Search Results (2,574)

The analysis of proteins in stool samples can significantly enhance the study of mammalian physiology and disease. In this study, we investigated the fecal proteome of clinically healthy dogs (n = 26) by a label-free proteomics approach to evaluate the impact of breed differences. The dogs were divided into two groups (n = 13 each) based on their breed, specifically Weimaraner and Dalmatian, the former known for their possible susceptibility to gastrointestinal disease. Quantitative and qualitative differences between the two experimental groups were identified based on analyses performed on pooled biological samples. The overall fecal proteome profile comprised 58 proteins, of which 37 were common, while comparative proteomics analysis detected 15 proteins with different abundances. Notably, the fecal proteome of Weimaraners showed an over-representation of proteins such as pantetheinase, which promotes inflammatory reactions; ferritin heavy chain and hemoglobin, possibly associated with gut ulceration and/or rectal bleeding typical of IBD; and anionic trypsin, implicated in inflammatory bowel disease. Notably, in Dalmatians, despite the absence of specific predispositions, some proteins associated with chronic enteropathy (e.g., carboxypeptidase B and serine protease 1) were also over-represented. Additionally, some proteins linked to breed variation included enzymes associated with “protein digestion and absorption” and “glycolysis and gluconeogenesis”. These findings suggest, for the first time, that the variable breed is a factor that may potentially influence the fecal proteome in dogs. Full article

Next-generation sequencing has overturned the dogma of biliary sterility, revealing low-biomass microbiota along the gut–biliary axis with metabolic and immunologic effects. This review synthesizes evidence on composition, function, and routes of colonization across benign and malignant disease. In cholelithiasis, Proteobacteria- and Firmicutes-rich consortia provide β-glucuronidase, phospholipase A₂, and bile salt hydrolase, driving bile supersaturation, nucleation, and recurrence. In primary sclerosing cholangitis, primary biliary cholangitis, and autoimmune hepatitis, intestinal dysbiosis and disturbed bile acid pools modulate pattern recognition receptors and bile acid signaling (FXR, TGR5), promote Th17 skewing, and injure cholangiocytes; bile frequently shows Enterococcus expansion linked to taurolithocholic acid. Distinct oncobiomes characterize cholangiocarcinoma subtypes; colibactin-positive Escherichia coli and intratumoral Gammaproteobacteria contribute to DNA damage and chemoresistance. In hepatocellular carcinoma, intratumoral microbial signatures correlate with tumor biology and prognosis. We critically appraise key methodological constraints—sampling route and post-sphincterotomy contamination, antibiotic prophylaxis, low biomass, and heterogeneous analytical pipelines—and outline a translational agenda: validated microbial/metabolomic biomarkers from bile, tissue, and stent biofilms; targeted modulation with selective antibiotics, engineered probiotics, fecal microbiota transplantation, and bile acid receptor modulators. Standardized protocols and spatial, multi-omic prospective studies are required to enable risk stratification and microbiota-informed therapeutics. Full article

Background/Objectives: Fecal peritonitis following penetrating abdominal trauma is a serious condition that often results in sepsis and organ failure. The aim of our study was to develop a novel conscious porcine model of sepsis and organ dysfunction caused by multiple penetrating injuries to the small and large intestines. Methods: Twelve female Yorkshire pigs (average weight 50.6 ± 6.5 kg) were divided into two groups: Penetrating Abdominal Trauma (PAT) (n = 8) and Control (n = 4). All surgical procedures were performed under anesthesia with adequate analgesia. In the PAT group, the small and large intestines were punctured, and feces mixed with saline were introduced into the abdominal cavity to induce peritonitis. The Control group received sham surgery with only saline solution. The animals were observed in a conscious state over a period of 72 h, vital parameters were recorded, and blood samples were taken regularly. We adapted a pig-specific SOFA score and developed pig-specific SIRS criteria and NEWS2 score to assess organ function. The model was validated by independent investigators. Results: The survival rate in the PAT group was 75%, with an average survival time of 58.5 h, while all animals in the Control group survived to euthanasia. Monitoring showed pathophysiological changes, such as tachycardia, leucopenia, and thrombocytopenia, indicative of sepsis and organ dysfunction. Blinded investigators independently confirmed the model’s validity. Conclusions: A new swine model of penetrating abdominal trauma and sepsis has been successfully developed that demonstrates significant physiological and immunologic changes comparable to human sepsis. This new model provides a realistic platform for future research into sepsis, its diagnostics, and the evaluation of therapeutic strategies. Full article

