In the present study, the immuno-enhancing effect of Eucommia ulmoides leaf polysaccharide (ELP) was investigated in immunosuppressed mice induced by cyclophosphamide (CTX). To evaluate the immune enhancement mechanism of ELP, the immunoregulation effect of ELP was evaluated in vitro and in vivo. ELP is primarily composed of arabinose (26.61%), galacturonic acid (25.1%), galactose (19.35%), rhamnose (16.13%), and a small amount of glucose (12.9%). At 1000~5000 μg·mL⁻¹, ELP could significantly enhance the proliferation and the phagocytosis of macrophages in vitro. Additionally, ELP could protect immune organs, reduce pathological damage, and reverse the decrease in the hematological indices. Moreover, ELP significantly increased the phagocytic index, enhanced the ear swelling response, augmented the production of inflammatory cytokines, and markedly up-regulated the expression of IL-1β, IL-6, and TNF-α mRNA levels. Furthermore, ELP improved phosphorylated p38, ERK1/2, and JNK levels, suggesting that MAPKs might be involved in immunomodulatory effects. The results provide a theoretical foundation for exploring the immune modulation function of ELP as a functional food.

Swine influenza virus (SIV) circulates worldwide, posing substantial economic loss and disease burden to humans and animals. Vaccination remains the most effective way to prevent SIV infection and transmission. In this study, we evaluated the protective efficacy of a recombinant, baculovirus-insect cell system-expressed bivalent nanoparticle SIV vaccine in mice challenged with drifted swine influenza H1N1 and H3N2 viruses. After a prime-boost immunization, the bivalent nanoparticle vaccine (BNV) induced high levels of hemagglutination inhibition (HAI) antibodies, virus-neutralization (VN) antibodies, and antigen-specific IgG antibodies in mice, as well as more efficient cytokine levels. The MF59 and CPG1 adjuvant could significantly promote both humoral and cellular immunity of BNV. The MF59 adjuvant showed a balanced Th1/Th2 immune response, and the CPG1 adjuvant tended to show a Th1-favored response. The BALB/c challenge test showed that BNV could significantly reduce lung viral loads and feces viral shedding, and showed fewer lung pathological lesions than those in PBS and inactivated vaccine groups. These results suggest that this novel bivalent nanoparticle swine influenza vaccine can be used as an efficacious vaccine candidate to induce robust immunity and provide broad protection against drifted subtypes in mice. Immune efficacy in pigs needs to be further evaluated.

Bovine enteroviruses (BEVs) are emerging pathogens with poorly understood evolutionary dynamics and zoonotic potential. Here, we report the discovery of a novel recombinant BEV strain, HeN-2022, isolated from cattle in China. Genomic analysis revealed that HeN-2022 is a primary hybrid of BEV-E1 (VG527, Ireland) and BEV-E4 (GX1901, China), with recombination breakpoints in the VP1 gene and 5′ UTR. Divergence dating traced its origin to 1991, predating closely related strains. Experimental infection in sheep demonstrated asymptomatic viral shedding (peak at 5 dpi) and robust neutralizing antibody responses, highlighting the potential cross-species adaptability. These findings underscore recombination as a potential key driver of BEV evolution and emphasize the need for global surveillance to address emerging livestock pathogens.

This study was conducted to elucidate the intestinal damage induced by the IPEC-J2 cell culture-passaged PDCoV. The results showed that PDCoV disrupted the intestinal structure and increased intestinal permeability, causing abnormalities in mucosal pathology. Additionally, PDCoV induced an imbalance in the intestinal flora and disturbed its stability. Microbial community profiling revealed bacterial enrichment (e.g., Proteobacteria) and reduction (e.g., Firmicutes and Bacteroidetes) in the PDCoV-inoculated piglet model. In addition, metabolomics analysis indicated that 82 named differential metabolites were successfully quantified, including 37 up-regulated and 45 down-regulated metabolites. Chenodeoxycholic acid, sphingosine, and oleanolic aldehyde levels were reduced in PDCoV-inoculated piglets, while phenylacetylglycine and geranylgeranyl-PP levels were elevated. Correlation analysis indicated a negative correlation between Escherichia-Shigella and choline, succinic acid, creatine, phenyllactate, and hippuric acid. Meanwhile, Escherichia-Shigella was positively correlated with acetylcholine, L-Glutamicacid, and N-Acetylmuramate. Roseburia, Lachnospiraceae_UCG-010, Blautia, and Limosilactobacillus were negatively and positively correlated with sphingosine, respectively. These data suggested PDCoV-inoculated piglets exhibited significant taxonomic perturbations in the gut microbiome, which may result in a significantly altered metabolomic profile.

