Search Results (610)

Porcine Teschovirus (PTV) is a highly prevalent pathogen within swine populations, primarily associated with encephalitis, diarrhea, pneumonia, and reproductive disorders in pigs, thereby posing a significant threat to the sustainable development of the pig farming industry. In this study, a novel strain of PTV was isolated from the feces of a pig exhibiting symptoms of diarrhea, utilizing PK-15 cell lines. The structural integrity of the viral particles was confirmed via transmission electron microscopy, and the viral growth kinetics and characteristics were evaluated in PK-15 cells. High-throughput sequencing facilitated the acquisition of the complete viral genome, and subsequent phylogenetic analysis and full-genome alignment identified the strain as belonging to the PTV 2 genotype. Further investigation revealed that infection with the PTV-GXLZ2024 strain induces phosphorylation of the eukaryotic translation initiation factor 2α (eIF2α) in PK-15 cells, indicating activation of the unfolded protein response (UPR) through the PERK pathway, with minimal involvement of the IRE1 or ATF6 pathways. Notably, ATF4 protein expression was progressively downregulated throughout the infection, while downstream CHOP protein levels remained unchanged, indicating an incomplete UPR induced by PTV-GXLZ2024. Furthermore, PERK knockdown was found to enhance the replication of PTV-GXLZ2024. This study provides critical insights into the molecular mechanisms underlying PTV pathogenesis and establishes a foundation for future research into its evolutionary dynamics and interactions with host organisms. Full article

Background/Objectives: The pandemic caused by SARS-CoV-2 boosted the development of different vaccine models. In parallel, yeasts stand out as a vaccine platform in healthcare biotechnology. Species such as Saccharomyces cerevisiae and Pichia pastoris can express heterologous proteins, which are capable of inducing specific antibodies and can perform as an attractive vaccine vehicle with immunomodulating properties due to their cell wall composition. Furthermore, the yeast surface display system facilitates antigen presentation to immune cells. We developed an oral vaccine based on P. pastoris displaying a synthetic antigen composed of Spike and Nucleocapsid epitopes. Methods: The vaccine was administered to BALB/c mice. Systemic immune response was measured through antibody detection in blood samples, and mucosal immunity was assessed via IgA levels in feces. Histopathological analysis of intestinal and gastric tissues was also conducted. Results: The yeast-based vaccine elicited a humoral immune response, reflected in the production of neutralizing antibodies and elevated levels of IgG2a and IgG2. No structural alterations or pathological changes were observed in gastrointestinal tissues. Conclusions: This study demonstrates the feasibility of using P. pastoris as an oral vaccine delivery system, supporting previous findings with other yeast species such as Saccharomyces cerevisiae, and highlighting its potential in developing effective mucosal vaccines. Full article

This study aimed to investigate the effects of dietary resveratrol supplementation on fermentation characteristics, microbial diversity, and community composition of feces from Hu sheep. A total of 20 three-month-old Hu sheep with similar body weights (20.62 ± 0.51 kg) were randomly divided into the control group (fed a basal diet, CON) and the treatment group (fed a basal diet supplemented with resveratrol at 100 mg/kg of feed, RES), with 10 sheep in each group, and lasted for 75 days. Feces were collected from each sheep at twenty-four time points for fecal fermentation characteristics determination and microbial analysis. The results showed that the pH value was higher in the RES group than in the CON group (p < 0.05), while the concentration of ammonia nitrogen was lower, showing a 10.6% reduction compared to the CON group (p = 0.013). The richness, Shannon index, and inverse Simpson index of fecal microbiota were higher in the CON group than in the RES group (p < 0.05). The relative abundances of Planctomycetota, Bacteroides, Alistipes, and NK4A214 group were higher in the CON group than in the RES group (p < 0.05). Additionally, the relative abundance of the glycan biosynthesis and metabolism pathway was higher in the CON group (p < 0.05). The relative abundance of Prevotella was lower in the CON group than in the RES group (p < 0.05). Principal co-ordinate analysis (PCoA) revealed no overlap between the two groups, and analysis of similarities (ANOSIM) showed significant differences between the CON and RES groups (R = 0.4560, p = 0.012). Linear discriminant analysis effect size (LEfSe) analysis identified 27 microbial biomarkers, with the RES group having more beneficial bacteria and the CON group having more potentially harmful bacteria. The study demonstrated that dietary resveratrol supplementation reduced the concentration of ammonia nitrogen in feces, decreased microbial diversity, and increased the abundance of beneficial bacteria. The findings of this research provide a post-digestion perspective for evaluating the application of resveratrol in ruminant production. Full article

