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. 2017 Jun 30:8:1226.
doi: 10.3389/fmicb.2017.01226. eCollection 2017.

Functional Characterization of Novel VSports app下载 - Faecalibacterium prausnitzii Strains Isolated from Healthy Volunteers: A Step Forward in the Use of F. prausnitzii as a Next-Generation Probiotic

Rebeca Martín  1 , Sylvie Miquel  1   2 , Leandro Benevides  1   3 , Chantal Bridonneau  1 , Véronique Robert  1 , Sylvie Hudault  1 , Florian Chain  1 , Olivier Berteau  1 , Vasco Azevedo  3 , Jean M Chatel  1 , Harry Sokol  1   4   5 , Luis G Bermúdez-Humarán  1 , Muriel Thomas  1 , Philippe Langella  1
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"V体育官网入口" Functional Characterization of Novel Faecalibacterium prausnitzii Strains Isolated from Healthy Volunteers: A Step Forward in the Use of F. prausnitzii as a Next-Generation Probiotic

Rebeca Martín et al. Front Microbiol. .

Abstract

Faecalibacterium prausnitzii is a major member of the Firmicutes phylum and one of the most abundant bacteria in the healthy human microbiota. F. prausnitzii depletion has been reported in several intestinal disorders, and more consistently in Crohn's disease (CD) patients. Despite its importance in human health, only few microbiological studies have been performed to isolate novel F. prausnitzii strains in order to better understand the biodiversity and physiological diversity of this beneficial commensal species. In this study, we described a protocol to isolate novel F. prausnitzii strains from feces of healthy volunteers as well as a deep molecular and metabolic characterization of these isolated strains. These F. prausnitzii strains were classified in two phylogroups and three clusters according to 16S rRNA sequences and results support that they would belong to two different genomospecies or genomovars as no genome sequencing has been performed in this work. Differences in enzymes production, antibiotic resistance and immunomodulatory properties were found to be strain-dependent. So far, all F. prausnitzii isolates share some characteristic such as (i) the lack of epithelial cells adhesion, plasmids, anti-microbial, and hemolytic activity and (ii) the presence of DNAse activity. Furthermore, Short Chain Fatty Acids (SCFA) production was assessed for the novel isolates as these products influence intestinal homeostasis. Indeed, the butyrate production has been correlated to the capacity to induce IL-10, an anti-inflammatory cytokine, in peripheral blood mononuclear cells (PBMC) but not to the ability to block IL-8 secretion in TNF-α-stimulated HT-29 cells, reinforcing the hypothesis of a complex anti-inflammatory pathway driven by F. prausnitzii. Altogether, our results suggest that some F. prausnitzii strains could represent good candidates as next-generation probiotic. VSports手机版.

Keywords: Faecalibacterium; commensal; immune-modulatory properties; molecular and metabolic characterization; probiotic V体育安卓版. .

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Figures (VSports)

Figure 1
Figure 1
Negative screening for isolation of new Extremely Oxygen Sensitive (EOS) strains from human healthy feces.
Figure 2
Figure 2
Growth profile of F. prausnitzii strains. (A) OD600nm determination after 20 h growth in YBHI supplemented medium and (B) determination of viable bacteria: the CFU/mL numeration in the same cultures. Each measurement have been done at least in triplicate.
Figure 3
Figure 3
Phylogenetic tree of F. prausnitzii strains based on 16S rRNA gene sequences. The tree was constructed with the MEGA6 software package using the Maximum Likelihood method. The bootstrap values above 70% are shown next to the branches. The F. prausnitzii isolates incorporated in this study have circles besides. The black circles represent the cultured strains and white circles represent uncultured isolates. Colors (purple, blue, and green) and letters (A, B, and C) indicate the tree groups with high bootstrap values, formed by our cultured strains.
Figure 4
Figure 4
Scanning electron microscopy images of F. prausnitzii strains phylogroup II. Strains were grown in YBHI liquid medium 20 h. Scale bars indicate 2 μm. Arrows indicates “swelling.”
Figure 5
Figure 5
SCFA metabolism of F. prausnitzii strains in vitro. (A) Acetate and butyrate concentrations after 20 h growth of strain in YBHI supplemented medium. The concentration of control media was subtracted for each measurement have been done at least in triplicate. (B) Correlation between OD600nm and butyrate production.
Figure 6
Figure 6
Immuno-modulation capacities of F. prausnitzii strains in vitro. (A) IL-8 production in HT-29 TNF-α stimulated cells. Experiments have been done at least in triplicate. Results are expressed as IL-8/ protein (pg/mg) and have been normalized using as reference value the IL-8 produced after the co-incubation with PBS as a negative control. (B) IL-10 production in peripheral blood mononuclear cells. Experiments have been done at least in triplicate. Results are expressed as IL-10 concentration (pg/mL). Significant differences from the control (YBHI) was specified as: *p < 0.05 and ***p < 0.001.
Figure 7
Figure 7
Adhesion to mucin of F. prausnitzii strains. Experiments have been done in triplicate. The adhesion values have been normalized using Lactobacillus rhamnosus GG (LGG) a positive control know by their good adhesion properties to mucin (50). Results are presented by the mean and the standard deviation.
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