Journal: Microbiology Spectrum

Article Title: Sub-inhibitory concentrations of fosfomycin enhance Staphylococcus aureus biofilm formation by a sarA -dependent mechanism

doi: 10.1128/spectrum.01521-25

Figure Lengend Snippet: Effect of sub-inhibitory concentrations of fosfomycin on the initial adhesion ability of S. aureus . ( A ) Adhesion densities of S. aureus Newman strain at 30 and 120 minutes under different fosfomycin concentrations (0, 0.5, and 1 µg/mL). ( B ) Adhesion densities of S. aureus N315 strain under the same conditions. Adhesion is quantified as colony-forming units (CFU) per cm². Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA. ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns (not significant) P > 0.05.

Article Snippet: Fosfomycin (cat. no: HY-B1075A) was obtained from MedChemExpress (Shanghai, China) and dissolved in sterile deionized water.

Techniques:

Journal: Microbiology Spectrum

Article Title: Sub-inhibitory concentrations of fosfomycin enhance Staphylococcus aureus biofilm formation by a sarA -dependent mechanism

doi: 10.1128/spectrum.01521-25

Figure Lengend Snippet: Effect of sub-inhibitory concentrations of fosfomycin on biofilm formation ability of S. aureus . Crystal violet staining and quantification of biofilms formed by S. aureus Newman ( A ) and N315 ( B ) strains under sub-inhibitory concentrations of fosfomycin on the biofilm formation ability of S. aureus . Statistical analysis was performed using one-way ANOVA. ( C ) Effect of fosfomycin (1/4 MIC) on biofilm formation of various clinical S. aureus isolates. The concentration of fosfomycin at 1/4 MIC was calculated based on the MIC value determined for each strain, as listed in . Data are presented as mean ± SEM. Statistical analysis was performed using an unpaired, two-tailed Student’s t-test. *** P < 0.001, **** P < 0.0001.

Article Snippet: Fosfomycin (cat. no: HY-B1075A) was obtained from MedChemExpress (Shanghai, China) and dissolved in sterile deionized water.

Techniques: Staining, Concentration Assay, Two Tailed Test

Journal: Microbiology Spectrum

Article Title: Sub-inhibitory concentrations of fosfomycin enhance Staphylococcus aureus biofilm formation by a sarA -dependent mechanism

doi: 10.1128/spectrum.01521-25

Figure Lengend Snippet: Confocal laser scanning microscopy analysis of S. aureus biofilm formation under sub-inhibitory concentrations of fosfomycin. Representative images of biofilms formed by S. aureus Newman and N315 strains with and without 1 µg/mL fosfomycin treatment. Bacteria were stained with SYTO9 (green, live cells) and propidium iodide (PI, red, dead cells). Images show magnification, SYTO9 channel, PI channel, and merged views for each condition.

Article Snippet: Fosfomycin (cat. no: HY-B1075A) was obtained from MedChemExpress (Shanghai, China) and dissolved in sterile deionized water.

Techniques: Confocal Laser Scanning Microscopy, Bacteria, Staining

Journal: Microbiology Spectrum

Article Title: Sub-inhibitory concentrations of fosfomycin enhance Staphylococcus aureus biofilm formation by a sarA -dependent mechanism

doi: 10.1128/spectrum.01521-25

Figure Lengend Snippet: Effect of sub-inhibitory concentrations of fosfomycin on major biofilm components of S. aureus . ( A ) Dot blot analysis of PIA production in Newman and N315 strains under different fosfomycin concentrations (0, 0.5, and 1 µg/mL). ( B ) Evaluation of gray values of PIA using ImageJ software. WGA: wheat germ agglutinin. Statistical analysis was performed using one-way ANOVA. ( C ) Visual assessment of S. aureus aggregation in Newman and N315 strains with and without 1 µg/mL fosfomycin treatment. ( D ) Quantitative analysis of S. aureus aggregation. ( E ) Autolysis assay of S. aureus Newman and N315 strains treated with or without 1 µg/mL fosfomycin. The autolysis rate is represented by the decrease in optical density (OD 600 ) over time. Data points represent mean ± SEM. ( F ) Quantification of eDNA release in Newman and N315 strains with and without 1 µg/mL fosfomycin treatment. eDNA concentration is normalized to OD 600 and expressed as ng/μL/OD 600 . ( G ) Quantification of biofilm biomass in Newman, N315, and five clinical MRSA isolates. Biofilms were first induced by 1/4 MIC fosfomycin for 24 hours, where the 1/4 MIC for each strain was calculated based on its individual MIC value, as shown in . After induction, biofilms were treated with different concentrations of proteinase K (10, 20, and 40 µg/mL) for 4 hours. Biofilm biomass was measured by crystal violet staining and expressed as OD 600 . Bars represent mean ± SEM. Statistical analysis was performed using one-way ANOVA. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Fosfomycin (cat. no: HY-B1075A) was obtained from MedChemExpress (Shanghai, China) and dissolved in sterile deionized water.

Techniques: Dot Blot, Software, Concentration Assay, Staining

Journal: Microbiology Spectrum

Article Title: Sub-inhibitory concentrations of fosfomycin enhance Staphylococcus aureus biofilm formation by a sarA -dependent mechanism

doi: 10.1128/spectrum.01521-25

Figure Lengend Snippet: The mechanism by which sub-inhibitory concentrations of fosfomycin enhance S. aureus biofilm formation. ( A ) Effects of sub-inhibitory concentrations of fosfomycin on the expression of biofilm-related genes of S. aureus . Relative fold changes in gene expression in Newman (left) and N315 (right) strains treated with 1 µg/mL fosfomycin compared to untreated controls. Statistical analysis was performed using an unpaired, two-tailed Student’s t-test. ( B ) Confirmation of the sarA deletion (Δ sarA ) in the Newman strain. Agarose gel electrophoresis (left; GL DNA Marker 1000, Accurate Biology) and Sanger sequencing results (right) are shown. ( C ) Crystal violet staining of biofilms formed by Newman and Δ sarA , Δ sarA -C (Δ sarA complemented with the sarA gene) strains under different fosfomycin concentrations (0, 0.5, and 1 µg/mL) in 96-well plates. ( D ) Quantification of biofilm biomass at OD 600 after acetic acid dissolution. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA. ( E ) Initial adhesion assays of Newman, Δ sarA , Δ sarA -C (Δ sarA complemented with the sarA gene) strains with and without 1 µg/mL fosfomycin treatment. Results are expressed as percentages of adhesion. Statistical analysis was performed using an unpaired, two-tailed Student’s t-test. ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns (not significant) P > 0.05.

Article Snippet: Fosfomycin (cat. no: HY-B1075A) was obtained from MedChemExpress (Shanghai, China) and dissolved in sterile deionized water.

Techniques: Expressing, Gene Expression, Two Tailed Test, Agarose Gel Electrophoresis, Marker, Sequencing, Staining, Dissolution