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Molecular evidence for homologous strains of infectious spleen and kidney necrosis virus (ISKNV) genotype I infecting inland freshwater cultured Asian sea bass (Lates calcarifer) in Thailand

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Abstract

Infectious spleen and kidney necrosis virus (ISKNV) is a fish-pathogenic virus belonging to the genus Megalocytivirus of the family Iridoviridae. In 2018, disease occurrences (40-50% cumulative mortality) associated with ISKNV infection were reported in grown-out Asian sea bass (Lates calcarifer) cultured in an inland freshwater system in Thailand. Clinical samples were collected from seven distinct farms located in the eastern and central regions of Thailand. The moribund fish showed various abnormal signs, including lethargy, pale gills, darkened body, and skin hemorrhage, while hypertrophied basophilic cells were observed microscopically in gill, liver, and kidney tissue. ISKNV infection was confirmed on six out of seven farms using virus-specific semi-nested PCR. The MCP and ATPase genes showed 100% sequence identity among the virus isolates, and the virus was found to belong to the ISKNV genotype I clade. Koch’s postulates were later confirmed by challenge assay, and the mortality of the experimentally infected fish at 21 days post-challenge was 50-90%, depending on the challenge dose. The complete genome of two ISKNV isolates, namely KU1 and KU2, was recovered directly from the infected specimens using a shotgun metagenomics approach. The genome length of ISKNV KU1 and KU2 was 111,487 and 111,610 bp, respectively. In comparison to closely related ISKNV strains, KU1 and KU2 contained nine unique genes, including a caspase-recruitment-domain-containing protein that is potentially involved in inhibition of apoptosis. Collectively, this study indicated that inland cultured Asian sea bass are infected by homologous ISKNV strains. This indicates that ISKNV genotype I should be prioritized for future vaccine research.

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Acknowledgments

This research was supported by funding from the Center of Excellence on Agricultural Biotechnology, Science and Technology Postgraduate Education and Research Development Office, Office of Higher Education Commission, Ministry of Education (AG-BIO/PERDO-CHE; grant ID. AG-BIO/61-003-010) and Thailand Research Fund under the Research Grant for New Scholar program (grant no. MRG6180054). The authors acknowledge the Center of Excellence for Shrimp Molecular Biology and Biotechnology (CENTEX Shrimp), Mahidol University, for providing technical support.

Funding

This research was supported by funding from the Center of Excellence on Agricultural Biotechnology, Science and Technology Postgraduate Education and Research Development Office, Office of Higher Education Commission, Ministry of Education (AG-BIO/PERDO-CHE; grant ID. AG-BIO/61-003-010) and Thailand Research Fund under the Research Grant for New Scholar program (grant no. MRG6180054).

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Authors and Affiliations

  1. Center for Agricultural Biotechnology, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 73140, Thailand

    Pattarawit Kerddee

  2. Center of Excellence on Agricultural Biotechnology: (AG-BIO/PERDO-CHE), Bangkok, 10900, Thailand

    Pattarawit Kerddee

  3. Fish Infectious Diseases Research Unit (FID RU), Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Nguyen Dinh‐Hung

  4. Department of Food, Agriculture and Bioresources, School of Environment, Resources and Development, Asian Institute of Technology, Pathum Thani, 12120, Thailand

    Ha Thanh Dong

  5. Department of Marine Biosciences, Tokyo University of Marine Science and Technology, Konan, Minato, 4-5-7, Tokyo, 108-8477, Japan

    Ikuo Hirono

  6. Department of Aquaculture, Faculty of Fisheries, Kasetsart University, Bangkok, 10900, Thailand

    Chayanit Soontara, Nontawith Areechon, Prapansak Srisapoome & Pattanapon Kayansamruaj

  7. Center of Excellence in Aquatic Animal Health Management, Faculty of Fisheries, Kasetsart University, Bangkok, 10900, Thailand

    Prapansak Srisapoome & Pattanapon Kayansamruaj

Authors
  1. Pattarawit Kerddee
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  2. Nguyen Dinh‐Hung
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  3. Ha Thanh Dong
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  7. Prapansak Srisapoome
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Contributions

Pattarawit Kerddee: Carried out the experiment, wrote the manuscript. Nguyen Dinh‐Hung: Performed analysis (disease diagnosis). Ha Thanh Dong: Performed the analysis (histopathology, cell culture and qPCR). Ikuo Hirono: Contributed analysis tool (GF cell line). Chayanit Soontara: Performed analysis (disease diagnosis and experimental challenge). Nontawith Areechon: Conceived of overall direction and planning. Prapansak Srisapoome: Collected the samples from natural outbreaks. Pattanapon Kayansamruaj: Designed experiment, wrote the manuscript.

Corresponding author

Correspondence to Pattanapon Kayansamruaj.

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The animal use protocol followed in this study was approved by the Institutional Animal Care and Use Committee, Faculty of Fisheries, Kasetsart University (permit ID: ACKU61-FIS-055).

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Kerddee, P., Dinh‐Hung, N., Dong, H.T. et al. Molecular evidence for homologous strains of infectious spleen and kidney necrosis virus (ISKNV) genotype I infecting inland freshwater cultured Asian sea bass (Lates calcarifer) in Thailand. Arch Virol 166, 3061–3074 (2021). https://doi.org/10.1007/s00705-021-05207-7

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  • DOI: https://doi.org/10.1007/s00705-021-05207-7

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