Project description:Fermented foods harbor beneficial microbes that support gut health and metabolic balance, offering natural probiotic strategies for managing conditions such as hyperuricemia through modulation of the gut microbiota and their derived metabolites. We demonstrate that Eurotium cristatum (E. cris), a dominant probiotic fungus in Fu brick tea, can directly utilize uric acid (UA) as a carbon source and ameliorate hyperuricemia progression by reprogramming the gut microbiome and metabolome, thereby alleviating NLRP3-mediated inflammation and pyroptosis in renal tissues. Oral administration of E. cristatum in hyperuricemic mice upregulated the expression of ATP-binding cassette subfamily G member 2 (ABCG2) in both the kidney and intestine, as well as the intestinal tight junction protein ZO-1, suggesting enhanced UA excretion and maintenance intestinal permeability. Microbiota analysis revealed that E. cristatum reversed dysbiosis by enriching beneficial bacteria (Coprococcus, Ruminococcus) while reducing pathogenic taxa (Turicibacteraceae, Clostridiaceae). Correlation analyses further indicated that these changes were linked to the restoration of the host's co-metabolite profile, characterized by enhanced purine degradation (decreased xanthine/hypoxanthine) and control amino acid metabolism (regulation of the urea cycle, histidine metabolism and the branched-chain amino acid pathway), which synergistically promoted UA clearance and reduced systemic metabolic burden. Notably, fecal microbiota transplantation in the intervention group replicated these effects, confirming that these benefits depend on the remodeling of the microbiota. Furthermore, we confirmed that the microbiome remodeling induced by E. cristatum improved the urea cycle, histidine metabolism and contributed to the suppression of renal macrophage infiltration and NLRP3/GSDMD-mediated pyroptosis. In summary, this study elucidates how a food-derived probiotic fungus reshapes the gut microbiota and its metabolome to provide renal protection by promoting UA catabolism, the urea cycle and histidine metabolism, thereby offering new perspectives for probiotic intervention strategies.
Project description:Fermented foods harbor beneficial microbes that support gut health and metabolic balance, offering natural probiotic strategies for managing conditions such as hyperuricemia through modulation of the gut microbiota and their derived metabolites. We demonstrate that Eurotium cristatum (E. cris), a dominant probiotic fungus in Fu brick tea, can directly utilize uric acid (UA) as a carbon source and ameliorate hyperuricemia progression by reprogramming the gut microbiome and metabolome, thereby alleviating NLRP3-mediated inflammation and pyroptosis in renal tissues. Oral administration of E. cristatum in hyperuricemic mice upregulated the expression of ATP-binding cassette subfamily G member 2 (ABCG2) in both the kidney and intestine, as well as the intestinal tight junction protein ZO-1, suggesting enhanced UA excretion and maintenance intestinal permeability. Microbiota analysis revealed that E. cristatum reversed dysbiosis by enriching beneficial bacteria (Coprococcus, Ruminococcus) while reducing pathogenic taxa (Turicibacteraceae, Clostridiaceae). Correlation analyses further indicated that these changes were linked to the restoration of the host's co-metabolite profile, characterized by enhanced purine degradation (decreased xanthine/hypoxanthine) and control amino acid metabolism (regulation of the urea cycle, histidine metabolism and the branched-chain amino acid pathway), which synergistically promoted UA clearance and reduced systemic metabolic burden. Notably, fecal microbiota transplantation in the intervention group replicated these effects, confirming that these benefits depend on the remodeling of the microbiota. Furthermore, we confirmed that the microbiome remodeling induced by E. cristatum improved the urea cycle, histidine metabolism and contributed to the suppression of renal macrophage infiltration and NLRP3/GSDMD-mediated pyroptosis. In summary, this study elucidates how a food-derived probiotic fungus reshapes the gut microbiota and its metabolome to provide renal protection by promoting UA catabolism, the urea cycle and histidine metabolism, thereby offering new perspectives for probiotic intervention strategies.
Project description:5 human fecal gut samples, collected and prepared for standard MudPIT data collection from healthy volunteers, searched with the ComPIL database. x3 replicates each
Project description:The aim of this study was to test the hypothesis that replenishing the microbiota with a fecal microbiota transplant (FMT) can rescue a host from an advanced stage of sepsis. We developed a clinically-relevant mouse model of lethal polymicrobial gut-derived sepsis in mice using a 4-member pathogen community (Candida albicans, Klebsiella oxytoca, Serratia marcescens, Enterococcus faecalis) isolated from a critically ill patient. In order to mimic pre-operative surgical patient condition mice were exposed to food restriction and antibiotics. Approximately 18 hours prior to surgery food was removed from the cages and the mice were allowed only tap water. Each mouse received an intramuscular Cefoxitin injection 30 minutes prior to the incision at a concentration of 25 mg/kg into the left thigh. Mice were then subjected to a midline laparotomy, 30% hepatectomy of the left lateral lobe of the liver and a direct cecal inoculation of 200 µL of the four pathogen community. On postoperative day one, the mice were administered rectal enema. Mice were given either 1 ml of fecal microbiota transplant (FMT) or an autoclaved control (AC). This was again repeated on postoperative day two. Mice were then followed for mortality. Chow was restored to the cages on postoperative day two, approximately 45 hours after the operation. The injection of fecal microbiota transplant by enema significantly protected mice survival, reversed the composition of gut microflora and down-regulated the host inflammatory response. The cecum, left lobe of the liver, and spleen were isolated from mice for microarray processing with three or more replicates for six expermental conditions: non-treated control, SAHC POD1, SAHC.AC POD2, SAHC.FMT POD2, SAHC.AC POD7, SAHC.FMT POD7
Project description:A metaproteomics analysis was conducted on the infant fecal microbiome to characterize global protein expression in 8 samples obtained from infants with a range of early-life experiences. Samples included breast-, formula- or mixed-fed, mode of delivery, and antibiotic treatment and one set of monozygotic twins. Although label-free mass spectrometry-based proteomics is routinely used for the identification and quantification of thousands of proteins in complex samples, the metaproteomic analysis of the gut microbiome presents particular technical challenges. Among them: the extreme complexity and dynamic range of member taxa/species, the need for matched, well-annotated metagenomics databases, and the high inter-protein sequence redundancy/similarity between related members. In this study, a metaproteomic approach was developed for assessment of the biological phenotype and functioning, as a complement to 16S rRNA sequencing analysis to identify constituent taxa. A sample preparation method was developed for recovery and lysis of bacterial cells, followed by trypsin digestion, and pre-fractionation using Strong Cation Exchange chromatography. Samples were then subjected to high performance LC-MS/MS. Data was searched against the Human Microbiome Project database, and a homology-based meta-clustering strategy was used to combine peptides from multiple species into representative proteins. Bacterial taxonomies were also identified, based on species-specific protein sequences, and protein metaclusters were assigned to pathways and functional groups. The results obtained demonstrate the applicability of this approach for performing qualitative comparisons of human fecal microbiome composition, physiology and metabolism, and also provided a more detailed assessment of microbial composition in comparison to 16S rRNA.