A highly divergent Encephalomyocarditis virus isolated from nonhuman primates in Singapore
Open accessAbstract
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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Within the genome and 3D phylogenies, the EMCV-3 isolates clustered with EMCV-2 and associated viruses with strong support.
“However, within the genome and 3D phylogenies, these EMCV-3 isolates clustered with EMCV-2 and associated viruses with strong support (Figs 2A and 4B), as previously noted [27,44].”
From Zoological transmission of encephalomyocarditis virus in the United States: Virus evolution, host ecology, and capsid antigenicity derived from an outbreak (2026) · cites this paper as [27]
EMCV outbreaks in zoos, involving a myriad of mammal species, have been reported from Australia, Asia, Africa, Europe, and North America, making the virus a worldwide problem in zoological settings.
“EMCV outbreaks in zoos – involving a myriad of mammal species such as primates, ungulates, carnivores, rodents, and marsupials – have been reported from Australia, Asia, Africa, Europe, and North America, making the virus a worldwide problem in zoological settings [16,22–29].”
From Zoological transmission of encephalomyocarditis virus in the United States: Virus evolution, host ecology, and capsid antigenicity derived from an outbreak (2026) · cites this paper as [27]
The proposed novel serotype EMCV-3, comprised of orangutan isolates from Singapore, was less divergent and clustered with the large EMCV-1 group in both capsid and polyprotein phylogenies.
“The proposed novel serotype (EMCV-3) comprised of orangutan isolates (Sing-M100-02 and M105-02) from Singapore [27] was less divergent and clustered with the large EMCV-1 group in both capsid (Fig 3) and polyprotein (Fig 4A) phylogenies.”
From Zoological transmission of encephalomyocarditis virus in the United States: Virus evolution, host ecology, and capsid antigenicity derived from an outbreak (2026) · cites this paper as [27]
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- Authors
- Yeo, D. S.-Y., Lian, J. E., Fernandez, C. J., Lin, Y.-N., Liaw, J. C.-W., Soh, M.-L., Lim, E. A.-S., Chan, K.-P., Ng, M.-L., Tan, H.-C., Oh, S., Ooi, E.-E., & Tan, B.-H.
- Affiliations
- DSO National Laboratories (SG)
- Nanyang Technological University (SG)
- Agri-Food and Veterinary Authority of Singapore (SG)
- Singapore General Hospital (SG)
- National University of Singapore (SG)
- Mandai Wildlife Group (SG)
- Corresponding author
- Tan, Boon-Huan
- Venue
- Virology Journal
- Published
- 2013-08-02
- Volume
- 10
- Issue
- 1
- Pages
- 248-248
- ISSN
- 1743-422X (ISSN-L)
- PMC
- PMC3750836
- OpenAlex
- W2141918624
- Cited by
- 31
- References
- 49
- Citations by year
- 2026: 5 · 2025: 4 · 2024: 2 · 2023: 2 · 2022: 3 · 2021: 2 · 2020: 3 · 2018: 2 · 2017: 4 · 2016: 2 · 2015: 1 · 2014: 1
- FWCI
- 0.91
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Virology Journal (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- National University of Singapore (submittedVersion)
- Funders
- Agri-Food and Veterinary Authority
- Topics
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Viral Infections and Immunology Research
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
- Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
- Keywords
- encephalomyocarditis virus, non-human primates, orangutans, myocarditis, viral divergence, picornavirus, VP1 gene, 3D polymerase gene, phylogenetic analysis, zoo animals, neutralizing antibodies, host range, wildlife disease outbreak, virus isolation, serological survey
- MeSH terms
- Animals, Animals, Zoo, Chlorocebus aethiops, Encephalomyocarditis virus, Heart, Lung, Molecular Sequence Data, Phylogeny, RNA, Viral, Singapore, Vero Cells, Viral Proteins, Virion, Virus Cultivation, Cluster Analysis, Genome, Viral, Sequence Homology, Sequence Analysis, DNA, Microscopy, Electron, Transmission, Pongo
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