Background: African swine fever (ASF) is a highly contagious and often deadly disease that poses a major threat to swine production worldwide. The lack of a commercially available vaccine underscores the critical need for innovative immunization strategies to combat ASF. Methods: Six ASFV antigenic proteins (K78R, A104R, E120R, E183L, D117L, and H171R) were fused with the Lactiplantibacillus plantarum WCFS1 surface anchor LP3065 (LPxTG motif) to generate recombinant Lactiplantibacillus plantarum NC8 (rNC8) strains. The surface expression was confirmed using immunofluorescence and Western blotting assays. Additionally, the dendritic cell-targeting peptides (DCpep) were co-expressed with each antigen protein. Mice were immunized at a dosage of 10⁹ colony-forming units (CFU) per strain per mouse via intragastric (I.G.), intranasal (I.N.), and intravenous (I.V.) routes. The bacterial mixture was heat-inactivated by boiling for 15 min to destroy viable cells while preserving antigenic structures. I.V. administration caused no hypersensitivity, confirming the method’s safety and effectiveness. Results: Following I.G. administration, rNC8-E120R, rNC8-E183L, rNC8-K78R, and rNC8-A104R induced significant levels of secretory immunoglobulin A (sIgA) in fecal samples, whereas rNC8-H171R and rNC8-D117L failed to induce a comparable response. Meanwhile, rNC8-D117L, rNC8-K78R, and rNC8-A104R also elicited significant levels of sIgA in bronchoalveolar lavage fluid (BALF). Following I.N. immunization, rNC8-E120R, rNC8-K78R, and rNC8-A104R significantly increased sIgA levels in both fecal and BALF immunization. In contrast, I.V. immunization with heat-inactivated rNC8-K78R and rNC8-A104R induced robust serum IgG titers, whereas the remaining antigens elicited minimal or insignificant responses. Flow cytometry analysis revealed expanded CD3⁺CD4⁺ T cells in mice immunized via the I.N. and I.G. and CD3⁺CD4⁺ T cells only in those immunized via the I.N. route. Th1 responses were also significant in the sera of mice immunized via the I.G. and I.N. routes. Conclusions: The rNC8 multiple-antigen cocktail elicited strong systemic and mucosal immune responses, providing a solid foundation for the development of a probiotic-based vaccine against ASF. Full article

Recent studies have increasingly emphasized the regulatory potential of plant-derived polysaccharides on gut microbial composition and metabolic function. Despite this growing interest, investigations focusing specifically on the simulated digestion and fermentation properties of arecanut polysaccharide (PAP1b) remain limited. In this work, we employed the standardized INFOGEST 2.0 protocol to mimic the oral, gastric, and intestinal digestion of PAP1b, followed by 48 h anaerobic fermentation using pooled human fecal samples from healthy adult donors. PAP1b treatment led to a progressive decrease in pH and a substantial elevation in SCFAs levels, notably acetic, propionic, and butyric acids. Simultaneously, PAP1b significantly promoted the growth of SCFA-producing microbial taxa, particularly members of the Firmicutes phylum such as Lachnospiraceae, Lachnoclostridium, Bilophila, and Phascolarctobacterium, while markedly suppressing Bacteroidota populations. Metabolomic analysis further indicated that PAP1b intake enhanced bile acid metabolism, suggesting its potential as a prebiotic candidate for improving intestinal health. Full article

