Pseudorabies virus (PRV) variants have caused substantial economic losses in the swine industry in China since 2011. To surveil the genetic variation in PRV field strains, here, two novel variant strains of PRV were isolated from Shanxi Province in central China and were designated SX1910 and SX1911. To identify the genetic characteristics of the two isolates, their complete genomes were sequenced, and phylogenetic analysis and sequence alignment revealed that field PRV variants have undergone genetic variations; notably, the protein-coding sequences UL5, UL36, US1 and IE180 exhibited extensive variation and contained one or more hypervariable regions. Furthermore, we also found that the glycoproteins gB and gD of the two isolates had some novel amino acid (aa) mutations. Importantly, most of these mutations were located on the surface of the protein molecule, according to protein structure model analysis. We constructed a mutant virus of SX1911 with deletion of the gE and gI genes via CRISPR/Cas9. When tested in mice, SX1911-ΔgE/gI-vaccinated mice were protected within a comparable range to Bartha-K61-vaccinated mice. Additionally, a higher dose of inactivated Bartha-K61 protected the mice from lethal SX1911 challenge, while a lower neutralization titer, higher viral load and more severe microscopic lesions were displayed in Bartha-K61-vaccinated mice. These findings highlight the need for continuous monitoring of PRV and novel vaccine development or vaccination program design for PRV control in China.

(1) The aim of this study was to elucidate the role of the Gallus gallus avian β-defensin 11 (AvBD11) in the immune response induced by the avian influenza virus H9N2. (2) AvBD11 was expressed using E. coli, and the effects of different concentrations of AvBD11 on cytokine expression in the ex vivo and in vivo erythrocytes of chickens infected with the avian influenza subtype H9N2 were detected by using fluorescence quantification. (3) The results showed that cytokine expression varied among the test groups compared to the control group in the in vitro assay at 2, 6, and 10 h. Lipopolysaccharide induced TNF factor (LITAF) and Interferon-γ (IFN-γ) were significantly increased in the AvBD11 group with the addition of the final concentration of 15 μg/mL at 6 h. At 10 h, Interleukin-1β (IL-1β) and IFN-γ were both more significantly increased in the 15 and 10 μg/mL groups than in the H9N2 group alone. In the in vivo test, IFN-γ and Interleukin-10 (IL-10) were significantly increased in the high-dose group than in the H9N2 group at 3 d and 7 d. (4) In conclusion, the ability of AvBD11 to induce the expression of more cytokines by chicken erythrocytes in a short period of time suggests that it is not only an antimicrobial peptide but also a possible immunomodulator.

PCR-based techniques play a crucial role in genotyping and genetic screening in fungal biology. Rapid access to nucleic acids for these reactions can significantly improve the efficiency of fungal analysis, especially when multiple samples need to be tested. In this study, we introduced a simple and rapid method for detecting small amounts of fungal DNA or RNA, named the One-Step method, and confirmed its applicability across various experimental scenarios for fungal detection. The method involves scraping a small quantity of spores or mycelium into sterile water, followed by heat shock, vortexing, and centrifugation to obtain a supernatant that serves as a template for the PCR reaction. Notably, nucleic acids were successfully extracted using the One-Step method from four different genera of fungi-Neurospora crassa, Aspergillus fumigatus, Fusarium oxysporum, and Schizosaccharomyces pombe, as well as from two mycovirus-containing strains, yielding reliable results in PCR identification. However, the nucleic acids of Cryptococcus neoformans were not successfully extracted using the One-Step method, possibly due to the challenge of cleaving its polysaccharide capsule. Taken together, the One-Step method significantly reduces nucleic acids extraction time while enhancing strain screening efficiency in four different fungi, indicating a broad applicability in fungal biology.

Parainfluenza virus 5 (PIV5), now reclassified as Orthorubulavirus mammalis, is a negative-sense, single-stranded RNA virus belonging to the genus Orthorubulavirus of the family Paramyxoviridae