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Diseases of Aquatic Organisms

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DAO 142:1-11 (2020)  -  DOI: https://doi.org/10.3354/dao03528

Betanodavirus infection in Kuhlia rupestris and Ambassis marianus and isolation of betanodavirus from infected pond water

Kalpana Agnihotri1,*, Roger Chong1,2, Darren Underwood1, Carol Kistler1, Michael Hutchison3

1Biosecurity Sciences Laboratory, Biosecurity Queensland, Department of Agriculture and Fisheries, Queensland Government, Block 12, Health and Food Sciences Precinct 39, Coopers Plains, Queensland 4108, Australia
2Commonwealth Scientific and Industrial Research Organisation (CSIRO) Queensland Bioscience Precinct, 306 Carmody Road, St Lucia, Queensland 4067, Australia
3Bribie Island Research Centre, Department of Agriculture and Fisheries, Queensland Government, 144 North Street, Woorim, Queensland 4507, Australia
*Corresponding author:

ABSTRACT: This is the first report of betanodavirus infection in 2 species of finfish, Kuhlia rupestris (jungle perch) and Ambassis marianus (estuary perchlet). This report also describes isolation of betanodavirus from infected pond water using the SSN-1 cell line. Histopathology of K. rupestris larvae revealed vacuolation in the eye and brain, which was confirmed using betanodavirus-specific immunohistochemistry. The eye and brain from A. marianus and betanodavirus isolated from pond water were confirmed using real-time PCR and Sanger sequencing. High throughput sequencing was used to obtain betanodavirus sequences from paraffin blocks containing infected K. rupestris. The phylogenetic analysis of betanodavirus RNA1 and RNA2 sequences from all 3 sources were associated with the red-spotted grouper nervous necrosis virus (RGNNV) genotype. The RNA1 nucleotide sequence from jungle perch showed 100% identity with the betanodavirus water isolate and 99.37% identity with A. marianus. Furthermore, we have demonstrated the usefulness of combining recovery of viable virus from environmental samples through fish cell line culture with PCR testing as a means of validating the efficacy of chlorination to eradicate betanodavirus from the pond environment.


KEY WORDS: Kuhlia rupestris · Ambassis marianus · Betanodavirus · High throughput sequencing · Phylogeny


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Cite this article as: Agnihotri K, Chong R, Underwood D, Kistler C, Hutchison M (2020) Betanodavirus infection in Kuhlia rupestris and Ambassis marianus and isolation of betanodavirus from infected pond water. Dis Aquat Org 142:1-11. https://doi.org/10.3354/dao03528

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