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In vitro propagation of infectious hypodermal hematopoietic necrosis virus (Penaeus stylirostris penstyldensovirus) in PmLyO-Sf9 cells

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Abstract

Infectious hypodermal hematopoietic necrosis virus (IHHNV/PstDVI) was isolated and propagated in the hybrid shrimp-insect cell line PmLyO-Sf9. A few hours after inoculation with an infected tissue extract or virus suspension, cytopathic changes could be observed in the cell line, including clustering, enlargement, syncytium formation, granulation, vacuole formation, tapering, irregularities in the plasma membrane with extended tails, detachment, cell death, and accumulation of cellular debris. Expression of viral genes, the presence of virions, and cytological changes observed using transmission electron microscopy suggested replication of the virus in these cells. The virus was purified by ultracentrifugation, negatively stained, and examined using an electron microscope, and the purified virus was found to be infectious both in vitro and in vivo. This development opens avenues for the study of the basic molecular mechanism of IHHNV infection, pathogenesis, and replication, which is much needed for developing an antiviral strategy in aquaculture.

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All data supporting the findings of this study are available within the paper and its supplementary data.

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Acknowledgements

This research was undertaken with financial support from the Department of Biotechnology, Government of India (project no. Ref: BT/PR12660/AAQ/3/710/2014 dated 11.7.2017). We acknowledge the Sophisticated Analytical Instrumentation Facility (SAIF), All India Institute of Medical Sciences, New Delhi, for electron microscope support.

Funding

This research was undertaken with financial support from the Department of Biotechnology, Government of India (project no. Ref: BT/PR12660/AAQ/3/710/2014 dated 11.7.2017).

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Authors and Affiliations

  1. National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Kochi, India

    Dominic Vithayathil Diya Dominic, Thavarool Puthiyedathu Sajeevan, Ahna Ameer, Abdulkhader Muneer, Linu Eldho, Bhaskaran Sathyabhama Anoop, Puthumana Jayesh, Valsamma Joseph & Isaac Sarojini Bright Singh

  2. Department of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology, Kochi, India

    Rosamma Philip

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  1. Dominic Vithayathil Diya Dominic
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Contributions

DD: Carried out the practical work and prepared the original draft manuscript. STP: Principal Investigator of the project in which the work was done. AA, MA, LE, and ABS: Assisted the first author in executing the work. JP: Validation and editing. VJ: Assisted the corresponding author. RP: Co-investigator and supervision. ISBS: Conceptualized the work, supervised, and prepared the manuscript in the final form.

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Correspondence to Isaac Sarojini Bright Singh.

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The authors declare no competing interests.

Compliance with ethical standards

Human subjects were not used in this study. Shrimp postlarvae and adults were either collected from farms or maintained under controlled conditions following standard protocols for the maintenance of crustaceans. All procedures involving shrimp were in accordance with the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA).

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Dominic, D.V.D., Sajeevan, T.P., Ameer, A. et al. In vitro propagation of infectious hypodermal hematopoietic necrosis virus (Penaeus stylirostris penstyldensovirus) in PmLyO-Sf9 cells. Arch Virol 168, 211 (2023). https://doi.org/10.1007/s00705-023-05827-1

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