Title:In vitro validation of camphene as a potential antiviral agent against betanodavirus causing viral nervous necrosis in barramundi
VOLUME: 19
Author(s):Ruby Singh, Prachi Srivastava*, Deepika Anand and Pani K. Prasad
Affiliation:Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow-227105,, Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow-227105,, ICAR- Central Institute of Fisheries Education, Versova, Mumbai, Maharashtra-400061,, ICAR- Central Institute of Fisheries Education, Versova, Mumbai, Maharashtra-400061
Keywords:Viral nervous necrosis, Barramundi, Phytochemical, Molecular docking, Molecular Dynamic simulation,
Camphene.
Abstract:Introduction: Viral infections are major threat to aquaculture industry throughout the world. Betanodavirus is
one of the most infectious virus that causes highest mortality in larval and juvenile stages of Lates calcarifer commonly
known as Barramundi.
Methods: It is a single stranded positive sense RNA virus and causes viral nervous necrosis (VNN).
VNN is caused by RNA virus which gets transmitted both horizontally and vertically so the most effective method against
this virus is to vaccinate the fish, however vaccination becomes difficult since the disease is associated with the outbreaks
in larval and juvenile stages which are not that much immunocompetent. In our previous in silico studies we proved the
stability of camphene as a better phytochemical agent.
Results: In continuation to prove the authenticity of Camphene as
potential antiviral agent against betanodavirus, its in vitro validation was performed. Sea bass kidney cell line (SISK) was
selected for carrying out the in vitro studies and cytotoxicity studies of Camphene in the SISK was was done by MTT
assay. Based on the analysis of MTT assay different dosage of camphene were selected viz.,(0.2, 0.5, 1, 1.5, 2, 2.5, 5, 10,
20, 30 μg/ml). The SISK cells were infected with virus inoculum (200μl). Further the antiviral activity of Camphene on
infected SISK cells by Betanodavirus was elucidated with the help of quantitative Real time PCR(qPCR) on the 3rd and 5th
day of infection.
Conclusion: Analysis of results depictecd that the dose of camphene 2 to 10 μg/ml is the safest dose
against Betanodavirus. Hence this is aptly revealed that camphene can be used as potential antiviral agent against
Betanodavirus.
