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Cyprinid herpesvirus 2 (CyHV-2) is the pathogen of herpesviral hematopoietic necrosis (HVHN), causing the severe economic losses in farmed gibel carp (Carassius gibelio). Further exploration of the genome structure and potential molecular pathogenesis of CyHV-2 through complete genome sequencing, comparative genomics, and molecular characterization is required. Herein, the genome of a CyHV-2 YC-01 strain isolated from diseased gibel carp collected in Yancheng, Jiangsu Province, China was sequenced, then we analyzed the genomic structure, genetic properties, and molecular characterization. First, the complete YC-01 genome comprises 275,367 bp without terminal repeat (TR) regions, with 151 potential open reading frames (ORFs). Second, compared with other representative published strains of the genus Cyvirus, several evident variations are found in YC-01, particularly the orientation and position of ORF25 and ORF25B. ORF107 and ORF156 are considered as potential molecular genetic markers for YC-01. ORF55 (encoding thymidine kinase) might be used to distinguish YC-01 and ST-J1 from other CyHV-2 isolates. Third, phylogenetically, YC-01 clusters with the members of the genus Cyvirus (together with the other six CyHV-2 isolates). Fourth, 43 putative proteins are predicted to be functional and are mainly divided into five categories. Several conserved motifs are found in nucleotide, amino acid, and promoter sequences including cis-acting elements identification of YC-01. Finally, the potential virulence factors and linear B cell epitopes of CyHV-2 are predicted to supply possibilities for designing novel vaccines rationally. Our results provide insights for further understanding genomic structure, genetic evolution, and potential molecular mechanisms of CyHV-2.

Crucian carp (Carassius auratus gibelio), an extensively cultivated freshwater fish, was one of the model species for the study of fish immunology. Polyclonal antibodies were advantageous molecular tools for studying teleost immune system. Specifically, polyclonal antibodies reacting with immunoglobulins (Ig) were used successfully in studies of the teleost fishes. In the present study, we produced polyclonal antibody against CH2 domains of crucian carp IgM, and measured the in vivo dynamics of IgM mRNA caused by CyHV-2 infection. The recombinant protein IgM with relative molecular weight about 53 KD was correctly expressed in prokaryotic cells. The specificity of the polyclonal antibody was evaluated by Western blotting and results revealed that the antibody not only specifically recognized crucian carp serum but also cross-reacted with grass carp serum. Quantitative RT-PCR analysis demonstrated the expression of IgM mRNA changed significantly after CyHV-2 infection. The expression of IgM in the kidney increased and reached a maximum at 6 h post-infection (hpi), while dropped to a low level at 5 days post-infection (dpi). In conclusion, the expression of IgM was significantly upregulated in the kidney of crucian carp infected with CyHV-2, indicating that IgM played a potential role in systemic immunity against viral infection. Polyclonal antibody against crucian carp IgM had certain clinical relevance, which might provide insight into the early stage of virus infection and prevention of the disease.

Hydrophobic interactions are widely applied in diverse fields, yet the effect of the degree of hydrophobicity on the interactions between hydrophobic surfaces is still unaddressed, limiting the development of functional hydrophobic surfaces. In this study, the interfacial interactions and configuration of hydrophobic chains on surfaces with ranging hydrophobicity are surveyed by surface forces apparatus. Our findings revealed that with partial hydrophobicity, a long-range electrical double-layer repulsive force pertains between surfaces, while between fully hydrophobized surfaces, only steric hindrance is observed, and the hydrophobic chains maintain configuration even after long time compression. Our findings provide useful implications for understanding hydrophobic interactions.