Background: Feces and cecum content are commonly involved in metabolomic analysis to understand the gut metabolic profile of the host, while, in fact, they are different. Feces represent the terminal excretory product after extensive host enzymatic digestion, absorption, and significant modification by the distal gut microbiota. In contrast, cecum content reflects the localized, in situ metabolic microenvironment at that specific site. However, it is worth noting that feces are the most accessible sample type for non-invasive studies, which could be considered proxies for cecum content in some specific cases. Unfortunately, the validity of fecal samples as an alternative to cecum content has rarely been assessed. Methods: The current study attempted to illustrate the distinct metabolomic and microbiota features of feces and cecum content in eight animals (mouse, pig, chicken, duck, rabbit, Gansu yak, Sichuan yak, and sheep) by means of ¹H-NMR and 16S rRNA, respectively. Results: A total of 116 molecules were characterized in feces and cecum content samples. Among them, 22 molecules were shared in all groups. Taking advantage of the univariate analysis, twenty-seven of the quantified molecules were significantly different between feces and cecum content, mainly pertaining to amino acids and organic acids. Moreover, in terms of mammals and non-mammals, short-chain fatty acids could be considered the main factor discriminating the metabolomic profiles between feces and cecum content. Furthermore, to better understand the mechanism of their metabolomic differences, 16S rRNA sequencing analysis was performed on feces and cecum content samples of mice, which is the most widely used animal model. The result showed that the Ace, Shannon, and Sobs indexes in feces were significantly higher than those of cecum content (p < 0.05). At the phylum and genus levels, the microbiota structures of feces and cecum content were similar, while the relative abundances of their microbiota exhibited distinct features. Conclusions: The present study could reduce this gap in information by characterizing, for the first time, the metabolomic differences between feces and cecum content using ¹H-NMR. Moreover, this study is meant as a reference guide for researchers wishing to apply a metabolomics approach to the gut of the host. Full article

Mixed legume–grass pastures may enhance nitrogen recycling via litter and excreta compared to unfertilized grass monocultures. This study evaluated litter biomass, litter deposition rate, and the chemical and isotopic composition of Urochloa decumbens litter in monoculture and mixed pasture intercropped with Arachis pintoi cv. Belmonte at five planting spacings (0.40, 0.50, 0.60, 0.70, and 0.80 m) in a Ferralsol. Additionally, isotopic analysis of sheep feces under grazing was conducted across the dry season. The experiment was conducted according to a split-plot scheme, with spacings in the plots and the periods or years in the subplots, in a randomized block design, with four replications. Litter biomass was not significantly influenced by planting spacing; however, the litter deposition rate was substantially greater in mixed pastures, reaching up to 77.2 kg ha⁻¹ day⁻¹ in the second year. Isotopic analysis revealed that up to 39% of the litter carbon was derived from C3 plants (Arachis pintoi), while nitrogen concentration ranged from 8.3 g kg⁻¹ in monoculture to 12.9 g kg⁻¹ at 0.40 m spacing. Spatial arrangement was critical for optimizing nutrients dynamic. Narrower planting spacings (0.40–0.50 m) increased the proportion of Arachis pintoi and enhanced litter deposition rates, improving nitrogen inputs and cycling within mixed Urochloa decumbens. Full article

