Cyprinid herpesvirus 2 (CyHV-2), a member of the Alloherpesviridae family belonging to the genus Cyprinivirus, is a fatal contagious aquatic pathogen that affects goldfish (Carassius auratus) and crucian carp (Carassius carassius). Although crucian carp and goldfish belong to the genus Carassius, it is unclear whether they are susceptible to the same CyHV-2 isolate. In addition, the origin of the crucian carp-derived CyHV-2 virus isolate remains unclear. CyHV-2 SH01 was isolated during herpesviral hematopoietic necrosis disease (HVHN) outbreaks in crucian carp at a local fish farm near Shanghai. CyHV-2 SH01 was confirmed by PCR and Western blot analysis of kidney, spleen, muscle, and blood tissue from the diseased crucian carp. Moreover, histopathological and ultra-pathological analyses revealed pathological changes characteristic of CyHV-2 SH01 infection in the tissues of the diseased crucian carp. In the present study, goldfish and crucian carp were challenged with CyHV-2 SH01 to elucidate viral virulence. We found that CyHV-2 SH01 could cause rapid and fatal disease progression in goldfish and crucian carp 24 h post-injection at 28 °C. Experimental infection of goldfish by injection indicated that the average virus titer in the kidney of the goldfish was 103.47 to 103.59 copies/mg. In addition, tissues exhibited the most prominent histopathological changes (cellular wrinkling and shrinkage, cytoplasmic vacuolation, fusion of the gill lamellae, and hepatic congestion) in CyHV-2 SH01-infected goldfish and crucian carp. Thus, crucian carp and goldfish showed a high sensitivity, with typical symptoms, to HVHN disease caused by CyHV-2 SH01.

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The freshwater crayfish Procambarus clarkii is native to North America and Mexico, and it was introduced to China in 1929. The production and consumption of P. clarkii in China are the highest worldwide, reaching 208.96 million tons in 2020. The white spot syndrome virus (WSSV) is a major pathogen that affects shrimp, crayfish, crabs and lobsters, and it has caused widespread loss to the P. clarkii industry. Epigallocatechin‐3‐gallate (EGCG), a small‐molecule compound, has a multitude of biological functions and the ability to bind to the 37 kDa/67 kDa laminin receptor (LamR). EGCG has potential antiviral effects against WSSV. In this study, we evaluated the potential anti‐WSSV applications of EGCG in P. clarkii. We demonstrated that various concentrations (10 μg/g·bw, 20 μg/g·bw and 40 μg/g·bw) of EGCG can suppress WSSV infection in P. clarkii. Histopathological examination revealed no characteristic pathological changes due to EGCG administration in P. clarkii tissues. Furthermore, pharmacokinetics studies of EGCG in P. clarkii revealed its rapid absorption (Tmax = 2 h), and the peak concentrations of EGCG were 73.78 µg/g in the liver and 24.87 µg/g in the muscle. Our results indicate the high potential applications of EGCG against WSSV in P. clarkii.

Background Cyprinid herpesvirus 2 (CyHV-2) is a pathogenic fish virus belonging to family Alloherpesviridae. The CyHV-2 gene encoding thymidine kinase (TK) is an important virulence-associated factor. Therefore, we aimed to investigate the biological function of open reading frame 55 (ORF55) in viral replication. Methods Purified CyHV-2 ORF55 protein was obtained by prokaryotic expression, and the interacting peptide was screened out using phage display. Host interacting proteins were then predicted and validated. Results ORF55 was efficiently expressed in the prokaryotic expression system. Protein and peptide interaction prediction and dot-blot overlay assay confirmed that peptides identified by phage display could interact with the ORF55 protein. Comparing the peptides to the National Center for Biotechnology Information database revealed four potential interacting proteins. Reverse transcription quantitative PCR results demonstrated high expression of an actin-binding Rho-activating protein in the latter stages of virus-infected cells, and molecular docking, cell transfection and coimmunoprecipitation experiments confirmed that it interacted with the ORF55 protein. Conclusion During viral infection, the ORF55 protein exerts its biological function through interactions with host proteins. The specific mechanisms remain to be further explored.