The prevalence of porcine rotavirus A (PoRVA) in China has increased significantly, threatening the swine industry. Pigs serve as “mixing vessels” for rotaviruses, facilitating reassortment between human and animal strains, leading to the emergence of zoonotic variants. In this study, two PoRVA strains, XXW2023 (G9P[7]) and HD2023 (G1P[7]), were isolated from diarrheic piglets in Guangdong Province, China. Genomic analysis revealed that both strains were human-porcine reassortants, with VP1, VP3, and NSP1 genes closely related to human rotaviruses. Intragenic recombination was identified in the VP4 and VP6 genes. Pathogenicity was evaluated in 7-day-old mice and 1-day-old piglets. Both strains caused persistent diarrhea in mice and severe watery diarrhea, intestinal lesions, and death within 48 h in piglets. Systemic infection was confirmed, with viral replication detected in the lungs. Infectious virus titers, VP6 antigen, and NSP4 were detected in lung tissues, providing evidence of active replication in the respiratory tract. The strains exhibited distinct tissue tropism, with XXW2023 being enterotropic and HD2023 showing pulmonary tropism. Viral RNA and antigen levels in the lungs of HD2023-infected piglets exceeded those in their intestines, and the infectious virus titer in their lungs was significantly higher than that in the lungs of XXW2023-infected piglets. These findings demonstrate that reassortant rotavirus strains infect the respiratory tract, extending the conventional view of rotavirus as a strictly enteric pathogen. The emergence of these highly pathogenic, phenotypically divergent human-porcine reassortants underscores their zoonotic risk, highlighting the need for enhanced surveillance and reconsideration of vaccine coverage.

XPR1 is the sole known protein that transports inorganic phosphate (Pi) out of cells in metazoans, dynamically regulated by inositol polyphosphates (InsP) signaling and protein interactions to maintain cellular phosphate homeostasis. While InsP-mediated regulation is well-characterized, the mechanistic role of direct protein interactors remains poorly understood. Here, we elucidate the structural and functional interplay between XPR1 and its regulatory partner KIDINS220 using cryo-EM, live-cell FLIM-FRET, and functional transport assays. We demonstrate that KIDINS220 stabilizes an unconventional conformation of the XPR1 SPX domain characterized by a ~ 180° rotation. This rearrangement establishes dual InsP 6 -anchored interfaces, allosterically repositioning the SPX N-terminus toward the XPR1 C-terminal region and trapping the transporter in an inactive state. FLIM-FRET profiling visualizes and confirms that KIDINS220 promotes a more compact and conformationally homogeneous population of XPR1 in live cells. Under InsP 6 -bound conditions, KIDINS220 enforces TM9b closure and cytoplasmic C-plug occlusion, obstructing the Pi permeation pathway. Upon Pi sensing, TM9b transitions to an open conformation, yet persistent C-plug engagement—mediated by hydrophobic and electrostatic networks—prevents substrate translocation. Functional validation demonstrates that KIDINS220 suppresses XPR1-mediated Pi transport, establishing its role as a dual-functional regulator: a trafficking chaperone and a membrane-localized brake. We propose a cooperative gating model where Pi acts as both chemical substrate and allosteric trigger to overcome KIDINS220-imposed inhibition. This work defines a tiered regulatory paradigm integrating protein scaffolding, ligand sensing, and dynamic gating for precision control of phosphate homeostasis.

