Journal: Journal of Bacteriology
Article Title: Genome-wide analysis of Enterococcus faecalis genes that facilitate interspecies competition with Lactobacillus crispatus
doi: 10.1128/jb.00438-24
Figure Lengend Snippet: L. crispatus antagonism of E. faecalis is facilitated in a contact-independent manner. ( A ) CFU recovered from E. faecalis OG1RF and L. crispatus VPI 3199 grown statically for 72 h in MRS media as single-species biofilm, either in separate wells or in the same well separated by a Transwell membrane insert that prevents physical contact between cells. L. crispatus biofilm is grown on the flat surface of the well in the tissue culture plate, whereas E. faecalis biofilm is seeded on the surface of the Transwell membrane insert. The dotted line indicates the limit of detection (LOD), CFU < 42.5. ( B ) Representative images of spot antagonism assay showing growth inhibition of E. faecalis when L. crispatus macrocolony biofilms were established at the same time (T0), 24 h, 48 h, or 72 h before inoculating E. faecalis . CFU recovered from E. faecalis OG1RF growth after 24 h in MRS media mixed with 72 h cell-free biofilm supernatant isolated from single-species and dual-species biofilms at an equal ratio, either ( C ) pH-unadjusted or ( D ) adjusted to pH 6.5 to mirror the MRS media. For A, C, and D, data points represent 9–12 biological replicates, collated from at least three repeated experiments. Statistical analysis was performed using the Brown-Forsythe ANOVA test with Welch’s correction. Error bars represent the standard error of the mean. ** P ≤ 0.01, **** P ≤ 0.0001.
Article Snippet: To investigate whether L. crispatus -mediated killing was contact-dependent or independent, biofilms were allowed to form in a six-well tissue culture-treated polystyrene plate with a Transwell insert (0.4 μm polyester membrane; Lot #20623011) (Corning, New York, USA).
Techniques: Membrane, Inhibition, Isolation
