Journal: bioRxiv

Article Title: Characterization and description of Clostridium filamentum ETTB, a novel gut bacterium with TLR5 modulating properties

doi: 10.1101/2025.02.28.640726

Figure Lengend Snippet: A. Gram staining of overnight cultures grown in LYBHI broth. B. Scanning electron microscopy showing the morphology of C. saudiense JCC and C. filamentum isolates. For acquiring SEM pictures, electron high tension (EHT) of 2.00 KV and magnifications of 7.00 KX (upper panel B) and 25.00 KX (bottom panel B) were used. Scale bars represent 5 µm length for panel A and 2 µm length for panel B.

Article Snippet: These isolates were designated as Clostridium filamentum ETTB1, ETTB2, and ETTB3, with ETTB3 as the type strain for the novel species C. filamentum deposited at DSMZ with name Clostridium sp. DSM 115107 and CCUG with the name CCUG 77667.

Techniques: Staining, Electron Microscopy

Journal: bioRxiv

Article Title: Characterization and description of Clostridium filamentum ETTB, a novel gut bacterium with TLR5 modulating properties

doi: 10.1101/2025.02.28.640726

Figure Lengend Snippet: A. Phylogenetic tree based on genome sequences showing the relationship of the C. filamentum ETTB1, ETTB2 and ETTB3 with other Clostridium species. B. Pangenome analysis highlighting genomic functional differences with C. saudiense JCC.

Article Snippet: These isolates were designated as Clostridium filamentum ETTB1, ETTB2, and ETTB3, with ETTB3 as the type strain for the novel species C. filamentum deposited at DSMZ with name Clostridium sp. DSM 115107 and CCUG with the name CCUG 77667.

Techniques: Functional Assay

Journal: bioRxiv

Article Title: Characterization and description of Clostridium filamentum ETTB, a novel gut bacterium with TLR5 modulating properties

doi: 10.1101/2025.02.28.640726

Figure Lengend Snippet: A. Growth curves in LYBHI broth for the bacteria incubated anaerobically for 24 hours in a Coy chamber at 37°C. B and C. Representative heatmaps of 3 independent experiments evaluating enzymatic activities ( B ) and substrate utilization for C. saudiense JCC and C. filamentum ETTB1, ETTB2 and ETTB3 using API ZYM, API 20A and rapid ID 32 STREP ( C ). D. Profiles of short-chain fatty acids produced in LYBHI broth by cultures grown anaerobically for 18 hours in a Coy chamber at 37°C (as described in A). Data are presented as mean ± S.E.M. n= 4-6 independent experiments. One-way ANOVA with post-hoc Tukey’s test: *p< 0.05 , **p< 0.01 , ***p< 0.001 and ****p< 0.0001 .

Article Snippet: These isolates were designated as Clostridium filamentum ETTB1, ETTB2, and ETTB3, with ETTB3 as the type strain for the novel species C. filamentum deposited at DSMZ with name Clostridium sp. DSM 115107 and CCUG with the name CCUG 77667.

Techniques: Bacteria, Incubation, Produced

Journal: bioRxiv

Article Title: Characterization and description of Clostridium filamentum ETTB, a novel gut bacterium with TLR5 modulating properties

doi: 10.1101/2025.02.28.640726

Figure Lengend Snippet: A. Representative images of C. saudiense JCC and C. filamentum ETTB1 and ETTB2 incubated in presence or absence of Caco-2 cells for 6 hours in anaerobic conditions in a Coy chamber at 37°C. B. Gene expression in Caco-2 cells co-cultured for 6 hours with C. saudiense JCC and C. filamentum ETTB1 and ETTB2. Scale bars represent 5 µm length. Data are presented as mean ± S.E.M. n= 5 independent experiments. One-way ANOVA with post-hoc Tukey’s test: *p< 0.05 , **p< 0.01 and ****p< 0.0001 .

Article Snippet: These isolates were designated as Clostridium filamentum ETTB1, ETTB2, and ETTB3, with ETTB3 as the type strain for the novel species C. filamentum deposited at DSMZ with name Clostridium sp. DSM 115107 and CCUG with the name CCUG 77667.

Techniques: Incubation, Gene Expression, Cell Culture

Journal: bioRxiv

Article Title: Characterization and description of Clostridium filamentum ETTB, a novel gut bacterium with TLR5 modulating properties

doi: 10.1101/2025.02.28.640726

Figure Lengend Snippet: A. Gene expression in Caco-2 cells incubated for 6 hours with C. filamentum ETTB1 and ETTB2 conditioned media (CM) or proteinase K-treated conditioned media (CM-PK). B. Amino acid sequence of the fliC protein encoded by the fliC gene of C. filamentum ETTB3. The TLR5 binding motif is highlighted in orange. C. Upper panel: gene expression in Caco-2 cells with or without TLR5 inhibition by an anti-TLR5 antibody (iTLR5) in the presence of 1 µg of recombinant fliC from C. filamentum ETTB3 or Salmonella typhimurium ( S. typh .). Lowe panel: gene expression in Caco-2 cells with or without TLR5 inhibition by an anti-TLR5 antibody (iTLR5) after incubation for 6 hours with live C. filamentum ETTB1 and ETTB2 or 1 µg of recombinant fliC from Salmonella typhimurium ( S. typh .). Data are presented as mean ± S.E.M. n= 4-5 independent experiments. Two-way ANOVA with post-hoc Šidák test for panel A . Pairwise t-test for panel C . *p< 0.05 , **p< 0.01 , ***p< 0.001 and ****p< 0.0001 .

Article Snippet: These isolates were designated as Clostridium filamentum ETTB1, ETTB2, and ETTB3, with ETTB3 as the type strain for the novel species C. filamentum deposited at DSMZ with name Clostridium sp. DSM 115107 and CCUG with the name CCUG 77667.

Techniques: Gene Expression, Incubation, Sequencing, Binding Assay, Inhibition, Recombinant

Journal: bioRxiv

Article Title: Characterization and description of Clostridium filamentum ETTB, a novel gut bacterium with TLR5 modulating properties

doi: 10.1101/2025.02.28.640726

Figure Lengend Snippet: A. Relative abundance of C. filamentum ETTB3 in mothers (n=100) and infants at 0, 4 months and 12 months of age (n=100) . B. Relative abundance of C. filamentum ETTB3 in a Swedish cohort of healthy individuals (S3PW) sampled 4 times over one year (n=75 individuals; 300 samples). V1, V2, V3 and V4 represent the four time points when fecal samples were assessed . C. Relative abundance of C. filamentum ETTB3 in the S3WP cohort (each dot represents the average of V1-V4 relative abundances in each individual) and in Hadza hunter-gatherers of Tanzania (n= 27) . Kruskal-Wallis with post-hoc Dunn’s test for panel A and non-parametric Mann-Whitney test for panel C . Adjusted P values are presented.

Article Snippet: These isolates were designated as Clostridium filamentum ETTB1, ETTB2, and ETTB3, with ETTB3 as the type strain for the novel species C. filamentum deposited at DSMZ with name Clostridium sp. DSM 115107 and CCUG with the name CCUG 77667.

Techniques: MANN-WHITNEY