The physiological parameters of Tibetan piglets have been found to be significantly influenced by environmental factors particularly during the weaning period and in the context of large fluctuations in ambient temperature in plateau regions. This study was designed to examine the effects of different ambient temperatures on gut microbiota diversity, serum antioxidant status, and growth performance of weaned Tibetan piglets. The study comprised of n = 40 weaned Tibetan piglets that were randomly allocated to five temperature-controlled groups with increase in temperature as BC4 (18 °C), BD4 (22–24 °C), BE4 (25–27 °C), BF4 (28–30 °C), and BG4 (31–33 °C). The growth performance was calculated by average daily weight gain (ADG) while serum antioxidant capacity was assessed through malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and glutathione peroxidase (GSH-Px) levels. Fecal samples were taken and through 16 S rRNA gene sequencing the gut microbiota profiling was checked. Microbial α- and β-diversity indices, community composition, and taxa-specific shifts were evaluated by temperature-dependent changes. Results indicated that moderate increase in ambient temperature (25 27 °C) significantly improved the growth performance with higher ADG in the BE4 group. The serum antioxidant markers SOD, T-AOC, and GSH-Px level were higher than MDA concentrations, indicating enhanced oxidative resistance. The beneficial bacterial taxa such as Paraprevotella, Limosilactobacillus, and Paralachnospira flourished and change the gut microbiota diversity and structure, whereas potentially harmful taxa (Bifidobacterium_388775 and Ligilactobacillus) were decreased at the moderate temperatures. The LEfSe and multiple t-tests determine the gut microbial ecology when temperature-dependent shifts in microbial composition indicate a regulatory effect of ambient temperature. It is concluded that the optimal growth performance and enhanced serum antioxidant capacity in weaned Tibetan piglets maintained by moderate raise of ambient temperature (25 27 °C) to support a balanced gut microbiota. These findings indicated that during the weaning period temperature played an important role in regulating oxidative status and gut microbial diversity.

Yaks (Bos grunniens) are important livestock on the Qinghai–Tibetan Plateau, but the relationship between Blastocystis positivity, fecal bacterial communities, and microbial metabolites remains unclear. This cross-sectional study compared fecal bacterial profiles and short-chain fatty acid (SCFA) concentrations in six Blastocystis-positive and six Blastocystis-negative diarrheic yaks from Lhasa, Tibet Autonomous Region, China. Bacterial communities were characterized by 16S rRNA gene amplicon sequencing, and fecal SCFAs were quantified by gas chromatography–mass spectrometry. Compared with negative yaks, positive yaks had lower concentrations of acetic acid (p < 0.01), propionic acid (p < 0.01), butyric acid (p = 0.000381), valeric acid (p < 0.01), caproic acid (p = 0.00359), and isobutyric acid (p < 0.01), whereas isovaleric acid did not differ between groups (p = 0.310). A total of 2,751,028 raw sequences were generated, of which 2,620,949 passed quality filtering. ACE (p = 0.0068), Chao1 (p = 0.0071), Faith’s phylogenetic diversity (p = 0.0019), observed ASVs (p = 0.0064), Pielou’s evenness (p = 0.011), Shannon diversity (p = 0.008), and Simpson diversity (p = 0.037) were lower in positive yaks. The groups also differed in bacterial community composition, and several taxa previously associated with fiber degradation or SCFA formation had lower relative abundances in positive yaks. PICRUSt2 indicated differences in predicted microbial functional potential. These findings indicate that Blastocystis positivity was associated with altered fecal bacterial communities and reduced concentrations of several SCFAs in diarrheic yaks. Because of the small sample size, cross-sectional design, and potential confounding by diarrhea and other unmeasured factors, causal relationships cannot be established.

