Fecal Microbiota Transplantation from Healthy Piglets Ameliorates Intestinal Inflammation in Mice by Modulating Recipient Metabolism

Fecal Microbiota Transplantation from Healthy Piglets Ameliorates Intestinal Inflammation in Mice by Modulating Recipient Metabolism | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Fecal Microbiota Transplantation from Healthy Piglets Ameliorates Intestinal Inflammation in Mice by Modulating Recipient Metabolism Qinjin Li, Yuqing Wang, Jingqiang Li, Yan Gao, Zhifeng Wu, Xiang Tan, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7536850/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 12 Jan, 2026 Read the published version in BMC Microbiology → Version 1 posted 17 You are reading this latest preprint version Abstract Fecal Microbiota Transplantation (FMT) has been clinically applied to treat host intestinal inflammation, such as inflammatory bowel disease (IBD). Research in livestock indicates that the gut microbiota of healthy piglets differs from that of diarrheic piglets, playing a crucial role in regulating intestinal immune development. However, the potential of FMT derived from healthy piglets to alleviate intestinal inflammation in recipients and the underlying mechanisms remain unexplored. This study utilized FMT from healthy piglets to intervene in a dextran sulfate sodium (DSS)-induced intestinal inflammation model in germ-free Kunming (KM) mice, investigating its effects on intestinal barrier function and inflammatory levels. As anticipated, the results demonstrated that FMT significantly alleviated DSS-induced intestinal inflammation. This was evidenced by reduced weight loss and lower disease activity index (DAI) scores. Furthermore, FMT improved intestinal barrier integrity, maintained homeostasis of host inflammatory cytokines, and markedly attenuated oxidative stress. Untargeted metabolomics analysis further revealed that FMT significantly increased the abundance of multiple anti-inflammatory metabolites, including 3-Methoxytyramine-betaxanthin and Sialorphin. Concurrently, FMT upregulated relevant metabolic pathways, notably Betalain Biosynthesis. Correlation analysis indicated a close association between FMT-elevated anti-inflammatory metabolites (e.g., 3-Methoxytyramine-betaxanthin) and improved markers of intestinal inflammation. This study provides novel insights into the mechanism by which pig-derived gut microbiota alleviates host intestinal inflammation through modulation of host metabolism. Healthy piglet microbiota Fecal microbiota transplantation Germ-free mice Inflammatory Bowel Disease Metabolome Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction The gut microbiota plays a critical role in maintaining immune homeostasis, regulating immune responses, and preventing pathogen invasion through its interactions with the host immune system [ 1 ]. This microbial ecosystem contributes significantly to microecological homeostasis, which is closely associated with immune regulatory functions and immune tolerance, exerting profound impacts on both livestock production and human health [ 2 , 3 ]. Recent studies have increasingly demonstrated that gut microbiota dysbiosis may be closely linked to the pathogenesis of various inflammatory and immune-mediated diseases [ 4 , 5 ]. Conversely, targeted modulation of host microbiota structure and function, such as through fecal microbiota transplantation (FMT) may provide novel therapeutic strategies for dysbiosis-associated disorders [ 6 – 8 ]. Intestinal health is fundamental to growth performance and disease resistance in livestock. Weaned piglets experiencing diarrhea develop intestinal inflammation, which significantly impairs subsequent growth performance. Post-weaning diarrhea (PWD) is a prevalent challenge, often leading to intestinal inflammation that significantly compromises subsequent growth performance and feed efficiency [ 9 – 11 ]. Studies comparing the gut microbiota of healthy post-weaning pigs with that of diarrheic piglets exhibiting intestinal inflammation have revealed substantial differences in microbial structure and function. [ 12 , 13 ]. This suggests a potential therapeutic role for healthy pig-derived microbiota in ameliorating intestinal inflammation [ 14 ]. However, the efficacy and underlying mechanisms through which healthy post-weaning pig microbiota exerts its anti-inflammatory effects remain largely undefined and warrant further investigation. The DSS-induced intestinal inflammation model represents a classical methodology for investigating intestinal inflammatory mechanisms [ 2 , 15 , 16 ]. This model effectively recapitulates key features of intestinal inflammation in livestock animals and human inflammatory bowel disease (IBD). The use of germ-free (GF) mice with defined microbial backgrounds facilitates the elimination of interference from native microbiota, an approach particularly critical for evaluating the anti-inflammatory efficacy of pig-derived microbiota [ 17 ]. This study employs healthy piglet-derived microbiota as the research subject. Through its transplantation into DSS-induced GF mouse models, we investigate its impact on host intestinal inflammation and barrier function, while aiming to elucidate the mechanistic basis by which healthy piglet microbiota maintains intestinal homeostasis from the perspective of metabolic regulation. This work aims to reveal the significant role of healthy piglet-derived gut microbiota in host immunomodulation via host metabolic pathways, concurrently laying theoretical groundwork for developing microbiota-based anti-inflammatory strategies. Materials and methods FMT Preparation Fresh faecal samples were collected from healthy Yorkshire piglets (35-day-old, n=3, Quanzhou Green Source Agriculture Co., Ltd) using a sterile faecal collector, and samples were transported to the laboratory under dry ice conditions, and the feces from the interior of the mid-section were homogenized under anaerobic conditions (80% N 2 , 10% H 2 , 10% CO 2 ) and mixed thoroughly with saline glycerol buffer (15% glycerol concentration) at a 1:10 (m/v) ratio. After complete dissolution, samples were pre-filtered through sterile gauze followed by final filtration through a 100-μm membrane [18]. Experimental Animals and Treatments GF female KM mice (8-10 weeks old) were obtained from the Germ-Free Animal Platform at Shanghai Tenth People's Hospital. Mice were maintained in a sterile isolator under controlled conditions: temperature 25 ± 2°C, relative humidity 45-60%, and a 12-h light/dark cycle (lights on 06:30-18:30), and had free access to sterilized food and water. This study investigated the impact of FMT derived from healthy Yorkshire piglets. Recipient mice received daily oral gavage of 100 μL bacterial suspension or saline control, followed by administration of 3% DSS. Experimental groups and timelines are detailed in Figure 1A . Body weight was recorded daily throughout the experiment, calculated as the percentage change relative to baseline (day 0) [19]. The mice were euthanized by cervical dislocation without anesthesia, a method approved by the North Stockholm Ethics Committee (permission N31/14). Upon experiment completion, fecal samples were collected within the isolator immediately prior to euthanasia. This study was reviewed and approved by the Laboratory Animal Welfare Ethics Committee of Shanghai Tenth People's Hospital, all procedures complied with the Shanghai Tenth People's Hospital Guide for the Care and Use of Laboratory Animals (Animal Ethics Approval No.: SHDSYY-2025-T0088-02). Disease Activity Index The Disease Activity Index (DAI) comprises a weight loss score, a fecal bleeding score, and stool characteristic assessments, as detailed in Table 1 [20, 21]. Briefly, the DAI was calculated based on weight change (no change = 0; 1-5% = 1; 5-10% = 2; 10-15% = 3; >15% = 4), fecal bleeding (normal colored stool = 0; brown stool = 1; red stool = 2; bloody stool = 3; heavy bleeding = 4), and stool consistency (normal stool, good shape = 0; soft stool, soft stool adhering to the anus = 1-2; diarrhea, adherent anal = 3-4), and the overall DAI score was obtained by averaging these three individual scores. Table 1. Disease Activity Index Score Weight loss (%) Stool consistency Bloody stool score 0 None Normal Normal colored stool 1 1-5 Loose stool Brown stool 2 5-10 Loose stool Reddish stool 3 10-15 Diarrhea Bloody stool 4 > 15 Diarrhea Gross bleeding Histologic Analysis of Mice Colon Distal colon segments from mice in each group were fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned into 4 μm-thick slices. Sections were stained with hematoxylin-eosin (H&E), immunohistochemistry (IHC) and immunofluorescence (IF), and images were acquired using a microscope (Nikon Eclipse 80i, Japan). H&E-stained sections were evaluated for inflammatory cell infiltration and tissue damage. Intestinal damage was assessed based on infection degree, extent of infection, crypt damage, and mucosal involvement [22] ( see Table 2) . The expression of ZO-1 (Proteintech Group, Inc. 21773-1-AP) and occludin (Proteintech Group, Inc. 27260-1-AP) were detected by IHC, and Image Pro Plus 6.0 (Media Cybernetics, Inc.) was utilized for statistical analysis of mean optical density. The expression of MUC2 (Boster Biological Technology co.Itd. A01212) were detected by IF, sections were incubated overnight with primary antibody against MUC2, followed by incubation with fluorescently-conjugated secondary antibody (HRP enzyme) and DAPI staining for nuclei. Slides were mounted with antifade mounting medium and examined by High-Resolution Slide Scanner (3DHISTECH Ltd. Pannoramic MIDI). Table 2. Histological grading of colitis Grade Inflammation Extent Crypt damage Percent involvement 0 None None 0 1 Slight Mucosa Basal 1/3 damage 1%-33% 2 Moderate Mucosa and Submucosa Basal 2/3 damage 34%-66% 3 Severe Transmural Entire crypt and epithelium lost 67%-100% Linked Immunosorbent Assay (ELISA) All ELISA assays were performed strictly according to the manufacturer's protocol. Concentrations of interleukin-1β (IL-1β, YJ301814), tumor necrosis factor-α (TNF-α, YJ002095), interleukin-10 (IL-10, YJ037873), myeloperoxidase (MPO, YJ002070), Superoxide Dismutase (SOD, YJ001998) and Malondialdehyde (MDA, YJ544883) in the serum samples were quantified using an enzyme-linked immunosorbent assay (ELISA) kit (Shanghai enzyme-linked biotechnology, Shanghai, China). Metabolomics analysis Metabolites Extraction The LC/MS system for metabolomics analysis is composed of Waters Acquity I-Class PLUS ultra-high performance liquid tandem Waters Xevo G2-XS QTof high resolution mass spectrometer. The column used is purchased from Waters Acquity UPLC HSS T3 column (1.8μm 2.1*100mm). Positive ion mode: mobile phase A: 0.1% formic acid aqueous solution; mobile phase B: 0.1% formic acid acetonitrile. Negative ion mode: mobile phase A: 0.1% formic acid aqueous solution; mobile phase B: 0.1% formic acid acetonitrile. Injection volume 2μL. LC-MS/MS Analysis Waters Xevo G2-XS QTOF high resolution mass spectrometer can collect primary and secondary mass spectrometry data in MSe mode under the control of the acquisition software (MassLynx V4.2, Waters). In each data acquisition cycle, dual-channel data acquisition can be performed on both low collision energy and high collision energy at the same time. The low collision energy is off, the high collision energy range is 10~40V, and the scanning frequency is 0.2 seconds for a mass spectrum. The parameters of the ESI ion source are as follows: Capillary voltage: 2500V (positive ion mode) or -2000V (negative ion mode); cone voltage: 30V; ion source temperature: 100°C; desolvent gas temperature 500°C; backflush gas flow rate: 50L/ h; Desolventizing gas flow rate: 800L/h. Data preprocessing and annotation The raw data collected using MassLynx V4.2 is processed by Progenesis QI software for peak extraction, peak alignment and other data processing operations, based on the Progenesis QI software online METLIN database and self-built library for identification. Data analysis After normalizing the original peak area information with the total peak area, the follow-up analysis was performed. Principal component analysis and Spearman correlation analysis were used to judge the repeatability of the samples within group and the quanlity control samples. The identified compounds are searched for classification and pathway information in KEGG and HMDB databases. According to the grouping information, calculate and compare the difference multiples, T test was used to calculate the difference significance pvalue of each compound. The R language package ropls was used to perform OPLS-DA modeling, and 200 times permutation tests was performed to verify the reliability of the model. The VIP value of the model was calculated using multiple cross-validation. The method of combining the difference multiple, the P value and the VIP value of the OPLS-DA model was adopted to screen the differential metabolites. The screening criteria are FC>1, P value1. The difference metabolites of KEGG pathway enrichment significance were calculated using hypergeometric distribution test. Statistical Methods Data analysis was performed using GraphPad Prism version 6 (GraphPad Software, San Diego, CA). Student T Test was selected for statistical analysis between two groups. Comparisons among data from more than two groups were conducted using one-way analysis of variance (ANOVA), followed by Tukey's multiple comparison test. All data are presented as means ± standard error of the mean (SEM). A P value of ≤ 0.05 was considered statistically significant. Results FMT Significantly Alleviates DSS-Induced Colitis Symptoms in Mice To investigate the impact of microbiota from healthy piglet donors on an intestinal inflammation model, mice were treated with either a fecal suspension from healthy piglets (FMT) or saline for 7 days, followed by 3% dextran sulfate sodium (DSS) administration for an additional 7 days (Fig. 1 A ) . Throughout the experimental period, mice in the DSS group exhibited significantly weight loss compared to the FMT + DSS group (Fig. 1 B). Recipients of FMT showed lower disease activity index (DAI) scores than the DSS group (Fig. 1 C). H&E staining revealed substantial histopathological damage in the intestines of DSS-treated mice, and the histological score of the DSS group was significantly higher than that of the FMT + DSS group (FMT + DSS vs. DSS, P = 0.022), indicating that FMT markedly attenuated DSS-induced intestinal injury (Fig. 1 D, E). Representative data on colon length demonstrated that the FMT + DSS group exhibited significantly attenuated colon shortening compared to the DSS group (FMT + DSS vs. DSS, P = 0.019) (Fig. 1 F, G). Collectively, these results indicate that prior colonization with FMT from healthy piglets alleviates DSS-induced colitis symptoms. FMT Modulates Oxidative Stress and Maintains Inflammatory Cytokines Homeostasis in Mice Myeloperoxidase (MPO) activity in the intestine serves as a reliable indicator of IBD severity [ 23 , 24 ]. Additionally, intestinal oxidative stress levels are significantly elevated during IBD, accompanied by dysregulation of pro-inflammatory cytokines (IL-1β, TNF-α, [ 25 , 26 ]) and anti-inflammatory cytokine IL-10 [ 27 ]. Therefore, we measured colitis marker MPO and oxidative stress markers Superoxide Dismutase (SOD) and Malondialdehyde (MDA) in mice