The mode and outcome of fish–virus interactions are influenced by many abiotic factors, among which water temperature is especially important in poikilothermic fish. Rare minnow Gobiocypris rarus is a eurythermal small cyprinid fish that is sensitive to infection with genotype II grass carp reovirus (GCRV). HSP70, a conservative and key player in heat shock response, is previously identified as an induced pro-viral factor during GCRV infection in vitro. Here, rare minnow was subjected to heat shock treatment (HST), 1 h treatment at 32 °C followed by reverting to a normal temperature of 24 °C, and subsequently challenged with GCRV-II at a dosage of 1 × LD50. The effect of HST on GCRV virulence in vivo was evaluated by calculating virus-associated mortality and viral load in both dead and survival fish. The results revealed that HST enhanced the mortality of rare minnow infected with GCRV; the fact that viral loads in the tissue samples of HST-treated fish were significantly higher than those in samples of the control group at 6, 8 d p.i. reflected a faster infection process due to HST. Quantitative gene expression analysis was further employed to show that the expression levels of Hsp70 in intestine and liver tissues from the HST group declined faster than muscle tissue after HST. HST W/O GCRV challenge upregulated proinflammatory cytokines such as MyD88 and Nf-κB, which was in consistence with the inflammation observed in histopathological analysis. This study shed light on the complexity of the interaction between fish abiotic and biotic stress response, which suggested that HST, an abiotic stress, could enhance the virulence of GCRV in Gobiocypris rarus that involved modulating the gene expression of host heat shock, as well as a pro-inflammatory response.

Hydration plays a crucial role in cement composites, but the traditional methods for measuring hydration heat face several limitations. In this study, we propose a machine learning-based approach to predict hydration heat at specific time points for three types of cement composites: ordinary Portland cement pastes, fly ash cement pastes, and fly ash–metakaolin cement composites. By adjusting the model architecture and analyzing the datasets, we demonstrate that the optimized artificial neural network model not only performs well during the learning process but also accurately predicts hydration heat for various cement composites from an extra dataset. This approach offers a more efficient way to measure hydration heat for cement composites, reducing the need for labor- and time-intensive sample preparation and testing. Furthermore, it opens up possibilities for applying similar machine learning approaches to predict other properties of cement composites, contributing to efficient cement research and production.

Cyprinid herpesvirus 2 (CyHV-2), a member of the Alloherpesviridae family belonging to the genus Cyprinivirus, was initially isolated from goldfish (Carassius auratus) and has been recently emerging as a virulent pathogen for cultured prussian carp (Carassius auratus gibelio) world-wide. In this study, a novel and effective method for concentration and quantification of live CyHV-2 virions from water was successfully established through coupling the iron flocculation with real time qPCR assay. Then, the shedding intensity of CyHV-2 in fish-tank water from artificially-challenged goldfish (25 fish/20 L) was monitored continuously for 7 days on a daily basis through quantitating viral genomic copy numbers by qPCR, and the maximum shedding level was determined to be 105 copies/L. Horizontal transmission research system was established by inoculating healthy goldfish in water spiked with serial dilution of CyHV-2 virions ranging from 107 to 10³ copies/L. Our results indicated that water-borne CyHV-2 efficiently caused the infection of tested goldfish even in a concentration of 10³ copies/L, and the overall transmission efficacy was not linearly correlated with the level of input virus in the fish tank. Commercial disinfectant Composite Sodium Chlorite Powder (CSCP) has been widely applied in aquaculture to control microbial infection through direct spill in the water, and its effect in inactivating the CyHV-2 infectivity remains unknown. We further determined that the EC50 of CSCP against 3.89 TCID50/mL CyHV-2 was close to 15.625 µg/mL in vitro, and application of CSCP in a level as high as 60 µg/mL (the safety concentration of CSCP for goldfish) couldn't protect goldfish from CyHV-2 challenge through immersion. Thus, the disinfectant CSCP was regarded as none-effective for blocking CyHV-2 transmission in water during epidemic. Overall, our data provided quantitative data to demonstrate the shedding intensity of CyHV-2 in water, and CSCP was shown to be not effective in blocking water-borne horizontal transmission of CyHV-2 in goldfish. The virus-concentration protocol and virus-inhibition assay established here also paved the way for evaluating more commercial disinfectants in their effects in blocking water-borne horizontal transmission of CyHV-2. [ABSTRACT FROM AUTHOR]