Zoonotic food- and waterborne protozoan parasites (FWPPs) pose a significant global public health risk, causing substantial morbidity via contaminated fresh produce, water, and meat. Despite their notable impact, surveillance and detection technologies remain inadequate for high-priority protozoans such as Cryptosporidium spp. and Toxoplasma gondii, as current World Health Organization (WHO) and Food and Agriculture Organization (FAO) guidelines primarily focus on bacterial pathogens. This review evaluates the global burden of Cryptosporidium spp. and T. gondii, and highlights the limitations of conventional detection methods, justifying the forward-looking perspective on biosensors' applications in detecting protozoan parasites (PPs), and future strategies in this regard. The complex nature and varied transmission routes of these parasites, along with challenges such as culturing, sample preparation, and morphological similarities, complicate their detection by conventional methods like microscopy, serology, and molecular assays. Additionally, these limitations include time-intensive protocols, infrastructure requirements, cost, and lack of portability, which restrict their suitability for rapid, on-site detection. Recent advances in biosensor technology may offer rapid, sensitive, and accurate on-site detection of FWPPs, driving a paradigm shift toward a smart food safety system. This review highlights the potential of emerging biosensor technologies, especially electrochemical, optical, and piezoelectric (gravimetric) biosensors, for the detection of Cryptosporidium spp. and T. gondii in food and water. Integrating biosensors with nanotechnology, artificial intelligence, point-of-care systems and microfluidics to create portable, cost-effective biosensors may revolutionize food safety surveillance, mitigating the impact of FWPPs, and aligning with Hazard Analysis and Critical Control Points (HACCP) priorities to safeguard public health.

Malaria has long been a significant global health concern, listed as a high-priority disease by several global health agencies, despite of several control measures have been put in place. Most widely utilized treatment options for malaria include chloroquine, artemisinin-based combination therapy (ACT), and quinine. However, challenges, such as drug resistance, misdiagnosis, and limited treatment efficacy remain major concerns. Despite ongoing efforts, the development of an effective malaria vaccine is still debatable. Many existing malaria treatments have drawbacks, such as low water solubility, poor bioavailability, and a rise in drug-resistant parasites owing to their non-judicious use, which contributes to increased malaria cases and fatalities. Nanotechnology presents a promising approach to safer and more effective malaria therapy and control. Nanoparticles offer several advantages over conventional treatments, including high drug-loading capacity, targeted delivery, improved biocompatibility, and reduced toxicity in host cells. Green nanotechnology-based antimalarial therapies have demonstrated potential therapeutic benefits, enhanced safety, and cost-effectiveness compared to traditional treatments, ultimately improving patient compliance and treatment outcomes. In this review paper, we discussed non-conventional breakpoints in the malarial life cycle, traditional herbal remedies for malaria, and nanoparticle-based delivery systems. Additionally, we reviewed the antimalarial effects of herbal nano-formulations, their pharmacological and therapeutic potential, drug-resistant malaria, preventive strategies, vector control using green nanomaterials, and the challenges associated with plant-based nanotechnologies. This review suggests nanotechnology-based therapeutics as promising candidates to treat malaria with significant room for applications and commercialization potential in the longer run.

Yak has ability to adapt the harsh environments like extremely cold, anoxic, strong ultraviolet rays and shortage of pasture. It is observed that the nutritional supply during the preweaning period is highly associated with the development of the gastrointestinal tract and immunity of animals. In this study, twenty-one male yak calves were divided into control group (DFC), starter feed group 1 (DFO), and starter feed group 2 (DFT) to study the growth of early weaning Chawula yak calves. Calves in group DFC were free-ranged, DFO group were fed with alfalfa (1.4 kg/head/day) and starter feed 1 (1.4 kg/head/day), and DFT group were fed with alfalfa ( 1.4 kg/head/day) and starter feed 2 ( 1.4 kg/head/day) for 6 weeks. The body weight of yak calves in DFT was significantly higher than that of control yaks in DFC (P < 0.05), and the net weight growth rate in DFO (P < 0.001) and DFT (P < 0.0001) were both obviously higher than that in the DFC group. The chest girth (bust) in group DFO (P < 0.05) and DFT (P < 0.05) were both markedly higher than that in the DFC group. The serum contents of T-AOC in DFC calves were markedly lower than DFO (P < 0.01) and DFT (P < 0.001) yaks. Also, T-AOC was obviously higher in DFT yaks than in DFO animals (P < 0.05). The levels of GSH-Px were significantly higher in DFO (P < 0.01) and DFT (P < 0.01) yaks than DFC yaks. High throughout sequencing achieved 391520, 356907 and 353763 filtered sequences in DFC, DFO and DFT yaks, and one phylum (Firmicutes B 370539) and thirty-seven genera (Phocaeicola A 858004, Cryptobacteroides, Evtepia, CAG-273, etc.) were identified as biomarkers in yak calves. We observed that starter feeds could promote the growth of early weaning Chawula yak calves by enhancing antioxidant capacity and regulating the gut microbiota.

