Zebrafish (Danio rerio) larvae as a model for real‐time studies of propagating VHS virus infection, tissue tropism and neutrophil activity

Moonika Haahr Marana, Jacob Günther Schmidt, Stéphane Biacchesi, Niels Lorenzen, Louise von Gersdorff Jørgensen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review


Viral haemorrhagic septicaemia virus (VHSV) is a negative‐sense single‐stranded RNA virus that infects more than 140 different fish species. In this study, zebrafish larvae were employed as in vivo model organisms to investigate progression of disease, the correlation between propagation of the infection and irreversibility of disease, cell tropism and in situ neutrophil activity towards the VHSV‐infected cells. A recombinant VHSV strain, encoding “tomato” fluorescence (rVHSV‐Tomato), was used in zebrafish to be able to follow the progress of the infection in the live host in real‐time. Two‐day‐old zebrafish larvae were injected into the yolk sac with the recombinant virus. The virus titre peaked 96 hr post‐infection in zebrafish larvae kept at 18°C, and correlated with 33% mortality and high morbidity among the larvae. By utilizing the transgenic zebrafish line Tg(fli1:GFP)Ψ1 with fluorescently tagged endothelial cells, we were able to demonstrate that the virus initially infected endothelial cells lining the blood vessels. By observing the rVHSV‐Tomato infection in the neutrophil reporter zebrafish line Tg(MPX:eGFP)i114 , we inferred that only a subpopulation of the neutrophils responded to the virus infection. We conclude that the zebrafish larvae are suitable for real‐time studies of VHS virus infections, allowing in vivo dissection of host–virus interactions at the whole organism level.
Original languageEnglish
JournalJournal of Fish Diseases
Issue number5
Pages (from-to)563-571
Number of pages9
Publication statusPublished - 2021


  • In vivo visualization
  • Infection kinetics
  • Mortality
  • Neutrophil response
  • Tissue tropism
  • Viral haemorrhagic septicaemia virus
  • Zebrafish


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