Cryptosporidium is a genus of protozoa that infects the gastrointestinal and respiratory epithelium of various host species. The aim of this study was to perform experimental infection in conventional mice with three Cryptosporidium species isolated from humans. The three Cryptosporidium species, namely Cryptosporidium canis, Cryptosporidium parvum, and Cryptosporidium ryanae, were obtained from fecal samples collected from patients hospitalized in an infectious disease hospital. The mice, from 10-day-old litters kept with their mother, were divided into three groups and orally infected with one of the Cryptosporidium species. The first oocysts were identified in the feces of the mice four days post-infection. The infection was successful with all three Cryptosporidium species, but the infection level (expressed as the number of oocysts per microscopic field) was low. The infection was detected using a rapid immunochromatographic test 40 days post-infection. Furthermore, starting on the 17th day after infection, the mothers also tested positive on the rapid immunochromatographic test, having been negative until that point. It was concluded that mice could represent a source of infection for the three Cryptosporidium species in other susceptible species, including humans. No behavioral changes or diarrhea were observed in any of the experimental cases. Full article

Antimicrobial resistance (AMR) poses an increasing threat to public health, with wildlife recognized as reservoirs and vectors of resistant bacteria. However, the role of wild species in the ecology of AMR remains insufficiently understood, highlighting the need to investigate resistant bacteria in these animals. This study focused on detecting and characterizing Escherichia coli obtained from 43 fecal samples of white storks (Ciconia ciconia), cattle egrets (Bubulcus ibis), and European hedgehogs (Erinaceus europaeus) admitted to a wildlife rehabilitation center in Portugal. Resistance profiles to twelve antibiotics and six virulence factors were characterized phenotypically. ESBL production was also tested. A total of 79 E. coli isolates were obtained from 39 out of 43 samples, and 75 were selected for further characterization. All isolates tested negative for ESBL production. Approximately 64% (n = 48/75) of isolates were resistant to at least one antibiotic, and 5.3% (n = 4/75) were multidrug-resistant. Most frequent resistances were to ampicillin (36%, n = 27/75), tetracycline (12%, n = 9/75), and chloramphenicol (8%), while all isolates were susceptible to meropenem, aztreonam, and third-generation cephalosporins. Most isolates (81.3%, n = 61/75) lacked virulence factors. These findings suggest that wildlife may act as a reservoir of resistant strains, emphasizing importance of AMR monitoring and the One Health approach. Full article

Background: In healthy adults, probiotic supplementation alone does not increase Urolithin A (UroA) and rarely increases butyrate, both microbiome-derived metabolites that influence key biological functions involved in regulating gastrointestinal symptoms. Accordingly, this clinical trial evaluated key biological functions of a multi-species synbiotic with 24 probiotic strains and a polyphenol-based prebiotic using capsule-in-capsule delivery technology. Methods: We conducted a randomized, placebo-controlled trial among healthy participants (n = 32). Participants were administered a daily synbiotic (53.6 billion AFU multi-species probiotic and 400 mg Indian pomegranate extract; DS-01) or matching placebo for 91 days. Samples were obtained at baseline Day 0, and Days 7, 14, 49, and 91. Endpoints included changes in fecal microbiome composition, urinary UroA, fecal butyrate, serum CRP, and safety. Results: The synbiotic significantly increased alpha-diversity of Bifidobacterium and Lactobacillus spp. at all timepoints, including at end-of-study (Day 91, p < 0.0001) and increased native beneficial microbes. UroA production was significantly increased in the synbiotic arm at short-term (Day 7, 12-fold, p < 0.02) and long-term (Day 91, 49-fold, p < 0.001) timepoints. A higher proportion of synbiotic participants were capable of converting polyphenols into UroA (Day 91, 100% vs. 44.4%; p < 0.01). Mechanistically, synbiotic participants showed an increased abundance of Lactobacillus species involved in UroA precursor metabolism and UroA-producing Gordonibacter species. The synbiotic also significantly increased fecal butyrate levels (p < 0.03), and butyrate-producing species, in low-baseline butyrate producers, through Day 91, and was associated with reduced systemic inflammation. Conclusions: This multi-species synbiotic significantly increases diversity and abundance of key beneficial bacteria, enhances UroA production and butyrate levels, and is associated with lowered systemic inflammation. This is the first synbiotic to increase both UroA and butyrate. Full article

