model antibiotic resistant strain enterococcus faecalis og1rf  (ATCC)

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 antagonizes the growth of enterococci. ( A ) Representative image of macro-colony biofilms on an MRS agar plate. ( B ) CFU recovered from E. faecalis OG1RF and L. crispatus VPI 3199 grown, respectively, either as single-species macro-colony biofilm or as dual-species ( E. faecalis + L. crispatus ) macro-colony biofilm for 24 h, 72 h, and 120 h post-incubation. ( C ) CFU recovered from selected E. faecalis and E. faecium strains, respectively, grown either as single-species macro-colony biofilm, or grown as dual-species macro-colony biofilms after 72 h incubation. ( D ) CFU recovered from E. faecalis OG1RF grown either as single-species macro-colony biofilm or grown respectively with selected L. crispatus strains as dual-species macro-colony biofilm after 72 h incubation. For B–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 standard error of the mean. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

Article Snippet: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques: Incubation

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: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques: Membrane, Inhibition, Isolation

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 antagonistic activity is enhanced in an aciduric environment. pH measurements from E. faecalis OG1RF and L. crispatus VPI 3199 grown statically between 24 and 120 h either as single-species biofilm or as dual-species ( E. faecalis + L. crispatus ) biofilm in MRS ( A ) or MRS supplemented with 300 mM MOPS buffer ( B ). Corresponding growth dynamics of E. faecalis ( C ) and L. crispatus ( D ) in MRS or MRS supplemented with 300 mM MOPS buffer. ( E ) CFU recovered from E. faecalis OG1RF and L. crispatus VPI 3199 grown, respectively, either as single-species macro-colony biofilm or as dual-species ( E. faecalis + L. crispatus ) macro-colony biofilm for 24 h, 72 h, and 120 h post-incubation in MRS agar supplemented with 300 mM MOPS. For A–D, data points represent 9–12 biological replicates, collated from at least three repeated experiments. For A and B, statistical analysis was performed using two-way ANOVA. For C and D, linear regression of the slope of the exponential growth phase was performed. For E, statistical analysis was performed using the Brown-Forsythe ANOVA test with Welch’s correction. Error bars represent the standard error of the mean. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

Article Snippet: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques: Activity Assay, Incubation

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: Select list of the most differentially expressed genes during co-culture with L. crispatus relative to E. faecalis single-species control

Article Snippet: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques: Binding Assay, Modification

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: Selected list of differentially abundant E. faecalis transposon mutants during co-culture with L. crispatus relative to E. faecalis single-species counterpart

Article Snippet: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques:

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: E. faecalis genes contribute to tolerance against L. crispatus antagonism. CFU recovered from E. faecalis OG1RF, its isogenic transposon mutants, and L. crispatus VPI 3199 grown, respectively, either as single-species macro-colony biofilm or as dual-species ( E. faecalis +L. crispatus ) macro-colony biofilm ( A and B ) for 72 h. CFU recovered from E. faecalis OG1RF, its isogenic LDH deletion, and complementation mutants, as well as L. crispatus VPI 3199 grown respectively either as single-species macro-colony biofilm or as dual-species ( E. faecalis +L. crispatus ) macro-colony biofilm ( C and D ) for 72 h. 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 ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

Article Snippet: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques:

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: Loss of ldh1 restores virulence of E. faecalis in a co-infection model of Galleria larvae. Percentage survival of G. mellonella larvae 72 h post-injection with either single-species controls; E. faecalis OG1RF or L. crispatus, and their heat-killed counterparts ( A ), or co-injected with both ( B ). Percentage survival of G. mellonella larvae 72 h post-injection with E. faecalis OG1RF parent strain and its isogenic LDH deletion and complementation strains alone ( C ), or their corresponding co-injected counterparts ( D ). In each Galleria infection experiment, 20 larvae were infected with one biological replicate of E. faecalis inoculant, with a total of three replicates per experiment. Data points represent 9 biological replicates of E. faecalis inoculum, repeated thrice ( n = 3) on non-consecutive days. Each curve represents a group of 180 larvae, individually injected with 10 5 CFUs of E. faecalis suspended in PBS at a final volume of 5 µL. Statistical analysis was performed using the log-rank (Mantel-Cox) test. **** P ≤ 0.0001.

Article Snippet: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques: Infection, Injection

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: Strains used in this study

Article Snippet: E. faecalis strains , OG1RF parent strain (ATCC 47077) , Laboratory strain, Rif R , Fus R , ( ) .

Techniques: Isolation