serum using ELISA, and analyzed FMT effects on inflammatory cytokines (IL-1β, TNF-α, IL-10). As shown in Fig. 2 , FMT significantly reduced MPO levels (Fig. 2 A, P < 0.0001) and decreased pro-inflammatory cytokines IL-1β and TNF-α (Fig. 2 D-E, P < 0.0001) compared to the DSS group. Conversely, FMT significantly elevated anti-inflammatory cytokine IL-10 levels (Fig. 2 F, FMT + DSS vs. DSS, P = 0.0011). Furthermore, FMT markedly increased SOD levels (Fig. 2 B, P < 0.0001) while reducing MDA content (Fig. 2 C, P = 0.0006) compared to the DSS group, demonstrating its efficacy in alleviating intestinal oxidative stress in the colitis model. Collectively, pre-colonization with donor microbiota from healthy piglets ameliorated colitis symptoms by reducing oxidative stress and inflammatory responses in colitic mice. FMT Significantly Improves Intestinal Barrier Function in Recipient Mice IBD is associated with intestinal epithelial barrier dysfunction. We therefore hypothesized that FMT from donor piglets could mitigates DSS-induced damage by preserving the intestinal barrier. The average optical density (AOD) of tight junction proteins ZO-1 and Occludin in colonic tissues was measured across three experimental groups using immunohistochemistry (IHC). As shown in Fig. 3 A-C, the AOD of ZO-1 ( P = 0.007) and Occludin ( P = 0.03) in the FMT + DSS group was significantly higher than that in the DSS group. Subsequent immunofluorescence staining of colonic tissues revealed that FMT substantially enhanced MUC2 expression in mice with colitis (Fig. 3 D). To sum up, FMT attenuated DSS-induced intestinal damage by maintaining barrier integrity. Metabolomic Analysis Reveals Fundamentally Distinct Metabolic Profiles Between FMT and DSS Groups Microbiota-host metabolic interactions may underlie FMT-mediated alleviation of intestinal inflammation through immunomodulation and oxidative stress mitigation. We performed LC/MS metabolomic profiling of serum samples from all three experimental groups. Intra-group correlation analysis demonstrated high biological reproducibility among replicates (Fig. 4 A). Both principal component analysis (PCA) and principal coordinates analysis (PCoA) revealed substantial separation between DSS and FMT + DSS groups (Fig. 4 B, C). Correlation heatmap analysis confirmed distinct clustering patterns corresponding to experimental treatments (Fig. 4 D). Volcano plot analysis (thresholds: |FC| ≥ 1.0, P ≤ 0.05, VIP ≥ 1) identified 1,125 differentially abundant metabolites between DSS and FMT + DSS groups, with 433 upregulated and 692 downregulated in the FMT + DSS group (Fig. 4 E). Pearson correlation analysis of these differential metabolites demonstrated opposite abundance trajectories between groups (Fig. 4 F). These results collectively indicate fundamentally distinct metabolic landscapes in FMT + DSS versus DSS-treated mice. Association Analysis of Differential Metabolites with Phenotypic, Inflammatory, and Gut Barrier Parameters Given the distinct metabolic profiles between DSS and FMT groups, we analyzed significantly upregulated metabolites in the FMT + DSS group to elucidate the therapeutic mechanisms of FMT. Compared to the DSS group, serum levels of 6-keto-PGF1α were significantly elevated in the FMT + DSS group ( Fig. 5 A ) . FMT substantially increased peptides including Valylglutamine, Ile-Gly-Ala-Val, Gly-Val, and Bz-Ile-Glu-Gly-Arg-pNA ( Fig. 5 B-E ) . Furthermore, multiple biomembrane-constructing glycerophospholipids such as PG (18:0_22:4 (7Z,10Z,13Z,16Z)) and PC (18:0/0:0) showed significantly increased abundance ( Fig. 5 F, G ). Additionally, metabolites including 3-Methoxytyramine-betaxanthin, Bis(glutathionyl)spermine disulfide, and Sialorphin were markedly upregulated in the FMT group ( Fig. 5 H-J ). Subsequently, we established associations between dominant differential metabolites in the FMT + DSS group and key host parameters through correlation analysis. Heatmap analysis revealed that the following metabolites exhibited significantly negative correlations with pro-inflammatory cytokines (IL-1β, TNF-α), oxidative stress marker MDA, disease activity index (DAI), and colonic inflammation markers: 3-Methoxytyramine-betaxanthin, PG(18:0_22:4(7Z,10Z,13Z,16Z)), Ile-Gly-Ala-Val, Valylglutamine, 6-keto-PGF1α, PC(18:0/0:0), Gly-Val, Sialorphin, and Bz-Ile-Glu-Gly-Arg-pNA. Conversely, these metabolites showed significantly positive correlations with the anti-inflammatory cytokine IL-10 and antioxidant enzyme SOD (Fig. 6 ). Collectively, these upregulated metabolites likely contribute to maintaining host cytokines homeostasis, protecting intestinal barrier integrity, and alleviating DSS-induced intestinal inflammation. FMT Significantly Modulates Host Metabolic Pathways Metabolites form dynamic interaction networks that drive essential biological pathways. Systematic analysis of these complex metabolic reactions provides comprehensive insights into FMT's mechanisms in combating colitis pathogenesis. Through KEGG pathway annotation (functionally defined KO level 3), we identified the top 20 most enriched metabolic pathways (Fig. 7 A), including 56 amino acid metabolism pathways (tryptophan metabolism etc.), 121 biosynthetic pathways for secondary metabolites, 29 digestive system pathways (primarily associated with bile secretion), 51 lipid metabolism pathways and 37 nucleotide metabolism pathways. Human Metabolome Database (HMDB) contains comprehensive information on small-molecule metabolites in the human. Top 20 metabolite classes (Fig. 7 B) account for 827 small-molecule metabolites involved in the Lipids and lipid-like molecules metabolic pathways (including Fatty Acyls), 17 small-molecule metabolites involved in the Nucleosides, nucleotides, and analogues metabolic pathways (including Pyrimidine nucleosides), and 286 small-molecule metabolites involved in the Organic acids and derivatives metabolic pathways (including Carboxylic acids and derivatives). Differential metabolites interact within the organism, forming distinct pathways. Using the KEGG database, the differential metabolites between the DSS group and the FMT + DSS group were annotated. Top 20 pathway entries with the highest number of annotated differential metabolites were selected (Fig. 7 C). Among these, 14 pathways related to amino acid metabolism (including tryptophan metabolism) showed significant differences between the two groups. 46 differential metabolic pathways were annotated as belonging to the Biosynthesis of other secondary metabolites. 15 differential metabolic pathways associated with bile secretion within the Digestive system were annotated. 25 and 14 differential metabolic pathways were annotated for Lipid metabolism and Nucleotide metabolism. Further analysis of Differential Abundance (DA) Score for the differential metabolic pathways between the DSS and FMT + DSS groups revealed that pathways such as Betalain biosynthesis and Methane metabolism were significantly upregulated in the FMT + DSS group (Fig. 7 D). In contrast, pathways including Pathways in cancer, Bile secretion, and Steroid hormone biosynthesis were significantly upregulated in the DSS group (Fig. 7 D). The differential metabolite enrichment network showed that all differential metabolites within the Betalain biosynthesis pathway were significantly upregulated in the FMT + DSS group (Fig. 7 E), suggesting that FMT may exert its anti-inflammatory effects by regulating multiple metabolic pathways in the host, including Betalain biosynthesis. Discussion Host-microbe homeostasis is closely associated with immune regulation and immune tolerance [ 28 ]. In recent years, modulating host microbiota structure and function by targeting the gut microbiota, such as FMT for microbial dysbiosis, provides novel strategies for related diseases [ 29 ]. The disease-resistant phenotype of piglets is closely linked to the structure of the host microbial community, holding significant implications for livestock production [ 2 ]. Differences in microbiota between healthy post-weaning piglets and diarrheic piglets with intestinal inflammation suggest that microbiota from healthy pigs may possess the ability to alleviate intestinal inflammation [ 30 ]. However, the efficacy and underlying mechanisms of healthy piglet-derived microbiota in regulating host intestinal inflammation remain unclear. In the present study, to explore the therapeutic effect of bacteria derived from healthy piglets on colitis, GF mice were colonized with donor microbiota from 35-day-old healthy piglets, followed by DSS-induced colitis. The experimental results demonstrate that FMT significantly alleviated DSS-induced body weight loss and DAI scores in mice. Histological staining revealed that FMT reduced intestinal inflammation levels, improved intestinal barrier function, and maintained homeostasis of cytokines in the colitis. Moreover, FMT also significantly downregulated the level of the colitis marker MPO [ 23 ], reduced the level of the oxidative stress biomarker MDA, and elevated the level of the antioxidant biomarker SOD [ 31 ]. Consistent with previous research identifying a cytokine dysregulation pattern in intestinal inflammation [ 32 , 33 ], this study demonstrates that FMT significantly modulates the host's cytokine profile in the DSS-induced colitis model. Specifically, FMT markedly increased serum IL-10 levels, the hallmark anti-inflammatory cytokine, thereby suppressing the host's pro-inflammatory immune response and alleviating mucosal damage symptoms in IBD [ 27 ]. Concurrently, FMT significantly decreased the serum concentrations of key pro-inflammatory mediators IL-1β and serum TNF-α [ 25 , 26 ], further substantiating its anti-inflammatory effect. Moreover, FMT effectively ameliorated markers of neutrophil infiltration and tissue damage, as evidenced by reduced MPO activity [ 23 ]. Oxidative stress is recognized as a potential mechanism in the pathophysiology of IBD ([ 34 ]). MDA serves as a common biomarker of oxidative stress, reflecting the extent of cell membrane damage. Organisms possess a primary antioxidant defense system, largely dependent on enzymes like SOD, to counteract oxidative stress [ 31 ]. In this study, FMT significantly reduced DSS-induced MDA levels and significantly increased host serum SOD levels, suggesting that FMT can significantly mitigate oxidative stress symptoms in the colitis model. IBD may result from the combined effects of microbial factors, intestinal mucosal barrier dysfunction, oxidative stress, and increased inflammatory mediators [ 34 ]. Research consistently shows that impaired intestinal epithelial barrier function is a hallmark of IBD [ 35 ]. This dysfunction involves downregulation of crucial components, including the MUC-2 and tight junction proteins [ 36 ]. In the present study, FMT intervention significantly upregulated the expression levels of ZO-1 and Occludin in colonic tissue and restored MUC2 expression, FMT from healthy pigs protects the intestinal barrier to alleviate DSS-induced damage. The interaction between microorganisms and the host via metabolism a potential mechanism by which FMT modulates host intestinal immunity and ameliorates inflammatory symptoms [ 37 , 38 ]. In the present study, untargeted metabolomics results revealed that FMT significantly modulated the composition of host serum metabolites. Furthermore, FMT markedly elevated the levels of multiple metabolites possessing anti-inflammatory properties. Specific amino acids and bioactive peptides have been demonstrated potential in alleviating intestinal inflammation symptoms in IBD patients [ 39 – 42 ]. Notably, FMT significantly increased the relative abundance of several peptide metabolites, including Valylglutamine; Ile Gly Ala Val; Gly Val; and Bz-Ile-glu-gly-arg-pna. This elevation may be associated with FMT's anti-inflammatory efficacy mediated through the provision of these peptides. Phosphatidylglycerol (PG) are a subset of glycerophospholipids [ 43 ], glycerophospholipids represent a diverse class of biological molecules that play vital roles in all living systems including cell membrane structure, vesicular transport, and intracellular signaling [ 44 ]. In this study, FMT significantly increased the levels of the metabolite PG (18:0/22:4(7Z,10Z,13Z,16Z)), this elevation may contribute to the restoration of cellular membrane architecture in various intestinal cell types of FMT recipient mice. A 30% phosphatidylcholine (PC)-containing lecithin in delayed intestinal release formulation improves clinical outcomes [ 45 ], and FMT significantly increased the levels of the metabolite phosphatidylcholine PC (18:0/0:0). 3-methoxytyramine-betaxanthin exhibited strong antioxidant activity [ 46 , 47 ]). In this study, FMT significantly increased the levels of the metabolite 3-methoxytyramine-betaxanthin, and this elevation may be associated with the attenuation of IBD symptoms. Sialorphin, a potent endogenous inhibitor of the opioid peptide-degrading enzymes neprilysin and aminopeptidase N. Notably, Systemic administration of Sialorphin has demonstrated efficacy in attenuating TNBS-induced colitis in mice, its anti-inflammatory actions are mediated through µ- and κ-opioid receptors [ 48 ]. The observed elevation in Sialorphin abundance likely contributes to the anti-inflammatory effects of FMT. The levels of the metabolites 6-keto-PGF1α and bis(glutathionyl)spermine disulfide were significantly elevated in the FMT group, while the mechanisms by which these metabolites contribute to IBD alleviation require further elucidation. Multiple anti-inflammatory metabolites significantly upregulated by FMT exhibited significantly negative correlations with pro-inflammatory cytokines (IL-1β, TNF-α), oxidative stress markers MDA [ 31 ], DAI scores, and colonic inflammation markers MPO [ 23 ]. Conversely, they showed significantly positive correlations with the anti-inflammatory cytokine IL-10 and the antioxidant marker SOD [ 31 ]. These findings further substantiate that FMT may exert protective effects against IBD, at least partially, through the upregulation of multiple beneficial anti-inflammatory metabolites. Metabolites within the host interact to form distinct metabolic pathways. Notably, 3-methoxytyramine-betaxanthin has been demonstrated to exhibit strong antioxidant activity [ 46 , 47 ]. In the present study, the betalain biosynthesis pathway was significantly upregulated in the FMT group. Furthermore, the differential metabolite enrichment network revealed that all key metabolites associated with betalain biosynthesis were significantly upregulated in the FMT + DSS group compared to the DSS group. These findings collectively propose that FMT may exert its anti-inflammatory effects, at least partially, through the coordinated modulation of host metabolic pathways, including betalain biosynthesis. Conversely, the Pathways in cancer were significantly upregulated in the DSS group, and this noteworthy observation suggests that DSS-induced colitis may confer a higher susceptibility to cancer development within the intestinal tract. However, this study has several limitations. Notably, the exploration of the precise underlying mechanisms remains insufficient. Future research should employ integrated multi-omics approaches, combining metagenomics and metabolomics, to further elucidate the specific mechanisms governing the interactions between pig-derived microbiota and host metabolic and immune regulation. Conclusion In summary, the gut microbiota plays a pivotal role in modulating host immune responses and inflammatory control. This study demonstrates that pig-derived microbiota alleviates host inflammation by modulating recipient metabolism. Collectively, our findings establish a theoretical foundation for developing microbiota-based anti-inflammatory strategies. Future research should prioritize investigating the therapeutic potential of pig-derived microbiota for clinical applications in veterinary medicine. Abbreviations DAI Disease Activity Index DSS Dextran Sulfate Sodium FMT Fecal Microbiota Transplantation IBD Inflammatory Bowel Disease IF Immunofluorescence IHC Immunohistochemistry GF Germ-free Declarations Availability of data and materials The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request. Ethics approval and consent to participate This study was reviewed and approved by the Laboratory Animal Welfare Ethics Committee of Shanghai Tenth People's Hospital, all procedures complied with the Shanghai Tenth People's Hospital Guide for the Care and Use of Laboratory Animals (Animal Ethics Approval No.: SHDSYY-2025-T0088-02). Conflicts of interest The authors have declared that there is no conflict of interest. Author Contributions YY designed the experiment. QL, YY, YW, ZW, JL, YG, XT, SF, WC, HZ performed the animal trials, sample collection, and data analysis. Donor faecal samples were provided by QG. YY and QL drafted the manuscript. HW revised the manuscript. All authors contributed to the manuscript and approved the submitted version. Funding This work was supported by the National Key Research and Development Program of China (2021YFA0805904). Acknowledgments We thank Quanzhou Lvzhiyuan Co., LTD for providing healthy piglet feces, which facilitated our study. Clinical trial number: Not applicable. Consent for publication Not applicable. Competing interests The authors declare that they have no competing interests. References Zheng D, Liwinski T, Elinav E. Interaction between microbiota and immunity in health and disease. Cell Res. 2020;30(6):492–506; doi: 10.1038/s41422-020-0332-7. Zhao X, Jiang L, Fang X, Guo Z, Wang X, Shi B, et al. Host-microbiota interaction-mediated resistance to inflammatory bowel disease in pigs. Microbiome. 