and Sing-M100-02 isolates. Viruses were concentrated from the tissue culture fluid of virus-infected Vero cells and negatively-stained. Magnification at x50,000. Scale bars represent 100 nm.

Mentions: The viruses in infected-Vero tissue culture fluid were negatively stained and examined with electron microscopy. The electron micrographs revealed numerous small round virus particles with smooth appearances in the range of 20 to 30 nm for both Sing-M105-02 and Sing-M100-02 isolates (Figure 1). The morphology and size observed are consistent with virus members belonging to the family of Picornaviridae[23].


A highly divergent Encephalomyocarditis virus isolated from nonhuman primates in Singapore.

Yeo DS, Lian JE, Fernandez CJ, Lin YN, Liaw JC, Soh ML, Lim EA, Chan KP, Ng ML, Tan HC, Oh S, Ooi EE, Tan BH - Virol. J. (2013)

Electron micrographs showing viruses from the Sing-M105-02 and Sing-M100-02 isolates. Viruses were concentrated from the tissue culture fluid of virus-infected Vero cells and negatively-stained. Magnification at x50,000. Scale bars represent 100 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750836&req=5

Figure 1: Electron micrographs showing viruses from the Sing-M105-02 and Sing-M100-02 isolates. Viruses were concentrated from the tissue culture fluid of virus-infected Vero cells and negatively-stained. Magnification at x50,000. Scale bars represent 100 nm.
Mentions: The viruses in infected-Vero tissue culture fluid were negatively stained and examined with electron microscopy. The electron micrographs revealed numerous small round virus particles with smooth appearances in the range of 20 to 30 nm for both Sing-M105-02 and Sing-M100-02 isolates (Figure 1). The morphology and size observed are consistent with virus members belonging to the family of Picornaviridae[23].

Bottom Line: In 2001 and 2002, fatal myocarditis resulted in the sudden deaths of four, two adult and two juvenile, orang utans out of a cohort of 26 in the Singapore Zoological Gardens.When compared with existing EMCV sequences in the VP1 and 3Dpol gene regions, the nucleotide divergence were at a maximum of 38.8% and 23.6% respectively, while the amino acid divergence were at a maximum of 33.9% and 11.3% respectively.The etiological agent responsible for the fatal myocarditis cases among two of the four orang utans in the Singapore Zoological Gardens was a highly divergent variant of EMCV.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: In 2001 and 2002, fatal myocarditis resulted in the sudden deaths of four, two adult and two juvenile, orang utans out of a cohort of 26 in the Singapore Zoological Gardens.

Methods: Of the four orang utans that underwent post-mortem examination, virus isolation was performed from the tissue homogenates of the heart and lung obtained from the two juvenile orang utans in Vero cell cultures. The tissue culture fluid was examined using electron microscopy. Reverse transcription and polymerase chain reaction with Encephalomyocarditis virus (EMCV)-specific primers targeting the gene regions of VP3/VP1 and 3D polymerase (3Dpol) confirmed the virus genus and species. The two EMCV isolates were sequenced and phylogenetic analyses of the virus genes performed. Serological testing on other animal species in the Singapore Zoological Gardens was also conducted.

Results: Electron microscopy of the two EMCV isolates, designated Sing-M100-02 and Sing-M105-02, revealed spherical viral particles of about 20 to 30 nm, consistent with the size and morphology of members belonging to the family Picornaviridae. In addition, infected-Vero cells showed positive immunoflorescence staining with antiserum to EMCV. Sequencing of the viral genome showed that the two EMCV isolates were 99.9% identical at the nucleotide level, indicating a similar source of origin. When compared with existing EMCV sequences in the VP1 and 3Dpol gene regions, the nucleotide divergence were at a maximum of 38.8% and 23.6% respectively, while the amino acid divergence were at a maximum of 33.9% and 11.3% respectively. Phylogenetic analyses of VP1 and 3Dpol genes further grouped the Sing-M100-02 and Sing-M105-02 isolates to themselves, away from existing EMCV lineages. This strongly suggested that Sing-M100-02 and Sing-M105-02 isolates are highly divergent variants of EMCV. Apart from the two deceased orang utans, a serological survey conducted among other zoo animals showed that a number of other animal species had neutralizing antibodies to Sing-M105-02 isolate, indicating that the EMCV variant has a relatively wide host range.

Conclusions: The etiological agent responsible for the fatal myocarditis cases among two of the four orang utans in the Singapore Zoological Gardens was a highly divergent variant of EMCV. This is the first report of an EMCV infection in Singapore and South East Asia.

Show MeSH
Related in: MedlinePlus