Primary cell culture of zebrafish (Danio rerio) as a material for developing H5N1 avian influenza vaccines

In recent decades, zebrafish (Danio rerio) has been widely used as a model of biological experiments not only in the immunology and oncology aspects but also experimental animals for infectious diseases in human and fishes. The virus used in this study was the H5N1 avian influenza reverse genetic virus obtained from the Professor Nidom Foundation, Surabaya, Indonesia. We revealed that TCID50 was 3.1623*10⁷/mL, then the virus was developed by inoculating into the primary cell culture and we performed HA test. Samples that showed positive results on the HA test then performed RNA extraction and confirmed by one-step RT-PCR. It can be seen from the presence of cytopathogenic effects (CPE) in primary cell cultures and also can be seen from HA titers in viral harvest samples from the primary cells of zebrafish (Danio rerio) that shows the same result of HA titers is 2⁸. In conclusion, the present study demonstrated that there is an alternative development of new primary cell sources which leads to halal aspects of the production of avian influenza H5N1 vaccines.