2022;10(1):115; doi: 10.1186/s40168-022-01303-1. Sekirov I, Russell SL, Antunes LC, Finlay BB. Gut microbiota in health and disease. Physiol Rev. 2010;90(3):859–904; doi: 10.1152/physrev.00045.2009. Kayama H, Okumura R, Takeda K. Interaction Between the Microbiota, Epithelia, and Immune Cells in the Intestine. Annu Rev Immunol. 2020;38:23–48; doi: 10.1146/annurev-immunol-070119-115104. Jin L, Shi X, Yang J, Zhao Y, Xue L, Xu L, et al. Gut microbes in cardiovascular diseases and their potential therapeutic applications. Protein Cell. 2021;12(5):346–59; doi: 10.1007/s13238-020-00785-9. Johnsen PH, Hilpusch F, Cavanagh JP, Leikanger IS, Kolstad C, Valle PC, et al. Faecal microbiota transplantation versus placebo for moderate-to-severe irritable bowel syndrome: a double-blind, randomised, placebo-controlled, parallel-group, single-centre trial. Lancet Gastroenterol Hepatol. 2018;3(1):17–24; doi: 10.1016/S2468-1253(17)30338-2. Islam J, Tanimizu M, Shimizu Y, Goto Y, Ohtani N, Sugiyama K, et al. Development of a rational framework for the therapeutic efficacy of fecal microbiota transplantation for calf diarrhea treatment. Microbiome. 2022;10(1):31; doi: 10.1186/s40168-021-01217-4. Wu R, Xiong R, Li Y, Chen J, Yan R. Gut microbiome, metabolome, host immunity associated with inflammatory bowel disease and intervention of fecal microbiota transplantation. J Autoimmun. 2023;141:103062; doi: 10.1016/j.jaut.2023.103062. Wu J, Wang J, Lin Z, Liu C, Zhang Y, Zhang S, et al. Clostridium butyricum alleviates weaned stress of piglets by improving intestinal immune function and gut microbiota. Food Chem. 2023;405(Pt B):135014; doi: 10.1016/j.foodchem.2022.135014. Zhou X, He Y, Chen J, Xiong X, Yin J, Liang J, et al. Colonic phosphocholine is correlated with Candida tropicalis and promotes diarrhea and pathogen clearance. NPJ Biofilms Microbiomes. 2023;9(1):62; doi: 10.1038/s41522-023-00433-0. Zhang J, Chen Y, Guo X, Li X, Zhang R, Wang M, et al. The gut microbial metabolite indole-3-aldehyde alleviates impaired intestinal development by promoting intestinal stem cell expansion in weaned piglets. J Anim Sci Biotechnol. 2024;15(1):150; doi: 10.1186/s40104-024-01111-7. Tao S, Fan J, Li J, Wu Z, Yao Y, Wang Z, et al. Extracellular vesicles derived from Lactobacillus johnsonii promote gut barrier homeostasis by enhancing M2 macrophage polarization. J Adv Res. 2025;69:545–63; doi: 10.1016/j.jare.2024.03.011. Gresse R, Chaucheyras-Durand F, Fleury MA, Van de Wiele T, Forano E, Blanquet-Diot S. Gut Microbiota Dysbiosis in Postweaning Piglets: Understanding the Keys to Health. Trends Microbiol. 2017;25(10):851–73; doi: 10.1016/j.tim.2017.05.004. Hu J, Ma L, Nie Y, Chen J, Zheng W, Wang X, et al. A Microbiota-Derived Bacteriocin Targets the Host to Confer Diarrhea Resistance in Early-Weaned Piglets. Cell Host Microbe. 2018;24(6):817–32 e8; doi: 10.1016/j.chom.2018.11.006. Panda SK, Peng V, Sudan R, Ulezko Antonova A, Di Luccia B, Ohara TE, et al. Repression of the aryl-hydrocarbon receptor prevents oxidative stress and ferroptosis of intestinal intraepithelial lymphocytes. Immunity. 2023;56(4):797–812 e4; doi: 10.1016/j.immuni.2023.01.023. Nayar S, Morrison JK, Giri M, Gettler K, Chuang LS, Walker LA, et al. A myeloid-stromal niche and gp130 rescue in NOD2-driven Crohn's disease. Nature. 2021;593(7858):275–81; doi: 10.1038/s41586-021-03484-5. Li J, Wei H. Establishment of an efficient germ-free animal system to support functional microbiome research. Sci China Life Sci. 2019;62(10):1400–3; doi: 10.1007/s11427-019-9832-9. Fouladi F, Glenny EM, Bulik-Sullivan EC, Tsilimigras MCB, Sioda M, Thomas SA, et al. Sequence variant analysis reveals poor correlations in microbial taxonomic abundance between humans and mice after gnotobiotic transfer. ISME J. 2020;14(7):1809–20; doi: 10.1038/s41396-020-0645-z. Souza DG, Senchenkova EY, Russell J, Granger DN. MyD88 mediates the protective effects of probiotics against the arteriolar thrombosis and leukocyte recruitment associated with experimental colitis. Inflamm Bowel Dis. 2015;21(4):888–900; doi: 10.1097/MIB.0000000000000331. Murthy SN, Cooper HS, Shim H, Shah RS, Ibrahim SA, Sedergran DJ. Treatment of dextran sulfate sodium-induced murine colitis by intracolonic cyclosporin. Dig Dis Sci. 1993;38(9):1722–34; doi: 10.1007/BF01303184. Nishiyama Y, Kataoka T, Yamato K, Taguchi T, Yamaoka K. Suppression of dextran sulfate sodium-induced colitis in mice by radon inhalation. Mediators Inflamm. 2012;2012:239617; doi: 10.1155/2012/239617. Dieleman LA, Palmen MJ, Akol H, Bloemena E, Pena AS, Meuwissen SG, et al. Chronic experimental colitis induced by dextran sulphate sodium (DSS) is characterized by Th1 and Th2 cytokines. Clin Exp Immunol. 1998;114(3):385–91; doi: 10.1046/j.1365-2249.1998.00728.x. Chadwick VS, Schlup MM, Ferry DM, Chang AR, Butt TJ. Measurements of unsaturated vitamin B12-binding capacity and myeloperoxidase as indices of severity of acute inflammation in serial colonoscopy biopsy specimens from patients with inflammatory bowel disease. Scand J Gastroenterol. 1990;25(12):1196–204; doi: 10.3109/00365529008998554. Klebanoff SJ, Coombs RW. Viricidal effect of polymorphonuclear leukocytes on human immunodeficiency virus-1. Role of the myeloperoxidase system. J Clin Invest. 1992;89(6):2014–7; doi: 10.1172/JCI115810. Yi LT, Wang XY, Zhou L, Cheng J, Xu GH, Zhu JX. Polysaccharides from Black Mulberry Attenuate Colitis through Gut Microbiota Mediated TNF-alpha/pNF-kappaB/ICAM-1 Signaling Pathway. J Agric Food Chem. 2025;73(23):14314–32; doi: 10.1021/acs.jafc.5c01870. Neurath MF. Strategies for targeting cytokines in inflammatory bowel disease. Nat Rev Immunol. 2024;24(8):559–76; doi: 10.1038/s41577-024-01008-6. Shouval DS, Biswas A, Goettel JA, McCann K, Conaway E, Redhu NS, et al. Interleukin-10 receptor signaling in innate immune cells regulates mucosal immune tolerance and anti-inflammatory macrophage function. Immunity. 2014;40(5):706–19; doi: 10.1016/j.immuni.2014.03.011. Yang T, Hu X, Cao F, Yun F, Jia K, Zhang M, et al. Targeting symbionts by apolipoprotein L proteins modulates gut immunity. Nature. 2025; doi: 10.1038/s41586-025-08990-4. Tian HL, Wang L, Ma CL, Yang B, Li L, Ye C, et al. [Fecal microbiota transplantation for the treatment of intestinal disorders: An analysis of treatment of 15 000 patients]. Zhonghua Wei Chang Wai Ke Za Zhi. 2025;28(3):296–303; doi: 10.3760/cma.j.cn441530-20250114-00025. Li J, Feng S, Wang Z, He J, Zhang Z, Zou H, et al. Limosilactobacillus mucosae-derived extracellular vesicles modulates macrophage phenotype and orchestrates gut homeostasis in a diarrheal piglet model. NPJ Biofilms Microbiomes. 2023;9(1):33; doi: 10.1038/s41522-023-00403-6. Jomova K, Raptova R, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, et al. Reactive oxygen species, toxicity, oxidative stress, and antioxidants: chronic diseases and aging. Arch Toxicol. 2023;97(10):2499–574; doi: 10.1007/s00204-023-03562-9. Zhu W, Ren L, Zhang L, Qiao Q, Farooq MZ, Xu Q. The Potential of Food Protein-Derived Bioactive Peptides against Chronic Intestinal Inflammation. Mediators Inflamm. 2020;2020:6817156; doi: 10.1155/2020/6817156. Moschen AR, Tilg H, Raine T. IL-12, IL-23 and IL-17 in IBD: immunobiology and therapeutic targeting. Nat Rev Gastroenterol Hepatol. 2019;16(3):185–96; doi: 10.1038/s41575-018-0084-8. Li L, Qiu N, Meng Y, Wang C, Mine Y, Keast R, et al. Preserved egg white alleviates DSS-induced colitis in mice through the reduction of oxidative stress, modulation of infl ammatory cytokines, NF-κB, MAPK and gut microbiota composition. Food Science and Human Wellness. 2022;12(1):312–23; doi: 10.1016/j.fshw.2022.07.021. Mehandru S, Colombel JF. The intestinal barrier, an arbitrator turned provocateur in IBD. Nat Rev Gastroenterol Hepatol. 2021;18(2):83–4; doi: 10.1038/s41575-020-00399-w. Fu Y, Guzior DV, Okros M, Bridges C, Rosset SL, Gonzalez CT, et al. Balance between bile acid conjugation and hydrolysis activity can alter outcomes of gut inflammation. Nat Commun. 2025;16(1):3434; doi: 10.1038/s41467-025-58649-x. Liu C, Du MX, Xie LS, Wang WZ, Chen BS, Yun CY, et al. Gut commensal Christensenella minuta modulates host metabolism via acylated secondary bile acids. Nat Microbiol. 2024;9(2):434–50; doi: 10.1038/s41564-023-01570-0. Liu M, Ma J, Xu J, Huangfu W, Zhang Y, Ali Q, et al. Fecal microbiota transplantation alleviates intestinal inflammatory diarrhea caused by oxidative stress and pyroptosis via reducing gut microbiota-derived lipopolysaccharides. Int J Biol Macromol. 2024;261(Pt 1):129696; doi: 10.1016/j.ijbiomac.2024.129696. Xu Q, Hu M, Li M, Hou J, Zhang X, Gao Y, et al. Dietary Bioactive Peptide Alanyl-Glutamine Attenuates Dextran Sodium Sulfate-Induced Colitis by Modulating Gut Microbiota. Oxid Med Cell Longev. 2021;2021:5543003; doi: 10.1155/2021/5543003. He D, Zeng W, Wang Y, Xing Y, Xiong K, Su N, et al. Isolation and characterization of novel peptides from fermented products of Lactobacillus for ulcerative colitis prevention and treatment. Food Science and Human Wellness. 2022;11(6):1464–74; doi: https://doi.org/10.1016/j.fshw.2022.06.003. Zhao Y, Xue P, Lin G, Tong M, Yang J, Zhang Y, et al. A KPV-binding double-network hydrogel restores gut mucosal barrier in an inflamed colon. Acta Biomater. 2022;143:233–52; doi: 10.1016/j.actbio.2022.02.039. Gravina AG, Prevete N, Tuccillo C, De Musis C, Romano L, Federico A, et al. Peptide Hp(2-20) accelerates healing of TNBS-induced colitis in the rat. United European Gastroenterol J. 2018;6(9):1428–36; doi: 10.1177/2050640618793564. Struzik ZJ, Weerts AN, Storch J, Thompson DH. Stereospecific synthesis of phosphatidylglycerol using a cyanoethyl phosphoramidite precursor. Chem Phys Lipids. 2020;231:104933; doi: 10.1016/j.chemphyslip.2020.104933. Dowhan W, Bogdanov M. Chapter 1 Functional roles of lipids in membranes. Elsevier; 2002. Stremmel W, Vural H, Evliyaoglu O, Weiskirchen R. Delayed-Release Phosphatidylcholine Is Effective for Treatment of Ulcerative Colitis: A Meta-Analysis. Dig Dis. 2021;39(5):508–15; doi: 10.1159/000514355. Cai Y, Sun M, Corke H. Antioxidant activity of betalains from plants of the amaranthaceae. J Agric Food Chem. 2003;51(8):2288–94; doi: 10.1021/jf030045u. Madadi E, Mazloum-Ravasan S, Yu JS, Ha JW, Hamishehkar H, Kim KH. Therapeutic Application of Betalains: A Review. Plants (Basel). 2020;9(9); doi: 10.3390/plants9091219. Salaga M, Mokrowiecka A, Jacenik D, Cygankiewicz AI, Malecka-Panas E, Kordek R, et al. Systemic Administration of Sialorphin Attenuates Experimental Colitis in Mice via Interaction With Mu and Kappa Opioid Receptors. J Crohns Colitis. 2017;11(8):988–98; doi: 10.1093/ecco-jcc/jjx043. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 12 Jan, 2026 Read the published version in BMC Microbiology → Version 1 posted Editorial decision: Revision requested 29 Oct, 2025 Reviews received at journal 28 Oct, 2025 Reviews received at journal 27 Oct, 2025 Reviews received at journal 23 Oct, 2025 Reviews received at journal 22 Oct, 2025 Reviewers agreed at journal 21 Oct, 2025 Reviewers agreed at journal 21 Oct, 2025 Reviewers agreed at journal 19 Oct, 2025 Reviewers agreed at journal 19 Oct, 2025 Reviewers agreed at journal 19 Oct, 2025 Reviewers agreed at journal 22 Sep, 2025 Reviewers agreed at journal 21 Sep, 2025 Reviewers invited by journal 19 Sep, 2025 Editor assigned by journal 19 Sep, 2025 Editor invited by journal 18 Sep, 2025 Submission checks completed at journal 18 Sep, 2025 First submitted to journal 18 Sep, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7536850","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":522189273,"identity":"80c3787c-426a-49f7-a798-e8e95736b9d1","order_by":0,"name":"Qinjin Li","email":"","orcid":"","institution":"Fujian Agriculture and Forestry University","correspondingAuthor":false,"prefix":"","firstName":"Qinjin","middleName":"","lastName":"Li","suffix":""},{"id":522189274,"identity":"749d9cb4-f4db-42fb-b008-292944508500","order_by":1,"name":"Yuqing Wang","email":"","orcid":"","institution":"Clinical Medicine Scientific and Technical Innovation Park, Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Yuqing","middleName":"","lastName":"Wang","suffix":""},{"id":522189275,"identity":"1e32bbe1-2ec9-4c6f-8b00-45eaa2a2c9f6","order_by":2,"name":"Jingqiang Li","email":"","orcid":"","institution":"Clinical Medicine Scientific and Technical Innovation Park, Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Jingqiang","middleName":"","lastName":"Li","suffix":""},{"id":522189276,"identity":"a96e3779-cb25-4009-8261-8872ece44e65","order_by":3,"name":"Yan Gao","email":"","orcid":"","institution":"Clinical Medicine Scientific and Technical Innovation Park, Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Yan","middleName":"","lastName":"Gao","suffix":""},{"id":522189277,"identity":"684e91ff-eded-4f3f-9096-8cb30bc9afa8","order_by":4,"name":"Zhifeng Wu","email":"","orcid":"","institution":"Huazhong Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Zhifeng","middleName":"","lastName":"Wu","suffix":""},{"id":522189278,"identity":"d44c026d-a5ba-4226-8751-e99c37595ee8","order_by":5,"name":"Xiang