Mycotoxin cocontamination is a severe threat to health and economic security worldwide. The mycotoxins aflatoxin B₁ (AFB₁), zearalenone (ZEN), deoxynivalenol, T-2 toxin, fumonisin B₁, and ochratoxin A are of particular concern due to their substantial toxicity. Lac-W is a laccase with the unique property of degrading these six mycotoxins in the absence of redox mediators. Nevertheless, their degradation rates are low. This work aims to improve the ability of Lac-W to degrade these six mycotoxins and to elucidate its detoxification mechanism. Including redox mediators increased the Lac-W degradation efficiency drastically, and completely degraded AFB₁ and ZEN within one hour. Additionally, Lac-W-AS has good temperature, pH, and ions adaptability in ZEN degradation. Lac-W-AS reduced the ZEN toxicity because ZEN degradation products significantly restored the bioluminescence intensity of Vibrio fischeri. A Lac-W-AS-mediated oxidation product of ZEN was structurally characterized as 15-OH-ZEN by UHPLC-MS/MS. Linear sweep voltammetry showed that AS affected the potential of Lac-W and accelerated the oxidation of ZEN. Finally, the combination of mediators (acetosyringone and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonate)) improved the degradation rate of mycotoxins. This work highlights that the combination of Lac-W with mediators serves as a good candidate for degrading multi-mycotoxin contaminants in food and feedstuff.

The porcine pseudorabies virus (PRV) is one of the most devastating pathogens and brings great economic losses to the swine industry worldwide. Viruses are intracellular parasites that have evolved numerous strategies to subvert and utilize different host processes for their life cycle. Among the different systems of the host cell, the cytoskeleton is one of the most important which not only facilitate viral invasion and spread into neighboring cells, but also help viruses to evade the host immune system. RhoA is a key regulator of cytoskeleton system that may participate in virus infection. In this study, we characterized the function of RhoA in the PRV replication by chemical drugs treatment, gene knockdown and gene over-expression strategy. Inhibition of RhoA by specific inhibitor and gene knockdown promoted PRV proliferation. On the contrary, overexpression of RhoA or activation of RhoA by chemical drug inhibited PRV infection. Besides, our data demonstrated that PRV infection induced the disruption of actin stress fiber, which was consistent with previous report. In turn, the actin specific inhibitor cytochalasin D markedly disrupted the normal fibrous structure of intracellular actin cytoskeleton and decreased the PRV replication, suggesting that actin cytoskeleton polymerization contributed to PRV replication in vitro. In summary, our data displayed that RhoA was a host restriction factor that inhibited PRV replication, which may deepen our understanding the pathogenesis of PRV and provide further insight into the prevention of PRV infection and the development of anti-viral drugs.

Pseudorabies virus (PRV), a highly pathogenic alphaherpesvirus, poses a potential threat to public health and safety due to its broad host range and risk of cross-species transmission. Viruses have evolved multiple strategies to exploit host factors for entry into and survival in host cells. Drebrin is an actin-binding protein that restricts rotavirus entry by inhibiting dynamin-mediated endocytosis. However, its role and mechanism in DNA virus infection, particularly in herpesviruses, remain unexplored. In this study, we investigated the role of Drebrin in PRV infection using pharmacological inhibition (BTP−2) and CRISPR-Cas9-mediated gene knockout. Both the Drebrin inhibitor BTP−2 and gene knockout significantly suppressed PRV replication. Intriguingly, Drebrin exhibited stage-specific effects on the viral life cycle: its inhibition enhanced viral internalization during early infection but impaired viral replication at later stages, suggesting that Drebrin plays a complex role in the regulation of PRV infection. PRV infection partially disrupted actin stress fibers and caused an increase in cell size. Drebrin knockout also altered the host-cell morphology, reduced the cell surface area, and induced actin cytoskeleton rearrangement, which was further modulated in PRV-infected cells. In summary, our data demonstrate that Drebrin functions as a critical host factor governing the entire PRV life cycle by regulating actin cytoskeleton reorganization.

This study successfully isolated a novel bovine enterovirus strain from a bovine fecal sample, which was designated as Sichuan/SQ/20. The isolate showed typical enterovirus morphology under electron microscopy. Phylogenetic analysis showed that this strain exhibits the closest genetic relationship with the HeN-YR91 and JPN/TottoriU-31 strains, and all three belong to the BEV-F1 genosubtype. Subsequently, comprehensive investigations were conducted on the biological characteristics of this virus, both in vivo and in vitro . In vitro characterization revealed that viral replication commenced at 3 h post-infection (hpi) in Madin-Darby bovine kidney cells, reaching peak at 48 hpi with a virus titer of 1 × 10 ^8.73 TCID ₅₀ /0.1 mL. Cytopathic effects initially appeared at 12 hpi. A 12-minute treatment at 55°C was sufficient to completely inactivate the virus. In vivo analysis revealed that significant pathological changes were specifically observed in the spleen, with no lesions observed in other organs. Immunofluorescence assay detected specific fluorescent signals in the liver, spleen, and small intestine, which were consistent with the PCR results. These findings provide a scientific foundation for vaccine design and antiviral drug screening, as well as for the development of effective prevention and control strategies. IMPORTANCE: Bovine enterovirus (BEV) is an important pathogen causing calf diarrhea and has been detected in the feces of calves with diarrhea, although its pathogenicity remains unclear. This study systematically established an isolation and identification protocol for BEV, characterized its physicochemical properties, and further investigated the pathogenicity and tissue tropism of the isolated strain in mice. These findings establish crucial baseline data for future vaccine development and therapeutic intervention strategies.