Canine parvovirus 2 (CPV-2) remains a leading cause of acute infectious gastroenteritis with high global morbidity in dogs. While murine neutralizing monoclonal antibodies (mAbs) are widely used for antiviral therapy, their efficacy is limited by immune rejection in canine recipients. Here, we developed an efficient single B-cell cloning platform to generate canine-derived neutralizing mAbs against CPV-2 and characterized their germline gene usage patterns. Specifically, using biotinylated CPV-2 virions as bait, CPV-2-binding B cells were singly isolated via fluorescence-activated cell sorting (FACS) from peripheral blood mononuclear cells of immunized dogs. The heavy and light chain variable region (VH/VL) sequences were amplified through nested RT-PCR from single B cells, and cloned into canine immunoglobulin heavy/light chain (IgH/IgL) expression vectors. A total of 22 canine-derived mAbs were successfully expressed and purified from suspended ExpiCHO-S cells, and 20 of which demonstrated CPV-2-binding reactivity in enzyme-linked immunosorbent assay (ELISA) or indirect immunofluorescence assay (IFA). Among these, 13 mAbs exhibited neutralizing activity (IC50 < 25 μg/mL) against a CPV-2c strain in F81 cells by virus micro-neutralization assays. Notably, the clone B11 showed potent virus neutralization activity with an IC50 of 0.06 μg/mL. Furthermore, germline gene usage analysis revealed preferential utilization of IGHV3-5, IGHD1, and IGHD3 in the heavy chain, and IGLV1-46, IGLV1-48, and IGLJ4/9 in the light chain in these CPV-2-specific canine antibodies. These canine-derived mAbs show promise for clinical diagnostics and therapeutics, overcoming the limitations of murine antibodies. Our platform establishes a framework for developing canine mAbs against other pathogens.

The escalating challenge of antimicrobial resistance has spurred interest in probiotics as alternatives for combating bacterial infections. This study aimed to isolate and characterize probiotic Lactobacillus johnsonii (L. johnsonii) from yak feces with protective efficacy against acute Escherichia coli (E. coli) infection. In vitro, DY2 supernatant inhibited the growth of E. coli. In vivo, mice pretreated orally with DY2 (1 × 109 CFU/mL) for 21 days before E. coli challenge exhibited significantly reduced weight loss (p < 0.001), lower bacterial translocation in the intestines (p < 0.001), and normalized organ indices (p < 0.05) compared to untreated infected controls. DY2 modulated host immune and oxidative responses by significantly lowering serum levels of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6; p < 0.001 to p < 0.05) and malondialdehyde (MDA; p < 0.001), while elevating levels of the anti-inflammatory IL-10 (p < 0.05) and antioxidant enzymes (SOD, GSH-Px, T-AOC; p < 0.001 to p < 0.01). Histologically, DY2 preserved intestinal mucosal integrity, with reduced villus shortening and inflammatory infiltration (p < 0.001 for villus length in key segments). 16S rRNA sequencing of intestinal microbiota revealed enhanced α-diversity (p < 0.05 to p < 0.001), community stability, and enrichment of beneficial genera such as Butyricimonas in DY2-treated mice. Conclusively, Lactobacillus johnsonii DY2 protects against acute E. coli infection via anti-inflammatory, antioxidant, gut barrier strengthening, and microbiota-modulating activities. Yak-derived lactobacilli are promising probiotics with excellent antibacterial properties.