Tan","email":"","orcid":"","institution":"Huazhong Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Xiang","middleName":"","lastName":"Tan","suffix":""},{"id":522189279,"identity":"d7179e78-3d80-4d54-82d9-336e6400a503","order_by":6,"name":"Shuaifei Feng","email":"","orcid":"","institution":"Huazhong Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Shuaifei","middleName":"","lastName":"Feng","suffix":""},{"id":522189280,"identity":"dd135b89-11f8-4d3f-a246-6c8bb21a4f20","order_by":7,"name":"Wei Cheng","email":"","orcid":"","institution":"Huazhong Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Wei","middleName":"","lastName":"Cheng","suffix":""},{"id":522189281,"identity":"1c409bc7-049c-4d21-b2dc-b1391639b3b2","order_by":8,"name":"Hang Zhang","email":"","orcid":"","institution":"Huazhong Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Hang","middleName":"","lastName":"Zhang","suffix":""},{"id":522189282,"identity":"3d8c92a6-a3ae-41d7-b0e3-5fca10ace3c0","order_by":9,"name":"Qianfu Gan","email":"","orcid":"","institution":"Fujian Agriculture and Forestry University","correspondingAuthor":false,"prefix":"","firstName":"Qianfu","middleName":"","lastName":"Gan","suffix":""},{"id":522189283,"identity":"9bebfb12-ad95-49b6-9466-5c7695a099fe","order_by":10,"name":"Hong Wei","email":"","orcid":"","institution":"Clinical Medicine Scientific and Technical Innovation Park, Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Hong","middleName":"","lastName":"Wei","suffix":""},{"id":522189284,"identity":"99dda695-8d0c-4f1c-8293-37c22880c1c1","order_by":11,"name":"Yapeng Yang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9UlEQVRIiWNgGAWjYJCCAx8MbOT42ZsPHP5RISEnT4QOxoMzCtKMJXuOJR5mOGNhbNhAWAvzYZ4PhxM33MgxPszYVpHIcICAeoMbuQcOzjBgZmw4c8DgcOE8iQTGBuaHj27g1ZKXAPQLGzNje0PC4ZnbJPLYGdiMjXPwaskxANrCw8bMc+DAAd5tEsWMDTxs0oS0HOYxkOBhk0hsOMA7B0QSpwWoRyKZ4TBvAxFaJM+8SwA6LAGo5xjDwRnHJIwNmwn4he947uEPH/78r99/vP/zhw81dXLy7M0PH+PTonABQ5YZj3IQkO8/Q0DFKBgFo2AUjAIAdKtas3YkzA4AAAAASUVORK5CYII=","orcid":"","institution":"Fujian Agriculture and Forestry University","correspondingAuthor":true,"prefix":"","firstName":"Yapeng","middleName":"","lastName":"Yang","suffix":""}],"badges":[],"createdAt":"2025-09-04 13:53:34","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7536850/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7536850/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12866-025-04590-4","type":"published","date":"2026-01-12T16:28:54+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":92593615,"identity":"47a79fbf-69da-4f79-b477-26e44c179a81","added_by":"auto","created_at":"2025-10-01 12:37:57","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":148953,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.docx","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/e50b7baa023cf63b607b549f.docx"},{"id":92593620,"identity":"532b12c0-f114-453a-a621-d318b26b34b5","added_by":"auto","created_at":"2025-10-01 12:37:57","extension":"tif","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":20718140,"visible":true,"origin":"","legend":"","description":"","filename":"Fig1.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/46290b4689e7972c34413080.tif"},{"id":92593619,"identity":"ad079ad8-6d52-4f8d-bd87-ef767dcfc06c","added_by":"auto","created_at":"2025-10-01 12:37:57","extension":"tif","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":6155106,"visible":true,"origin":"","legend":"","description":"","filename":"Fig2.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/313817e3507adf459953a9f3.tif"},{"id":92595164,"identity":"e0a4366b-0724-4277-ab23-137d208e9025","added_by":"auto","created_at":"2025-10-01 12:53:58","extension":"tif","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":40691456,"visible":true,"origin":"","legend":"","description":"","filename":"Fig3.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/c8c7c79225f714f8b5014e96.tif"},{"id":92594265,"identity":"478d61e8-986e-4b94-bcc8-c81b175a6710","added_by":"auto","created_at":"2025-10-01 12:45:57","extension":"tif","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":12095808,"visible":true,"origin":"","legend":"","description":"","filename":"Fig4.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/fcd1dba3a280f6cc26c71ca7.tif"},{"id":92594269,"identity":"4b3cb408-de33-4df4-a495-e398e10ca3b7","added_by":"auto","created_at":"2025-10-01 12:45:58","extension":"tif","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":5889098,"visible":true,"origin":"","legend":"","description":"","filename":"Fig5.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/a250afe9f8e29bdbcdc50515.tif"},{"id":92594266,"identity":"c1863750-aa54-4928-8afc-cd065a37eb87","added_by":"auto","created_at":"2025-10-01 12:45:58","extension":"tif","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":994418,"visible":true,"origin":"","legend":"","description":"","filename":"Fig6.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/16538b920ad7284e2231619e.tif"},{"id":92594271,"identity":"59e1a72c-2903-493f-9379-37bbbc89ff7d","added_by":"auto","created_at":"2025-10-01 12:45:59","extension":"tif","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":8719384,"visible":true,"origin":"","legend":"","description":"","filename":"Fig7.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/2ce0f044b2d461a1b6b9347d.tif"},{"id":92593617,"identity":"2d5d4ebe-5fbb-4abb-9bfd-27dd49c3587b","added_by":"auto","created_at":"2025-10-01 12:37:57","extension":"json","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":12049,"visible":true,"origin":"","legend":"","description":"","filename":"3ad2c3b67b1c4e6189520f0a328a0592.json","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/6c8b076348dc5cc6dae5bfc0.json"},{"id":92593618,"identity":"56a4efa5-5ac2-4d1d-b3b7-e5cb6551cb73","added_by":"auto","created_at":"2025-10-01 12:37:57","extension":"xml","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":140497,"visible":true,"origin":"","legend":"","description":"","filename":"3ad2c3b67b1c4e6189520f0a328a05921enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/f6591ff0dd79f354107894fd.xml"},{"id":92593631,"identity":"3057dee3-80e2-48ef-baef-e1a734532aaf","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"tif","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":20718140,"visible":true,"origin":"","legend":"","description":"","filename":"Fig1.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/09cc0c2583f80d39d6b4c8ec.tif"},{"id":92593624,"identity":"9d34a95e-e15e-4985-a39b-e0ef9e196836","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"tif","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":6155106,"visible":true,"origin":"","legend":"","description":"","filename":"Fig2.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/5a88f6166a52e0b1fbe52313.tif"},{"id":92594270,"identity":"cff18457-066b-41a0-9ed9-b18e9dbbe78f","added_by":"auto","created_at":"2025-10-01 12:45:58","extension":"tif","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":40691456,"visible":true,"origin":"","legend":"","description":"","filename":"Fig3.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/7be700aa3ebd86e6dcfcba7d.tif"},{"id":92593636,"identity":"205a43fb-c310-4f0e-b672-b92f96d4897a","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"tif","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":12095808,"visible":true,"origin":"","legend":"","description":"","filename":"Fig4.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/ef9bd93c1a87f878fbeead6d.tif"},{"id":92593640,"identity":"a3fde567-3631-4f18-be52-acaec0522d5c","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"tif","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":5889098,"visible":true,"origin":"","legend":"","description":"","filename":"Fig5.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/75f9ad9c7f07260aa2c9c554.tif"},{"id":92593646,"identity":"dddb2857-ea55-493b-9094-6f69b75cbec5","added_by":"auto","created_at":"2025-10-01 12:37:59","extension":"tif","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":994418,"visible":true,"origin":"","legend":"","description":"","filename":"Fig6.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/0324eb2fcf3b50165d2e19ba.tif"},{"id":92593641,"identity":"2a5e014a-4f01-4b19-8a5c-e4a5f630aef6","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"tif","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":8719384,"visible":true,"origin":"","legend":"","description":"","filename":"Fig7.tif","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/14150dcd934029f7bfc31d02.tif"},{"id":92594272,"identity":"109647ba-ff70-4e02-9fa2-544eda6f36dc","added_by":"auto","created_at":"2025-10-01 12:45:59","extension":"png","order_by":17,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1032894,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig1.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/2a67c7f43dc058a720f3b310.png"},{"id":92595165,"identity":"fc075acf-943e-4152-a030-c3b47600cde0","added_by":"auto","created_at":"2025-10-01 12:53:58","extension":"png","order_by":18,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":393100,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig2.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/73df95946a3c5ed933314e37.png"},{"id":92595166,"identity":"e542dc1f-4501-414a-b847-e553326cfd4f","added_by":"auto","created_at":"2025-10-01 12:53:59","extension":"png","order_by":19,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2336675,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig3.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/c6713cb272732d65d9da5c8f.png"},{"id":92593633,"identity":"ccca158e-93ba-4614-8979-e6c4295e4dd0","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"png","order_by":20,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1509666,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig4.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/9d7372b21d51c3baef2026bf.png"},{"id":92594268,"identity":"3b43b54d-f840-4e12-b71f-cc08b90392cf","added_by":"auto","created_at":"2025-10-01 12:45:58","extension":"png","order_by":21,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":715864,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig5.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/9120234f517495b6f3ef7130.png"},{"id":92593643,"identity":"96b70c1b-19ae-43b0-9107-999746ca1e18","added_by":"auto","created_at":"2025-10-01 12:37:59","extension":"png","order_by":22,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":139991,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig6.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/5bde1aea17d17cbdc80d1bda.png"},{"id":92593638,"identity":"dff92841-dcb1-42a5-a894-8667e512a3eb","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"png","order_by":23,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":388203,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig7.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/2d27d4b06f80bc6c363e25d5.png"},{"id":92593634,"identity":"79b72296-d51e-4b4b-b4d5-4a3c669ef9f1","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"xml","order_by":24,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":139371,"visible":true,"origin":"","legend":"","description":"","filename":"3ad2c3b67b1c4e6189520f0a328a05921structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/0111e73a694f1ded9360058d.xml"},{"id":92593627,"identity":"5d85ba65-74ef-45b5-a9c6-2628a6e7ff92","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"html","order_by":25,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":150090,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/cc257bd9beac1d699edeef7e.html"},{"id":92593625,"identity":"a0cdba92-1cc1-401d-8324-89901ad4aa3c","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":40827778,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFMT significantly alleviates DSS-induced colitis symptoms in mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Study design; (B) Body weight evolution; (C) DAI score; (D) H\u0026amp;E staining of colon tissue (100×); (E) Histological score; (F) Colon length; (G) Representative colon. *\u003cem\u003eP\u003c/em\u003e ≤ 0.05; **\u003cem\u003eP\u003c/em\u003e ≤ 0.01; ***\u003cem\u003eP\u003c/em\u003e ≤ 0.001; ****\u003cem\u003eP\u003c/em\u003e ≤ 0.0001, data are represented as mean ± SEM.\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/ce9c79c6ea469714e4fc7bc9.png"},{"id":92593613,"identity":"4972f4a5-4d25-4e8b-8efa-259093e462d2","added_by":"auto","created_at":"2025-10-01 12:37:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":18234947,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFMT modulates oxidative stress and maintains cytokine homeostasis in colitis models.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Serum MPO; (B) Serum SOD; (C) Serum MDA; (D) Serum IL-1β; (E) Serum TNF-α; (F) Serum IL-10. **\u003cem\u003eP\u003c/em\u003e ≤ 0.01; ***\u003cem\u003eP\u003c/em\u003e ≤ 0.001; ****\u003cem\u003eP\u003c/em\u003e ≤ 0.0001, data are represented as mean ± SEM.\u003c/p\u003e","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/87d856c0b11f850fa5df89d9.png"},{"id":92593629,"identity":"5d571bdc-87ef-492b-bcbb-5d201e48229e","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":62329013,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of FMT by four distinct groups on the intestinal barrier in DSS-induced colitis.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eImmunohistochemistry for ZO-1 and occludin (A) in each group (100μm, n=3-4); (B, C) Average optical density; (D) Representative images for MUC2 of colon. Red represents MUC2, blue shows DAPI. Scale bars, 100 μm. *\u003cem\u003eP\u003c/em\u003e ≤ 0.05; **\u003cem\u003eP\u003c/em\u003e ≤ 0.01, data are represented as mean ± SEM.\u003c/p\u003e","description":"","filename":"Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/28947498c8a6d77c83a8b1d5.png"},{"id":92593616,"identity":"da2546fc-7421-4679-bb8e-2227d12ff120","added_by":"auto","created_at":"2025-10-01 12:37:57","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":13444940,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMetabolomics analysis showed that the metabolic profiles of the FMT group were significantly different from those of the DSS group.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Biological replicate assessment between samples within groups; (B) PCA analysis; (C) PCoA analysis; (D) Metabolite correlations between the three groups; (E) Differential metabolites between the two groups were demonstrated by volcano diagrams (DSS VS. FMT+DSS, Based on parameters: FC ≥ 1, \u003cem\u003eP\u003c/em\u003evalue ≤ 0.05, VIP ≥ 1); (F) Correlation of Top20 differential metabolites between the two groups (DSS VS. FMT+DSS). The correlation is highest at 1 for a perfect positive correlation (red) and lowest at -1 for a perfect negative correlation (blue), the figure shows the correlation of the Top20 differential metabolites by T-test of \u003cem\u003eP\u003c/em\u003e value.\u003c/p\u003e","description":"","filename":"Fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/3fa972e152445e29f55dbc2b.png"},{"id":92593626,"identity":"8fad6008-e20d-4b07-8aab-f8fcd95e4b75","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":21200788,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAnalysis of significantly up-regulated differential metabolites in the FMT group.