Coastal fecal contamination is a global public health concern, particularly due to waterborne protozoan parasites such as Giardia duodenalis and Blastocystis sp. Concepcion Bay (Chile) is an important recreational and productive area in Chile. Nevertheless, it is impacted by two submarine outfalls and a rural sewage treatment plant, which may contribute to fecal pollution. This study evaluated the presence of waterborne parasites in Aulacomya atra mussels intended for human consumption. The mussels were collected from three sectors of the bay: northern, central, and southern. A total of 600 mussels were analyzed as accumulators using PCR targeting SSU-rDNA and β-giardin genes for the detection of Blastocystis sp. and G. duodenalis, respectively. Additionally, thermotolerant coliforms were quantified using the most probable number (MPN) method. Both parasites and coliforms were detected in all sectors, with the southern zone showing the highest number of positive samples, indicating a localized public health concern. This is the first report of these protozoa in mussels from Concepción Bay. The findings highlight the need for regulatory frameworks to control protozoan discharge and reduce pathogen transmission risks in coastal ecosystems, especially in areas with high recreational and economic activity, both in Chile and worldwide. Full article

Background/Objective: Probiotic and vitamin D supplements are widely studied in clinical and animal studies as potential treatments for inflammatory bowel disease. However, their potential synergistic or additive effect in ameliorating colitis development is still poorly understood. The aim of this study was to investigate the potential beneficial enhancement of combining a mixed-strain probiotic with vitamin D supplementation in a colitis animal model. Method: After 5 days of acclimation, C57BL/6 mice received Vivomixx probiotic (at least 1 × 10⁹ Colony-Forming Units) and/or vitamin D (5 IU/g) in drinking water and chow, respectively, for 7 days prior to intracolonic TNBS-induced colitis and until sacrifice. On day 10, animals were sacrificed, and colons were collected to assess colonic damage, cytokine and chemokine expression, total M1 macrophage phenotype, and neutrophil recruitment. Serum and fecal samples were collected to assess vitamin D levels and microbiome composition. Results: Administration of probiotic and vitamin D alone or combined decreased colonic damage and neutrophil recruitment and activity. This effect was associated with an increase in the active form of vitamin D in serum and mucosal barrier integrity. However, administration of probiotics and/or vitamin D did not modulate macrophage infiltration or the M1 pro-inflammatory phenotype. Conclusions: These results suggest that combined probiotic and vitamin D supplementation attenuates TNBS-induced colitis by targeting neutrophil infiltration while enhancing the mucosal barrier. This alternative approach may offer protective potential for IBD management. 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

Cryptosporidium is a major waterborne parasite that causes gastrointestinal illness. Conventional assays, including microscopy and immunological identification, often suffer from false positives or negatives due to non-specific binding or morphological differences between Cryptosporidium species. We developed a novel qPCR assay based on a Cryptosporidium-specific Conserved Signature Protein (CSP) to address the limitations of testing complex samples, including those from recreational waters. The CSP (hypothetical protein (cgd2_3830)) was identified as taxonomically unique to Cryptosporidium species. The CSP sequence and designed qPCR assay primers/probe demonstrated high specificity for the targeted Cryptosporidium species when tested against NCBI RefSeq databases. qPCR assay efficiency was determined as 95% and an R² value of 0.99, with a slope and intercept of −3.4 and 40.1, respectively. Additionally, the Lower Limit of Detection (ALLOD) was determined as three gene copies, suggesting the potential to detect even a single oocyst. No non-specific amplification products or primer dimers were observed when the qPCR assay was evaluated using recreational water, fecal solution, and wastewater, while spike-in-control tests indicated minimal interference with the sensitivity of the assay, highlighting application for testing complex environmental DNA extracts. These findings highlight the application of the novel CSP-based qPCR assay for the rapid and sensitive detection of Cryptosporidium sp., thereby circumventing the sequence variability and multi-copy limitations associated with existing molecular markers. This proof-of-concept study presents a diagnostic framework utilizing CSP-based markers for developing water quality monitoring strategies, with scope for expansion to other microbial pathogens and potential applications in clinical and food safety settings. Full article