This study evaluated the effects of traditional Chinese herbal medicine compounds (TCHMCs) on gut microbiota, maternal health, and calf performance in perinatal yaks. Thirty-two yaks with expected calving dates (two months pre-calving to one month post-calving) were randomly assigned to four groups (n = 8). Groups WA, WB, and WC received normal diets supplemented with different TCHMC formulations, while group WD served as the control. High-throughput sequencing revealed that TCHMC supplementation significantly increased gut microbial diversity and altered community composition. Compared with controls, supplemented yaks exhibited higher abundances of Bacteroidetes and enrichment of short-chain fatty acid (SCFA)-producing genera, including Phascolarctobacterium, Butyricicoccus, Bulleidia, and Blautia. Potentially beneficial taxa with anti-inflammatory properties, such as Adlercreutzia, Akkermansia, Odoribacter, Dehalobacterium, and Sporosarcina, were also enriched. Calves born to treated yaks demonstrated significantly greater birth weights, reduced serum levels of pro-inflammatory cytokines (TNF-α, IL-6), and improved antioxidant capacity, as indicated by increased T-AOC and SOD activity. Among the three formulations, TCHMCⅡ (WB) exerted the most pronounced effects, enhancing microbial shifts, maternal physiological resilience, and neonatal health outcomes. In conclusion, dietary supplementation with TCHMCs during the perinatal period modulated gut microbiota, reduced maternal inflammation, strengthened antioxidant defenses, and improved calf growth performance. These findings provide new evidence supporting the use of TCHMCs as practical nutritional interventions to improve reproductive performance and offspring vitality in yak husbandry under high-altitude conditions.

Foot-and-mouth disease virus (FMDV) is a highly contagious picornavirus that affects cloven-hoofed animals and carries significant economic implications for the global livestock industry. FMDV features two Leader (L) protein isoforms, Lab and Lb, differing at their amino termini by 28 amino acids (La region). Currently, the activity of La protein sequences has not been investigated. To address this issue, the comparison study of biological and functional roles of Lab and Lb was performed as the La region alone did not independently perform protein function. We found that Lab and Lb significantly regulated FMDV replication and pathogenicity, and their coexistence afforded optimal FMDV properties. Subsequently, we observed that both L isoforms cleaved eukaryotic translation initiation factor 4G (eIF4G) I, suppressed type I and type III interferon (IFN) expression, and exhibited marked cytotoxicity, indicating that they were all key components in FMDV's antagonism of host antiviral defenses. Finally, the subcellular distribution of Lab and Lb was detected. Despite dual localization in cytoplasmic and nuclear compartments, both isoforms displayed different spatial distribution patterns, and Lb induced more pronounced morphological changes to host cells than Lab. Furthermore, bioinformatics predicted that the La region might contain a non-classical secretory signal peptide, potentially facilitating Lab distribution to the cell membrane or extracellular space. Collectively, the primary encoding role of La region was to control the intracellular distribution of L protein, as opposed to regulating its functional activity. This study may help to deepen our understanding of why FMDV encoded two isoforms of L protein.

Classical swine fever virus (CSFV) is a highly contagious pathogen that causes substantial economic losses in swine production, yet the mechanisms governing its cellular entry remain poorly defined. Here, through a combination of pharmacological inhibition, genetic knockout/overexpression, and co-immunoprecipitation, we identify annexin A2 (ANXA2) as a pivotal host determinant of CSFV entry into PK-15 cells. Confocal microscopy revealed that CSFV infection rapidly induced the translocation of ANXA2 to the cell surface, and functional assays confirmed that its plasma membrane localization was essential for efficient infection. Mechanistically, the viral envelope protein E2 specifically interacts with the heterotetrameric A2t complex (ANXA2/S100A10), establishing this complex as a functional cellular receptor for CSFV. ANXA2 also maintains clathrin at the plasma membrane and facilitates clathrin-dependent viral entry, as evidenced by inhibitor and siRNA knockdown studies. Intriguingly, ANXA2 depletion redirects viral entry to a caveolin-1-dependent pathway. We further demonstrate that ANXA2 competitively binds the C-terminal domain of caveolin-1, a site that overlaps with the binding region for the viral Erns protein, thereby suppressing the caveolin-1 route in wild-type cells. In summary, this study identifies the A2t complex as a CSFV receptor and reveals a crucial regulatory role for ANXA2 in viral entry pathway selection. These findings provide new insights into CSFV infection and a rationale for host-directed antiviral strategies.