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) 6 keto-PGF1 alpha; (B) Valylglutamine; (C) Ile Gly Ala Val; (D) Gly Val; (E) Bz-Ile-glu-gly-arg-pna; (F) PG (18_0_22_4 (7Z,10Z,13Z,16Z)); (G) PC (18:0/0:0); (H) 3-Methoxytyramine-betaxanthin; (I) Bis (glutathionyl) spermine disulfide; (J) Sialorphin.\u003c/p\u003e","description":"","filename":"Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/8f71382f7434eb97cb45dfb4.png"},{"id":92593637,"identity":"d599db9a-31c1-4760-a14c-94362772f8fe","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":3880951,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCorrelation analysis of differential metabolite in FMT+DSS group\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003ewith cytokines and DAI score.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Fig6.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/91c0eebcfbadc4b22c9a0df1.png"},{"id":92593632,"identity":"c3c8b39b-7fa5-414d-ab4d-7280761b00c9","added_by":"auto","created_at":"2025-10-01 12:37:58","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":10890053,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffects of FMT on metabolic pathways.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) All identified metabolites were annotated using the KEGG database, and the top20 items of information with the most metabolites annotated to KO pathway level3 were selected. The items under the same box in the figure represent the hierarchical categorization of KEGG pathway annotations, corresponding to KO pathway level2 and KO pathway level3. the length of the columns represents the number of metabolites annotated to this pathway; (B) The top 20 classes with the most annotations to metabolites were selected based on the HMDB database. The items under the same box in the figure represent the HMDB hierarchical classification information, corresponding to the super class and class information of the HMDB database. The length of the bar represents the number of metabolites annotated to that classification;\u003cstrong\u003e \u003c/strong\u003e(C) Information on the top20 items with the most annotations to differential metabolites in the pathway was selected based on the KEGG database, DSS VS. FMT+DSS. The items under the same box in the figure represent the hierarchical categorization of KEGG pathway annotations, corresponding to KO pathway level2 and KEGG pathway. The length of the bar represents the number of differential metabolites annotated to that pathway; (D) Differential abundance scores（DA Score） for differential metabolites (DSS VS. FMT+DSS). DA Score reflects the overall change of all metabolites in the metabolic pathway, with a score of 1 or -1 indicating a trend of up- or down-regulation of the expression of all annotated metabolites in the pathway. The size of the dot at the endpoint of the line segment indicates the number of differential metabolites in the pathway;\u003cstrong\u003e \u003c/strong\u003e(E) KEGG enrichment network diagram of differential metabolites. The yellowish nodes in the figure are pathways, the small nodes connected to them are specific metabolites annotated to that pathway, the size of the dots indicates the number of differential metabolites annotated to that pathway, and the shade of the color indicates that the differential multiplicity takes the value of log2.\u003c/p\u003e","description":"","filename":"Fig7.png","url":"https://assets-eu.researchsquare.com/files/rs-7536850/v1/3801c747b6df62210cd110e7.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Fecal Microbiota Transplantation from Healthy Piglets Ameliorates Intestinal Inflammation in Mice by Modulating Recipient Metabolism","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe gut microbiota plays a critical role in maintaining immune homeostasis, regulating immune responses, and preventing pathogen invasion through its interactions with the host immune system [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. This microbial ecosystem contributes significantly to microecological homeostasis, which is closely associated with immune regulatory functions and immune tolerance, exerting profound impacts on both livestock production and human health [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Recent studies have increasingly demonstrated that gut microbiota dysbiosis may be closely linked to the pathogenesis of various inflammatory and immune-mediated diseases [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Conversely, targeted modulation of host microbiota structure and function, such as through fecal microbiota transplantation (FMT) may provide novel therapeutic strategies for dysbiosis-associated disorders [\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIntestinal health is fundamental to growth performance and disease resistance in livestock. Weaned piglets experiencing diarrhea develop intestinal inflammation, which significantly impairs subsequent growth performance. Post-weaning diarrhea (PWD) is a prevalent challenge, often leading to intestinal inflammation that significantly compromises subsequent growth performance and feed efficiency [\u003cspan additionalcitationids=\"CR10\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Studies comparing the gut microbiota of healthy post-weaning pigs with that of diarrheic piglets exhibiting intestinal inflammation have revealed substantial differences in microbial structure and function. [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. This suggests a potential therapeutic role for healthy pig-derived microbiota in ameliorating intestinal inflammation [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. However, the efficacy and underlying mechanisms through which healthy post-weaning pig microbiota exerts its anti-inflammatory effects remain largely undefined and warrant further investigation.\u003c/p\u003e\u003cp\u003eThe DSS-induced intestinal inflammation model represents a classical methodology for investigating intestinal inflammatory mechanisms [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. This model effectively recapitulates key features of intestinal inflammation in livestock animals and human inflammatory bowel disease (IBD). The use of germ-free (GF) mice with defined microbial backgrounds facilitates the elimination of interference from native microbiota, an approach particularly critical for evaluating the anti-inflammatory efficacy of pig-derived microbiota [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. This study employs healthy piglet-derived microbiota as the research subject. Through its transplantation into DSS-induced GF mouse models, we investigate its impact on host intestinal inflammation and barrier function, while aiming to elucidate the mechanistic basis by which healthy piglet microbiota maintains intestinal homeostasis from the perspective of metabolic regulation. This work aims to reveal the significant role of healthy piglet-derived gut microbiota in host immunomodulation via host metabolic pathways, concurrently laying theoretical groundwork for developing microbiota-based anti-inflammatory strategies.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003e\u003cstrong\u003eFMT Preparation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFresh faecal samples were collected from healthy Yorkshire piglets (35-day-old, n=3, Quanzhou Green Source Agriculture Co., Ltd) using a sterile faecal collector, and samples were transported to the laboratory under dry ice conditions, and the feces from the interior of the mid-section were homogenized under anaerobic conditions \u0026nbsp; (80% N\u003csub\u003e2\u003c/sub\u003e, 10% H\u003csub\u003e2\u003c/sub\u003e, 10% CO\u003csub\u003e2\u003c/sub\u003e) and mixed thoroughly with saline glycerol buffer (15% glycerol concentration) at a 1:10 (m/v) ratio. After complete dissolution, samples were pre-filtered through sterile gauze followed by final filtration through a 100-\u0026mu;m membrane [18].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental Animals and Treatments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGF female KM mice (8-10 weeks old) were obtained from the Germ-Free Animal Platform at Shanghai Tenth People\u0026apos;s Hospital. Mice were maintained in a sterile isolator under controlled conditions: temperature 25 \u0026plusmn; 2\u0026deg;C, relative humidity 45-60%, and a 12-h light/dark cycle (lights on 06:30-18:30), and had free access to sterilized food and water. This study investigated the impact of FMT derived from healthy Yorkshire piglets. Recipient mice received daily oral gavage of 100 \u0026mu;L bacterial suspension or saline control, followed by administration of 3% DSS. Experimental groups and timelines are detailed in \u003cstrong\u003eFigure 1A\u003c/strong\u003e. Body weight was recorded daily throughout the experiment, calculated as the percentage change relative to baseline (day 0) [19]. The mice were euthanized by cervical dislocation without anesthesia, a method approved by the North Stockholm Ethics Committee (permission N31/14). Upon experiment completion, fecal samples were collected within the isolator immediately prior to euthanasia. This study was reviewed and approved by the Laboratory Animal Welfare Ethics Committee of Shanghai Tenth People\u0026apos;s Hospital, all procedures complied with the Shanghai Tenth People\u0026apos;s Hospital \u003cem\u003eGuide for the Care and Use of Laboratory Animals\u003c/em\u003e (Animal Ethics Approval No.: SHDSYY-2025-T0088-02).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisease Activity Index\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Disease Activity Index (DAI) comprises a weight loss score, a fecal bleeding score, and stool characteristic assessments, as detailed in \u003cstrong\u003eTable\u0026nbsp;1\u003c/strong\u003e [20, 21]. Briefly, the DAI was calculated based on weight change (no change = 0; 1-5% = 1; 5-10% = 2; 10-15% = 3; \u0026gt;15% = 4), fecal bleeding (normal colored stool = 0; brown stool = 1; red stool = 2; bloody stool = 3; heavy bleeding = 4), and stool consistency (normal stool, good shape = 0; soft stool, soft stool adhering to the anus = 1-2; diarrhea, adherent anal = 3-4), and the overall DAI score was obtained by averaging these three individual scores.\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"643\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 643px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1.\u003c/strong\u003e Disease Activity Index\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eScore\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eWeight loss (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eStool consistency\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eBloody stool score\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNormal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNormal colored stool\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e1-5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eLoose stool\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eBrown stool\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5-10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eLoose stool\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eReddish stool\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e10-15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eDiarrhea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eBloody stool\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026gt; 15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eDiarrhea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eGross bleeding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eHistologic Analysis of Mice Colon\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDistal colon segments from mice in each group were fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned into 4 \u0026mu;m-thick slices. Sections were stained with hematoxylin-eosin (H\u0026amp;E), immunohistochemistry (IHC) and\u0026nbsp;immunofluorescence (IF), and images were acquired using a microscope (Nikon Eclipse 80i, Japan). H\u0026amp;E-stained sections were evaluated for inflammatory cell infiltration and tissue damage. Intestinal damage was assessed based on infection degree, extent of infection, crypt damage, and mucosal involvement [22] \u003cstrong\u003e(\u003c/strong\u003esee\u003cstrong\u003e\u0026nbsp;Table 2)\u003c/strong\u003e. The expression of ZO-1 (Proteintech Group, Inc. 21773-1-AP) and occludin (Proteintech Group, Inc. 27260-1-AP) were detected by IHC, and Image Pro Plus 6.0 (Media Cybernetics, Inc.) was utilized for statistical analysis of mean optical density. The expression of MUC2 (Boster Biological Technology co.Itd. A01212) were detected by IF,\u0026nbsp;sections were incubated overnight with primary antibody against MUC2, followed by incubation with fluorescently-conjugated secondary antibody (HRP enzyme) and DAPI staining for nuclei. Slides were mounted with antifade mounting medium and examined by High-Resolution Slide Scanner (3DHISTECH Ltd.\u0026nbsp;Pannoramic MIDI).\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"652\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" style=\"width: 652px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 2.\u003c/strong\u003e Histological grading of colitis\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 54px;\"\u003e\n \u003cp\u003eGrade\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 98px;\"\u003e\n \u003cp\u003eInflammation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003eExtent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 201px;\"\u003e\n \u003cp\u003eCrypt damage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 139px;\"\u003e\n \u003cp\u003ePercent involvement\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 54px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 98px;\"\u003e\n \u003cp\u003eNone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 201px;\"\u003e\n \u003cp\u003eNone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 139px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 54px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 98px;\"\u003e\n \u003cp\u003eSlight\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003eMucosa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 201px;\"\u003e\n \u003cp\u003eBasal 1/3 damage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 139px;\"\u003e\n \u003cp\u003e1%-33%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 54px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 98px;\"\u003e\n \u003cp\u003eModerate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003eMucosa and Submucosa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 201px;\"\u003e\n \u003cp\u003eBasal 2/3 damage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 139px;\"\u003e\n \u003cp\u003e34%-66%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 54px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 98px;\"\u003e\n \u003cp\u003eSevere\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003eTransmural\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 201px;\"\u003e\n \u003cp\u003eEntire crypt and epithelium lost\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 139px;\"\u003e\n \u003cp\u003e67%-100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eLinked Immunosorbent Assay (ELISA)\u003cbr\u003e\u003c/strong\u003eAll ELISA assays were performed strictly according to the manufacturer\u0026apos;s protocol. Concentrations of interleukin-1\u0026beta; (IL-1\u0026beta;, YJ301814), tumor necrosis factor-\u0026alpha; (TNF-\u0026alpha;, YJ002095), interleukin-10 (IL-10, YJ037873), myeloperoxidase (MPO, YJ002070), Superoxide Dismutase (SOD, YJ001998) and Malondialdehyde (MDA, YJ544883) in the serum samples were quantified using an enzyme-linked immunosorbent assay (ELISA) kit (Shanghai enzyme-linked biotechnology, Shanghai, China).