Defined anaerobic co-culture of Bifidobacterium longum CICC25033, Clostridium butyricum CICC23847, and Limosilactobacillus reuteri CICC6131 used to produce mixed-species probiotic membrane vesicles (M-MVs). Hou et al. grew the strains singly, pairwise, and as a 1:1:1 three-strain combination in MRS broth, observed higher viable counts in co-culture and with cell-free supernatants from partner strains, isolated M-MVs from the supernatant of the co-culture, found a distinct M-MV proteomic profile with antioxidant activity, and loaded luteolin into the resulting M-MVs for in-vitro and DSS-induced mouse-colitis assays.
Taxonomy
| Taxon | Ontology ID | Functional Roles | Abundance |
|---|---|---|---|
| Bifidobacterium longum CICC25033 — ontology label: Bifidobacterium longum | NCBITaxon:216816 · TaxonMech · TraitMech | N/A | |
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| Clostridium butyricum CICC23847 — ontology label: Clostridium butyricum | NCBITaxon:1492 · TaxonMech · TraitMech | N/A | |
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| Limosilactobacillus reuteri CICC6131 — ontology label: Limosilactobacillus reuteri | NCBITaxon:1598 · TaxonMech · TraitMech | N/A | |
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Ecological Interactions
The graph shows only explicitly linked interaction participants. Community members without an asserted participant link are omitted; membership alone does not imply an interaction.
Three-Strain Co-Culture Growth Enhancement
CROSS_FEEDINGThe exact B. longum CICC25033, C. butyricum CICC23847 and L. reuteri CICC6131 pairwise and three-strain cultures showed higher viable counts, and 1% partner cell-free supernatant promoted monoculture growth, especially of L. reuteri. PARTIAL - These interventions support diffusible growth promotion and the authors putative cross-feeding interpretation, but do not identify a transferred nutrient or its necessity. Co-culture plate counts are not reported here as member-resolved counts, so increased total growth does not establish absence of antagonism toward every member.
Biological Processes:
- interspecies interaction (GO:0044419 · PathwayMech)
Evidence
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PMID:42604006 - SUPPORT (IN_VITRO)"viable cell counts in the co-cultured strains increased significantly"
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PMID:42604006 - PARTIAL (IN_VITRO)"This could be attributed to a cross-feeding relationship"
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PMID:42604006 - SUPPORT (IN_VITRO)"Each probiotic strain was cultured in MRS medium supplemented with 1% cell-free supernatant"
Associated Datasets
| Dataset | Type | Repository | Accession |
|---|---|---|---|
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M-MV and single-strain probiotic MV LC-MS/MS proteomics
ProteomeXchange accession for LC-MS/MS proteomics comparing proteins in mixed-species M-MVs and B. longum, C. butyricum, and L. reuteri mono-culture MVs. |
PROTEOMICS | OTHER | PXD071787 |
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iProX record for M-MV LC-MS/MS proteomics
iProX partner-repository accession for the same LC-MS/MS vesicle proteomics deposition. |
PROTEOMICS | OTHER | IPX0014605000 |
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Lut-MV DSS mouse-colitis 16S rDNA amplicon BioProject
NCBI BioProject containing raw 16S rDNA sequencing reads from intestinal microbiota analysis of mice treated with M-MVs, luteolin, or Lut-MVs. |
AMPLICON_16S | NCBI_BIOPROJECT | PRJNA1482721 |
Knowledge gaps & discussions (2)
KNOWLEDGE_GAP OPEN Does the M-MV-only methionine sulfoxide reductase MsrA protein causally drive the improved antioxidant and colitis-protective phenotype of mixed-species vesicles?
The LC-MS/MS dataset identified MsrA as unique to M-MVs and its known antioxidant role is directionally consistent with higher radical scavenging, but the study did not test an MsrA knockout, inhibition, or add-back design.
Attaches to: ecological_interactions, associated_datasets#PXD071787
1 proposed experiment(s)
Approach: genetic perturbation of probiotic MsrA followed by vesicle proteomics and activity assays
Decision: A specific loss of M-MV antioxidant and colitis-protective activity would support a causal MsrA contribution; retention of activity would point to other vesicle cargo.
1 evidence item(s)
we did not perform direct functional validation to establish causality
Documents the paper's stated absence of direct MsrA causal validation.
KNOWLEDGE_GAP OPEN Which selective mechanisms explain the measured community responses?
Preserve higher viable counts and the positive cell-free-supernatant assay; distinguish soluble growth promotion from traced nutritional exchange and aggregate from member-resolved growth. The graph still lacks the measured mixed-species membrane-vesicle production/properties outcome. One-record Edison dry run is complete; paid authorization and report inspection are pending before extension. Do not infer growth-to-vesicle-yield causation, MsrA necessity, or universally superior antioxidant activity: L-MVs exceed M-MVs in the reported ABTS assay. Existing MsrA knowledge gap remains open.
Attaches to: ecological_interactions#Three-Strain Co-Culture Growth Enhancement
External Resources
| Name | Repository | Resource ID |
|---|---|---|
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PubMed - multi-species probiotic-derived vesicles for luteolin delivery
PubMed record for the iScience study of B. longum/C. butyricum/L. reuteri M-MVs. |
OTHER | PMID:42604006 |
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Version-of-record DOI
DOI landing page for the iScience primary publication. |
OTHER | doi:10.1016/j.isci.2026.117105 |
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PMC open-access full text
Open-access PMC full text for the iScience primary publication. |
OTHER | PMC13476518 |
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Growth Media
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