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In vitro isolation and molecular identification of reptarenavirus in Malaysia

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Abstract

Boid inclusion body disease (BIBD) is a viral disease of boids caused by reptarenavirus. In this study, tissue from naturally infected boid snakes were homogenized and propagated in African Monkey kidney (Vero) and rat embryonic fibroblast (REF) cells. Virus replication was determined by the presence of cytopathic effect, while viral morphology was observed using transmission electron microscopy. Viral RNA was amplified using RT-PCR with primers specific for the L-segment of reptarenavirus; similarly, quantification of viral replication was done using qPCR at 24–144 h postinfection. Viral cytopathology was characterized by cell rounding and detachment in both Vero and REF cells. The viral morphology showed round-to-pleomorphic particles ranging from 105 to 150 nm which had sand-like granules. Sanger sequencing identified four closely associated reptarenavirus species from 15 (37.5 %) of the total samples tested, and these were named as follows: reptarenavirus UPM-MY 01, 02, 03, and 04. These isolates were phylogenetically closely related to the University Helsinki virus (UHV), Boa Arenavirus NL (ROUTV; BAV), and unidentified reptarenavirus L20 (URAV-L20). Comparison of deduced amino acid sequences further confirmed identities to L-protein of UHV, L-polymerase of BAV and RNA-dependent RNA polymerase of URAV-L20. Viral replication in Vero cells increased steadily from 24 to 72 h and peaked at 144 h. This is the first study in South East Asia to isolate and characterize reptarenavirus in boid snakes with BIBD.

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Acknowledgments

The authors are grateful for the funding received under the Universiti Putra Malaysia Postgraduate Grant Scheme (GP/IPS/2014/9438740) and the Ministry of Higher Education Malaysia, Fundamental Research Grant Scheme (FRGS/2014/STWN03/UPM/01/2).

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All authors contributed equally to this work and have read and approved the final manuscript.

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Authors and Affiliations

  1. Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Yusuf Abba, Hazilawati Hamzah, Yusuf Ilyasu, Faruku Bande, Mohd Azmi Mohd Lila & Mohamed Mustapha Noordin

  2. Department of Veterinary Preclinical Sciences, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Yusuf Abba & Yusuf Ilyasu

  3. Department of Veterinary Pathology, Faculty of Veterinary Medicine, University of Maiduguri, PMB 1069, Maiduguri, 600233, Borno, Nigeria

    Hasliza Hassim

  4. Department of Pathology, Faculty of Dentistry, Universiti Teknologi Mara, 47000, Sungai Buloh, Selangor Darul Ehsan, Malaysia

    Omar Emad Ibrahim

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  1. Yusuf Abba
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  2. Hasliza Hassim
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  3. Hazilawati Hamzah
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  4. Omar Emad Ibrahim
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  5. Yusuf Ilyasu
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  6. Faruku Bande
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  7. Mohd Azmi Mohd Lila
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  8. Mohamed Mustapha Noordin
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Corresponding authors

Correspondence to Yusuf Abba or Mohamed Mustapha Noordin.

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The authors declare no conflict of interest regarding this work.

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This article does not contain any studies performed on live animals by any of the authors. Snakes used for the study were carcasses brought in dead (BID) to the necropsy unit.

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Edited by Zhen F. Fu.

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Abba, Y., Hassim, H., Hamzah, H. et al. In vitro isolation and molecular identification of reptarenavirus in Malaysia. Virus Genes 52, 640–650 (2016). https://doi.org/10.1007/s11262-016-1345-7

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  • DOI: https://doi.org/10.1007/s11262-016-1345-7

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