Adamantoyl chloride inhibited replication of the largemouth bass virus via enhanced immunity and inhibition of apoptosis

Abstract

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 (IC50 = 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.