Nitazoxanide (NTZ), the only FDA-approved anti-Cryptosporidium drug, shows inconsistent efficacy in immunocompromised patients. This study employed HCT-8 cell-based cytotoxicity/parasite inhibition assays and an immunosuppressed murine model to explore the anti-C. parvum efficacy and mechanisms of combined allicin and Qingchang Huashi Formula (QHF). In vitro, WST-1-based cytotoxicity screening preceded inoculation of 4 × 104 C. parvum oocysts to assess parasitic inhibition. In vivo, sixty immunosuppressed ICR mice were divided into control (C), model-infected (M), paromomycin-treated (P), allicin-treated (A), QHF-treated (F), and allicin + QHF (AF)-treated. On day 1, groups M, P, A, F, and A + F were orally infected with 3 × 104 C. parvum oocysts each; on day 2, the P, A, F, and A + F groups received daily intragastric administration of the respective drugs for 8 days (1-8 dpi), before euthanasia and 16S rRNA sequencing of rectal feces. In vitro results showed a significant parasitic inhibition at 750 μg/mL (allicin): 1500 μg/mL (QHF) (p < 0.05). In vivo A + F combined treatment significantly reduced fecal C. parvum burden, attenuated inflammation (IL-6, IL-1β, and TNF-α), upregulated IL-10, strengthened antioxidant defense (SOD, GSH-Px, and T-AOC), and reduced MDA levels (p < 0.05). Combined treatment significantly ameliorated intestinal damage, upregulated tight junction proteins (occludin, claudin-1, and ZO-1) and NLRP6 expression, while inhibiting caspase-1 (p < 0.05). 16S rRNA sequencing revealed treatment significantly restored the abundance of beneficial flora, including Firmicutes_A, Bacteroidota, Actinobacteriota (phyla), and Ligilactobacillus, Bacteroides_H, Dwaynesavagella, and UBA3282 (genera), along with regulation of energy metabolic pathways and amino acid synthesis (p < 0.05). Conclusively, allicin and QHF ameliorated the C. parvum-induced damage through gut barrier restoration and microbiota modulation, holding great significance in both public health and livestock sectors.

Of the seven serotypes of foot-and-mouth disease virus (FMDV) strains circulating globally, serotype Asia1 has been effectively eradicated in China through systematic vaccination in livestock. The structural characteristics of serotype Asia1 may enhance its immunogenicity compared to other serotypes. Herein, we present a preliminary exploration of Asia1-binding B-cell receptor repertoire, containing 3571 clones, and identified 17 porcine-derived neutralizing monoclonal antibodies (pnAbs) from the top 33 high-frequency clonotypes. The majority of pnAbs (14/17) recognized the epitopes on VP2, with a common determinant at residue 72 (D) on the B-C loop; two pnAbs (2/17) recognized a novel epitope spanning VP2 and VP3; and the remaining one (1/17) bound to the C-terminus of VP1. Furthermore, the antigenic structures on VP2 and spanning VP2 and VP3 were respectively elucidated by determining the cryo-EM structures of FMDV serotype Asia1 in complexes with two pnAbs, PAS5 and PAS12. The light chain of PAS5, forming the majority of contact sites with the viral particle, focuses on the βB, B-C loop, βC and H-I loop of VP2, with key determinants at residues 68, 72 and 77 around the three-fold axis, corresponding to antigenic site 2. The contact sites of both VH and VL of PAS12 uncover a novel antigenic structure comprising the B-C, and H-I loops on VP2, and the B-B knob and βB on VP3, with key determinants at residue 73 on VP2 and 59 on VP3. Subsequently, site-directed competitive ELISA analysis of sera from primary and booster vaccinated pigs revealed a balanced antibody response profile, suggesting a potentially even immunodominance among antigenic site 2, VP1 G-H loop, and the novel antigenic structure spanning VP2 and VP3 on FMDV serotype Asia1. Compared to the focused immunodominance observed in other serotypes, this balanced antigenic recognition across VP1, VP2, and VP3 of FMDV serotype Asia1 reflects a diversified antibody response that may contribute to effective neutralization and protection.