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMetabolomics analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMetabolites Extraction\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe LC/MS system for metabolomics analysis is composed of Waters Acquity I-Class PLUS ultra-high performance liquid tandem Waters Xevo G2-XS QTof high resolution mass spectrometer. The column used is purchased from Waters Acquity UPLC HSS T3 column (1.8\u0026mu;m 2.1*100mm). Positive ion mode: mobile phase A: 0.1% formic acid aqueous solution; mobile phase B: 0.1% formic acid acetonitrile. Negative ion mode: mobile phase A: 0.1% formic acid aqueous solution; mobile phase B: 0.1% formic acid acetonitrile. Injection volume 2\u0026mu;L.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLC-MS/MS Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWaters Xevo G2-XS QTOF high resolution mass spectrometer can collect primary and secondary mass spectrometry data in MSe mode under the control of the acquisition software (MassLynx V4.2, Waters). In each data acquisition cycle, dual-channel data acquisition can be performed on both low collision energy and high collision energy at the same time. The low collision energy is off, the high collision energy range is 10~40V, and the scanning frequency is 0.2 seconds for a mass spectrum. The parameters of the ESI ion source are as follows: Capillary voltage: 2500V (positive ion mode) or -2000V (negative ion mode); cone voltage: 30V; ion source temperature: 100\u0026deg;C; desolvent gas temperature 500\u0026deg;C; backflush gas flow rate: 50L/ h; Desolventizing gas flow rate: 800L/h.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData preprocessing and annotation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe raw data collected using MassLynx V4.2 is processed by Progenesis QI software for peak extraction, peak alignment and other data processing operations, based on the Progenesis QI software online METLIN database and self-built library for identification.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter normalizing the original peak area information with the total peak area, the follow-up analysis was performed. Principal component analysis and Spearman correlation analysis were used to judge the repeatability of the samples within group and the quanlity control samples. The identified compounds are searched for classification and pathway information in KEGG and HMDB databases. According to the grouping information, calculate and compare the difference multiples, T test was used to calculate the difference significance pvalue of each compound. The R language package ropls was used to perform OPLS-DA modeling, and 200 times permutation tests was performed to verify the reliability of the model. The VIP value of the model was calculated using multiple cross-validation. The method of combining the difference multiple, the P value and the VIP value of the OPLS-DA model was adopted to screen the differential metabolites. The screening criteria are FC\u0026gt;1, \u003cem\u003eP\u003c/em\u003e value\u0026lt;0.05 and VIP\u0026gt;1. The difference metabolites of KEGG pathway enrichment significance were calculated using hypergeometric distribution test.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical Methods\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData analysis was performed using GraphPad Prism version 6 (GraphPad Software, San Diego, CA). Student T Test was selected for statistical analysis between two groups. Comparisons among data from more than two groups were conducted using one-way analysis of variance (ANOVA), followed by Tukey\u0026apos;s multiple comparison test. All data are presented as means \u0026plusmn; standard error of the mean (SEM). A \u003cem\u003eP\u003c/em\u003e value of \u0026le; 0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003eFMT Significantly Alleviates DSS-Induced Colitis Symptoms in Mice\u003c/h2\u003e\u003cp\u003e To investigate the impact of microbiota from healthy piglet donors on an intestinal inflammation model, mice were treated with either a fecal suspension from healthy piglets (FMT) or saline for 7 days, followed by 3% dextran sulfate sodium (DSS) administration for an additional 7 days (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA\u003cb\u003e)\u003c/b\u003e. Throughout the experimental period, mice in the DSS group exhibited significantly weight loss compared to the FMT\u0026thinsp;+\u0026thinsp;DSS group (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). Recipients of FMT showed lower disease activity index (DAI) scores than the DSS group (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). H\u0026amp;E staining revealed substantial histopathological damage in the intestines of DSS-treated mice, and the histological score of the DSS group was significantly higher than that of the FMT\u0026thinsp;+\u0026thinsp;DSS group (FMT\u0026thinsp;+\u0026thinsp;DSS vs. DSS, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.022), indicating that FMT markedly attenuated DSS-induced intestinal injury (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eD, E). Representative data on colon length demonstrated that the FMT\u0026thinsp;+\u0026thinsp;DSS group exhibited significantly attenuated colon shortening compared to the DSS group (FMT\u0026thinsp;+\u0026thinsp;DSS vs. DSS, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.019) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eF, G). Collectively, these results indicate that prior colonization with FMT from healthy piglets alleviates DSS-induced colitis symptoms.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003eFMT Modulates Oxidative Stress and Maintains Inflammatory Cytokines Homeostasis in Mice\u003c/h2\u003e\u003cp\u003eMyeloperoxidase (MPO) activity in the intestine serves as a reliable indicator of IBD severity [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Additionally, intestinal oxidative stress levels are significantly elevated during IBD, accompanied by dysregulation of pro-inflammatory cytokines (IL-1β, TNF-α, [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]) and anti-inflammatory cytokine IL-10 [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Therefore, we measured colitis marker MPO and oxidative stress markers Superoxide Dismutase (SOD) and Malondialdehyde (MDA) in mice serum using ELISA, and analyzed FMT effects on inflammatory cytokines (IL-1β, TNF-α, IL-10). As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, FMT significantly reduced MPO levels (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA, P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) and decreased pro-inflammatory cytokines IL-1β and TNF-α (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD-E, P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) compared to the DSS group. Conversely, FMT significantly elevated anti-inflammatory cytokine IL-10 levels (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eF, FMT\u0026thinsp;+\u0026thinsp;DSS vs. DSS, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0011). Furthermore, FMT markedly increased SOD levels (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB, P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) while reducing MDA content (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC, P\u0026thinsp;=\u0026thinsp;0.0006) compared to the DSS group, demonstrating its efficacy in alleviating intestinal oxidative stress in the colitis model. Collectively, pre-colonization with donor microbiota from healthy piglets ameliorated colitis symptoms by reducing oxidative stress and inflammatory responses in colitic mice.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003eFMT Significantly Improves Intestinal Barrier Function in Recipient Mice\u003c/h2\u003e\u003cp\u003eIBD is associated with intestinal epithelial barrier dysfunction. We therefore hypothesized that FMT from donor piglets could mitigates DSS-induced damage by preserving the intestinal barrier. The average optical density (AOD) of tight junction proteins ZO-1 and Occludin in colonic tissues was measured across three experimental groups using immunohistochemistry (IHC). As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA-C, the AOD of ZO-1 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.007) and Occludin (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.03) in the FMT\u0026thinsp;+\u0026thinsp;DSS group was significantly higher than that in the DSS group. Subsequent immunofluorescence staining of colonic tissues revealed that FMT substantially enhanced MUC2 expression in mice with colitis (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). To sum up, FMT attenuated DSS-induced intestinal damage by maintaining barrier integrity.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003eMetabolomic Analysis Reveals Fundamentally Distinct Metabolic Profiles Between FMT and DSS Groups\u003c/h2\u003e\u003cp\u003eMicrobiota-host metabolic interactions may underlie FMT-mediated alleviation of intestinal inflammation through immunomodulation and oxidative stress mitigation. We performed LC/MS metabolomic profiling of serum samples from all three experimental groups. Intra-group correlation analysis demonstrated high biological reproducibility among replicates (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). Both principal component analysis (PCA) and principal coordinates analysis (PCoA) revealed substantial separation between DSS and FMT\u0026thinsp;+\u0026thinsp;DSS groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB, C). Correlation heatmap analysis confirmed distinct clustering patterns corresponding to experimental treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD). Volcano plot analysis (thresholds: |FC| \u0026ge; 1.0, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05, VIP\u0026thinsp;\u0026ge;\u0026thinsp;1) identified 1,125 differentially abundant metabolites between DSS and FMT\u0026thinsp;+\u0026thinsp;DSS groups, with 433 upregulated and 692 downregulated in the FMT\u0026thinsp;+\u0026thinsp;DSS group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE). Pearson correlation analysis of these differential metabolites demonstrated opposite abundance trajectories between groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eF). These results collectively indicate fundamentally distinct metabolic landscapes in FMT\u0026thinsp;+\u0026thinsp;DSS versus DSS-treated mice.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\u003ch2\u003eAssociation Analysis of Differential Metabolites with Phenotypic, Inflammatory, and Gut Barrier Parameters\u003c/h2\u003e\u003cp\u003eGiven the distinct metabolic profiles between DSS and FMT groups, we analyzed significantly upregulated metabolites in the FMT\u0026thinsp;+\u0026thinsp;DSS group to elucidate the therapeutic mechanisms of FMT. Compared to the DSS group, serum levels of 6-keto-PGF1α were significantly elevated in the FMT\u0026thinsp;+\u0026thinsp;DSS group \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA\u003cb\u003e)\u003c/b\u003e. FMT substantially increased peptides including Valylglutamine, Ile-Gly-Ala-Val, Gly-Val, and Bz-Ile-Glu-Gly-Arg-pNA \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB-E\u003cb\u003e)\u003c/b\u003e. Furthermore, multiple biomembrane-constructing glycerophospholipids such as PG (18:0_22:4 (7Z,10Z,13Z,16Z)) and PC (18:0/0:0) showed significantly increased abundance \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eF, G\u003cb\u003e).\u003c/b\u003e Additionally, metabolites including 3-Methoxytyramine-betaxanthin, Bis(glutathionyl)spermine disulfide, and Sialorphin were markedly upregulated in the FMT group \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eH-J\u003cb\u003e).\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eSubsequently, we established associations between dominant differential metabolites in the FMT\u0026thinsp;+\u0026thinsp;DSS group and key host parameters through correlation analysis. Heatmap analysis revealed that the following metabolites exhibited significantly negative correlations with pro-inflammatory cytokines (IL-1β, TNF-α), oxidative stress marker MDA, disease activity index (DAI), and colonic inflammation markers: 3-Methoxytyramine-betaxanthin, PG(18:0_22:4(7Z,10Z,13Z,16Z)), Ile-Gly-Ala-Val, Valylglutamine, 6-keto-PGF1α, PC(18:0/0:0), Gly-Val, Sialorphin, and Bz-Ile-Glu-Gly-Arg-pNA. Conversely, these metabolites showed significantly positive correlations with the anti-inflammatory cytokine IL-10 and antioxidant enzyme SOD (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Collectively, these upregulated metabolites likely contribute to maintaining host cytokines homeostasis, protecting intestinal barrier integrity, and alleviating DSS-induced intestinal inflammation.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\u003ch2\u003eFMT Significantly Modulates Host Metabolic Pathways\u003c/h2\u003e\u003cp\u003eMetabolites form dynamic interaction networks that drive essential biological pathways. Systematic analysis of these complex metabolic reactions provides comprehensive insights into FMT's mechanisms in combating colitis pathogenesis. Through KEGG pathway annotation (functionally defined KO level 3), we identified the top 20 most enriched metabolic pathways (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eA), including 56 amino acid metabolism pathways (tryptophan metabolism etc.), 121 biosynthetic pathways for secondary metabolites, 29 digestive system pathways (primarily associated with bile secretion), 51 lipid metabolism pathways and 37 nucleotide metabolism pathways. Human Metabolome Database (HMDB) contains comprehensive information on small-molecule metabolites in the human. Top 20 metabolite classes (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eB) account for 827 small-molecule metabolites involved in the Lipids and lipid-like molecules metabolic pathways (including Fatty Acyls), 17 small-molecule metabolites involved in the Nucleosides, nucleotides, and analogues metabolic pathways (including Pyrimidine nucleosides), and 286 small-molecule metabolites involved in the Organic acids and derivatives metabolic pathways (including Carboxylic acids and derivatives).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eDifferential metabolites interact within the organism, forming distinct pathways. Using the KEGG database, the differential metabolites between the DSS group and the FMT\u0026thinsp;+\u0026thinsp;DSS group were annotated. Top 20 pathway entries with the highest number of annotated differential metabolites were selected (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eC). Among these, 14 pathways related to amino acid metabolism (including tryptophan metabolism) showed significant differences between the two groups. 46 differential metabolic pathways were annotated as belonging to the Biosynthesis of other secondary metabolites. 