Louis, MO, USA) at the dose rate of 150 mg/L of water and the caudal fin was excised after wiping with 70% ethanol. The goldfish was anesthetized using 3- aminobenzoic acid ethyl ester methanesulphonate (MS-222, Sigma-Aldrich, St. The fish was fed daily and water changes were performed on alternate days. However, no cell line from goldfish is available in India, therefore, the need of establishing a homologous cell line from goldfish was deemed necessary for the continuous propagation and study of CyHV-2.Ī healthy fantail goldfish weighing 20 g was purchased from a local commercial aquarium shop and kept in properly aerated and filtered fish tank. More than 15 cell lines have been developed globally from different goldfish tissues since 1964 ( Table 1). It has been reported that cell lines derived from homologous species provide sensitive tools for virus propagation ( Fryer & Lannan, 1994) and successful cultivation of CyHV-2 has been reported in cell lines developed from goldfish, namely GFTF ( Yan, Nie & Lu, 2011), GFF and SRTF ( Ito et al., 2013) and GiCB ( Ma et al., 2015). In an earlier study from India, the virus could not be propagated beyond 4th passage, despite producing cytopathic effects in CCKF cell line ( Sahoo et al., 2016). Earlier, a number of cell lines from other fish species, namely Epithelioma papulosum cyprini cells (EPC), fathead minnow cells (FHM) and koi fin cell lines (KF-1) have been used for CyHV-2 cultivation, but these cell lines did not support its continuous propagation beyond 5th passage ( Jeffery et al., 2007 Wang et al., 2012). As the surviving fish become lifelong carriers of latent infection without exhibiting any clinical signs of the disease ( Wang et al., 2012) and goldfish are one of the most traded ornamental fish globally, the international trade of such apparently healthy but latently infected goldfish is suspected to be responsible for the global spread of CyHV-2 ( Ito et al., 2013 Adamek et al., 2018).Ĭell lines are considered the gold standard for isolation and identification of viruses ( Landry, 2009). In India, CyHV-2 infection was associated with large-scale mortalities of goldfish in West Bengal ( Sahoo et al., 2016). The pathogen first reported as the cause of outbreaks in juvenile goldfish in 1992 from Japan ( Jung & Miyazaki, 1995), has subsequently been reported from more than 13 countries ( Adamek et al., 2018). Cyprinid herpesvirus-2 (CyHV-2), the etiological agent of herpesviral hematopoietic necrosis (HVHN) disease, is a pathogen of goldfish that has been associated with disease outbreaks in goldfish culture ( Jeffery et al., 2007).
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