Susceptibility of betanodavirus in a newly established vertebra‐derived cell line from Mosquitofish (Gambusia affinis)

Abstract

In the present study, a new cell line from the vertebra of mosquitofish Gambusia affinis was successfully established and characterized. The cell line is named as bone Gambusia affinis (BGA) and subcultured for more than 55 passages in Leibovitz's/L15 medium supplemented with 15% FBS at 28°C. The cell line has a modal chromosome number of 48. Molecular characterization of the partial sequence of the coi gene confirmed the origin of the BGA cell line from mosquitofish. These cells exhibited epithelial morphology confirmed by the cytokeratin marker. The BGA cells showed mineralization of their extracellular matrix when stained with alizarin red and von Kossa stain. BGA cells were found to be susceptible to RGNNV and SJNNV strains of betanodavirus (NNV) showing cytopathic effect with multiple vacuolations in the cells. The RT-PCR confirmed the betanodavirus infections in BGA cells. The SEM micrograph showed the morphological changes observed in the cell during virus infection. The in vivo challenge experiment also showed the viral replicating efficiency in the Gambusia affinis with increasing viral titre. Thus, our present results show that the BGA cell line is a useful tool for isolating betanodavirus and could be used to investigate bone cell differentiation and extracellular matrix mineralization. K E Y W O R D S betanodavirus, cell line, Gambusia affinis, mineralization, vertebra S U PP O RTI N G I N FO R M ATI O N Additional supporting information may be found online in the Supporting Information section. How to cite this article: Jyotsna, Vijayakumar P, Mani R, Raja S, Dhas TS. Susceptibility of betanodavirus in a newly established vertebra-derived cell line from Mosquitofish (Gambusia affinis).

Establishment and characterization of a liver cell line, ALL, derived from yellowfin sea bream, Acanthopagrus latus, and its application to fish virology

“…So far, Epithelioma papulosum cyprini , BF‐2, rainbow trout ( Oncorhynchus mykiss ) gonad and rainbow trout gill cell lines have been reported to be susceptible to VHSV (Gorgoglione et al., 2020). Although many cell lines derived from various fishes were susceptible to RGNNV (Jyotsna et al., 2020; Liu et al., 2018; Mai et al., 2018; Tu et al., 2016; Zhang, Jia et al., 2019), no fish cell line originated from A. latus was generated. Our results show that ALL cell line was susceptible to these three fish viruses, indicating that ALL cell line is an ideal in vitro platform for fish virology study.…”