15 differential metabolic pathways associated with bile secretion within the Digestive system were annotated. 25 and 14 differential metabolic pathways were annotated for Lipid metabolism and Nucleotide metabolism. Further analysis of Differential Abundance (DA) Score for the differential metabolic pathways between the DSS and FMT\u0026thinsp;+\u0026thinsp;DSS groups revealed that pathways such as Betalain biosynthesis and Methane metabolism were significantly upregulated in the FMT\u0026thinsp;+\u0026thinsp;DSS group (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eD). In contrast, pathways including Pathways in cancer, Bile secretion, and Steroid hormone biosynthesis were significantly upregulated in the DSS group (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eD). The differential metabolite enrichment network showed that all differential metabolites within the Betalain biosynthesis pathway were significantly upregulated in the FMT\u0026thinsp;+\u0026thinsp;DSS group (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eE), suggesting that FMT may exert its anti-inflammatory effects by regulating multiple metabolic pathways in the host, including Betalain biosynthesis.\u003c/p\u003e\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eHost-microbe homeostasis is closely associated with immune regulation and immune tolerance [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. In recent years, modulating host microbiota structure and function by targeting the gut microbiota, such as FMT for microbial dysbiosis, provides novel strategies for related diseases [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. The disease-resistant phenotype of piglets is closely linked to the structure of the host microbial community, holding significant implications for livestock production [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Differences in microbiota between healthy post-weaning piglets and diarrheic piglets with intestinal inflammation suggest that microbiota from healthy pigs may possess the ability to alleviate intestinal inflammation [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. However, the efficacy and underlying mechanisms of healthy piglet-derived microbiota in regulating host intestinal inflammation remain unclear. In the present study, to explore the therapeutic effect of bacteria derived from healthy piglets on colitis, GF mice were colonized with donor microbiota from 35-day-old healthy piglets, followed by DSS-induced colitis. The experimental results demonstrate that FMT significantly alleviated DSS-induced body weight loss and DAI scores in mice. Histological staining revealed that FMT reduced intestinal inflammation levels, improved intestinal barrier function, and maintained homeostasis of cytokines in the colitis. Moreover, FMT also significantly downregulated the level of the colitis marker MPO [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], reduced the level of the oxidative stress biomarker MDA, and elevated the level of the antioxidant biomarker SOD [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eConsistent with previous research identifying a cytokine dysregulation pattern in intestinal inflammation [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e], this study demonstrates that FMT significantly modulates the host's cytokine profile in the DSS-induced colitis model. Specifically, FMT markedly increased serum IL-10 levels, the hallmark anti-inflammatory cytokine, thereby suppressing the host's pro-inflammatory immune response and alleviating mucosal damage symptoms in IBD [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Concurrently, FMT significantly decreased the serum concentrations of key pro-inflammatory mediators IL-1β and serum TNF-α [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e], further substantiating its anti-inflammatory effect. Moreover, FMT effectively ameliorated markers of neutrophil infiltration and tissue damage, as evidenced by reduced MPO activity [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Oxidative stress is recognized as a potential mechanism in the pathophysiology of IBD ([\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]). MDA serves as a common biomarker of oxidative stress, reflecting the extent of cell membrane damage. Organisms possess a primary antioxidant defense system, largely dependent on enzymes like SOD, to counteract oxidative stress [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. In this study, FMT significantly reduced DSS-induced MDA levels and significantly increased host serum SOD levels, suggesting that FMT can significantly mitigate oxidative stress symptoms in the colitis model.\u003c/p\u003e\u003cp\u003eIBD may result from the combined effects of microbial factors, intestinal mucosal barrier dysfunction, oxidative stress, and increased inflammatory mediators [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Research consistently shows that impaired intestinal epithelial barrier function is a hallmark of IBD [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. This dysfunction involves downregulation of crucial components, including the MUC-2 and tight junction proteins [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. In the present study, FMT intervention significantly upregulated the expression levels of ZO-1 and Occludin in colonic tissue and restored MUC2 expression, FMT from healthy pigs protects the intestinal barrier to alleviate DSS-induced damage.\u003c/p\u003e\u003cp\u003eThe interaction between microorganisms and the host via metabolism a potential mechanism by which FMT modulates host intestinal immunity and ameliorates inflammatory symptoms [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. In the present study, untargeted metabolomics results revealed that FMT significantly modulated the composition of host serum metabolites. Furthermore, FMT markedly elevated the levels of multiple metabolites possessing anti-inflammatory properties. Specific amino acids and bioactive peptides have been demonstrated potential in alleviating intestinal inflammation symptoms in IBD patients [\u003cspan additionalcitationids=\"CR40 CR41\" citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Notably, FMT significantly increased the relative abundance of several peptide metabolites, including Valylglutamine; Ile Gly Ala Val; Gly Val; and Bz-Ile-glu-gly-arg-pna. This elevation may be associated with FMT's anti-inflammatory efficacy mediated through the provision of these peptides. Phosphatidylglycerol (PG) are a subset of glycerophospholipids [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], glycerophospholipids represent a diverse class of biological molecules that play vital roles in all living systems including cell membrane structure, vesicular transport, and intracellular signaling [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. In this study, FMT significantly increased the levels of the metabolite PG (18:0/22:4(7Z,10Z,13Z,16Z)), this elevation may contribute to the restoration of cellular membrane architecture in various intestinal cell types of FMT recipient mice. A 30% phosphatidylcholine (PC)-containing lecithin in delayed intestinal release formulation improves clinical outcomes [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e], and FMT significantly increased the levels of the metabolite phosphatidylcholine PC (18:0/0:0). 3-methoxytyramine-betaxanthin exhibited strong antioxidant activity [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]). In this study, FMT significantly increased the levels of the metabolite 3-methoxytyramine-betaxanthin, and this elevation may be associated with the attenuation of IBD symptoms. Sialorphin, a potent endogenous inhibitor of the opioid peptide-degrading enzymes neprilysin and aminopeptidase N. Notably, Systemic administration of Sialorphin has demonstrated efficacy in attenuating TNBS-induced colitis in mice, its anti-inflammatory actions are mediated through \u0026micro;- and κ-opioid receptors [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The observed elevation in Sialorphin abundance likely contributes to the anti-inflammatory effects of FMT. The levels of the metabolites 6-keto-PGF1α and bis(glutathionyl)spermine disulfide were significantly elevated in the FMT group, while the mechanisms by which these metabolites contribute to IBD alleviation require further elucidation.\u003c/p\u003e\u003cp\u003eMultiple anti-inflammatory metabolites significantly upregulated by FMT exhibited significantly negative correlations with pro-inflammatory cytokines (IL-1β, TNF-α), oxidative stress markers MDA [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], DAI scores, and colonic inflammation markers MPO [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Conversely, they showed significantly positive correlations with the anti-inflammatory cytokine IL-10 and the antioxidant marker SOD [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. These findings further substantiate that FMT may exert protective effects against IBD, at least partially, through the upregulation of multiple beneficial anti-inflammatory metabolites.\u003c/p\u003e\u003cp\u003eMetabolites within the host interact to form distinct metabolic pathways. Notably, 3-methoxytyramine-betaxanthin has been demonstrated to exhibit strong antioxidant activity [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. In the present study, the betalain biosynthesis pathway was significantly upregulated in the FMT group. Furthermore, the differential metabolite enrichment network revealed that all key metabolites associated with betalain biosynthesis were significantly upregulated in the FMT\u0026thinsp;+\u0026thinsp;DSS group compared to the DSS group. These findings collectively propose that FMT may exert its anti-inflammatory effects, at least partially, through the coordinated modulation of host metabolic pathways, including betalain biosynthesis. Conversely, the Pathways in cancer were significantly upregulated in the DSS group, and this noteworthy observation suggests that DSS-induced colitis may confer a higher susceptibility to cancer development within the intestinal tract.\u003c/p\u003e\u003cp\u003eHowever, this study has several limitations. Notably, the exploration of the precise underlying mechanisms remains insufficient. Future research should employ integrated multi-omics approaches, combining metagenomics and metabolomics, to further elucidate the specific mechanisms governing the interactions between pig-derived microbiota and host metabolic and immune regulation.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, the gut microbiota plays a pivotal role in modulating host immune responses and inflammatory control. This study demonstrates that pig-derived microbiota alleviates host inflammation by modulating recipient metabolism. Collectively, our findings establish a theoretical foundation for developing microbiota-based anti-inflammatory strategies. Future research should prioritize investigating the therapeutic potential of pig-derived microbiota for clinical applications in veterinary medicine.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eDAI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 440px;\"\u003e\n \u003cp\u003eDisease Activity Index\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eDSS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 440px;\"\u003e\n \u003cp\u003eDextran Sulfate Sodium\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eFMT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 440px;\"\u003e\n \u003cp\u003eFecal Microbiota Transplantation\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eIBD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 440px;\"\u003e\n \u003cp\u003eInflammatory Bowel Disease\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eIF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 440px;\"\u003e\n \u003cp\u003eImmunofluorescence\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eIHC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 440px;\"\u003e\n \u003cp\u003eImmunohistochemistry\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 440px;\"\u003e\n \u003cp\u003eGerm-free\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was reviewed and approved by the Laboratory Animal Welfare Ethics Committee of Shanghai Tenth People\u0026apos;s Hospital, all procedures complied with the Shanghai Tenth People\u0026apos;s Hospital \u003cem\u003eGuide for the Care and Use of Laboratory Animals\u003c/em\u003e (Animal Ethics Approval No.: SHDSYY-2025-T0088-02).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have declared that there is no conflict of interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eYY designed the experiment. QL, YY, YW, ZW, JL, YG, XT, SF, WC, HZ performed the animal trials, sample collection, and data analysis. Donor faecal samples were provided by QG. YY and QL drafted the manuscript. HW revised the manuscript. All authors contributed to the manuscript and approved the submitted version.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the National Key Research and Development Program of China (2021YFA0805904).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Quanzhou Lvzhiyuan Co., LTD for providing healthy piglet feces, which facilitated our study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial number:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eZheng D, Liwinski T, Elinav E. Interaction between microbiota and immunity in health and disease. Cell Res. 2020;30(6):492\u0026ndash;506; doi: 10.1038/s41422-020-0332-7.\u003c/li\u003e\n\u003cli\u003eZhao X, Jiang L, Fang X, Guo Z, Wang X, Shi B, et al. Host-microbiota interaction-mediated resistance to inflammatory bowel disease in pigs. Microbiome. 2022;10(1):115; doi: 10.1186/s40168-022-01303-1.\u003c/li\u003e\n\u003cli\u003eSekirov I, Russell SL, Antunes LC, Finlay BB. Gut microbiota in health and disease. Physiol Rev. 2010;90(3):859\u0026ndash;904; doi: 10.1152/physrev.00045.2009.\u003c/li\u003e\n\u003cli\u003eKayama H, Okumura R, Takeda K. Interaction Between the Microbiota, Epithelia, and Immune Cells in the Intestine. Annu Rev Immunol. 2020;38:23\u0026ndash;48; doi: 10.1146/annurev-immunol-070119-115104.\u003c/li\u003e\n\u003cli\u003eJin L, Shi X, Yang J, Zhao Y, Xue L, Xu L, et al. Gut microbes in cardiovascular diseases and their potential therapeutic applications. Protein Cell. 2021;12(5):346\u0026ndash;59; doi: 10.1007/s13238-020-00785-9.\u003c/li\u003e\n\u003cli\u003eJohnsen PH, Hilpusch F, Cavanagh JP, Leikanger IS, Kolstad C, Valle PC, et al. Faecal microbiota transplantation versus placebo for moderate-to-severe irritable bowel syndrome: a double-blind, randomised, placebo-controlled, parallel-group, single-centre trial. Lancet Gastroenterol Hepatol. 2018;3(1):17\u0026ndash;24; doi: 10.1016/S2468-1253(17)30338-2.\u003c/li\u003e\n\u003cli\u003eIslam J, Tanimizu M, Shimizu Y, Goto Y, Ohtani N, Sugiyama K, et al. Development of a rational framework for the therapeutic efficacy of fecal microbiota transplantation for calf diarrhea treatment. Microbiome. 2022;10(1):31; doi: 10.1186/s40168-021-01217-4.\u003c/li\u003e\n\u003cli\u003eWu R, Xiong R, Li Y, Chen J, Yan R. Gut microbiome, metabolome, host immunity associated with inflammatory bowel disease and intervention of fecal microbiota transplantation. J Autoimmun. 