Supplementation of Bacillus cereus Enhances Anti-Colitis Effects of Triple Probiotics in Mice: Correlative Evidence for a Gut Microbiota-Spermidine-Related Axis in Quadruple-Probiotic Protection.
Adding Bacillus cereus to a triple-probiotic mixture (BLE) to form a quadruple formulation (BLEB) exerted superior dose-dependent anti-colitis effects compared with BLE, with ecological synergy reshaping gut microbiota and increasing microbiota-related spermidine, partially contributing to enhanced mucosal protection.
Key Findings
Results
The quadruple-probiotic formulation BLEB demonstrated superior dose-dependent anti-colitis effects compared with the triple-probiotic formulation BLE in DSS-induced colitis mice.
Acute dextran sulfate sodium (DSS)-induced colitis mouse models were used to compare therapeutic effects of BLE and BLEB.
Disease phenotype outcomes were evaluated alongside intestinal epithelial barrier integrity and splenic Treg/Th17 immune homeostasis.
The superiority of BLEB over BLE was described as dose-dependent.
The addition of facultative-aerobic Bacillus cereus to the oxygen-sensitive strains Bifidobacterium infantis, Lactobacillus acidophilus, and Enterococcus faecalis formed the BLEB formulation.
Results
BLEB treatment enriched beneficial Lactobacillaceae and restored luminal polyamine metabolites including spermidine in colitis mice.
Dynamic 16S rRNA sequencing was used to characterize gut microbial alterations.
Untargeted intestinal metabolomics was used to characterize metabolic alterations.
BLEB specifically enriched Lactobacillaceae-affiliated taxa in the gut microbiota.
Restoration of luminal polyamine metabolites, including spermidine, was observed following BLEB treatment.
Results
Spearman correlation analysis revealed significant positive correlations between Lactobacillaceae-affiliated genera and polyamine levels.
Spearman correlation was applied for microbiota-metabolite association analysis.
The correlation linked Lactobacillaceae-affiliated genera specifically to polyamine metabolite levels.
This analysis provided evidence for a gut microbiota-spermidine-related regulatory axis.
The correlative findings were described as supporting the concept of an ecological synergy introduced by B. cereus reshaping the gut microbiota.
Results
Spermidine reversed DSS-provoked transcriptional perturbations in DSS-injured Caco-2 cells and upregulated glutathione-centered antioxidant-related gene transcription.
Transcriptomic profiling was performed on DSS-injured Caco-2 cells to examine molecular responses to spermidine.
Spermidine treatment reversed transcriptional perturbations induced by DSS injury in vitro.
Spermidine upregulated the transcription of glutathione-centered antioxidant-related genes.
These in vitro findings provided mechanistic evidence linking microbiota-derived spermidine to mucosal epithelial protection.
Background
The triple-probiotic mixture BLE contains highly oxygen-sensitive strains prone to viability loss during gastrointestinal transit, which motivated the addition of facultative-aerobic B. cereus.
BLE comprises Bifidobacterium infantis, Lactobacillus acidophilus, and Enterococcus faecalis, described as 'highly oxygen-sensitive strains prone to viability loss during gastrointestinal transit.'
B. cereus is characterized as a facultative-aerobic bacterium.
The rationale for adding B. cereus was to provide ecological synergy that may protect the oxygen-sensitive strains and reshape gut microbiota.
The resulting four-strain combination was designated BLEB.
Splenic Treg/Th17 immune homeostasis was evaluated as part of the assessment of BLEB's therapeutic effects.
Treg/Th17 balance is a key immunological parameter relevant to inflammatory bowel disease.
This immune modulation was assessed alongside intestinal epithelial barrier integrity and disease phenotype outcomes.
The findings were obtained in the acute DSS-induced colitis mouse model.
Conclusions
The study provides pre-clinical evidence supporting the development of aerobic-anaerobic combined probiotic cocktails for auxiliary UC therapy.
The authors describe their findings as 'pre-clinical evidence supporting the development of aerobic-anaerobic combined probiotic cocktails for auxiliary UC therapy.'
The ecological synergy introduced by B. cereus was proposed to partially contribute to the enhanced mucosal protection conferred by BLEB.
The mechanism was described as operating through a 'gut microbiota-spermidine-related axis.'
The study used both in vivo (mouse colitis model) and in vitro (Caco-2 cells) approaches.
What This Means
This research suggests that adding a specific bacterium called Bacillus cereus to an existing three-strain probiotic mixture significantly improves its ability to reduce symptoms of ulcerative colitis in mice. The original three-strain mixture (called BLE) contains bacteria that are sensitive to oxygen and may lose effectiveness during digestion. By adding Bacillus cereus, which can survive in both oxygen-rich and oxygen-poor environments, the researchers created a four-strain combination (BLEB) that outperformed the original mixture in reducing intestinal inflammation, protecting the gut lining, and restoring immune balance.
The study found that BLEB changed the composition of gut bacteria in colitis mice, particularly increasing beneficial Lactobacillaceae bacteria, and also restored levels of a natural compound called spermidine in the gut. Statistical analysis showed that these Lactobacillaceae bacteria were positively linked to spermidine levels, suggesting the bacteria may help produce or maintain this compound. When tested in laboratory gut cells damaged by the same chemical used to induce colitis in mice, spermidine reversed the damage and activated genes involved in protecting cells from oxidative stress, specifically genes related to glutathione, one of the body's key antioxidant systems.
This research suggests that combining oxygen-tolerant and oxygen-sensitive probiotic strains may create a more effective probiotic therapy for inflammatory bowel conditions like ulcerative colitis. The proposed mechanism involves a chain of events where the added bacterium reshapes the gut microbial community, which in turn increases spermidine production, which then helps protect the gut lining from inflammation. These findings are from animal and cell-based experiments, so further research would be needed to determine whether similar effects occur in humans.
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Li J, Du X, Zhao J, Li W, Wang Q, Zhang L, et al.. (2026). Supplementation of Bacillus cereus Enhances Anti-Colitis Effects of Triple Probiotics in Mice: Correlative Evidence for a Gut Microbiota-Spermidine-Related Axis in Quadruple-Probiotic Protection.. Nutrients. https://doi.org/10.3390/nu18193164