Using wheat flour milling (WFM) co-products in pig diets may reduce feed cost. Still, energy digestibility is lower for WFM co-products than for feed grains. Inadequate information exists about their fermentation characteristics and the relationship between digestible energy (DE) value and chemical characteristics or in vitro energy digestibility. The objectives were to (1) determine the chemical characteristics, in vitro and in vivo DE values, and energy digestibility of WFM co-products in growing pigs; (2) determine their in vitro microbial fermentation characteristics, and (3) establish relationships between in vivo DE value of WFM co-products and their chemical composition, fermentation characteristics, or in vitro digestibility values. Across Canada, 94 WFM co-products were sampled and characterized for their chemical composition and in vitro dry matter (DM) and energy digestibility using pepsin, pancreatin, and a multi-enzyme complex containing arabinase, β-glucanase, hemicellulase, xylanase, and cellulase. The in vivo energy, DM digestibility and DE value of 9 WFM co-products (2 shorts, 5 millrun, 1 middling, and 1 bran) were determined using a corn-based diet and 40 growing pigs in two periods to obtain 8 observations per diet. After in vitro digestion, the 9 WFM co-product samples were subjected to microbial fermentation using fresh fecal inoculum in a cumulative gas-production technique. The WFM co-products had a high content of crude fiber (up to 7.9% in shorts, 9.9% in millrun, 7.1% in middlings, and 12.0% in bran) and crude protein (CP; up to 27.8% in shorts, 20.0% in millrun, 22.1% in middlings, 15.9% in bran). The DE values ranged from 2.84 to 3.74 Mcal/kg DM among WFM co-products. Among chemical characteristics, neutral detergent fiber was the best predictor (R² = 0.81) for in vivo DE value, followed by crude fiber (R² = 0.78), and acid detergent fiber (R² = 0.72). The in vitro DE values predicted (R² = 0.80) in vivo DE values of 9 WFM co-products. Based on principal component analysis, total gas and short-chain fatty acid production varied among WFM co-products and was associated with the CP content of WFM co-products. In conclusion, WFM co-products contain high crude protein and have a high DE value for growing pigs but vary substantially in nutritional value. Full article

Monitoring surface water quality offers a clear understanding of its parameters over time and space. The Ibrahim River, one of the main rivers in Lebanon, was monitored over one hydrological year, from March 2021 to April 2022. Samples were collected from seven stations in the watershed, once every two weeks. A total of 504 samples were then analyzed for pH, conductivity, turbidity, total dissolved solids, dissolved oxygen, biochemical oxygen demand, dissolved nitrate, dissolved potassium, dissolved chloride, total alkalinity, fecal coliforms, and total coliforms. Principal Component Analysis (PCA) was able to highlight two principal components (PCs), representing spatial and temporal variations, identifying areas of pollution and the influence of flow on water quality. The adapted Water Quality Index (WQI) confirmed the PCA trend with an overall average for the entire watershed of 83.70 ± 4.97, indicating a “good” water quality. Full article

Mycotoxins are common feed contaminants that can affect the health, immune function, and productivity of ruminants by causing oxidative stress and organ dysfunction. In this field study, the effects of a phytogenic multicomponent mycotoxin detoxifier on oxidative status, liver function, udder health, and productive parameters were investigated in dairy ewes. One hundred clinically healthy ewes were randomly assigned to either a control group or a treatment group, with the latter receiving 1.5 kg/ton of the detoxifier over a 90-day period during lactation. The detoxifying agent contained adsorptive clays as well as phytogenic ingredients such as silymarin and curcumin, which are known for their hepatoprotective and antioxidant properties. Blood, milk, and colostrum samples were collected and analyzed for oxidative stress markers (TBARS and protein carbonyl (CARBS)), total antioxidant capacity (TAC), liver enzymes (ALT, AST, and ALP), and milk quality parameters (fat, protein, and solid content). Clinical assessments included mastitis scoring, udder inflammation, and fecal consistency. The treated ewes showed a statistically significant reduction in blood plasma and milk oxidative stress markers and liver enzyme levels while at the same time improving the fat and solid content of the milk. The incidence and severity of mastitis, udder reddening, and lactation abnormalities were lower in the treatment group. Brix refractometry indicated improved colostrum quality in the treated ewes. These results suggest that the detoxifier improved the oxidative balance, liver function, and overall health and productivity of dairy ewes under field conditions, supporting its use as a practical nutritional measure. Full article