MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14164/m_MTBLS14164_LC-MS_positive_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14164/m_MTBLS14164_LC-MS_negative_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14164/s_MTBLS14164.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14164/a_MTBLS14164_LC-MS_negative_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14164/a_MTBLS14164_LC-MS_positive_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14164/i_Investigation.txtprimaryOK200intestinal contentsftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14164<p>Metabolites were annotated based on the self-built database MWDB (Wuhan Metware Biotechnology Co., Ltd., Wuhan, China). Metabolite identification was based on the accurate mass of metabolites, MS2 fragments, MS2 fragments isotope distribution and retention time (RT). Through the company's self-built intelligent secondary spectrum matching method, the secondary spectrum and RT of metabolites in our samples were matched intelligently with the secondary spectrum and RT of the company's database one by one. The MS tolerance and MS2 tolerance were set to 2 ppm and 5 ppm, respectively. Metabolites that did not have standard products were compared with MS2 spectra in public databases or literature. Some of the metabolites without standard secondary spectra were inferred based on experience.</p>mass spectrometry assay<p>The sample stored at-80 °C refrigerator was thawed on ice. A 400 μL solution (Methanol : Water = 7:3, V/V) containing internal standard was added into 20 mg sample, and vortexed for 3 min. The sample was sonicated in an ice bath for 10 min and vortexed for 1 min, and then placed in-20 °C for 30 min. The sample was then centrifuged at 12000 rpm for 10 min (4 °C). And the sediment was removed, then centrifuged the supernatant at 12000 rpm for 3 min (4 °C). A 200 μL aliquots of supernatant were transferred for LC-MS analysis.</p>Sus scrofa<p>Metabolite quantification was carried out via the MRM mode of the QQQ mass spectrometer. In the MRM mode, the quadrupole first searched for precursor ions (parent ions) of target substances while screening any ions derived from substances of different molecular weights to eliminate their interference preliminarily. The precursor ions were fragmented via induced ionization in the collision chamber to form many fragment ions, which were then filtered through QQs to select single-fragment ions with the desired characteristics while eliminating the interference from nontarget ions; this step leads to increased precision and repeatability of the quantification results. After the metabolite mass spectrometry data were obtained for the different samples, all the mass spectrum peaks were subjected to area integration. To compare the differences in the content of each metabolite for each of the detected metabolites from the different samples, we corrected the mass spectral peaks detected from the different samples for each metabolite on the basis of information on metabolite Rt and peak type, which ensured the accuracy of the qualitative and quantitative analyses.</p>Grouphttps://www.ebi.ac.uk/metabolights/MTBLS1416432122186@qq.com<p>After slaughter, the abdominal cavity was opened, and intestinal segments (jejunum, ileum, cecum, and colon) were ligated at both ends to avoid cross contamination. The intestinal contents were gently squeezed into sterile cryogenic tubes, immediately snap frozen in liquid nitrogen, and then stored at -80 for subsequent analysis of intestinal microbiota and metabolites.</p>MetaboLightsPublicMetabolomicsLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseMetabolomicsIntestinal barrierImmune functionLactobacillus reuteriuntargeted analysisGut MicrobiotaThermo Scientific Vanquish Flex UHPLC SystemThermo Scientific Q Exactive HF-X75-1000Suckling pigletsintestinal contentsSus scrofa<p>All samples were for two LC/MS methods. One aliquot was analyzed using positive ion conditions and waseluted from T3column(Waters ACQUITYPremierHSST3Column1.8µm,2.1mm*100mm) using 0.1 % formic acid in water as solvent A and 0.1 % formic acid in acetonitrile as solvent B in the following gradient: 5 to 20 % in 2 min, increased to 60 % in the following 3 mins, increased to 99 % in 1 min and held for 1.5 min, then come back to 5 % mobile phase B witnin 0.1 min, held for 2.4 min. The analytical conditions were as follows, column temperature, 40 °C; flow rate, 0.4 mL/min; injection volume, 4 μL; Another aliquot was using negative ion conditions and was the same as the elution gradient of positive mode.</p>Early-Life Intervention with Lactobacillus reuteri Enhances Intestinal2 Barrier Function and Resilience in Suckling Piglets via Modulation of3 Gut Microbiota and Metabolites.MetabolomicsIntestinal barrierImmune functionLactobacillus reuteriuntargeted analysisThermo Scientific Vanquish Flex UHPLC SystemGut MicrobiotaThermo Scientific Q Exactive HF-X75-1000Suckling pigletsintestinal contentsSus scrofaCollege of Animal Science & Technology, Guangxi UniversityQin Luo<p>All the methods alternated between full scan MS and data dependent MSn scans using dynamic ex clusion. MS analyses were carried out using electrospray ionization in the positive ion mode and negative ion mode using full scan analysis over m/z 75-1000 at 35000 resolution. Additional MS settings are: ion spray voltage, 3.5 KV or 3.2 KV in positive or negative modes, respevtively; Sheath gas (Arb), 30; Aux gas, 5; Ion transfer tube temperature, 320 °C; Vaporizer temperature, 300 °C; Collision energy, 30,40,50 V; Signal Intensity Threshold, 1*e6 cps; Top N vs Top speed, 10; Exclusion duration, 3s.</p>falseEarly-Life Intervention with Lactobacillus reuteri Enhances Intestinal2 Barrier Function and Resilience in Suckling Piglets via Modulation of3 Gut Microbiota and MetabolitesEarly-life nutritional interventions are pivotal for promoting piglet growth and health, particularly in reducing weaning-associated disorders. This study investigated the effects of early-life Lactobacillus reuteri (L.reuteri) supplementation on growth performance, immune function, intestinal morphology, gut microbiota, ileal metabolites, and barrier function in suckling piglets. Neonatal piglets were administered L.reuteri for either 3 or 7 days post-birth. Results indicated that L.reuteri supplementation significantly increased weaning weight and average daily gain (ADG) while markedly reducing diarrhea incidence. Immune and antioxidant capacities were enhanced, evidenced by elevated serum and ileal IgG, IL-4, IL-10, T-SOD, T-AOC, and GSH-Px levels, alongside decreased pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and MDA. Histological analysis revealed improved intestinal architecture, characterized by increased ileal villus height and reduced crypt depth. 16S rRNA sequencing and metabolomic analyses showed that L.reuteri reshaped the gut microbiota by expanding beneficial Lactobacillus species and suppressing potential pathogens (Streptococcus, Pasteurellaceae), while modulating ileal metabolites involved in amino acid and energy metabolism. Multi-omics integration highlighted coordinated interactions between microbial composition and metabolites linked to improved health outcomes. Furthermore, the expression of tight junction proteins (Occludin, Claudin-3, ZO-1) and mucins (MUC-1, MUC-2) was significantly upregulated, indicating strengthened intestinal barrier integrity. Collectively, these findings demonstrate that early-life intervention with L.reuteri confers comprehensive benefits on suckling piglet health through immune enhancement, antioxidant protection, microbiota remodeling, metabolic regulation, and barrier reinforcement, supporting its potential as a practical strategy to improve early-life resilience and mitigate weaning-associated disorders in swine production.2026-03-272026-03-27MTBLS14164