The emergence of multidrug-resistant Salmonella poses a significant threat to global public health and food safety, necessitating the urgent search for new strategies to replace conventional antibiotics. Phages are viruses that can directly target bacteria and have garnered attention in recent years for their development as antibiotic alternatives. In this study, 4458 samples were collected from farms, supermarkets, and human feces, yielding 65 strains of Salmonella, which were serotyped using multiplex PCR. Subsequently, a lytic phage was isolated and identified using the dominant serotype of Salmonella as the host bacterium. We further explored the biological characteristics of this phage through host range, growth properties, and genomic analysis. Finally, we analyzed the potential of the phage to block the cross-host transmission of Salmonella, combining PFGE Salmonella classification, strain sources, and phage lytic phenotypes. The results showed that phage gmqsjt-1 could lyse 69.23% (45/65) of Salmonella, of which 75.56% (34/45) were resistant strains. The optimal multiplicity of infection (MOI) for gmqsjt-1 was 0.01, with a latent period of about 10 min, maintaining high activity within the temperature range of 30 to 60 °C and pH range of 2 to 13. No virulence or resistance genes were detected in the gmqsjt-1 genome, which carries two tail spike proteins (contain FAD binding_2 superfamily, the Tail spike TSP1/Gp66 N-terminal domain, and the Pectin lyase fold) and a holin–lysozyme–spanin lytic system. Phylogenetic classification indicates that phage gmqsjt-1 belongs to a new genus and species of an unnamed family within the class Caudoviricetes. PFGE classification results show a high genetic relationship among human, farm animal, and food source Salmonella, and the comprehensive lytic phenotype reveals that phage gmqsjt-1 can lyse Salmonella with high genetic correlation. These results suggest that this novel lytic Salmonella phage has the potential to inhibit cross-host transmission of Salmonella, making it a promising candidate for developing alternative agents to control Salmonella contamination sources (farms), thereby reducing the risk of human infection with Salmonella through ensuring food system safety. Full article

Background and Objectives: Feces-induced peritonitis (FIP), a clinically relevant model of polymicrobial sepsis, induces systemic inflammation and acute lung injury (ALI). Methylene blue (MB), a phenothiazine-based compound, exhibits vasoregulatory, antioxidant, and anti-inflammatory properties in the context of sepsis. This study aimed to evaluate the protective effects of MB on pulmonary injury in a rat model of FIP-induced sepsis. Materials and Methods: Forty male Wistar rats were randomly assigned to four groups: control, FIP, FIP + Saline, and FIP + MB. MB was administered intraperitoneally at a dose of 20 mg/kg, 1 h after FIP induction. At 24 h post-induction, plasma levels of inflammatory markers [interleukin-6 (IL-6), interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP)], oxidative stress marker [malondialdehyde (MDA)], metabolic indicator [lactic acid], and vascular signaling marker [cyclic guanosine monophosphate (cGMP)] were measured. Lung injury was evaluated through histopathological analysis and thoracic computed tomography (CT)-based Hounsfield unit (HU) quantification, while pulmonary function was assessed via arterial blood gas analysis, including arterial oxygen pressure (PaO₂) and carbon dioxide pressure (PaCO₂). Results: FIP induction led to significant increases in plasma levels of IL-6, IL-1β, TNF-α, CRP, MDA, cGMP, and lactic acid, accompanied by elevated CT attenuation (HU) values and a marked reduction in arterial PaO₂ and PaCO₂. MB treatment significantly decreased the levels of IL-6, IL-1β, TNF-α, CRP, MDA, lactic acid, and cGMP, improved PaO₂, and attenuated both histopathological lung injury and CT-assessed parenchymal density. No significant differences were observed in PaCO₂ among the groups. Conclusions: MB mitigates inflammation, oxidative damage, and pulmonary dysfunction in FIP-induced sepsis. Further studies are warranted to optimize dosing and timing and to evaluate long-term outcomes. Full article

