Fig 1.
Impact of lactoferrin on the growth of the S7 consortium.
Growth curves of the S7 consortium and difference (Δ) in the log of 16S rRNA gene copy number of S7 members after incubation with (A) apo (LF-A), (B) native (LF-N) and (C) holo (LF-H) lactoferrin forms for a 24 h-period. The growth curves values are baseline corrected and representing mean values of four replicates. The log change is relative to that of the S7 without LF and is represented as the ∆ log copies/mL. Concentrations of LF-A of 0.5 and 5 mg/mL, LF-N of 0.5 and 5 mg/mL, and LF-H of 0.5 and 50 mg/mL. Strains are indicated by their genus capital letter as follows: A, Alistipes putredinis, B, Barnesiella intestinihominis, C, Coprococcus catus, D, Dorea longicatena, E, Eubacterium rectale (reclassified as Agathobacter rectalis), F, Faecalibacterium prausnitzii, and R, Roseburia hominis.
Fig 2.
Impact of lactoferrin on the growth of the individual S7 strains and S7 consortium.
Growth curves of the individual S7 strains and S7 consortium after incubation with holo lactoferrin (LF-H) for a 24 h-period. The growth curves values are baseline corrected and representing mean values of four replicates. Strains are indicated by their genus capital letter as follows: A, Alistipes putredinis, B, Barnesiella intestinihominis, C, Coprococcus catus, D, Dorea longicatena, E, Eubacterium rectale (now Agathobacter rectalis), F, Faecalibacterium prausnitzii, and R, Roseburia hominis.
Fig 3.
Composition of the healthy and frail microbiota at baseline.
Bar graphs of relative abundance at the phylum (A) and family (B) level for the healthy and frail subjects’ microbiota samples with and without the LF-H supplementation at time point 0 h. Each column represents a sample.
Fig 4.
Compositional differences between the faecal microbiota from healthy and frail older donors at time point 0 h.
Boxplot of α-diversity analysis based on observed ASV (A), Simpson index (B) and Shannon index (C) of healthy and frail microbiota samples with and without the LF-H supplementation. Statistical significance was determined for the boxplots using Dunn’s Kruskal-Wallis Multiple Comparisons. Different letters (a, b, c) in the plots indicate statistically significant differences between groups (p < 0.05). Principal Component Analysis (PCoA) of β-diversity (Bray–Curtis dissimilarity) at the ASV level of the healthy and frail microbiota samples with and without the LF-H supplementation (D). Pair-wise comparison p-value included in S5 Table.
Fig 5.
Composition of the healthy and frail microbiota after 24 h-fermentation.
Bar graph demonstrating relative abundance at the phylum (A) and family (B) level for the healthy and frail subject microbiota samples with and without the LF-H supplementation at time point 24 h.
Fig 6.
Compositional differences between the faecal microbiota from healthy and frail older donors at time point 24 h.
Boxplot of α-diversity analysis based on observed ASV (A), Simpson index (B) and Shannon index (C) of healthy and frail microbiota samples with and without the LF-H supplementation. Statistical significance was determined for the boxplots using Dunn’s Kruskal-Wallis multiple comparisons. Different letters (a, b, c) in the plots indicate statistically significant differences between groups (p < 0.05). Principal Component Analysis (PCoA) of β-diversity (Bray–Curtis dissimilarity and Unweighted UniFrac) at the ASV level of the healthy (D, E) and frail (F, G) microbiota samples with and without the LF-H supplementation.
Fig 7.
Differential taxa between the untreated and treated healthy and frail samples after 24 h-fermentation.
DeSeq2 analysis of differential taxa between the microbiota of healthy (A) and frail (B) donors with and without the LF-H supplementation. * p < 0.05, ** p < 0.01, *** p < 0.001.