College of Animal Science and Technology, Northwest A & F University, Yangling, Shaanxi, 712100, China.
Guangxi Key Laboratory of Aquatic Biotechnology and Modern Ecological Aquaculture, Guangxi Academy of Sciences, Nanning, China.
Fish Shellfish Immunol. 2023 Nov;142:109167. doi: 10.1016/j.fsi.2023.109167. Epub 2023 Oct 15.
The largemouth bass virus (LMBV) is a commonly encountered pathogen in aquaculture and presents significant challenges to development of the largemouth bass industry due to the lack of effective treatment methods. Here, the inhibitory potential and underlying mechanisms of adamantoyl chloride (AdCl) against LMBV were assessed both in vitro and in vivo. The results showed that AdCl (IC = 72.35 μM) significantly inhibited replication of LMBV in epithelioma papulosum cyprini (EPC) cells. The results of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide and cytopathic effect (CPE) assays confirmed that AdCl inhibited replication of LMBV in EPC cells and significantly reduced the CPE effect, respectively. As a potential mechanism, AdCl inhibited apoptosis as determined by fluorescence and transmission electron microscopy. The results of flow cytometry showed that the apoptosis rate was decreased by 69 % in the AdCl-treated group as compared to the LMBV-infected group. Additionally, AdCl inhibited viral release. In vivo, the survival rate was 16.2 % higher in the AdCl-treated group as compared to the LMBV-infected group (26.9 % vs. 10.7 %, respectively). Additionally, the results of quantitative reverse transcription polymerase chain reaction (RT-qPCR) showed that AdCl significantly reduced the viral load of the fish liver, spleen, and kidneys at 3, 6, and 9 days postinfection. In addition, RT-qPCR analysis found that AdCl upregulated expression of immune-related genes to suppress replication of LMBV. Collectively, these results confirmed the anti-LMBV activities of AdCl for use in the aquaculture industry.
大口黑鲈病毒（LMBV）是水产养殖中常见的病原体，由于缺乏有效的治疗方法，给大口黑鲈产业的发展带来了巨大挑战。本研究评估了金刚烷氯（AdCl）在体外和体内对 LMBV 的抑制潜力和潜在机制。结果表明，AdCl（IC=72.35μM）显著抑制了鲤鱼上皮瘤细胞（EPC）中 LMBV 的复制。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐（MTT）和细胞病变效应（CPE）试验结果证实，AdCl 抑制了 EPC 细胞中 LMBV 的复制，并分别显著降低了 CPE 效应。作为一种潜在的机制，AdCl 通过荧光和透射电子显微镜抑制了细胞凋亡。流式细胞术结果表明，与 LMBV 感染组相比，AdCl 处理组的细胞凋亡率降低了 69%。此外，AdCl 抑制了病毒的释放。在体内，AdCl 处理组的存活率比 LMBV 感染组高 16.2%（分别为 26.9%和 10.7%）。此外，定量逆转录聚合酶链反应（RT-qPCR）结果表明，AdCl 显著降低了感染后 3、6 和 9 天鱼肝脏、脾脏和肾脏的病毒载量。此外，RT-qPCR 分析发现，AdCl 上调了免疫相关基因的表达，以抑制 LMBV 的复制。综上所述，这些结果证实了 AdCl 对水产养殖行业具有抗 LMBV 活性。