Research is required to determine whether the coinfections by Fasciola spp. and other parasite species result from poor rural hygiene or reflect underlying epidemiological patterns and causes. Therefore, the role of fascioliasis is analyzed concerning coinfection complexity, risk of multiparasitism, parasite associations, pathogenic implications and their multifactorial causes. Helminth and protozoan coinfections are studied in 2575 previously untreated individuals from four rural hyperendemic areas (Northern Bolivian Altiplano, Peruvian Altiplano and Cajamarca valley, and the Egyptian Nile Delta). This cross-sectional study was conducted from January 2011 to December 2023. Coinfections were coprologically assessed by the merthiolate–iodine–formalin and formol–ether concentration techniques. Infection intensity was measured as eggs/gram of feces (epg) with the Kato–Katz technique. Parasite and coinfection prevalences were stratified by age, sex and geographical location. High mixed infections, fascioliasis prevalences and very low non-coinfected Fasciola-infected subjects were associated with the following regions: Bolivian Altiplano, 96.5%, 16.8% and 3.5%; Peruvian Altiplano, 100%, 24.6% and 0%; Cajamarca valley, 98.7%, 21.4% and 1.8%; Nile Delta, 84.1%, 13.0% and 15.9%. Transmission routes and human infection sources underlie fascioliasis associations with protozoan and other helminth infections. Prevalence pattern of protozoan–helminth coinfections differed between Fasciola-infected individuals and individuals not infected with Fasciola, presenting higher prevalences in individuals with fascioliasis. Multiparasitism diagnosed in Fasciola-infected subjects included coinfections by up to nine parasite species, eight protozoan species, and five helminth species. The most prevalent pathogenic protozoan was Giardia intestinalis. The most prevalent helminth species differ according to environmental conditions. Several parasites indicate fecal environmental contamination. When the fascioliasis burden increases, the total number of parasite species also increases. The fascioliasis risk increases when the total helminth species number/host increases. Fasciola-infected subjects may present a modification in the clinical phenotypes of coinfecting parasitic diseases. Fascioliasis coinfection factors include transmission ways and immunological, environmental and social aspects. Coinfections must be considered when assessing the health impact of fascioliasis, including the analysis of the fascioliasis effects on malnutrition and physical/intellectual child development. Fascioliasis-control schemes should, therefore, integrate control measures mainly against other helminthiases. Full article

Rhabditis (Rhabditella) axei is a predominantly free-living nematode commonly found in sewage systems and decomposing organic matter. While primarily saprophytic, it has been documented as an opportunistic pathogen in human urinary and gastrointestinal tracts. The Chinese red panda (Ailurus styani), a rare and protected species in China, has not previously been reported as a host for Rhabditis (Rhabditella) spp. infections. This study reports the first documented occurrence of R. axei in red panda feces, unambiguously confirmed through integrative taxonomic approaches combining morphological and molecular analyses. The nematodes exhibited key morphological features consistent with R. axei, including a cylindrical rhabditiform esophagus, sexually dimorphic tail structures, and diagnostic spicule morphology. Molecular analysis based on 18S-ITS-28S rDNA sequencing confirmed their identity, showing >99% sequence similarity to R. axei reference strains (GenBank: PP135624.1, PP135622.1). Phylogenetic reconstruction using 18S rDNA and ITS rDNA sequences placed the isolate within a well-supported R. axei clade, clearly distinguishing it from related species such as R. blumi and R. brassicae. The findings demonstrate the ecological plasticity of R. axei as a facultative parasite capable of infecting non-traditional hosts and further highlight potential zoonotic risks associated with environmental exposure in captive wildlife populations. Our results emphasize the indispensable role of molecular diagnostics in accurately distinguishing morphologically similar nematodes within the Rhabditidae family, while providing essential baseline data for health monitoring in both in situ and ex situ conservation programs for this endangered species. Full article

The gross benefit of feeding multi-species probiotics has been reported, but the effect on the gut microbiota in pre-weaned dairy calves has not been elucidated. To address this gap, a randomized controlled trial was conducted in California, USA, to investigate the effect of feeding probiotics on the fecal microbiota of pre-weaned dairy calves. A total of 30 neonatal calves were randomly assigned to either the probiotic (PRO) or control (CON) treatment. Fecal samples were collected at four age timepoints: days 7, 14, 21, and 42. Fecal bacterial population was analyzed using 16S rRNA amplicon sequencing. Differential abundance analysis was conducted to investigate the difference between the PRO and CON treatments, and diarrheic and non-diarrheic calves in each PRO and CON group. The PRO group had decreased Clostridium perfringens and Fusobacterium varium compared to the CON at 7 days of age. At 7 days of age, diarrheic calves in CON had more abundant F. varium compared to non-diarrheic calves, but there was no difference between diarrheic and non-diarrheic calves in the PRO group. In conclusion, probiotics administration decreased the population of pathogenic bacteria in feces from pre-weaned dairy calves on Day 7 of age. However, the treatment did not have an impact on bacterial diversity. These results suggest that the administration of probiotics has the potential to control gastrointestinal pathogens. Full article

