The probiotic enhances donor microbiota stability and improves the efficacy of fecal microbiota transplantation for treating colitis

Probiotic pretreatment with L. plantarum GR-4 enhances donor microbiota stability and functionality, improving fecal microbiota transplantation (FMT) efficacy in a DSS-induced IBD mouse model. Modified FMT (MFMT) increases butyrate levels, boosts beneficial bacterial colonization, improves gut barrier function, and alleviates colitis symptoms more effectively than standard FMT. This probiotic-based strategy offers a novel approach to optimizing FMT outcomes for inflammatory bowel disease. • Probiotic-modified donor microbiota boosts short-chain fatty acids. • Probiotic modification enhances donor microbes’ gut adaptation. • Modified FMT outperforms FMT and 5-ASA in reducing colitis in mice. • Modified FMT increases anti-inflammatory and immunomodulatory metabolites. • Modified FMT improves implantation and protects critical strain survival. The stability and metabolic functionality of donor microbiota are critical determinants of fecal microbiota transplantation (FMT) efficacy in inflammatory bowel disease (IBD). While probiotics show potential to enhance microbiota resilience, their role in optimizing donor microbiota for FMT remains underexplored. This study investigated whether pretreatment of donor microbiota with L. plantarum GR-4 could improve FMT outcomes in a DSS-induced colitis model by modulating microbial stability, metabolic activity, and host-microbiome interactions. Donor mice received L. plantarum GR-4 for 3 weeks to generate modified FMT (MFMT). DSS-colitis mice were treated with MFMT, conventional FMT, or 5-aminosalicylic acid (5-ASA). Multi-omics analyses and functional assays (stress resistance, engraftment efficiency) were used to evaluate therapeutic mechanisms. GR-4 pretreatment conferred three key advantages to donor microbiota: Ecological stabilization: 1. GR-4-driven acidification (pH 3.97 vs. 4.59 for LGG, p < 0.0001) enriched butyrogenic Butyricicoccus (73 % butyrate increase, p < 0.05) and improved stress resistance to bile acids/gastric conditions (1.25 × survival vs. FMT). 2. Metabolic reprogramming: GR-4 metabolized 25.3 % of tryptophan (vs. 10.3 % for LGG) to generate immunomodulatory indoles (ILA, IAA), activating aryl hydrocarbon receptor (AHR) signaling and upregulating anti-inflammatory IL-10/IL-22. 3. Bile acid remodeling: MFMT restored sulfolithocholic acid and β-MCA levels, outperforming FMT in resolving DSS-induced dysregulation. MFMT achieved an 83 % remission rate (vs. 50 % for FMT), enhanced gut barrier integrity, and reversed colitis-associated metabolic dysregulation (e.g., elevated spermidine, 7-sulfocholic acid). Probiotic preconditioning improved donor engraftment by 1.25 × and enriched success-associated taxa ( Sporobacter , Butyricimonas ), while suppressing pathogens ( Clostridium papyrosolvens ). L. plantarum GR-4 optimizes donor microbiota via pH-driven niche engineering, immunometabolic reprogramming, and bile acid modulation, addressing key limitations of conventional FMT. The multi-targeted efficacy of MFMT, evidenced by superior remission rates and metabolic restoration, establishes this approach as a translatable strategy for IBD therapy. This study establishes probiotic-enhanced FMT as a paradigm for precision microbiome interventions.