Porcine viral diarrhea has always been one of the main obstacles to the healthy development of the pig industry in China with its variety of pathogens and complexity of co-infections. Analysis of the dominant mixed-infection model is a fundamental step in boosting the prevention and control of porcine diarrhea. In this study, 3256 porcine fecal samples were collected from 17 pig herds in Shanghai, China, from 2015 to 2023 to identify novel pathogenic infection patterns. The results confirmed that porcine astrovirus (PAstV), porcine sapelovirus (PSV), and porcine epidemic diarrhea virus (PEDV) were the top three agents with positive rates of 28.47%, 20.71%, and 20.23%, respectively. Porcine rotavirus (PoRV) and transmissible gastroenteritis virus (TGEV) accounted for only 8.12% and 1.12%, respectively. Importantly, mixed infection rates were high and complicated. The double infection rate was higher than that of a single infection. Next, the mixed-infection model of PEDV and emerging diarrheal pathogens was explored. The predominant dual-infection models were PEDV/PKoV (porcine kobuvirus) (14.18%), PEDV/PAstV (10.02%), and PEDV/PSV (9.29%). The predominant triple infection models were PEDV/PKoV/PAstV (18.93%), PEDV/PSV/PAstV (10.65%), and PEDV/PKoV/PSV (7.10%). The dominant quadruple-infection model was PEDV/PAstV/PSV/PKoV (46.82%). In conclusion, PEDV is mainly mix-infected with PAstV, PSV, and PKoV in clinical settings. Furthermore, multiple-factor logistic regression analysis confirmed that PAstV, PKoV, bovine viral diarrhea virus (BVDV), and PEDV were closely related to porcine diarrhea. PEDV/PKoV, PEDV/porcine sapovirus (PoSaV), PKoV/BVDV, PoSaV/BVDV, and porcine deltacoronavirus (PDCoV)/PoSaV had great co-infection dominance, which will be helpful for porcine co-infection research.

Porcine sapovirus (PoSaV) is one of the most significant pathogens causing piglet diarrhea, and one with limited genetic characterization. In this study, the prevalence, infection pattern, and genetic evolution of porcine sapovirus were elucidated in detail. The positive rate of PoSaV was 10.1% (20/198), with dual, triple, and quadruple infections of 45%, 40%, and 5%, respectively. To further explore the viral composition in the PoSaV-positive diarrhea feces, metagenomic sequencing was carried out. The results confirmed that RNA viruses accounted for a higher proportion (55.47%), including the two primary viruses of PoSaV (21.78%) and porcine astrovirus (PAstV) (24.54%) in the tested diarrhea feces samples. Afterward, a full-length sequence of the PoSaV isolate was amplified and named SHCM/Mega2023, and also given the identifier of GenBank No. PP388958. Phylogenetic analysis identified the prevalent PoSaV strain SHCM/Mega2023 in the GIII genogroup, involving a recombinant event with MK962338 and KT922089, with the breakpoint at 2969–5132 nucleotides (nt). The time tree revealed that the GIII genogroup exhibits the widest divergence time span, indicating a high likelihood of viral recombination. Moreover, SHCM/Mega2023 had three nucleotide “RPL” insertions at the 151–153 nt site in the VP2 gene, compared to the other GIII strains. Further selective pressure calculations demonstrate that the whole genome of the SHCM/Mega2023 strain was under purifying selection (dN/dS < 1), with seven positively selected sites in the VP1 protein, which might be related to antigenicity. In conclusion, this study presents a novel genomic evolution of PoSaV, offering valuable insights into antigenicity and for vaccine research.