Background/Objectives: The novel compound 221s (2,9), derived from danshensu and ACEI-active proline, exhibits antihypertensive effects (50/35 mmHg SBP/DBP reduction in SHRs) with potential cough mitigation. However, its excretion kinetics remain unstudied. This study investigates 221s (2,9) elimination in rats to bridge this knowledge gap. Methods: Excretion of unchanged 221s (2,9) was quantified in urine, feces, and bile of Sprague-Dawley rats after oral administration (30 mg/kg). Concentrations of unchanged 221s (2,9) in all matrices were quantified using developed UPLC-MS/MS that underwent methodological validation. Excretion amount, excretion velocity, and accumulative excretion rate of 221s (2,9) were calculated. Results: Urinary excretion exhibited rapid elimination kinetics, reaching peak cumulative excretion rates (138.81 ± 15.56 ng/h) at 8 h post-dosing and plateauing by 48 h (cumulative excretion: 1479.81 ± 155.7 ng). Fecal excretion displayed an accelerated elimination phase between 4 and 8 h (excretion rate: 7994.29 ± 953.75 ng/h), followed by a sustained slow-release phase, culminating in a cumulative output of 36,726.31 ± 5507 ng at 48 h. Biliary excretion was minimal and ceased entirely by 24 h. Notably, total recovery of unchanged drug across all matrices remained below 1% (urine: 0.020 ± 0.021%; feces: 0.73 ± 0.069%; bile: 0.00044 ± 0.00002%) at 72 h. Conclusions: This study provides the first definitive excretion data for 221s (2,9). Quantitative analysis via a validated UPLC-MS/MS method revealed that fecal excretion is the principal elimination pathway for unchanged 221s (2,9) in rats, with direct excretion of the parent compound accounting for <1% of the administered dose over 72 h. Future studies will employ extended pharmacokinetic monitoring and concurrent UPLC-MS/MS analysis of the parent drug and phase II conjugates to resolve the observed mass imbalance and elucidate contributions to total elimination. Full article

Background/Objectives: Ulcerative colitis (UC) is characterized by the interplay between immune responses and dysbiosis in disease development. Aiming to provide additional insights into disease development and potential treatment strategies, the present study investigates the local effect of oral treatment with polysaccharides obtained from Auricularia auricula’s submerged culture in an experimental model of DSS-induced colitis and its impact on lesion resolution. Methods: The structure and monosaccharide composition of Auricularia polysaccharides were characterized through Nuclear Magnetic Resonance (NMR). To evaluate the effect of this polysaccharide on the murine model, wild-type and Dectin-1 knockout mice were treated or not with the exopolysaccharide (EPS) while under DSS consumption. During the experimental period, feces samples were collected to evaluate microbial shifts during disease development, and, finally, the colonic tissue was analyzed to assess the inflammatory process and cytokine production. Results: The EPS composition showed a polymeric mixture of glucans and fucogalactomannans. The treatment of the wild-type DSS-induced colitis group improved the inflammatory response by increasing gut–homeostatic cytokines, such as interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-α). The Dectin-1 KO mice group did not show the same enhancement after EPS treatment. The microbiome analysis revealed a difference in the genotype, and the treatment modified the DSS microbiome modulation, with nine and four ASVs in WT and Dectin-1 KO mice, respectively. Conclusions: The EPS treatment demonstrated therapeutic potential in treating inflammatory intestinal diseases by modulating cytokine secretion and microbiota composition, which is dependent on the Dectin-1 receptor’s carbohydrate recognition. Full article

