Duck Tembusu virus (DTMUV) is an emerging flavivirus that has inflicted significant economic losses on China’s poultry industry. Rapid and accurate detection of DTMUV is crucial for effective prevention and control measures. In this study, we developed a novel, rapid visual detection assay that combines reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) with the CRISPR/Cas12a system for on-site detection of DTMUV. Our results demonstrate that this assay can sensitively and specifically detect the specific DNA plasmids containing the DTMUV NS3 gene within 100 min, with a limit of detection as low as 19.3 copies/μL. We successfully applied the RT-LAMP-CRISPR/Cas12a assay to diagnose DTMUV in eight duck embryos and 11 chicken embryonic fibroblast samples, and the results obtained with direct visualization by the naked eye were consistent with those obtained using real-time RT-PCR. Overall, our RT-LAMP-CRISPR/Cas12a assay is a reliable, sensitive, specific, and user-friendly method that holds great promise for early on-site detection of DTMUV in clinical samples, facilitating timely interventions and improved disease management in the poultry industry.

Duck Tembusu virus (DTMUV), a novel positive-sense RNA virus, has caused significant economic losses in the poultry industry of Eastern and Southeast Asia since its outbreak in 2010. Furthermore, the rapid transmission and potential zoonotic nature of DTMUV pose a threat to public health safety. In this study, a 4D-DIA quantitative proteomics approach was employed to identify differentially expressed cellular proteins in DTMUV-infected DF-1 cells, which are routinely used for virus isolation and identification for DTMUV, as well as the development of vaccines against other poultry viruses. One hundred fifty-seven differentially expressed cellular proteins were identified, including 84 upregulated and 73 downregulated proteins at 48 h post-infection, among which CXCL8, DDX3X, and TRPV2 may play crucial roles in viral propagation. Notably, for the upregulated protein TRPV2, the DTMUV replication was inhibited in TRPV2-low-expressing DF-1 cells. In summary, our research represents the application of 4D-DIA quantitative proteomics to analyze the proteomic landscape of DTMUV-infected poultry cells. These findings may provide valuable insights into understanding the interaction mechanism between DTMUV and poultry cells, as well as the identification of disease-resistant host factors in poultry breeding research.

This study aimed to analyze the associations between the composition and function of the gut microbiome in weaned piglets of different breeds and Fucosyltransferase 1 (FUT1) genotypes. Ear tissue and fecal samples were collected from 163 piglets (61 Durocs, 39 Yorkshires, and 63 Landraces) for genotyping and 16S rRNA sequencing. Three genotypes were identified in the FUT1 gene: AA, GA, and GG, with the resistant AA genotype observed only in Duroc. α-diversity analysis revealed that Landrace pigs had the highest microbial diversity, whereas pigs with the AA genotype displayed the lowest diversity. At the phylum level, Bacillota and Bacteroidota were the dominant phyla across all pig breeds and genotypes. At the genus level, significant differences among different groups were observed in microbes such as Lactobacillus and Segatella. PICRUSt2-based functional prediction revealed a higher relative abundance of predicted functions related to xenobiotics biodegradation and metabolism, digestive system, nucleotide metabolism, and other metabolic processes in the AA genotype group. These findings identify breed- and genotype-associated variation in the fecal microbiota of weaned piglets and provide a foundation for future studies investigating whether FUT1-associated microbial differences are related to intestinal microbial function and resistance to F18-positive enterotoxigenic Escherichia coli.