Jeong Ye-Jin, Kim Kwang-Il
Department of Aquatic Life Medicine, Pukyong National University, Busan 48513, Republic of Korea.
Biology (Basel). 2023 Jun 7;12(6):829. doi: 10.3390/biology12060829.
The detection of megalocytiviruses, especially the infectious spleen and kidney necrosis virus (ISKNV), in ornamental fish has increased with the rapid growth of the ornamental fish industry. In this study, dwarf gourami fin (DGF) cells derived from the caudal fin of the dwarf gourami (), which is highly susceptible to red sea bream iridovirus (RSIV) and ISKNV, were established and characterized. The DGF cells were grown at temperatures ranging from 25 °C to 30 °C in Leibovitz's L-15 medium supplemented with 15% fetal bovine serum and were subcultured for more than 100 passages, predominantly with epithelial-like cells. DGF cells had a diploid chromosome number of 2 = 44. Although the initial purpose of this study was to establish a cell line for the causative agents of red sea bream iridoviral disease (RSIV and ISKNV), DGF cells were also susceptible to rhabdoviruses (viral hemorrhagic septicemia virus, hirame rhabdovirus, and spring viraemia of carp virus), exhibiting a significant cytopathic effect characterized by cell rounding and lysis. Additionally, viral replication and virion morphology were confirmed using virus-specific conventional polymerase chain reaction and transmission electron microscopy. Furthermore, both RSIV and ISKNV were replicated at high concentrations in DGF cells compared to other cell lines. Notably, the DGF cells maintained a monolayer during ISKNV infection, indicating the possibility of persistent infection. Thus, DGF can be used for viral diagnosis and may play a critical role in advancing our understanding of ISKNV pathogenesis.
随着观赏鱼产业的迅速发展，观赏鱼中巨细胞病毒尤其是传染性脾肾坏死病毒（ISKNV）的检出率有所增加。在本研究中，建立并鉴定了源自侏儒 Gourami（）尾鳍的侏儒 Gourami 鳍（DGF）细胞，该细胞对真鲷虹彩病毒（RSIV）和 ISKNV 高度敏感。DGF 细胞在补充有 15%胎牛血清的 Leibovitz's L - 15 培养基中于 25℃至 30℃的温度下生长，并传代培养超过 100 代，主要为上皮样细胞。DGF 细胞的二倍体染色体数为 2n = 44。尽管本研究的最初目的是建立用于真鲷虹彩病毒病病原体（RSIV 和 ISKNV）的细胞系，但 DGF 细胞也易受弹状病毒（病毒性出血性败血症病毒、牙鲆弹状病毒和鲤春病毒血症病毒）感染，表现出以细胞变圆和裂解为特征的明显细胞病变效应。此外，使用病毒特异性常规聚合酶链反应和透射电子显微镜确认了病毒复制和病毒粒子形态。此外，与其他细胞系相比，RSIV 和 ISKNV 在 DGF 细胞中均能高浓度复制。值得注意的是，DGF 细胞在 ISKNV 感染期间维持单层，表明存在持续感染的可能性。因此，DGF 可用于病毒诊断，并可能在推进我们对 ISKNV 发病机制的理解方面发挥关键作用。
