Journal: bioRxiv

Article Title: Contact dependent suppression of Clostridioides difficile sporulation by enterococci requires the endocarditis and biofilm associated pilus

doi: 10.64898/2026.04.22.718763

Figure Lengend Snippet: (A-D) Monoculture and coculture of C. difficile in liquid SMC anaerobically for 48 hours with E. feacalis OG1RF , Enterococcus durans, Enterooccus faecalis, Enterococus faeicum, or Enterococus hirae followed by plating on selective media. n = 3-11 biological replicates combined from 4 independent expeirments. (A) Enterococci CFUs (B) Total C. difficile CFUs (C) Heat-resistant C. difficile spore CFUs (D) C. difficile sporulation efficiency. (E-H) Monoculture and coculture of Enterococcus saccharolyticus and C. difficile in liquid SMC anerobically for 48 hours followed by plating on selective medium. n = 8-9 biological repicates combined from 3 independent experiments (E) E. saccharolyticus CFUs. (F) Total C. difficile CFUs. ( G) Heat-resistant C. difficile spore CFUs. (H) Percent sporulation efficieincy. Statistics in A, E, F, G, H: t-test with Welch’s correction, statistics in B, C, D: Kruskal-Wallis One-way ANOVA with Dunn’s multiple comparisions test. LoD = Limit of Detection

Article Snippet: Samples inoculated in the well were plated on both BHI-CC-TA and Pfizer Selective Enterococcus Agar to confirm no bacteria migrated through the insert filter.

Techniques:

Journal: bioRxiv

Article Title: Contact dependent suppression of Clostridioides difficile sporulation by enterococci requires the endocarditis and biofilm associated pilus

doi: 10.64898/2026.04.22.718763

Figure Lengend Snippet: A-D) C. difficile and VRE were inoculated in transwells as indicated in the diagram on each x-axis. V = VRE, C = C. difficile. Transwells were incubated for 48 hours and plated on selective media. A) Total VRE CFUs. B) Total C. difficile CFUs. C) Heat resistant C. difficile spore CFUs. D) Percent sporulation efficiency. n = 6 biological replicates. Statistics: Pairwise comparisons for the effects of the presence and absence of a transwell: Unpaired Welch’s t-test. Multiple comparisons for differences in CFUs across conditions: Kruskall-Wallace One-way ANOVA with Dunn’s correction. E-G) Macrocolony migration assay, spots were inoculated on BHI agar and were incubated for 5 days followed by sampling the inside and periphery of the colony and selective plating. CD = C. difficile , 43076 = E. saccarolyticus E) Enterococcus CFUs, F) Total C. difficile CFUs, G) C. difficile heat resistant spore CFUs, H) Percent sporulation efficiency. Each data point represents one biological replicate combined from 3 experiments. Statistics: Kruskall-Wallace One-way ANOVA with Dunn’s correction. I) Representative images of 5-day single and dual species macrocolonies.

Article Snippet: Samples inoculated in the well were plated on both BHI-CC-TA and Pfizer Selective Enterococcus Agar to confirm no bacteria migrated through the insert filter.

Techniques: Incubation, Migration, Sampling

Journal: Veterinary Sciences

Article Title: Isolation and Characterization of Vancomycin-Resistant Enterococcus faecium from Cattle: Antimicrobial Resistance, Virulence Genes, and Pathogenicity

doi: 10.3390/vetsci12090880

Figure Lengend Snippet: Identification and molecular characterization of E. faecium isolates. ( A ) Colony morphology on Enterococcus-selective agar showing brown-black colonies with brown halos. ( B ) Colony morphology on blood agar with small, round, slightly raised, grayish-white, translucent colonies. ( C ) Gram staining showing Gram-positive cocci, occurring singly or in pairs. ( D ) PCR amplification of the ddl gene showing a 176 bp product specific to E. faecium ; Lanes 1–3 in correspond to isolates SCQ3, SCQ4, and SCQ11, respectively. ( E ) 16S rRNA gene amplification producing an approximately 1500 bp band; Lanes 1–3 in correspond to isolates to SCQ3, SCQ4, and SCQ11, respectively. ( F ) Neighbor-joining phylogenetic tree constructed using MEGA X from aligned 16S rRNA sequences, illustrating genetic relationships between isolates and reference E. faecium strains from NCBI. Bootstrap values (n = 1000 replicates) shown at branches; scale bar indicates evolutionary distance.

Article Snippet: Cultures were then streaked onto Pfizer enterococcus selective agar plates (Pfizer Inc., Shanghai, China) and incubated at 37 °C for 18–24 h. Colonies appeared as brown/black with brown halos, and the colony morphology was observed and recorded.

Techniques: Staining, Amplification, Construct

Journal: bioRxiv

Article Title: LicD-Mediated Cell Wall Decoration Governs Phage Sensitivity in Enterococcus faecalis Clinical Isolates

doi: 10.1101/2025.05.13.653012

Figure Lengend Snippet: Antibiofilm activity of Enterococcus phages . (a) Minimum biofilm eradication concentration presented as percent reduction in the biofilm biomass compared to non-treated control using phage titres ranging from (9-5) log 10 PFU/mL. (b) antibiofilm activity of Enterococcus phages against host ATCC-700802 and clinical isolates 8010 and 8050. Data presented as mean ± SD n=3. ns: non-significant, p > 0.05, *p < 0.05; **p < 0.01, ***p < 0.001 one-way ANOVA followed by multiple comparisons tests.

Article Snippet: Briefly, 100 µL of the collected samples were spread on Difco™ m Enterococcus selective Agar (Thermo Fisher Scientific Australia Pty Ltd., Scoresby, VIC, Australia) and incubated at 37°C.

Techniques: Activity Assay, Concentration Assay, Control

Journal: bioRxiv

Article Title: LicD-Mediated Cell Wall Decoration Governs Phage Sensitivity in Enterococcus faecalis Clinical Isolates

doi: 10.1101/2025.05.13.653012

Figure Lengend Snippet: Time-kill kinetics of Enterococcus bacteriophages. (a) Growth pattern of E. faecalis ATTC 700802 incubated with individual phages APTC-Efa.10, Efa.16 and Efa.20 (MOI) in the range between (0.01-100). (b) Growth pattern of E. faecalis ATTC 700802 incubated with 1:1 phage cocktail at different MOIs (0.01-100).

Article Snippet: Briefly, 100 µL of the collected samples were spread on Difco™ m Enterococcus selective Agar (Thermo Fisher Scientific Australia Pty Ltd., Scoresby, VIC, Australia) and incubated at 37°C.

Techniques: Incubation