Chromium has become increasingly hazard due to its strong carcinogenicity; effective therapeutic options for managing such injuries remain limited. We examined the effect of the polysaccharide of Xizang Coriolus versicolor (CV) on spleen damage in mice exposed to chromium. Mice (n=30) were divided into KCH, KMH, and KYH groups. Group KMH and KYH were induced by K2Cr2O7 (15 mg/kg), and KYH was administered with 50 mg/kg CV polysaccharide for 35 days. We found that the weight of mice in the KMH group was significantly lower than in the KCH (P<0.05) and KYH (P<0.05) on the 35(th) day, while the spleen index in KMH was higher than that of animals in other groups (P<0.05). H&E and Sirius red staining showed that hexavalent chromium led to serious atrophy of white pulp and acini lienal, blurry marginal zone, and infiltration of inflammatory cells in mice; however, mice supplemented with CV polysaccharide had clear red and white pulp, fewer inflammatory cells, and decreased fibrosis. CV polysaccharide decreased serum IL-6 (P<0.01), IL-1 beta (P<0.001), TNF-alpha (P<0.001), and MDA (P<0.001), while increasing IL-10 (P<0.01), T-AOC (P<0.001), and GSH-Px (P<0.001) in mice. Microbiota sequencing achieved 1,383,401 filtered sequences and found two phyla and nine genera with significant differences among the three groups. Genera were Lactococcus A 343473, Gemella, and Bacillus P 294101. In summary, we confirmed that the polysaccharide of Xizang Coriolus versicolor could alleviate spleen damage in mice exposed to chromium by regulating inflammatory response, antioxidant capacity, and gut microbiota.

Diarrhea is a common cause posing significant economic losses in sheep farming, primarily caused by various infectious pathogens, among which Escherichia coli is one of the key etiological agents. However, limited data exist on the antimicrobial resistance profiles and virulence-associated factors of E. coli isolated from diarrheic sheep in Ningxia, China. In this study, a total of 32 fecal samples were collected from diarrheic sheep at farms in Ningxia to investigate the resistance phenotypes and virulence gene profiles of E. coli. The 16S rRNA gene amplification and phylogenetic analysis were used to identify bacterial isolates. E. coli recovered from 59.4% (19/32) of the samples. Antimicrobial susceptibility was determined by the disk diffusion method, antibiotics included beta-lactams, quinolones, aminoglycosides, tetracyclines, polymyxins, and sulfonamides. The eaeH gene was detected in all isolates (19/19); several strains also harbored virulence genes associated with enteroaggregative (EAEC), enteropathogenic (EPEC), and enterohemorrhagic (EHEC) E. coli. Antimicrobial susceptibility revealed that all isolates were resistant to ampicillin and trimethoprim-sulfamethoxazole. In conclusion, these findings indicate a high prevalence of virulence traits and multidrug resistance among the ovine E. coli isolates from Ningxia, underscoring the urgent need to strengthen antimicrobial stewardship and pathogen surveillance in the region.