Background: Diarrhea induced by an excessive amount of fat is a prevalent gastrointestinal disorder. Currently, there are limited animal models and treatment options for diarrhea associated with fat. This study aims to develop a mouse model of high-fat-associated diarrhea using glyceryl-trioleate (GTO) and evaluate the potential of montmorillonite clay (MMT) in mitigating this condition. Methods: GTO was administered to mice at different doses through oral gavage to induce diarrhea. Clay was treated through oral gavage to evaluate its anti-diarrhea effect. Fecal conditions were monitored. Intestinal tissues were subjected to histological examination to assess structural integrity. The total fecal bile acids were evaluated using a bile acid assay kit to determine the mechanism of action. Results: The results showed that a diarrhea model was established by administering GTO at 2000 mg/kg. When the animals were treated with clay, diarrhea incidence and severity were decreased significantly in a dose-dependent manner. Compared to the untreated group receiving GTO alone, clay co-administration at 2000 mg/kg reduced diarrhea scores by approximately 48%, while the higher dose of 4000 mg/kg achieved an 83% reduction. Fecal bile acid analysis showed that diarrhea is associated with total bile acid levels in the feces. Histological exams showed that diarrhea is associated with tissue inflammation in the colon. Conclusions: This study showed that GTO administration induced diarrhea in mice, and clay effectively alleviates fat-induced diarrhea through modulation of fecal bile acid composition. These findings suggest that this model can be used to evaluate diarrhea associated with excessive amounts of fat and clay that can be further tested for diarrhea attenuation. Full article

Grazing is a free-range farming model commonly practiced in low-external-input agricultural systems. The widespread use of veterinary antibiotics in livestock farming has led to significant environmental accumulation of antibiotic residues and antibiotic resistance genes (ARGs), posing global health risks. This study investigated the antibiotic residues, bacterial community, ARG profiles, and mobile genetic elements (MGEs) in cattle feces from three provinces in western China (Ningxia, Xinjiang, and Inner Mongolia) under grazing modes. The HPLC-MS detection showed that the concentration of tetracycline antibiotics was the highest in all three provinces. Correlation analysis revealed a significant negative correlation between antibiotic residues and the diversity and population abundance of intestinal microbiota. However, the abundance of ARGs was directly proportional to antibiotic residues. Then, the Sankey analysis revealed that the ARGs in the cattle fecal samples were concentrated in 15 human pathogenic bacteria (HPB) species, with 9 of these species harboring multiple drug resistance genes. Metagenomic sequencing revealed that carbapenemase-resistant genes (bla_KPC and bla_VIM) were also present in considerable abundance, accounting for about 10% of the total ARGs detected in three provinces. Notably, Klebsiella pneumoniae strains carrying bla_CTX-M-55 were detected, which had a possibility of IncFII plasmids harboring transposons and IS19, indicating the risk of horizontal transfer of ARGs. This study significantly advances the understanding of the impact of antibiotic residues on the fecal microbiota composition and ARG profiles in grazing cattle from northwestern China. Furthermore, it provides critical insights for the development of rational antibiotic usage strategies and comprehensive public health risk assessments. Full article

Vibrios are widespread in marine environments, and their persistence is often linked to natural reservoirs such as filter-feeding bivalves. This study investigated the capacity of the Mediterranean mussel, Mytilus galloprovincialis, to act as a reservoir of Vibrio harveyi using a GFP-tagged strain in controlled experiments. Mussels (shell length 4–6 cm) were exposed to V. harveyi gfp in estuarine and seawater at 12 °C and 20 °C over six days. Bacterial accumulation in gills, digestive gland, and gonads, as well as in feces and pseudofeces, was quantified, and the immune response following microbial challenge was assessed by histopathological analysis. Mussels actively removed V. harveyi from the water, but not completely. Vibrios were rapidly accumulated in organs, with the highest densities in the digestive gland (up to 10⁷–10⁸ CFU g⁻¹), and substantial bacterial loads detected in biodeposits (1.55–3.77 × 10⁷ CFU g⁻¹). Salinity had a greater effect than temperature on bacterial accumulation, with consistently higher counts in seawater assays. Concurrently with bacterial accumulation, mussels activated their immune system, as evidenced by the detection of granulocytomas and hemocytic infiltrations. Overall, these results demonstrate that M. galloprovincialis accumulates V. harveyi in tissues and biodeposits, serving as a natural reservoir for this bacterium. Full article