2023;141:103062; doi: 10.1016/j.jaut.2023.103062.\u003c/li\u003e\n\u003cli\u003eWu J, Wang J, Lin Z, Liu C, Zhang Y, Zhang S, et al. Clostridium butyricum alleviates weaned stress of piglets by improving intestinal immune function and gut microbiota. Food Chem. 2023;405(Pt B):135014; doi: 10.1016/j.foodchem.2022.135014.\u003c/li\u003e\n\u003cli\u003eZhou X, He Y, Chen J, Xiong X, Yin J, Liang J, et al. Colonic phosphocholine is correlated with Candida tropicalis and promotes diarrhea and pathogen clearance. NPJ Biofilms Microbiomes. 2023;9(1):62; doi: 10.1038/s41522-023-00433-0.\u003c/li\u003e\n\u003cli\u003eZhang J, Chen Y, Guo X, Li X, Zhang R, Wang M, et al. The gut microbial metabolite indole-3-aldehyde alleviates impaired intestinal development by promoting intestinal stem cell expansion in weaned piglets. J Anim Sci Biotechnol. 2024;15(1):150; doi: 10.1186/s40104-024-01111-7.\u003c/li\u003e\n\u003cli\u003eTao S, Fan J, Li J, Wu Z, Yao Y, Wang Z, et al. Extracellular vesicles derived from Lactobacillus johnsonii promote gut barrier homeostasis by enhancing M2 macrophage polarization. J Adv Res. 2025;69:545\u0026ndash;63; doi: 10.1016/j.jare.2024.03.011.\u003c/li\u003e\n\u003cli\u003eGresse R, Chaucheyras-Durand F, Fleury MA, Van de Wiele T, Forano E, Blanquet-Diot S. Gut Microbiota Dysbiosis in Postweaning Piglets: Understanding the Keys to Health. Trends Microbiol. 2017;25(10):851\u0026ndash;73; doi: 10.1016/j.tim.2017.05.004.\u003c/li\u003e\n\u003cli\u003eHu J, Ma L, Nie Y, Chen J, Zheng W, Wang X, et al. A Microbiota-Derived Bacteriocin Targets the Host to Confer Diarrhea Resistance in Early-Weaned Piglets. Cell Host Microbe. 2018;24(6):817\u0026ndash;32 e8; doi: 10.1016/j.chom.2018.11.006.\u003c/li\u003e\n\u003cli\u003ePanda SK, Peng V, Sudan R, Ulezko Antonova A, Di Luccia B, Ohara TE, et al. Repression of the aryl-hydrocarbon receptor prevents oxidative stress and ferroptosis of intestinal intraepithelial lymphocytes. Immunity. 2023;56(4):797\u0026ndash;812 e4; doi: 10.1016/j.immuni.2023.01.023.\u003c/li\u003e\n\u003cli\u003eNayar S, Morrison JK, Giri M, Gettler K, Chuang LS, Walker LA, et al. A myeloid-stromal niche and gp130 rescue in NOD2-driven Crohn\u0026apos;s disease. Nature. 2021;593(7858):275\u0026ndash;81; doi: 10.1038/s41586-021-03484-5.\u003c/li\u003e\n\u003cli\u003eLi J, Wei H. Establishment of an efficient germ-free animal system to support functional microbiome research. Sci China Life Sci. 2019;62(10):1400\u0026ndash;3; doi: 10.1007/s11427-019-9832-9.\u003c/li\u003e\n\u003cli\u003eFouladi F, Glenny EM, Bulik-Sullivan EC, Tsilimigras MCB, Sioda M, Thomas SA, et al. Sequence variant analysis reveals poor correlations in microbial taxonomic abundance between humans and mice after gnotobiotic transfer. ISME J. 2020;14(7):1809\u0026ndash;20; doi: 10.1038/s41396-020-0645-z.\u003c/li\u003e\n\u003cli\u003eSouza DG, Senchenkova EY, Russell J, Granger DN. MyD88 mediates the protective effects of probiotics against the arteriolar thrombosis and leukocyte recruitment associated with experimental colitis. Inflamm Bowel Dis. 2015;21(4):888\u0026ndash;900; doi: 10.1097/MIB.0000000000000331.\u003c/li\u003e\n\u003cli\u003eMurthy SN, Cooper HS, Shim H, Shah RS, Ibrahim SA, Sedergran DJ. Treatment of dextran sulfate sodium-induced murine colitis by intracolonic cyclosporin. Dig Dis Sci. 1993;38(9):1722\u0026ndash;34; doi: 10.1007/BF01303184.\u003c/li\u003e\n\u003cli\u003eNishiyama Y, Kataoka T, Yamato K, Taguchi T, Yamaoka K. Suppression of dextran sulfate sodium-induced colitis in mice by radon inhalation. Mediators Inflamm. 2012;2012:239617; doi: 10.1155/2012/239617.\u003c/li\u003e\n\u003cli\u003eDieleman LA, Palmen MJ, Akol H, Bloemena E, Pena AS, Meuwissen SG, et al. Chronic experimental colitis induced by dextran sulphate sodium (DSS) is characterized by Th1 and Th2 cytokines. Clin Exp Immunol. 1998;114(3):385\u0026ndash;91; doi: 10.1046/j.1365-2249.1998.00728.x.\u003c/li\u003e\n\u003cli\u003eChadwick VS, Schlup MM, Ferry DM, Chang AR, Butt TJ. Measurements of unsaturated vitamin B12-binding capacity and myeloperoxidase as indices of severity of acute inflammation in serial colonoscopy biopsy specimens from patients with inflammatory bowel disease. Scand J Gastroenterol. 1990;25(12):1196\u0026ndash;204; doi: 10.3109/00365529008998554.\u003c/li\u003e\n\u003cli\u003eKlebanoff SJ, Coombs RW. Viricidal effect of polymorphonuclear leukocytes on human immunodeficiency virus-1. Role of the myeloperoxidase system. J Clin Invest. 1992;89(6):2014\u0026ndash;7; doi: 10.1172/JCI115810.\u003c/li\u003e\n\u003cli\u003eYi LT, Wang XY, Zhou L, Cheng J, Xu GH, Zhu JX. Polysaccharides from Black Mulberry Attenuate Colitis through Gut Microbiota Mediated TNF-alpha/pNF-kappaB/ICAM-1 Signaling Pathway. J Agric Food Chem. 2025;73(23):14314\u0026ndash;32; doi: 10.1021/acs.jafc.5c01870.\u003c/li\u003e\n\u003cli\u003eNeurath MF. Strategies for targeting cytokines in inflammatory bowel disease. Nat Rev Immunol. 2024;24(8):559\u0026ndash;76; doi: 10.1038/s41577-024-01008-6.\u003c/li\u003e\n\u003cli\u003eShouval DS, Biswas A, Goettel JA, McCann K, Conaway E, Redhu NS, et al. Interleukin-10 receptor signaling in innate immune cells regulates mucosal immune tolerance and anti-inflammatory macrophage function. Immunity. 2014;40(5):706\u0026ndash;19; doi: 10.1016/j.immuni.2014.03.011.\u003c/li\u003e\n\u003cli\u003eYang T, Hu X, Cao F, Yun F, Jia K, Zhang M, et al. Targeting symbionts by apolipoprotein L proteins modulates gut immunity. Nature. 2025; doi: 10.1038/s41586-025-08990-4.\u003c/li\u003e\n\u003cli\u003eTian HL, Wang L, Ma CL, Yang B, Li L, Ye C, et al. [Fecal microbiota transplantation for the treatment of intestinal disorders: An analysis of treatment of 15 000 patients]. Zhonghua Wei Chang Wai Ke Za Zhi. 2025;28(3):296\u0026ndash;303; doi: 10.3760/cma.j.cn441530-20250114-00025.\u003c/li\u003e\n\u003cli\u003eLi J, Feng S, Wang Z, He J, Zhang Z, Zou H, et al. Limosilactobacillus mucosae-derived extracellular vesicles modulates macrophage phenotype and orchestrates gut homeostasis in a diarrheal piglet model. NPJ Biofilms Microbiomes. 2023;9(1):33; doi: 10.1038/s41522-023-00403-6.\u003c/li\u003e\n\u003cli\u003eJomova K, Raptova R, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, et al. Reactive oxygen species, toxicity, oxidative stress, and antioxidants: chronic diseases and aging. Arch Toxicol. 2023;97(10):2499\u0026ndash;574; doi: 10.1007/s00204-023-03562-9.\u003c/li\u003e\n\u003cli\u003eZhu W, Ren L, Zhang L, Qiao Q, Farooq MZ, Xu Q. The Potential of Food Protein-Derived Bioactive Peptides against Chronic Intestinal Inflammation. Mediators Inflamm. 2020;2020:6817156; doi: 10.1155/2020/6817156.\u003c/li\u003e\n\u003cli\u003eMoschen AR, Tilg H, Raine T. IL-12, IL-23 and IL-17 in IBD: immunobiology and therapeutic targeting. Nat Rev Gastroenterol Hepatol. 2019;16(3):185\u0026ndash;96; doi: 10.1038/s41575-018-0084-8.\u003c/li\u003e\n\u003cli\u003eLi L, Qiu N, Meng Y, Wang C, Mine Y, Keast R, et al. Preserved egg white alleviates DSS-induced colitis in mice through the reduction of oxidative stress, modulation of infl ammatory cytokines, NF-\u0026kappa;B, MAPK and gut microbiota composition. Food Science and Human Wellness. 2022;12(1):312\u0026ndash;23; doi: 10.1016/j.fshw.2022.07.021.\u003c/li\u003e\n\u003cli\u003eMehandru S, Colombel JF. The intestinal barrier, an arbitrator turned provocateur in IBD. Nat Rev Gastroenterol Hepatol. 2021;18(2):83\u0026ndash;4; doi: 10.1038/s41575-020-00399-w.\u003c/li\u003e\n\u003cli\u003eFu Y, Guzior DV, Okros M, Bridges C, Rosset SL, Gonzalez CT, et al. Balance between bile acid conjugation and hydrolysis activity can alter outcomes of gut inflammation. Nat Commun. 2025;16(1):3434; doi: 10.1038/s41467-025-58649-x.\u003c/li\u003e\n\u003cli\u003eLiu C, Du MX, Xie LS, Wang WZ, Chen BS, Yun CY, et al. Gut commensal Christensenella minuta modulates host metabolism via acylated secondary bile acids. Nat Microbiol. 2024;9(2):434\u0026ndash;50; doi: 10.1038/s41564-023-01570-0.\u003c/li\u003e\n\u003cli\u003eLiu M, Ma J, Xu J, Huangfu W, Zhang Y, Ali Q, et al. Fecal microbiota transplantation alleviates intestinal inflammatory diarrhea caused by oxidative stress and pyroptosis via reducing gut microbiota-derived lipopolysaccharides. Int J Biol Macromol. 2024;261(Pt 1):129696; doi: 10.1016/j.ijbiomac.2024.129696.\u003c/li\u003e\n\u003cli\u003eXu Q, Hu M, Li M, Hou J, Zhang X, Gao Y, et al. Dietary Bioactive Peptide Alanyl-Glutamine Attenuates Dextran Sodium Sulfate-Induced Colitis by Modulating Gut Microbiota. Oxid Med Cell Longev. 2021;2021:5543003; doi: 10.1155/2021/5543003.\u003c/li\u003e\n\u003cli\u003eHe D, Zeng W, Wang Y, Xing Y, Xiong K, Su N, et al. Isolation and characterization of novel peptides from fermented products of Lactobacillus for ulcerative colitis prevention and treatment. Food Science and Human Wellness. 2022;11(6):1464\u0026ndash;74; doi: https://doi.org/10.1016/j.fshw.2022.06.003.\u003c/li\u003e\n\u003cli\u003eZhao Y, Xue P, Lin G, Tong M, Yang J, Zhang Y, et al. A KPV-binding double-network hydrogel restores gut mucosal barrier in an inflamed colon. Acta Biomater. 2022;143:233\u0026ndash;52; doi: 10.1016/j.actbio.2022.02.039.\u003c/li\u003e\n\u003cli\u003eGravina AG, Prevete N, Tuccillo C, De Musis C, Romano L, Federico A, et al. Peptide Hp(2-20) accelerates healing of TNBS-induced colitis in the rat. United European Gastroenterol J. 2018;6(9):1428\u0026ndash;36; doi: 10.1177/2050640618793564.\u003c/li\u003e\n\u003cli\u003eStruzik ZJ, Weerts AN, Storch J, Thompson DH. Stereospecific synthesis of phosphatidylglycerol using a cyanoethyl phosphoramidite precursor. Chem Phys Lipids. 2020;231:104933; doi: 10.1016/j.chemphyslip.2020.104933.\u003c/li\u003e\n\u003cli\u003eDowhan W, Bogdanov M. Chapter 1 Functional roles of lipids in membranes. Elsevier; 2002.\u003c/li\u003e\n\u003cli\u003eStremmel W, Vural H, Evliyaoglu O, Weiskirchen R. Delayed-Release Phosphatidylcholine Is Effective for Treatment of Ulcerative Colitis: A Meta-Analysis. Dig Dis. 2021;39(5):508\u0026ndash;15; doi: 10.1159/000514355.\u003c/li\u003e\n\u003cli\u003eCai Y, Sun M, Corke H. Antioxidant activity of betalains from plants of the amaranthaceae. J Agric Food Chem. 2003;51(8):2288\u0026ndash;94; doi: 10.1021/jf030045u.\u003c/li\u003e\n\u003cli\u003eMadadi E, Mazloum-Ravasan S, Yu JS, Ha JW, Hamishehkar H, Kim KH. Therapeutic Application of Betalains: A Review. Plants (Basel). 2020;9(9); doi: 10.3390/plants9091219.\u003c/li\u003e\n\u003cli\u003eSalaga M, Mokrowiecka A, Jacenik D, Cygankiewicz AI, Malecka-Panas E, Kordek R, et al. Systemic Administration of Sialorphin Attenuates Experimental Colitis in Mice via Interaction With Mu and Kappa Opioid Receptors. J Crohns Colitis. 2017;11(8):988\u0026ndash;98; doi: 10.1093/ecco-jcc/jjx043.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Healthy piglet microbiota, Fecal microbiota transplantation, Germ-free mice, Inflammatory Bowel Disease, Metabolome","lastPublishedDoi":"10.21203/rs.3.rs-7536850/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7536850/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eFecal Microbiota Transplantation (FMT) has been clinically applied to treat host intestinal inflammation, such as inflammatory bowel disease (IBD). Research in livestock indicates that the gut microbiota of healthy piglets differs from that of diarrheic piglets, playing a crucial role in regulating intestinal immune development. However, the potential of FMT derived from healthy piglets to alleviate intestinal inflammation in recipients and the underlying mechanisms remain unexplored. This study utilized FMT from healthy piglets to intervene in a dextran sulfate sodium (DSS)-induced intestinal inflammation model in germ-free Kunming (KM) mice, investigating its effects on intestinal barrier function and inflammatory levels. As anticipated, the results demonstrated that FMT significantly alleviated DSS-induced intestinal inflammation. This was evidenced by reduced weight loss and lower disease activity index (DAI) scores. Furthermore, FMT improved intestinal barrier integrity, maintained homeostasis of host inflammatory cytokines, and markedly attenuated oxidative stress. Untargeted metabolomics analysis further revealed that FMT significantly increased the abundance of multiple anti-inflammatory metabolites, including 3-Methoxytyramine-betaxanthin and Sialorphin. Concurrently, FMT upregulated relevant metabolic pathways, notably Betalain Biosynthesis. Correlation analysis indicated a close association between FMT-elevated anti-inflammatory metabolites (e.g., 3-Methoxytyramine-betaxanthin) and improved markers of intestinal inflammation. This study provides novel insights into the mechanism by which pig-derived gut microbiota alleviates host intestinal inflammation through modulation of host metabolism.\u003c/p\u003e","manuscriptTitle":"Fecal Microbiota Transplantation from Healthy Piglets Ameliorates Intestinal Inflammation in Mice by Modulating Recipient Metabolism","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-01 12:37:52","doi":"10.21203/rs.3.rs-7536850/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-29T04:39:12+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-28T14:19:39+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-28T03:07:06+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-24T01:19:40+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-23T03:02:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"147191436892119763401957154350362524338","date":"2025-10-21T18:05:14+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"112604067587802706191938657648167593626","date":"2025-10-21T09:47:16+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"122859440633248635255004000748319917274","date":"2025-10-20T01:54:08+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"184265895077601974738116874865039442820","date":"2025-10-19T12:36:20+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"258316884633352136824196423711501466787","date":"2025-10-19T09:56:15+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"44787329922036396017056136435515913112","date":"2025-09-22T11:53:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"172276718888021360212530382178903829260","date":"2025-09-22T02:06:04+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-19T10:42:16+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-19T07:32:29+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-09-18T17:58:56+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-18T13:44:52+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Microbiology","date":"2025-09-18T12:14:01+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2bb0aa34-3026-43bc-b6d5-0f71d73ab778","owner":[],"postedDate":"October 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-01-19T17:02:48+00:00","versionOfRecord":{"articleIdentity":"rs-7536850","link":"https://doi.org/10.1186/s12866-025-04590-4","journal":{"identity":"bmc-microbiology","isVorOnly":false,"title":"BMC Microbiology"},"publishedOn":"2026-01-12 16:28:54","publishedOnDateReadable":"January 12th, 2026"},"versionCreatedAt":"2025-10-01 12:37:52","video":"","vorDoi":"10.1186/s12866-025-04590-4","vorDoiUrl":"https://doi.org/10.1186/s12866-025-04590-4","workflowStages":[]},"version":"v1","identity":"rs-7536850","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7536850","identity":"rs-7536850","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}