The whole life cycle of the highly pathogenic foot-and-mouth disease virus (FMDV) significantly depends on the host determinants to achieve its infection. ATG16L1 is well known to be required to form the autophagosomes membrane at the early steps of autophagy, while its non-autophagic roles in FMDV infection remain unclear. We found that following entry, FMDV O/Fujian/CHA/5/99 trafficked to early endosomes (EEs) and the trans-Golgi network (TGN), bypassing late endosomes (LEs) /lysosome and recycling endosomes (REs). This specific intracellular distribution mirrored the vesicular sorting pathway involving ATG16L1 that had been reported previously. Further analyses showed that ATG16L1 increased the internalization of FMDV and recruited EEs to facilitate the initial phase of FMDV infection. However, ATG16L1 degraded FMDV 2BC protein in the Golgi via its non-autophagic function to inhibit late stages of FMDV infection. To counteract this, membrane-associated 2BC interacted with ATG16L1 and mediated its reduction via the caspase pathway, thereby sustaining FMDV replication. In conclusion, our evidence suggested that ATG16L1 played dual roles in regulating the life cycle of FMDV.

IntroductionAlthough polysaccharides from Psoralea corylifolia L. (PPs) have been reported to possess immune-stimulatory effects, their precise mechanisms of action remain unclear.MethodsIn this study, the potential mechanism of PPs in alleviating CTX-induced immunosuppression was investigated by analyzing the gut microbiota, metabolomics, and immune parameters in mice.ResultsThe results showed that PPs significantly alleviated CTX-induced immunosuppression, as evidenced by increased immune organ indices, improved intestinal mucosalintegrity, elevated serum levels of IL-6 and TNF-α, enhanced activities of ACP, LDH, SOD, and GSH-Px, and reduced MDA content. Western blot analysis indicated that PPs activated the NF-kB and MAPK signaling pathways and upregulated the expression of intestinal tight junction proteins (Claudin-1, Occludin, and ZO-1). Immunohistochemical results further revealed that PPs modulated the numbers of CD4+ and CD8+ T cells in the small intestine. Based on 16S rDNA sequencing and untargeted metabolomics analysis, PPs promoted the proliferation of Lachnospiraceae_NK4A136_group and Lactobacillus, while reducing the abundance of Prevotellaceae_UCG-001, f:Lachnospiraceae_Unclassified, and Alloprevotella, thereby ameliorating metabolic disorders and counteracting CTX-induced immunosuppression. Spearman’s correlation coefficient analysis indicated significant associations among gut microbiota, serum metabolites, and immune as well as antioxidant indicators.DiscussionThese results suggested that PPs enhanced the immune response in immunocompromised mice by boosting antioxidant capacity, improving the intestinal barrier, modulating gut microbiota structure, and correcting metabolic disturbances.

Eimeria (E.) tenella, the causative agent of avian coccidiosis, employs apical complex proteins like RON2 for host cell invasion. While EtRON2 has been well-studied, its paralogs remain poorly characterized. This study investigated EtRON2L1-1, a RON2 homolog, for its structural features, stage-specific expression, and vaccine potential. Bioinformatic analysis revealed EtRON2L1-1 shares only 23.46% amino acid identity withEtRON2, lacks a signal peptide, and contains three transmembrane domains. Expression profiling revealed distinct transcriptional and translational regulations, with peak mRNA levels in sporulated oocysts and highest protein expression in sporozoites, indicative of post-transcriptional control. Immunofluorescence studies showed stage-dependent localization patterns: cytoplasmic distribution in free sporozoites transitioning to apical end during host cell invasion. In vitro neutralization assays established EtRON2L1-1 's involvement in invasion, with specific antibodies exhibiting dose-dependent inhibition (13% at 50 tg/mL to 42% at 400 tg/mL). Vaccination trials demonstrated that while 100 tg pCAGGS-EtRON2L1-1 significantly improved weight gain and reduced oocyst output compared to controls, its protective efficacy was inferior to pCAGGS-EtRON2L2, particularly in mitigating intestinal lesions. These findings characterize EtRON2L1-1 as a functionally distinct RON2 paralog involved in host cell invasion, with partial but promising vaccine potential that requires further optimization for effective coccidiosis control.